Volume 8 No. 1 June 2017 IF=4.69 NEWS REPORTS AWARDS AND PERSONALIA RESEARCH NEWS BOOK NEWS MYCOLENS CORRESPONDENCE FORTHCOMING MEETINGS ARTICLES

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1 THE GLOBAL MYCOLOGICAL JOURNAL Volume 8 No. 1 June 2017 IF=4.69 NEWS REPORTS AWARDS AND PERSONALIA RESEARCH NEWS BOOK NEWS MYCOLENS CORRESPONDENCE FORTHCOMING MEETINGS S

2 Colofon IMA Fungus Compiled by the International Mycological Association for the world s mycologists. Scope: All aspects of pure and applied mycological research and news. Aims: To be the flagship journal of the International Mycological Association. IMA FUNGUS is an international, peer-reviewed, open-access, full colour, fast-track journal. Frequency: Published twice per year (June and December). Articles are published online with final pagination as soon as they have been accepted and edited. ISSN E-ISSN (print) (online) Websites: www. imafungus.org d.hawksworth@nhm.ac.uk Volume 8 No. 1 June 2017 Cover: Microbotryum silenesinflatae on Silene uniflora from the Outer Hebrides (see Smith et al., p ). EDITORIAL BOARD Editor-in-Chief Prof. dr D.L. Hawksworth CBE, Department of Life Sciences, The Natural History Museum, Cromwell Road, London SW7 5BD, UK; Comparative Plant and Fungal Biology, Royal Botanic Gardens, Kew, Surrey TW9 3DS, UK; E- mail: d.hawksworth@nhm.ac.uk; d.hawksworth@kew.org Managing Editor Prof. dr P.W. Crous, Westerdijk Fungal Biodiversity Institute, P.O. Box 85167, 3508 AD Utrecht, The Netherlands; E- mail: p.crous@westerdijkinstitute.nl Layout Editors M.J. van den Hoeven-Verweij & M. Vermaas, Westerdijk Fungal Biodiversity Institute, P.O. Box 85167, 3508 AD Utrecht, The Netherlands; westerdijkinstitute.nl Associate Editors Dr T.V. Andrianova, M.G. Kholodny Institute of Botany, Tereshchenkivska Street 2, Kiev, MSP-1, 01601, Ukraine; tand@darwin.relc.com Prof. dr D. Begerow, Lehrstuhl für Evolution und Biodiversität der Pflanzen, Ruhr-Universität Bochum, Universitätsstrasse 150, Gebäude ND 03/174, 44780, Bochum, Germany; dominik.begerow@rub.de Prof. dr Mary Berbee, Department of Botany, University of British Columbia, # University Boulevard, Vancouver, BC V6T 1Z4, Canada; mary.berbee@gmail.com Dr S. Cantrell, Department of Plant Pathology and Crop Physiology, Louisiana State University, Agricultural Centre, 455 Life Sciences Building, Baton Rouge, LA 70803, USA; scantrel@suagm.edu Dr P.S. Dyer, School of Biology, Institute of Genetics, University of Nottingham, University Park, Nottingham NG7 2RD, UK; paul.dyer@nottingham.ac.uk Dr Ana Esperanza Franco Molano, Instituto de Biologí, a Universidad de Antioquia, A.A. 1226, Medellín, Colombia; anaesperanza@gmail.com Dr K. Hansen, Kryptogambotanik Naturhistoriska Riksmuseet, Box 50007, Stockholm, Sweden; karen. hansen@nrm.se Prof. dr David Hibbett, Biology Department, Clark University, Lasry Biological Science Center, 950 Main St., Worcester, MA 01610, USA; dhibbett@clarku.edu Prof. dr Xingzhong Liu, State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, No. 3 1st Beichen West Road, Chaoyang District, Beijing , P. R. China; liuxz@im.ac.cn Dr Janet Jennifer Divinagracia Luangsa-ard, National Center for Genetic Engineering and Biotechnology (BIOTEC), 113 Thailand Science Park, Phahonyothin Road, Khlong Nueng, Khlong Luang, Pathum Thani 12120, Thailand; jajen@biotec.or.th Prof. dr W. Meyer, Molecular Mycology Research Laboratory, CIDM, ICPMR, Level 3, Room 3114A, Westmead Hospital, Darcy Road, Westmead, NSW, 2145, Australia; w.meyer@usyd.edu.au Dr Chiharu Nakashima, Graduate school of Bioresources, Mie University, Kurima-Machiya 1577, Tsu, Mie, Japan; chiharu@bio.mie-u.ac.jp Dr Meritxell Riquelme, Department of Microbiology, Centro de Investigación Científica y de Educación Superior de Ensenada CICESE, Carretera Ensenada-Tijuana N. 3918, Ensenada Baja California, Mexico; riquelmemeritxell@gmail.com Prof. dr K.A. Seifert, Research Scientist / Biodiversity (Mycology and Botany), Agriculture & Agri-Food Canada, K.W. Neatby Building, 960 Carling Avenue, Ottawa, ON, K1A OC6, Canada; seifertk@agr.gc.ca Prof. dr J.W. Taylor, Department of Plant and Microbial Biology, University of California, 111 Koshland Hall, Berkeley, CA 94720, USA; jtaylor@berkeley.edu Prof. dr M.J. Wingfield, Forestry and Agricultural Research Institute (FABI), University of Pretoria, Pretoria 0002, South Africa; mike.wingfield@fabi.up.ac.za

3 WHO WILL DECIDE ON FUNGAL NOMENCLATURE? Concerns over fixing the application of scientific names, and limiting changes in names for non-scientific reasons, are widespread amongst biologists. Names are the one universal mechanism for correlating and communicating all categories of information on particular organisms. It is therefore critical that the application of names is regulated by pertinent international bodies. Next month in Shenzhen, China, a decision on what body should govern fungal nomenclature will be voted on. One consequence of fungi traditionally being studied within botany, was that their naming was controlled by the International Rules [Code from 1952] of Botanical Nomenclature. The provisions in the Code are now considered and modified at subsequent International Botanical Congresses (IBCs), normally held at sixyear intervals. That was not seen as a major issue for mycologists while such congresses had a substantial mycological component and attendance. However, IBCs became ever larger and more expensive, mycologists a decreasing proportion of participants, and lacked enough symposia to cover the needs of a blossoming of mycological research into the 1960s. Mycologists felt increasingly marginalized in IBCs, and found their scale an impediment to personto-person interactions. As commented by Simmons (2010: 21), disillusionment set in as mycology had outgrown the traditional relationship with botany, in both practical and scientific terms. In response to this situation, Geoffrey C. Ainsworth ( ), who had retired in 1968, initiated discussions to hold a separate International Mycological Congress (IMC). Having gained support from key mycological societies worldwide, this became a reality at Exeter (UK) in It was recognized at IMC1 that improvements in the nomenclatural rules relating to fungi were needed, including issues over the separate naming of morphs in pleomorphic fungi, the acceptability of cultures as type material, and starting point dates. A Nomenclature Secretariat was established to pursue these matters, with Richard P. Korf ( ) as Secretary. I recall Korf mentioning the idea of a separate Code for fungi at the inaugural meeting, but only as something not even to be considered; there was no dissent. The Secretariat tasked a series of committees with addressing the issues of concern, and they developed proposals to put to the 1981 IBC in Sydney, which were accepted at that Congress; the Secretariat was then dissolved, having completed its tasks. In general, IBCs have accepted proposals that were clearly supported by most mycologists, and particularly the Nomenclature Committee for Fungi (NCF). Nomenclatural matters have become an increasingly important aspect of IMCs, initially with debates on topics of concern, but since IMC9 in Edinburgh in 2010 have incorporated Nomenclature Sessions and questionnaires circulated to all Congress delegates. At IMC10, and subsequently at meetings in Amsterdam the following year, mycologists supported the transfer of decision making on issues only concerning fungi from IBCs to IMCs. This was discussed at the IBC held in Melbourne later in 2011, when a Special Subcommittee was formed to address how this should be done. That Committee has now reported, and the proposals made are to be voted on at the IBC in Shenzhen in July Key mycological committees support the proposals made (see pp. (9) (11) in this issue), but there is a concern they may not be accepted next month. The Nomenclature Section meetings at the IBC in Vienna in 2005 and Melbourne in 2011 were attended by just seven and ten mycologists, out of a total of 198 and 204 delegates, respectively. In contrast, at IMC10 in Bangkok in 2014, mycologists attended the Nomenclature Sessions, and 117 turned in ballot papers. At present mycologists do not favour leaving the current Code, re-named the International Code of Nomenclature for algae, fungi, and plants in They appreciate the care with which the rules are refined by botanists well-versed in the intricacies of nomenclature facing the same problems as mycologists, for example on issues related to electronic publication. At the same time, mycologists are uncomfortable with, and do not wish, decisions on matters relating soley to fungi to continue to be taken at IBC meetings when so few mycologists are present, and the Nomenclature Committee for Fungi (NCF) is elected. There is also a feeling that a six year interval between possible changes is too long when aspects of the subject are having to adapt to new approaches. As there will be two IMCs before the next IBC in 2023, this will lead to two decision-making opportunities before the next IBC. All mycologists unable to attend the Shenzhen meetings and who work in institutions that have institutional votes to cast are urged to encourage their representatives to support the proposals for a change in the control of matters relating specifically to fungi from IBCs to IMCs. If these proposals do not gain approval next month, it is unlikely that mycologists would be content with the issue being referred to another committee, charged with reporting at the 2023 IBC. IMC11 in Puerto Rico in 2018 would then face the challenge of deciding whether to reject that decision and operate independently a schism I would not wish to see. Simmons EG (2010) The Interntaional Mycological Association: its history in brief with summaries of its International Mycological Congresses and diverse international relationships. IMA Fungus 1: David L. Hawksworth Editor-in-Chief, IMA Fungus (d.hawksworth@kew.org) EDITORIAL VOLUME 8 NO. 1 (1)

4 NEWS Interested in hosting IMC12 (2022)? Under the Statutes of the IMA ( the deadline for receipt of pre-proposals from Member Mycological Organizations (MMOs) to host the next International Mycological Congress is 12 months before the date of the current IMC 15 July The pre-proposals will then be reviewed by the Executive Committee, and a vote to solicit full proposals from not fewer than two of the MMOs submitting pre-proposals is due not later than 10 months before the date of the current IMC 15 September Full proposals to host the next IMC must then be received by the Secretary- General for distribution to the Executive Committee not later than six months before the current IMC 15 January The venues and dates for the next IMC will then be voted on by the Executive Committee not later than three months before the current IMC. The President and Secretary-General will visit the proposed venue selected by the Executive Committee before final ratification by the Executive Committee. The final decision will then be announced to the General Assembly of the IMA, to be held at the upcoming IMC. For further information, or to submit a pre-proposal, contact the IMA Secretary-General, Pedro Crous (p.crous@ westerdijkinstitute.nl). CBS becomes the Westerdijk Fungal Biodiversity Institute Johanna Westerdijk. Photo courtesy of the Westerdijk Fungal Biodiversity Institute. The Centraalbureau voor Schimmelcultures (CBS-KNAW) was established in 1903 by Friederich A.F.C. Went ( ) in The Netherlands, with the mission to house the world s fungal strains. In 1907, at the tender age of 24, Johanna Westerdijk became its first director, a position she held until One hundred years ago, on the 10 th of February 1917, Johanna Westerdijk also became the first female professor in The Netherlands when the University of Utrecht appointed her as a professor in phytopathology and she gave her inaugural lecture at the university. Under her inspiring leadership as director of the CBS, and starting with a mere 80 living fungi, the collection evolved into the largest and most versatile fungal resource centre in the world, containing over different strains today. In honour of its first Director, and to reflect the legacy this institute is built on, the name of the CBS-KNAW changed to the Westerdijk Fungal Biodiversity Institute on the centenary of her professorship. Joanna was a renowned multi-talented scientist, a believer in equal opportunity and inclusion, and a globetrotter; a true homo universalis. She educated 56 PhD students, almost half of which were female, and thought fungi had the potential to contribute to the solution of some of the world s greatest challenges. The mission of the Institute remains the same today, to explore, culture and preserve. It continues to explore the world and collect new fungi, and investigate their characteristics to address relevant societal challenges. For the anniversary of Westerdijk s professorship, 2017 has been designated as the Westerdijk year in The Netherlands, where scientists celebrate her contribution to science. As a research institute of the Royal Dutch Academy of Arts and Sciences (KNAW), together with the universities of Utrecht and Amsterdam and other parties, the Institute will commemorate Johanna Westerdijk s contributions to science and women s position in science. Many events will take place during this year of celebration. The University Museum in Utrecht has prepared an exhibition about Westerdijk and phytopathology and The new logo of the Westerdijk Fungal Biodiversity Institute. mycological research in general. Institute researchers visit schools to show and tell young children about fungi, scientists hold lunchtime lectures on campus, and, best of all, a citizen science project has been initiated: World fame, a fungus with your name. Visitors to the exhibition and schools in the Utrecht region receive a special collection kit to take soil samples in their gardens, which they then submit to the Westerdijk Institute for analysis and identification. To date 241 soil samples have been received (and continue to flow) and are now being analysed. With this citizen science project, the Institute aims to involve the public in scientific research, and present the past and future of fungal research. During the research period, the results will be shown online ( westerdijkinstitute.nl/). When a new fungus is identified, it will be named after the submitter, hence the theme World fame, a fungus with your name. The newly discovered fungi will be added to the collection and can be used for future research by the global community, and simultaneously screened for potential new (2) IMA FUNGUS

5 NEWS Spring holiday at the Utrecht University Museum: staff of the Westerdijk Fungal Biodiversity Institute introduced children (and their parents or caretakers) to science. They could experience what it was like to be a technician working with fungi. No fungi were harmed in the process! Plates courtesy staff of the Westerdijk Institute. Citizen Science project "Wereldfaam, een schimmel met je naam" ("World fame, a fungus with your name") proved to be a great success with almost 250 soil samples being sent to the institute by visitors of the Utrecht University Museum (both young and old), eager to earn eternal fame. antibiotics. Plans are afoot to upscale this project to include all 7000 Dutch primary schools in With a new name comes a new logo. The new logo still represents the legacy of the former logo. With a name change, a number of other things change as well, most importantly the website and addresses. The website has changed to but www. cbs.knaw.nl will redirect automatically to the new website. The address has changed to name@westerdijkinstitute.nl, but s sent to the old address will still be forwarded to the new address. The international acronym to be used for the collections remains as CBS. From 28 August until 1 September 2017, the Institute is to host a symposium week ( nl/biolomicsnews.aspx?rec=7728), which will start with the 2 nd Symposium on Plant Biomass Conversion by Fungi, followed by one on the Leading Women in Fungal Biology, featuring leading female mycologists from around the world, and a final symposium Cryptic Speciation in Classifications. The week will close with the opening of the new Building of the Westerdijk Fungal Biodiversity Institute in Utrecht, and a fungal barbeque. During the week the Institute will also present new discoveries from the citizen science project in a special public engagement evening, Famous Fungi. Last but not least, the Institute will also launch its own beer, especially brewed for the occasion from the ex-type strain of Saccharomyces cerevisiae, Schoone Geest [Healthy Spirit], a word play on the Westerdijk motto which was chiselled in the lintel stone above the door to her laboratory: Werken en feesten vormt schoone geesten [Work and play forms a healthy spirit]. Another Westerdijk saying, was to avoid a dull life (or die) [Even a fungus dies from a dull life], and all mycologists who can are invited to join the party, get the T-shirt, drink the beer in a special Westerdijk cup, and talk about fungi! For further details, consult the Institute website ( westerdijkinstitute.nl/). VOLUME 8 NO. 1 (3)

6 NEWS DNA sequence data as types: a potential loophole in the rules discovered? Type illustration for Lawreymyces pulchellae. Reproduced from Lücking & Moncada (2017). The issue of whether or not to give formal scientific names to organisms only known from DNA sequences is becoming an increasing cause for debate. Proposals of how this issue might be addressed for fungi were first made by Hibbett et al. (2011). Subsequently, proposals to modify the International Code of Nomenclature for algae, fungi, and plants (McNeill et al. 2012) have been published (Hawksworth et al. 2016) and are due to be voted on at the International Botanical Congress, Shenzhen, China, this July. A species known only from sequence data, with no cultures or other specimens, was described last year as Hawksworthiomyces sequentia with only the actual base sequences of selected DNA regions given as the type (De Beer et al. 2016). Species names, however, are required to have a physical type, or in some cases an illustration, in order to be validly published, so that name does not meet the current criteria and must be ruled as invalid. Lücking & Moncada (2017), who found a new genus Lawreymyces with seven new species from sequences obtained from lichen thalli, argue that there is a loophole in the existing rules as illustrations are permitted as types of microscopic fungi when no material can be preserved. They therefore designated illustrations of the alignments for the new species as the types) and consider that this approach opens the door to the formal recognition of thousands of species of voucherless fungi detected through environmental sequencing techniques under the current Code. Whether this loophole will be considered acceptable is likely to be hotly debated, and questioned as the illustrations designated are a diagram showing alignments rather than the molecules themselves in situ. If the proposals on this matter are rejected in the upcoming congress, the Nomenclature Committee for Fungi (NCF) may well be asked to give a formal opinion on this particular case whether this is a loophole or not. If the opinion is in the affirmative, the issue will become whether the Code should be modified to permit or to stop such practices in the future. De Beer ZW, Marincowitz S, Duong TA, Kim J-J, Rodrigues A, Wingfield MJ (2016) Hawksworthiomyces gen. nov. (Ophiostomatales), illustrates the urgency for a decision on how to name novel taxa known only from environmental nucleic acid sequences (ENAS). Fungal Biology 120: Hawksworth DL, Hibbett DS, Kirk PM, Lücking R (2016) ( ) Proposals to permit DNA sequence data to serve as types of names of fungi. Taxon 65: Hibbett DS, Ohman A, Glotzer D, Nuhn M, Kirk PM, Nilsson RH (2011) Progress in molecular and morphological taxon discovery and options for formal classification of environmental samples. Fungal Biology Reviews 25: Lücking R, Moncada B (2017) Dismantling Marchandiomphalina into Agonimia (Verrucariaceae) and Lawreymyces gen. nov. (Corticiaceae): setting a precedent to the formal recognition of thousands of voucherless fungi based on type sequences. Fungal Diversity: DOI /s McNeill J, Barrie FR. Buck WR, Demoulin V, Greuter W, et al. (eds) (2012) International Code of Nomenclature for algae, fungi, and plants (Melbourne Code) adopted by the Eighteenth International Botanical Congress Melbourne, Australia, July [Regnum Vegetabile No. 154.] Königstein: Koeltz Scientific Books. Who were the pioneer women in taxonomic mycology? The forthcoming centenary of the appointment of Johanna Westerdijk as a professor at Utrecht University (to be celebrated at the Leading Women in Fungal Biology symposium at the newly christened Westerdijk Fungal Biodiversity Institute in August this year - see page (2) in this issue) prompts a consideration of her female predecessors in mycology around the world. Given the dispersed nature of mycological records, the relatively centralised and complete records of taxonomic mycology offer a practical means of investigating this subject. An initial survey of Authors of Fungal Names (Kirk 2003) and other sources reveals that only a handful of women described new fungal taxa prior to Such women include, in order of birth date: Catharina Helena Dörrien ( , Germany), Marie- Anne Libert ( , Belgium), Mary Elizabeth Banning ( , USA), Elisa Caroline Bommer (née Destrée, , Belgium) [who also published on fungi of The Netherlands under the name Caroline E. Destrée], Marietta Rousseau (née Hannon, , Belgium) and lichenologist Annie Lorrain Smith ( , UK; Ainsworth 1996). (4) IMA FUNGUS

7 Marie-Anne Libert, and Ascochyta padi 1, which she described in her exsiccate Plantae cryptogamicae, Arduenna, fasc. 2: no. 153 (1832). A somewhat larger group of women made early contributions to mycology as researchers, collectors, illustrators and popularizers of fungi. These include: prolific collector Josephine Kablíková ( , Czech Republic), Elisabetta Fiorini-Mazzanti ( , Italy), Anna Maria Hussey ( , UK) whose much sought-after Illustrations of British Mycology was issued in 1855, Anna Russell 1 The current name for this species is given as Blumeriella jaapii (Rehm) Arx 1961 in Species Fungorum ( but that may need revision since the separate naming of fungal morphs was discontinued in (née Worsley, , UK), Margaret Plues (ca , UK) author of the 1864 delightfully illustrated volume Rambles in Search of Flowerless Plants (with a second edition in 1865), Flora Martin (née Campbell, , Australia), myxomycete specialist Gulielma Lister ( , UK), Beatrix Potter ( , UK), and Helen Charlotte Isabella Gywnne-Vaughan (née Fraser, ; Ainsworth 1996) who prepared the well-used textbook The Structure and Development of Fungi in 1927 (2 nd edn, 1937). While a select group of women had access to higher education as early as the 1860s, the impact of systematic accredited study was not evident in the numbers of women participating in mycology until after the turn of the twentieth century. We have identified around 100 university educated female mycologists born prior to 1900 (some working on taxonomy, others on other facets of fungi). Detailed biographical accounts of many of this cohort of women graduates can be found in Creese ( ) and Ogilvie & Harvey (2000). Relatively well-documented individuals who made contributions to mycology include: Rose Stoppel ( ), first German female professor of botany, Ethel Mary Doidge ( ), first South African woman to obtain a doctorate ( Jacobs 2016), Flora Wambaugh Patterson ( : Rogers 1981), first female mycologist at the United States Department of Agriculture (USDA; Rossman 2002), Effie Almira Spalding (née Southworth, ), first female plant pathologist at USDA (Ristaino & Peterson 2002), and, of course, in The Netherlands, Johanna Westerdijk ( ), whose foundation of the Central Bureau voor Schimmelcultures (now the Westerdijk Fungal Biodiversity Institute) underpinned Dutch taxonomic mycology in the 20th century. We are preparing a historical analysis of early women taxonomic mycologists, with a focus on the group who were not university or college-educated, or those who, like Gulielma Lister (see above) in the UK, had some access to higher education but did not gain formal qualifications. This group were exceptional examples of women who challenged, and helped to change, cultural norms around the participation of women in science. We are interested to hear about women who were active pre-1900 who have been overlooked in mycological history, especially those who described new taxa, but also collectors and illustrators who may have been have been on the threshold of making taxonomic contributions (Maroske 2014). Our experience leads us most readily to women in the English speaking and Western world, but we are mindful that exceptional mycological women may have been active elsewhere. We especially welcome information on pioneering female taxonomic mycologists in Asia, Africa, and South America. Ainsworth GC (1996) Brief Biographies of British Mycologists. Stourbridge: British Mycological Society. Creese MRS, Creese TM ( ) Ladies in the Laboratory. 4 vols. Lanham, MD: Scarecrow Press. Jacobs A (2016) The life and times of Ethel Mary Doidge, a pioneer of South African mycology. IMA Fungus 7: (69) (70) Kirk PM (2003) Authors of Fungal Names. Version 2. AuthorsOfFungalNames.htm. Maroske S (2014) A taste for botanic science : Ferdinand Mueller s female collectors and the history of Australian botany. Muelleria 32: Ogilvie MB, Harvey JD (2000) The Biographical Dictionary of Women in Science: Pioneering Lives From Ancient Times to the Mid-20th Century. 2 vols. New York: Routledge. Ristaino J, Peterson P (2002) Effie A. Southworth, first woman plant pathologist hired at USDA. The Plant Health Instructor: DOI: / PHI-I Rogers DP (1981) A Brief History of Mycology in North America. Augmented edn. Amherst, MA: Mycological Society of America. Rossman AY (2002) Flora W. Patterson: the first woman mycologist at the USDA. The Plant Health Instructor: DOI: / PHI-I Sara Maroske and Tom W. May Royal Botanic Gardens Victoria, Melbourne, Australia (tom.may@rbg.vic.gov.au) NEWS VOLUME 8 NO. 1 (5)

8 NEWS Online game changer for tree health? Photos: M. Coleman. Computer games that save trees! Is that even possible? That s the aim of scientists involved in the PROTREE project funded by the Tree Health and Plant Biosecurity Initiative, and they have a big challenge to meet, with new threats to tree health arising all the time. A consortium of seven Scottish research institutes (Royal Botanic Garden Edinburgh; The James Hutton Institute; Forest Research; Scotland s Rural College, SRUC; University of Aberdeen; The University of Edinburgh; and Centre for Ecology and Hydrology) working with games designers Hyper Luminal Games. have come up with CALEDON. The game falls within the survival strategy genre and aims to raise awareness of tree health problems, and communicate how we might go about ensuring trees are more resilient in the future. Since the 1970s we have had an ever growing number of tree pests and diseases arriving in the UK. Sometimes they have spread under their own steam, but more often than not we humans unwittingly give them a helping hand through global trade. Infected timber and young trees slip through the net and before too long we have a new tree health problem on our hands. Should we shut down trade to protect our trees? No. We can t ignore globalisation if we value shared prosperity. We do need to reduce risks as much as possible, be vigilant, and adopt strategies of forest and tree management that encourage resilience. Accepting that tree health problems are here to stay highlights the need for a step change in how we deal with this inevitable aspect of forestry. So why use computer games to tackle tree health? The hope is that through a popular and engaging medium a wide range of people, including children, can be switched on to tree health and potential new approaches to dealing with the problems we face. Engaging a new generation of potential plant health professionals is a vital part of the solution. All too often the media portrays tree health as a disaster waiting to happen. The truth is more nuanced. In nature the diversity of species and genes means that populations cope with, and bounce back from, attack. What we need to do is acknowledge and work with the natural processes that keep trees healthy. Putting it another way we need to help nature to help itself. What does this mean in practice? Our forestry is heavily reliant on single species plantations, often with limited genetic variation between individuals. The wisdom of old sayings is worth taking note of here as this is a case of putting all our eggs in what we hope will be one highly productive basket. Unfortunately, this approach is also an ideal incubator for pests and diseases. The solution is diversity, and the hope is that CALEDON will help to get that message across. Educational games or games with a purpose are a growing niche in the market. The trick is not to lose the fun factor. The best educational games maintain a balance of reality, meaning and play. In CALEDON the player is a forest manager responsible for keeping a forest healthy and productive. Although the game follows the popular survival model, this aspect of the game is switched around so that survival becomes something that is all about the trees and not the player. The information needed to make informed decisions is built into an encyclopaedia that players can consult at any point in play. Further information is presented as tips after each turn. In this way the game achieves education by stealth and players learn without even realising. In the process of managing their virtual forest it will also become apparent to players what works and what does not. It is hoped that the features of the game that make it like a simulation will give it application within the forestry sector. It has potential to enable people to learn about a range of pests and diseases and to appreciate the role of diversity in building resilience. Another motivator in the game is money. Players can fell trees to generate income in order to be able to plant more trees and deal with problems that arise, such as pathogens or herds of deer. In this way the game gives a sense of the commercial side of forestry and the balancing of issues like financial sustainability and environmental benefit. As there is potentially so much to consider at each turn this is not a fast and furious game. Instead, the game is played at the pace determined by the player. As long as there is money in the bank a player can continue making management decisions and will only advance to the next turn when no more decisions are needed or the money has run out. Today, a small number of scientists are battling a growing number of tree health problems. By combining gaming with education, to educate widely, as well as to encourage a new generation of plant health scientists, we can have hope for the future. CALEDON is free of charge and can be played online at and is also available for ipad on the App Store. [The Editor-in-Chief thanks Joanne Taylor for drawing this initiative to his attention.] Max Coleman (m.coleman@rbge.ac.uk) (6) IMA FUNGUS

9 Fleming Penicillium disc sold for US$ NEWS A disc of the Penicillium labelled by Alexander Fleming ( ) as part of the original culture that led to the discovery of penicillin in 1928 was sold by auctioneers Bonhams on 1 March 2017 for US$ as part of a sale of various Fleming artefacts (Anon 2017), which also included letters and photographs. Similar discs have been sold in the past, and it appears that he made several such medallions as gifts. This is not the most such a disc has reached in a sale, however, as Pfizer is reported to have paid for one in As might have been anticipated, the species was named as P. notatum in the sale particulars and P. chrysogenum in the report cited, the correct identification as P. rubens having been overlooked (Houbraken et al. 2011) here as in a recent book focussing on the penicillin story (Rosen 2017; see also pp. (31) (32) in this issue). Anon (2017) Mould money. Nature 543: 155. Houbraken J, Frisvad JC, Samson RA (2011) Fleming s penicillin producing strain is not Penicillium chrysogenum but P. rubens. IMA Fungus 2: Rosen W (2017) Miracle Cure: the creation of antibiotics and the birth of modern medicine. New York: Viking. Australian fungi photographer s footage in Planet Earth II Fungal photographer Steve Axford, based in northern New South Wales, Australia, has produced some stunning video time-lapse of fungi that appears in Planet Earth II, the BBC nature documentary narrated by David Attenborough. The fungal footage is included in the Jungle episode, which was Hygrocybe anomala at Minyon Falls, New South Wales. Image: Steve Axford, reproduced with permission. first aired late in Iconic fungi included in the Planet Earth II segment include Anemone Stinkhorn Aseroe rubra and the luminous Mycena chlorophos. You can see more of Steve s fungal photography at: smugmug.com/ and examples of video footage at: watch?v=zpcark689zu. Inclusion of fungi footage in Planet Earth was reported on the Australian Broadcasting Commission news and in the Australia Wide documentary series. See: planet-earth-fungi-photographer-stephenaxford/ Fungi have always struggled for equal time in nature documentaries, especially in relation to their diversity and ecological significance. No matter how beautiful static images are hard to sell in documentaries. Bringing motion to fungi introduces drama, and hence human interest. Steve s fungi in motion show dramatic changes from egg to expanded stinkhorn and from primordium to mature mushroom, while capturing the evanescent nature of many fungal sporophores. The video footage also reveals fascinating details for the developmental biologist (for example, in some agarics, a circling motion when pilei are viewed from above, as the sporophore expands). In addition, there is a cast of tiny invertebrates rushing around, taking nibbles from sporophores. A key aspect of achieving high quality fungi video footage is avoiding unwanted movement of the subject. Steve uses a fungarium where he places substrate with primordia in a moist environment, so that he can follow the production of sporophores in a still atmosphere. Relatively inexpensive digital video cameras are readily available. Mycologists are encouraged to create quality video, of both macrofungi and microfungi, and upload to video sharing channels, to give fungi the prominence they deserve. Perhaps one day we will be watching a whole series entitled Planet Fungi something that is long overdue! Tom W. May (tom.may@rbg.vic.gov.au) VOLUME 8 NO. 1 (7)

10 Rimaconus coronatus NEWS Westerdijk calendar 2018 June Visit for cultures, publications and courses Rimaconus coronatus asci Photographer: Dan Mahoney; American Optical H20 trinocular microscope with 35 mm film camera. Sunday Monday Tuesday Wednesday Thursday Friday Saturday Leading Women in Fungal Biology symposium (30 31 August) in Utrecht. To this end we invite all female mycologists making photographs or micrographs to submit their most beautiful fungal illustrations. Photographs of fungi cultivated in the laboratory, or observed in nature will be considered. Illustrations should be identified by the species name. Images should be in landscape layout, at least 300 dpi (3600 x 2400 px) and in full colour. The publication of the 2018 calendar is scheduled for August 2017 and the submissions for the 2018 calendar are welcome until 31 July, Show us your fungi! In April 2013 the Westerdijk Fungal Biodiversity Institute (then still CBS-KNAW) launched its new (12 month) fungal calendar series, focusing on the beauty of fungal biodiversity. The next calendar is scheduled for August 2017, and will be handed out at the Submissions can either be sent to p.crous@ westerdijkinstitute.nl or For larger files we recommend using dropbox or any other service that will allow you to share large files. STOP PRESS! IMA Fungus was issued with its first Impact Factor by Thomson Reuters while this issue was going to press. We are pleased to inform readers that this is 4.69, placing IMA Fungus fourth in the mycology category. Papers from all issues from the first in 2010 IF = are being incorporated into the Web of Science. We thank all contributors and readers for making this achievement possible. (8) IMA FUNGUS

11 Mycologists' committees strongly support changes to the governance of fungal nomenclature REPORTS At the 10 th International Mycological Congress (IMC10) in Edinburgh in 2010, 86 % of mycologists responding to a questionnaire favoured that the nomenclature of fungi should continue to be governed by the then International Code of Botanical Nomenclature provided that matters relating only to fungi were decided at International Mycological rather than International Botanical Congresses. A suggestion supported by 71 % was that the name of the Code be changed to make clear it covered fungi (Norvell et al. 2010). At the 18 th International Botanical Congress (IBCXVIII) in Melbourne in 2011, mycologists were instrumental in getting the name of the Code changed to the International Code of Nomenclature for algae, fungi, and plants (McNeill et al. 2012), but the issue of governance was referred to a Special Subcommittee on Governance of the Code with Respect to Fungi, which was charged with reporting to the next IBC to be held in Shenzhen, China, in July In the interim, 93.6 % (104:7:6) of those responding to a questionnaire distributed at IMC11 in Bangkok in 2014 agreed that decisions relating to fungal nomenclature should be voted at IMCs and not IBCs (Redhead et al. 2014). In line with these sentiments, a detailed and carefully considered set of proposals was published by the Subcommittee (May et al. 2016) and these are now to be voted on in Shenzhen. Essentially, the proposals replicate procedures for altering the Code already in place at the Nomenclature Section of an IBC at a proposed formal Nomenclature Session of an IMC, charged with dealing with matters in the Code solely related to fungi. The recommendations of the Special Subcommittee were supported by 80 % of the Subcommittee membership and also endorsed by the International Mycological Association (May 2016). Proposals made to an upcoming IBC are considered by the General Committee on Nomenclature (GC) appointed by the previous IBC, commented on by the Rapporteurs for the Congress, and a guiding mail vote is requested. Only individual members of the International Association for Plant Taxonomy (IAPT), members of Committees appointed by IBCs (e.g. the Nomenclature Committee for Fungi, NCF), and those who had made formal proposals are allowed to participate in the mail vote. The International Commission on the Taxonomy of Fungi (ICTF) was pleased to note that the NCF had supported the proposals of the Subcommittee (11:5:2 1 ), but disappointed that the GC had voted against them (8:14:3) (Turland & Wiersema 2017). The results of the mail vote are expected to be reported in the June issue of Taxon. Given the extremely strong support among the mycological community for changes to the governance of fungal nomenclature, mycologists expect that the proposals from the Special Subcommittee will be considered with due care and respect at the Shenzhen Nomenclature Section emphasising to the Section that the fungi governance proposals relate only to matters specific to fungi. Notwithstanding the expectation of a positive outcome for fungi governance at Shenzhen, following a telephone conference on 6 March 2017, the ICTF considered it prudent for mycologists to start to consider how to respond either to the proposals being accepted; rejected; or referred to another Special Committee. Special Committees established at an IBC report to the next IBC, which in this case would be in 2023 and with no assurance that any proposals made would be accepted at the next IBC. The ICTF therefore established an ad hoc Working Group to consider the various options. The Working Group comprised interested members of the Commission, with the addition of several other mycologists: Lei Cai (China), Pedro W. Crous (The Netherlands), Z. Wilhelm De Beer (South Africa), David L. Hawksworth (UK, Convenor), Kevin D. Hyde (Thailand), Paul M. Kirk (UK), Robert Lücking (Germany), H. Thorsten Lumbsch (USA), Tom W. May (Australia), Andrew Miller (USA), Amy Rossman (USA), Conrad Schoch (USA), and Keith A. Seifert (Canada). This report has been prepared following exchanges amongst members of the ad hoc Working Group, and provides a consensus of its opinions. There was overwhelming support for: (a) Remaining within the framework of the existing Code, especially in view of the change in title and the adoption of organism-neutral language wherever possible, agreed in 2011; (b) fungal-only matters being under the control of IMCs not IBCs; and (c) fungal-only material controlled by IMCs being presented as a separate section within the body of the Code. The optimum outcome is therefore that the fungi governance proposals (with the amendment introduced below) are accepted at Shenzhen (Option 1 in Box 1). Nevertheless, four other options were identified during discussions (Box 1). While a few felt the idea of a separate MycoCode attractive, it was felt there was much to be gained by continuing to draw on the expertise and support of the wider botanical, mycological and phycological nomenclatural community. The BioCode option (Greuter et al. 1998) could have much to commend it for the future, but could not be implemented until there were comprehensive lists of names at all ranks to be accepted as validly published. An amendment to the already published fungi governance proposals made here, 1 Ballots are reported in the form yes : no : abstain. VOLUME 8 NO. 1 (9)

12 REPORTS Box 1: Possible options for the future governance of the nomenclature of fungi (1) Separate out all fungal-only provisions into a separate chapter in the Code to be modified only by decisions at IMCs, with the proposals necessary to enact this being accepted at Shenzhen. (2) If the proposals in (1) are not accepted at Shenzhen, present the proposals to the next IMC and should they be accepted there, announce any new changes agreed at IMCs through IMA/ICTF with an instruction they are to be followed by mycologists. (3) Delete all non-fungal provisions in the Code to be adopted in Shenzhen and issue it as a separate publication with fungal rather than plant examples under a title such as the International Code of Nomenclature for Fungi (based on the International Code of Nomenclature for algae, fungi and plants). (4) Develop an independent MycoCode, based on the current Code, but not following its precise arrangement and numbering. (5) Adopt the organism-neutral BioCode developed by the International Commission on Bionomenclature. providing additional clarity, is to collect all material in the Code relating to fungi into a single section; this being the matter that will be controlled by IMCs. As IBCs are on a six-year cycle and IMCs one of four years, in some cases there would be two IMCs between IBCs; for example, IMC11 and IMC12 will be in 2018 and 2022, and the XX th IBC in Making changes in the body of the Code between IBCs could potentially become rather messy and confusing to mycologists, and the Working Group saw great advantage in having all material relating to fungi in a single section. This concept was originally proposed by Werner Greuter (General Committee, Germany) on 7 April 2017 during discussions of the proposals at the Botanical Garden and Museum in Berlin. It was noted that there is already a precedent for having a particular section in the Code devoted to particular groups of organisms in Appendix I dealing with the names of hybrids, and this and perhaps other organisms with special rules (e.g. fossils) might also be brought together in separate sections in the existing Code; perhaps better indicated as Chapters rather than Appendices. In conclusion, this ad hoc Working Group suggests that the Nomenclature Section of the XIXth IBC, meeting in Shenzhen on July 2017, might favourably view the following proposal to be made from the floor during the Congress as a clarifying amendment to the set of proposals already in place concerning fungi governance (May et al. 2016): The Section instructs the Editorial Committee to bring together all material relating only to fungi into a separate section or chapter within the Code, and that this section be subject to modification only by International Mycological Congresses operating as proposed by the Special Subcommittee on the Governance of the Code with Respect to Fungi. The material comprises pertinent Articles, Notes, and Recommendations: Articles 13.1 (d) (starting point date), 14.3 and 56.3 (lists of names for protection or suppression), 15 (sanctioning), 42 (registration), 57.2 (take up of names typified by asexual morphs), and 59 (pleomorphic fungi). Of these, there is strong support for Art to be removed at Shenzhen (Hawksworth 2016). The parts of Preamble 8, addressing what organisms are treated as fungi for nomenclatural purposes, would also be better moved to the start of the new section or chapter. Fungal examples given elsewhere in the Code would remain in their current positions (which mycologists would need to consult on matters not only related to fungi), as would mentions in the Glossary. Similarly, the fungal entries in the Appendices would remain; although, for lists of fungi generated through Art , there are various options for delivery as discussed by Wiersema et al. (2017). The Working Group considers that this outcome would be in the best interests of the nomenclature of fungi. The proposal above is regarded by the Special Subcommittee on Governance of the Code with Respect to Fungi as a friendly amendment to their formal proposals, agreed by eight of the ten voting members (being those who supported the original published proposals of the Subcommittee). The proposal has also been circulated to the complete membership of the ICTF which strongly support the Working Group s conclusion (18:2:1), and also to the Executive Committee of the International Mycological Association (IMA) 19 members of which (100 % of those responding) voted in favour. The ICTF ad hoc Working Group is indebted to Sandra A. Knapp (Chair of the Shenzhen Nomenclatural Section) and Nicholas J. Turland (Rapporteur-General to the Section; Germany) for constructive and frank discussions and advice. Greuter W, Hawksworth DL, McNeill J, Mayo MA, Minelli A, et al. (1998) Draft BioCode (1997): the prospective international rules for the scientific names of organisms. Taxon 47: Hawksworth DL (2016) Proposals to clarify and enhance the naming of fungi under the International Code of Nomenclature for algae, fungi, and plants. IMA Fungus 6: May TW (2016) Report of the Special Subcommittee on Governance of the Code with Respect to Fungi. Taxon 65: May TW, de Beer ZW, Crous PW, Hawksworth DL, Liu X, et al. (2016) ( ) Proposals to amend the Code to modify its governance with respect to names of organisms treated as fungi. Taxon 65: McNeill J, Barrie FR, Buck WR, Demoulin V, Greuter W, et al. (eds) (2012) International Code of Nomenclature for algae, fungi, and plants (Melbourne Code) adopted by the Eighteenth (10) IMA FUNGUS

13 International Botanical Congress Melbourne, Australia, July [Regnum Vegetabile No. 154.] Königstein: Koeltz Scientific Books. Norvell LL, Hawksworth DL, Petersen RG, Redhead SA (2010) The IMC9 Edinburgh Nomenclature Sessions. Mycotaxon 113: ; IMA Fungus 1: Redhead SA, Demoulin V, Hawksworth DL, Seifert KA, Turland NH (2014) Fungal nomenclature at IMC10: report of the Nomenclature Sessions. IMA Fungus 5: Turland NJ, Wiersema JH (2017) Synopsis of proposals on nomenclature Shenzhen 2017: a review of the proposals concerning the International Code of Nomenclature for algae, fungi, and plants submitted to the XIX International Botanical Congress. Taxon 66: Wiersema JH, May TW, Turland NJ (2017) Report on corrections and future considerations for Appendices II VIII of the International Code of Nomenclature for algae, fungi, and plants. Taxon 66: in press. Andrew Miller (Secretary, ICTF; amiller7@illinois.edu) Lei Cai, Pedro W. Crous, Z. Wilhelm De Beer, David L. Hawksworth, Kevin D. Hyde, Paul M. Kirk, Robert Lücking, H. Thorsten Lumbsch, Tom W. May, Amy Y. Rossman, Conrad L. Schoch, and Keith A. Seifert REPORTS Programme planning continues for IMC 11 In the previous issue of IMA Fungus (7(2): (57) (59), December 2016), IMA Vice President and Chair of the Local Organizing Committee of the 11 th International Mycological Congress, Sharon A. Cantrell, provided an overview and the venue for this conference, to be held in San Juan, Puerto Rico on July Over the past six months, co-chairs Chris Schardl and Donald H. Pfister, have been leading the Scientific Programme Committee in the development of the scientific programme for this marquis event of the mycological calendar. An outline of the scientific programme is presented here. Please place this meeting on your calendar as your priority event for 2018! Keynote Speaker Paola Bonfante The keynote speaker for IMC11 is Paola Bonfante, a Professor of Plant Biology, University of Turin, Italy. Well-known for her work on microbe-plant interactions, Paola is a pioneer in the study of arbuscular mycorrhizal (AM) fungi and a leading figure in studying bacterial-fungal interactions, playing a pivotal role in the discovery of intrafungal bacteria in AM fungi. Her plenary presentation at the European Conference on Fungal Genetics last year was outstanding in preparation, delivery and breadth, including overviews of her work on evolution, ecology, cell biology and molecular biology. A recent profile of Paola and her research (Trends in Plant Science 19: , 2014) provides a personal portrait of this dynamic and internationally known researcher. The title of her keynote speech will be Fungi, plants, bacteria: a network of dialogues and interactions Plenary Speakers The scientific programme for IMC11 is divided into eight subject themes (Applications, Cell Biology, Ecology, Education, Environment, Evolution, Genomics and Pathology). Each morning and afternoon of the conference will feature a plenary address. Our speakers are distinguished researchers with broad mycological expertise, often covering multiple conference themes. Winners of the Young Mycologist Awards (see pp. (16) (17) in this issue of IMA Fungus) will make short presentations before each keynote address. Russell Cox Applications Theme: Russell Cox, Professor in the Institute of Organic Chemistry, Leibniz Universität, Hannover, Germany. Research in the Cox group focuses on the discovery, understanding and engineering of biosynthetic pathways in fungi. Their work fuses organic chemistry, microbiology, molecular biology and enzymology and focuses on secondary metabolites from the polyketide, peptide and terpene families. The title of his plenary talk will be Heterologous expression of secondary metabolite biosynthetic gene clusters as a tool for understanding and engineering fungal natural products Cell Biology Theme: Jesús Aguirre, Institute of Cellular Physiology, Universidad Nacional Autónoma de México. Jesús works with the model fungi Aspergillus nidulans and Neurospora crassa, to approach questions related to stress signaling and cell differentiation. The title of his plenary talk will be ROS Signaling and fungal development Ecology Theme: Tom Bruns, Department of Plant and Microbial Ecology, University of Jesús Aguirre VOLUME 8 NO. 1 (11)

14 REPORTS California Berkeley, USA. Tom Bruns work is in fungal ecology, especially the ecology of ectomycorrhizal and post-fire fungi. The title of his plenary talk is Experimental fungal communities: tools for testing theory and determining mechanisms. Environment Theme: Matthew Fisher, Imperial College London, UK. The title of his plenary speech will be Big data approaches to addressing big fungal problems. Evolution Theme: Priscilla Chaverri, University of Maryland, USA and University of Costa Rica. The title of her plenary speech will be Evolution of protective mutualism in plantfungal endosymbiosis. Tom Bruns Matthew Fisher Priscilla Chaverri Genomics Theme: Chengshu Wang, Director of the Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China, and President of the Mycological Society of China. The title of his plenary speech will be From one to many: fungal genomics and the future of population genetics. Pathology Theme: Anuradha Chowdhary, Vallabhbhai Patel Chest Institute, University of Delhi, India. The title of her plenary speech will be Fungal human pathogens: from obscure significance to impending disasters. Symposia and workshops at IMC11 Chengshu Wang Anuradha Chowdhary The list of symposia and their organizers is now being finalized. The preliminary list below reflects the symposia approved by the Scientific Programme Committee of IMC11. It gives a first look at what promises to be a dynamic agenda for this congress. Additional late-breaking symposia will be added over the next several months to address timely or emerging issues of interest to mycologists. Applications Theme Food mycology in the 21 st century: impacts on food security and safety Home life: the mycobiomes of built environments Fungi and fungal enzymes for a more sustainable world Challenges in the exploitation of beneficial fungal secondary metabolites Applications and molecular aspects of mycoparasitic fungi Fungi as biocontrol agents for sustainable agriculture Cell Biology Theme Membrane dynamics in fungal cells Morphogenesis and invasion (Fungal-host interactions) Light sensing in fungi Fungal Sexual Development and Exploitation Biology of the fungal pigmentation: advances and perspectives of the study of melanin in fungi. Ecology Theme Bringing the dark taxa into the light - prospects and challenges (Original proposal: Can DNA sequences be used as the sole identifying criterion for naming species?) Hot fungi in hot spots in a hot region Resolving uncharacterized symbiotic relationships: the delicate balance from mutualist to parasite A big puzzle to assemble: using taxonomy to unravel ecology and biogeography of ectomycorrhizal symbiosis in the tropics Fungal-bacterial interactions and functions of the fungal metaorganism Fungal communities and the functioning of forest ecosystems Marine Mycology Experimental Approaches to the Conservation of Rare Fungi (12) IMA FUNGUS

15 Education Theme Teaching mycology around the world: examples from South America, North America, Europe, Japan and Australia Bringing awareness to fungi for teachers and the general public Oral history for mycology Mycology and Outreach Boosting Diversity in Mycology Environment Theme Rhizobiomes their interactions with the hosts and function in a changing environment Fungi in a changing environment Polyextremotolerant fungi in natural and urban extreme environments Evolution and diversity of lichenization in the Basidiomycota Ethnomycology: Scientists and Shaman on Historic and Current Uses of Fungi Evolution Theme Evolutionary genomics Gondwana reunited! Fungal biogeography in the Southern Hemisphere Early fungi that changed the world: phylogenomic and fossil evidence. Lichens on Islands: evolution, endemism, and conservation Species limits in the age of genomics. Integrative approaches to understanding the diversity and function of the Boletales Genomics Theme Integrative approaches to understand the ecology and evolution of fungi Fungal pan-genomes Metagenomics: whole fungal genomes from complex samples Fusarium: The genomics of functional and ecological diversity Expanding the taxonomic context of genome sampled fungi Pathology Theme Threatening fungal plant pathogens for tropical countries: acting before the foes arrive Deciphering fungi-archaea/ bacteria interactions for biocontrol of soil-borne pathogens Fungal extracellular vesicles Molecular mechanisms of human fungal pathogenesis Breeding for resistance to fungal pathogens of crops IMC/ISHAM Symposium: Human pathogenic fungi, taxonomy and global emergence In addition to the symposia, many Special Interest Group meetings and Workshops are being planned during IMC11. The list is not yet finalized, but proposals include sessions on Ascomycetes, Bacterial-fungal Interactions, Forest Pathology, Fungal Conservation, Tropical Plant Pathogenic Microfungi, and endophytic Xylariaceae. Keith A. Seifert is thanked for assistance in putting this information together for IMA Fungus. Chris I. Schardl, Donald H. Pfister, and Sharon A. Cantrell Rodríguez (chris.schardl@uky.edu) REPORTS International Commission on the Taxonomy of Fungi (ICTF) The International Commission on the Taxonomy of Fungi (ICTF) has been quite active the past year holding online meetings, organizing lists of competing genera, and pushing forward several issues significant to the mycological community. An online meeting was held on 11 October 2016 via Adobe Connect to discuss the upcoming International Botanical Congress in Shenzhen in July, progress on the various lists of competing names, DNA-based nomenclature, administrative structure of the ICTF, and synchronization of the three name repositories. The ICTF voted to approve the following motion: Should the three current repositories for the registration of fungal names synchronize their data (at least weekly) by July 1, 2017? Otherwise, the ICTF recommends that the NCF 2 reduce the official repositories to only those that are fully synchronized or a single repository of their choosing if no synchronization occurs by July 1, This was seen as an essential step for ending several years of stagnation and noncompliance among the three repositories and a key issue that the ICTF hopes the NCF will soon act upon. Another online meeting was held on 6 March 2017 to approve the Sordariomycetes excluding Diaporthales, Hypocreales and Magnaporthales Working Group and to form ad hoc groups that are currently working on five key action items: (1) An official ICTF statement on IMC/ IBC governance 3 (2) Adjustments to current statutes regarding memberships, member turnover, and new a Vice-President position (3) Reassessment of the 2013 Without prejudice list of generic names article (4) Manuscript on PubMed keywords for papers containing novel taxa (5) Guidelines for the use of DNA as types in fungi. In addition, the ICTF is to host or co-host two symposia and a debate/discussion at IMC11 in 2018: Symposia: Expanding the taxonomic context of genome sampled fungi 2 NCF = Nomenclature Committee for Fungi, which is currently appointed by the Nomenclature Section of each six-yearly International Botanical Congress. 3 See pp. (9) (11) in this issue. VOLUME 8 NO. 1 (13)

16 REPORTS Bringing the dark taxa into the light - prospects and challenges Debate/Discussion Can DNA sequences be used as the sole identifying criterion for naming species? Meeting recordings can be found at: meetings; Working groups at: fungaltaxonomy.org/subcommissions/; and Lists of publications at: fungaltaxonomy.org/lists Conrad L. Schoch, ICTF Chair Andrew N. Miller, ICTF Secretary (amiller7@illinois.edu) 2 nd Chinese Lingzhi (Ganoderma lucidum) Conference On 9 12 September 2016, the 2 nd Chinese Lingzhi (Ganoderma lucidum) Conference, jointly hosted by the Chinese Academy of Engineering (CAE), China Chamber of Commerce of Foodstuffs and Native Produce (CFNA), Mycological Society of China (MSC), Jilin Agricultural University ( JAU), National Strategic Alliance of Technological Innovation of Edible & Medicinal Fungi Industry (NSATIEMFI), International Society of Medicinal Mushrooms (ISMM), and the Peoples Government of Longquan City was ceremoniously held in Longquan, Zhejiang Province the hometown of Chinese lingzhi production. The 229 th International Conference on Economic Fungi & Forum of the Chinese Academy of Engineering on Engineering Science of Economic Fungi, initiated by Li Yu, Chinese academician of engineering, foreign academician of the Russian Academy of Science, and President of ISMM, was held at the same time. Ten Academicians of the Chinese Academy of Engineering in ten different areas, covering plant germplasm, pomology, and mycology, as well as scientists concerned with edible fungi were invited to the event for in-depth discussions on lingzhi. The invitees included Lin Zhibin (Peking University Health Science Center), David L. Hawksworth (Royal Botanic Gardens (14) IMA FUNGUS

17 REPORTS Kew, etc.), Anthony J. Whalley (Liverpool John Moores University), Patricia Wiltshire (University of Southampton), Kakishima Makoto (University of Tsukuba), Ian R. Hall (New Zealand), and Shoji Ohga (Kyusu University). Clear water, green mountains are wealth as great as a mountain was the development philosophy of the conference, with quality control and construction of system of standards for evaluation as the theme, and to enhance quality of lingzhi products, shape health industry of lingzhi as the goal. The latest developments and technology of the lingzhi industry were focused on in the conference, in combination with the superior ecological resources available in Longquan with a pollution-free cultivation model. This occasion not only informed the world on the achievements of the ecological cultivation technology of Longquan s lingzhi, of which the city is especially proud, but also aimed to assist in upgrading and enhancing a healthy lingzhi industry in China. Scientists, researchers, and leaders of the industry from around the world were attracted to the conference to enjoy as many as 40 presentations in which specialists shared the latest development and achievements in different aspects of lingzhi production. These included policies, laws and regulations, breeding and production, quality control of products, evaluation standards, research and development and deep-processing products, marketing, brand building, and promotion. The 1 st Chinese Lingzhi Conference was in 2015, and the 2 nd again incorporated additional activities including: signing of a business, investment and talent introduction project; celebrating high school student winners in a poster competition; an advisory meeting of academicians and specialists for the strategic development of Longquan county s economy; an edible fungi resource utilization and fungal foray arranged by the country s technological system for edible fungi; a visit to the Meidi original ecological lingzhi cultivation base in Lanju Town, Longquan, the only ecological lingzhi farm in a forest in Zhejiang Province; the Meidi Lingzhi Culture Exhibition Hall; the Dendrobium officinale farm in Xijiezhou Village; and the opening ceremony of the Chinese Edible Fungi Trading Center in Lucai. At an award ceremony incorporated in the opening session of the conference, plates with the engravings Fungal Foray Base of the International Medicinal Mushrooms, Education and Practice Base of Jilin Agricultural University, Research and Education Base of the Engineering Research Centre of Chinese Ministry of Education for Edible and Medicinal Fungi, Demonstration Zone for Cultivation in Original Ecology of Chinese Lingzhi, and Longquan Service Station for Zhejiang Edible Fungi Association were presented to participating organizations. This was the third international forum on engineering related to mycology initiated by Li Yu, the first two being an International Top-level Forum on Engineering Science and Technology Development - Medicinal Mushrooms held during the 7 th International Conference on Medicinal Mushrooms in Beijing in 2013, and the 186 th Forum of the Chinese Academy of Engineering on Engineering Science Systematics and Ecology of Myxomycetes held during the 8 th International Conference on Systematics and Ecology of Myxomycetes held in Changchun in Chang- tian Li (changtianli@126.com) VOLUME 8 NO. 1 (15)

18 AWARDS AND PERSONALIA AWARDS Call for nominations for IMA Fellows 2018 IMA Fellows are mycologists who have made an outstanding contribution to the advancement of mycology at an international level, through service to the IMA, its Regional Committees, organization of international meetings or initiatives, or otherwise, as the Award Committees deems appropriate. The first nine IMA Fellows were elected at IMC10 in Bangkok (see IMA Fungus 5: (46), 2014), and nominations are now solicited. These may be made by individual mycologists or Regional Mycological Organizations (RMOs). Those from individuals should be sent to IMA Past-President Meredith M Blackwell (mblackwell@lsu.edu) who has been appointed by the IMA President to Chair the Awards Committee, while those from RMOs should be submitted to the IMA President, Keith Seifert (keith.seifert@agr. gc.ca). In both cases nominations should arrive by 1 December Nominations should include: (1) a letter of nomination and one supporting letter that addresses the stated criteria for nomination; (2) a curriculum vitae of the nominee; and (3) if available reference to web sites or additional material appropriate to support the nomination. Please incorporate all materials into a single document. The protocols relating to the appointment of IMA Fellows have been previously published (IMA Fungus 4: (40), 2013) but are presented here for convenience with key dates updated. Protocols for Nomination and Selection Eligibility for selection. To be eligible for nomination as IMA Fellow, a member of the IMA must, at the time of the International Mycological Congress where the award is to be made, have completed at least 11 years of service after the award of a PhD degree. [I.e. for IMC11 in 2018, a nominee must have received a PhD before 2007.] Numbers. As many as 12 IMA Fellows may be selected for induction at each IMC, one from each IMA Regional Mycological Member Organization, and as many as six from the IMA in general. Committees for Nomination and Selection (1) General IMA Fellows Award Committee. The IMA President shall appoint senior and distinguished mycologists to an IMA Fellows Committee consisting of at least two members and a chairperson. (2) IMA RMMO IMA Fellows Committees. Each IMA Regional Member Mycological Organization shall appoint senior and distinguished mycologists to an IMA RMMO Fellows Committee consisting of at least two members and a chairperson. Responsibilities of the IMA Fellows Committees (1) Timing of events. The Committees shall arrange for a call for nominations to be published in IMA Fungus in June of the year preceding the IMC with a deadline for receipt of nominations by December of that year [i.e. 1 December 2017]. The committees will vote to select the fellow(s) and report the results to the IMA President by February of the year of the IMC so that the new IMA Fellows can be notified in time to attend that year s IMC. (2) Nomination of candidates. The nominator shall submit, by 1 December of the year before the IMC [i.e. 1 December 2017], to the IMC Fellows Committee, a letter of nomination that addresses the criteria for selection, a current curriculum vitae for the candidate that contains information relating to the criteria for selection, and two supporting letters from mycologists familiar with the candidate. The IMC Fellows Committee shall distribute the nominations to the appropriate RMMO Fellows Committees by 5 December of the year before the ICM [i.e. 5 December 2017]. (3) Selection of candidates. Each IMA RMMO Fellows Committee shall, by 1 February of the year of the IMC [i.e. 1 February 2018], select one fellow from among their pool of nominees and forward to the IMA Fellows Committee the nomination material for all of the nominees for ratification of their choice by the IMA Fellows Committee. The IMA Fellows Committee shall select up to six fellows from their pool of nominees and report all selected Fellows to the IMA President before March 1st of the year of the IMC. Induction of IMA Fellows The IMA Fellows shall be inducted by the President of the IMA at the International Mycological Congress. IMA Young Mycologist Awards 2018 To honour the accomplishments of those who are the future of our field, the IMA Executive Committee initiated IMA Young Mycologist Awards in Six awards were made for IMC9, and six for IMC10 (see IMA Fungus 5: (47 48), 2014). The next Young Mycologist Awards are to be made at IMC11 in 2018, and aim to promote mycology by helping to advance young mycologists. The Awards for IMC11 will be presented by the President of the IMA at the Closing Ceremony of the Congress, and be accompanied by a certificate. The recipients will have the registration fee for the IMC waived. The protocols relating to the appointment of IMA Fellows have been previously published (IMA Fungus 2: (18) (19), 2010) but are reproduced here for convenience with key dates updated. The IMA established six IMA Young Mycologist Awards to mark outstanding research accomplishment by young mycologists from each of the six IMA Regional Mycological Organizations (RMOs). (16) IMA FUNGUS

19 Ethel Mary Doidge Medal - African Regional Mycological Member Organization Keisuke Tubaki Medal - Asian Regional Mycological Member Organization Daniel McAlpine Medal - Australasian Regional Mycological Member Organization Elias Magnus Fries Medal - European Regional Mycological Member Organization Carlos Luis Spegazzini Medal - Latin American Regional Mycological Member Organization Arthur Henry Reginald Buller Medal - North American Regional Mycological Member Organization Eligibility of at least two members (although the appointment of more members to represent the diversity of mycology in the region is encouraged) and a chair, who will serve from one International Mycological Congress to the next. To ensure a broad pool of nominees, nine months prior to the next International Mycological Congress, this committee will notify the members of the IMA Regional Mycological Member Organization of the award, solicit nominations and vote to select a candidate. To ensure a breadth of experience on the IMA Young Mycologist Award Regional Committee, the chair and members of this committee should be senior mycologists with distinguished records and should represent the diversity of Mycological Member Organizations and Sustaining Mycological Member Organizations from the region. The chair will vote only in the event of a tie vote by the other members. Responsibilities of the Award Regional Committees candidate, and a current curriculum vitae of the candidate. Candidates are expected to have contributed appreciably to mycology and they should have achieved international recognition based on several criteria: (a) The quality, innovation, thoroughness, and impact on science of their published research, with consideration given to the contribution of the nominee to multiauthored publications. (b) Service as editors of journals or as officers of societies. (c) Membership on national or international policy committees. (d) Invitations to present research at national or international meetings. (3) The committee will forward the nomination material for each nominee and the committee s choice for the award to the IMA Young Mycologist Awards Committee convened by the IMA President no more than one month following the deadline for nominations and no less than five months prior to the next IMC. The IMA Young Mycologist Awards Committee is charged with ratifying the choices made by the IMA Young Mycologist Award Regional Committees. AWARDS AND PERSONALIA To be eligible to be nominated for an IMA Young Mycologist Award, a member of the IMA must, at the time of the next International Mycological Congress, be within ten years of his or her PhD degree i.e. for IMC11 in 2018, a nominee must have received his or her PhD degree no earlier than Establishment of Award Regional Committees The President of each IMA Regional Mycological Member Organization is asked to establish an IMA Young Mycologist Award Regional Committee consisting (1) Nine months prior to the next International Mycological Congress and three months prior to the deadline for receiving nominations, members of each IMA Regional Mycological Member Organization shall be notified of the call for nominations for the relevant IMA Young Mycologist Award through the IMA website and the IMA Regional Mycological Member Organization website and through s sent to delegates from the region who attended the preceding International Mycological Congress. (2) Nomination of candidates for each IMA Young Mycologist Award shall consist of a letter of nomination, two letters of support from mycologists familiar with the The timeline for the IMA Young Mycologist Awards for IMC 2018 can be summarized as follows. The notification to the RMOs of the call for nominations shall be nine months prior to the IMC [i.e. 15 October 2017], the deadline for receipt of nominations by the IMA Young Mycologist Award Regional Committees shall be six months prior to the IMC [15 January 2018], and the receipt of the IMA Young Mycologist Award Regional Committee s choice by the IMA Young Mycologist Awards Committee shall be five months prior to the IMC [15 February 2018]. In this way, recipients can be notified of their award at least four months prior to the IMC to encourage their attendance at the IMC. BIRTHDAY GREETINGS Michael Corlett Asco specialist turns 80 IMA Fungus extends warm congratulations to Michael ( Mike ) Corlett in celebration of his 80 th birthday on 28 April Mike is an ascomycete specialist who spent his career with Agriculture & Agri-Food Canada (AAFC) as part of the mycology group now at the Ottawa Research and Development Centre. Mike was a student of John F. Morgan-Jones at the University of Toronto, much of whose research concerned ascomycete ontogeny. This tradition was continued by Mike, whose early studies focused on the development and ultrastructure of plant pathogenic fungi, and his work with VOLUME 8 NO. 1 (17)

20 AWARDS AND PERSONALIA AAFC progressed to the taxonomy of plant pathogenic Pleosporales and other bitunicates. His compilation of published names in Mycosphaerella (Mycologia Memoirs 18, 1991) was a milestone for researchers on this genus. Mike was an associate editor for the Canadian Journal of Plant Pathology and responsible for the series Fungi Canadenses. In the last few years of his career, Mike turned his attention to Alternaria, work that has become increasingly relevant as regulatory attention turns to mycotoxins produced by these moulds. Since his retirement in 1999, Mike has been a volunteer in Egon Horak Macromycete doyen the National Mycological Herbarium (DAOM), assisting with the digitization of data for legacy specimens, especially of hyphomycetes. Keith A. Seifert (keith.seifert@agr.gc.ca) Our best compliments on your 80 th birthday. We are pleased to see you still in the best of health and creative powers, and to be able to share your interest in the latest findings of molecular taxonomy helped by your steady kindness and especially your profound knowledge of mushroom morphology to find links with classical taxonomy. Egon is one of the last cosmopolitan taxonomists with a true global view of agarics, based on his own observations and personal collections. Innumerable field trips have brought him to almost all the exciting corners of our planet where new species could be expected. Once, he travelled together with Rolf Singer ( ) and Meinhard Moser ( ) to South America, and later with local mycologists many times to South-East Asia and North America. Among all the many places with a little known agaric funga, one country plays a very special role for him: New Zealand. Fascinated, Egonhas explored all parts of this country and described many new species. His special interest in the identification and description of New Zealand s fungi is still vividly evident in an extending list of major publications. Egon started his career as a student of Moser in Innsbruck, Austria. One of his first research projects concerned alpine macrofungi in a glacier forefield. This special interest in alpine agarics has been maintained throughout the years. Only recently he was involved in a study aiming to clarify the taxonomy of some alpine Inocybe species as part of a transatlantic co-operation. The position as a curator of the fungal collections at ETH in Zürich, which he held for most of his active tenure has been the perfect base for his aim to describe the whole range of species diversity in agarics. As a professor he has guided several students who focussed mostly on ecological aspects of macromycetes. And, most importantly, he Jack Rogers Mr Xylariaceae turning 80 gave them all the freedom they required to develop their own academic ideas. Egon is well-known for his precise line drawings of morphological details and his search and scrutiny of type specimen in order to clarify the taxonomy. We would like to see all these relevant drawings in a large reference book. Beatrice Senn-Irlet (beatrice.senn@wsl.ch) Jack David Rogers retired in January 2013 from Washington State University, Pullman, after 50 distinguished years on the WSU Faculty. He received his BS from Davis & Elkins College, West Virginia, and his MF from Duke University, North Carolina. He then moved to the University of Wisconsin where he was awarded a PhD for research on Hypoxylon pruinatum (later Entoleuca mammata), a major forest pathogen of quaking aspen in the Great Lakes States, under the supervision of John Berbee. This triggered a keen interest, and later a passion, for Xylariaceae. On his appointment as Assistant Professor at the WSU faculty in 1963, with positions in the Departments of Plant Pathology and Forestry and Range Management, he continued with his studies of Xylariaceae, at first mainly on chromosomes in Hypoxylon and later researching all aspects of the family. In his remarkable and distinguished career, Jack, together with a number of his many postgraduate students, established WSU as the world centre for studies on Xylariaceae. He was also largely responsible for developing the Mycological Herbarium at WSU making it a renowned worldclass facility for educational and research purposes. His mycological contributions and teaching skills have been recognized by numerous awards and honours over the (18) IMA FUNGUS

21 years from WSU, the Mycological Society of America, and the British Mycological Society. Jack was President of the MSA in 1977 and his Presidential Address The Xylariaceae: systematic, biological and evolutionary aspects delivered at Tampa in 1977 during IMC2 demonstrated his clear thoughts and knowledge of the family and is a must-read for anyone with an interest in these fungi. If Julian Miller were alive today he would have been delighted and impressed with A revision of the genus Hypoxylon by Yu-Ming Ju and Jack D. Rogers. Jack Yu-Ming and San Martin have now revised 14 genera of the family and together with other former students and colleagues published numerous papers on all aspects of Xylariaceae. I am privileged to have known Jack since the mid-1970s, and to have visited him and his family in Pullman on a number of occasions and to have received them at our home in North Wales. During his trips we visited the type locality for Nemania chestersii in Anglesey, collected Hypoxylon rutilum in Llanbedr and Euepixylon udum by Aber Falls, Gwynedd; all rare species.. We have enjoyed many hours discussing various aspects of Xylariaceae especially host specificity, habitat preferences, and biogeography. Considering Pullman is hardly tropical, Jack has a broad and detailed knowledge of the tropical species owing to his impressive global network of correspondents, collaborators and former students. He presented the Benefactors Lecture of the British Mycological Society at the Millenium Meeting of the Society at Liverpool John Moores University in April 2000 on Thoughts and musings on the Xylariaceae. This was not just an inspiring lecture, but provided a wonderful opportunity for the postgraduate mycology students as Jack gave generously of his time to discuss their work with them and to offer good expert advice. Jack has a wonderful sense of humour and, together with his wife Belle, they have endeared themselves to all who know them. He enjoyed hunting and fishing whenever he had the opportunity and the 17 lb Steelhead caught on the Clearwater River, Idaho in November 1982 (shown here) is true testimony to his success. The respect and esteem for Jack Rogers is deservingly demonstrated by his promotion to Regents Professor with the Eminent Faculty award, the highest honour a faculty member could receive at WSU. Mycologists worldwide thank you for your invaluable contributions and we all wish you a very happy 80 th birthday this 3 September Anthony J. Whalley (a.j.whalley@ljmu.ac.uk) AWARDS AND PERSONALIA IN MEMORIAM Martha A. Christensen ( ) Martha (left) with Ellen Hoekstra (centre) and Amelia Stolk (right) at the 2 nd International Penicillium and Aspergillus Workshop in Photo: Keith Seifert. Martha Christensen, a great mycological ecologist, died on 19 March 2017, at the age of 85. She is survived by her brother, a nephew and two nieces, three grandnieces, and one grandnephew. Martha s major contributions to science has been in two areas of mycological research, fungal ecology, especially concerning forest and grassland soils, and taxonomy, of the genera Aspergillus and Penicillium as focus points, but being an ecologist, she also enjoyed and worked with many other genera. She also established a very important and large collection of soil-borne fungi, emphasizing the importance of citing typical exemplars in her publications and maintaining them in a freeze-dried state for other researchers to explore. Even though an ecologist by heart, she realized how important it was to be able to give correct species names to all the microfungi she found, and so she devoted much time to identification, fungal systematics, and the description of new species from soil. On a personal level, she was a very warm outgoing person and she enthusiastically repeated cite-worthy expressions from anyone talking with her, and always had a lot of wise sentences from her own mouth or citations from people she admired. She had many friends, and either went fungus hunting or bird-watching, but one of her great additional interests was in classical music. Martha s PhD thesis from the University of Wisconsin was published in 1960 (Christensen 1960) and followed by an excellent publication in ecology (Christensen 1969). The fungi isolated were kept in her WSF (Wisconsin Soil Fungi) collection. She soon moved to the University of Wyoming in Laramie, and stayed there for most of her career becoming a professor, latterly emerita. She loved all the very interesting, beautiful and extremely different habitats in Wyoming (the Rocky Mountain Fungi (RMF) collection), and in addition she loved to travel, and sampled soil microfungi, for example in India, Namibia, Fair Isle (Scotland) and many other places (the World Tour (WT) Collection). She donated all these collections to the Westerdijk VOLUME 8 NO. 1 (19)

22 AWARDS AND PERSONALIA Fungal Biodiversity Institute (formerly Centraalbureau voor Schimmelcultures, CBS), as it was very important for her that other ecologists and taxonomists could compare their cultures to her large collection of soil fungi. A very important part of Martha s career was the sabbatical with Kenneth B. Raper ( ) in Wisconsin. When Raper was driving to the airport to meet her, Martha was quietly asking about Emericella, and Raper said in a very loud voice EMERICELLA, here we are using Aspergillus. Indeed, in the description of Aspergillus spectabilis (Chistensen et al. 1978), the taxon was headed by this sentence: Aspergillus spectabilis Christensen & Raper, sp. nov. status perfectus sit judicatus Emericella spectabilis Christensen, sp. nov. Raper simply did not want to describe any Emericella. Indeed, in the monograph on Aspergillus (Raper & Fennell 1965) he wrote in hand as part of a dedication to Martha: Up with Aspergillus, down with Neosartorya. So even though Martha applied the correct nomenclature at the time, she was happy to learn that in the one fungus one name system from 2011 onwards both Emericella and Neosartorya are now back in Aspergillus, commenting It would have pleased Dr Raper. Martha made many important contributions to especially Aspergillus and Penicillium taxonomy, and was very enthusiastic whenever an ecologically relevant taxonomic character was used. She also started discussions in technocoenosis, or organisms in domesticated landscapes. Martha was President of the Mycological Society of America in 1988, where she wrote a very fine presidential address (Christensen 1989; cited 123 times as of March 2017), and she received, among other prizes, the prestigious Johanna Westerdijk Award from CBS in Three species have been named in her honour: Penicillium christenseniae, P. marthae-christenseniae and Aspergillus christenseniae. Martha was lucky to have several very fine collaborators, including Ken Raper, Jack S. States and Dorothy E. Tuthill, and she also liked to discuss fungal ecology with her friends Juliet Frankland, and Sally Gochenaur. She was active also when she went back to the University of Wisconsin, and established new friendships there. She will be missed by mycologists, because of her great knowledge, intense interest in fungi and their ecology, and warm approach to other people. Christensen M (1960) The Soil Microfungi of Conifer-hardwood Forests in Wisconsin. PhD thesis. University of Wisconsin, Madison. Christensen M (1969) Soil microfungi of dry to mesic conifer-hardwood forests in northern Wisconsin. Ecology 50: Christensen M (1989) A view of fungal ecology. Mycologia 81: Christensen M, Raper KB, States JS (1978) Two new Aspergillus nidulans group members from Wyoming soils. Mycologia 70: Raper KB, Fennell DL (1965) The Genus Aspergillus. Baltimore: Williams & Wilkins. [See also IMA Fungus 4(1): (14) (2013).] Jens C. Frisvad (jcf@bio.dtu.dk) K. Walter Gams ( ) With great sadness, we inform readers of IMA Fungus of the passing of Walter Gams, a frequent contributor to these pages and mentor and colleague to many mycologists around the world. A polymath in the true sense of the word, Walter had expert knowledge in many fungal groups, was equally conversant with classical botany, fluent in many languages (for several generations of mycologists, he was the actual author of many of the Latin diagnoses accompanying new taxa), and widely respected for his insight into nomenclature. Born in Austria on 9 August 1934, Walter spent his early academic years with Klaus Domsch, with whom he wrote his first book Fungi in Agricultural Soils 1957, and then in 1980 the classic Compendium of Soil Fungi. Most of his career was spent at the Centraalbureau voor Schimmelcultures in Baarn (now the Westerdijk Institute in Utrecht), where he specialized in hyphomycetes and zygomycetes, writing his famous book on Acremonium in 1971, and culminating in Walter Gams waiting with colleagues celebrating his 80 th birthday with a tour of the Chayophraya River in Bangkok Thailand at IMC10. his collaboration with Seifert, Morgan- Jones and Kendrick on The Genera of Hyphomycetes published in 2011 after more than 20 years of effort. Walter had many students, and endowed a nonprofit foundation in Germany to support Research in mycological taxonomy and ecology, which provided financial support for many young mycologists to attend conferences, pay page charges, or participate in collecting trips. Walter travelled and taught widely and had friends all over the world, especially in Australia, Austria, the Czech Republic, Germany, Iran, Italy, and Japan. Hundreds of guests visited his home in Baarn, hosted by his wife Sophia and his two daughters Hede and Hilde. Following his retirement, he spent part of each year north-west of Rome in an apartment that was part of the hill top castle of Bomarzo, Italy, where he passed away on 9 April [Further information on Walter and his achievements can be found in IMA Fungus 3(1): (26) (2012) and 5(1): (13) (2014).] Keith A. Seifert (keith.seifert@agr.gc.ca) (20) IMA FUNGUS

23 Jack R. Laundon ( ) Lichenologist Jack Rodney Laundon was born at Kettering, Northamptonshire, UK, on 28 July He developed an interest in lichens while at school, and without any university training obtained a position at what is now The Natural History Museum London in 1952, remaining there until his retirement in He was especially fascinated by sterile crustose lichens, producing keys to those on the ground and bark, and produced detailed treatments of Chrysothrix, Leproloma, and Lepraria. A life member of the International Association for Plant Taxonomy (IAPT), Jack endeavoured to apply the then International Code of Botanical Nomenclature strictly, with major papers on Withering s neglected lichen names in 1984 and those described by James E. Smith, founder of the Linnean Society of London. Familiar names were often changed in a time species name conservation was not permitted; something not welcome with replaced names coming to be referred to as having been jacked. He was a major contributor to The Lichen Flora of Great Britain and Ireland (Natural History Museum, 1992), and with his wife translated the lichen text of H. Martin Jahns Collins Guide to the Ferns Mosses and Lichens of Britain and North and Central Europe (1983). He also prepared a popular well-illustrated book Lichens (Shire Natural History, 1986). He was a founder member of the British Lichen Society, editing its Bulletin from , serving as Secretary from , and becoming President for Jack was not, however, only a taxonomist and will also be remembered for his studies on lichen sociology and Vadim A. Mel nik ( ) changes over time in response to the effects of sulphur dioxide air pollution. In 1956 he started a meticulous survey of London s lichens, which resulted in detailed papers in 1967 and 1970 documenting species loss; the distributions of several species correlated with mean sulphur dioxide isopleths. Further, his studies in churchyards led to the recognition of relict lichen communities; species persisting on older memorials but unable to colonize newer ones. After sulphur dioxide levels fell markedly in the city, species started to return, and amongst his last publications was one in 2012 on lichens invading the city. Sadly, Jack was made redundant from the Museum during a restructuring when just 56 years old, but fortunately was able to continue to work from home and to use the libraries at the Royal Botanic Gardens Kew. He moved back to his home town of Kettering following the death of his wife in 2003, and died there following a long illness on 31 December [An obituary is in press in the Lichenologist and expected to appear later this year.] AWARDS AND PERSONALIA On 10 April 2017, Vadim Aleksandrovich Mel nik died in hospital about two months after a fall in the street. He lived a very busy life, fully devoted to mycology, rich in events and scientific discoveries, and he became one of the most authoritative world experts in the taxonomy of conidial fungi. Vadim, whose ancestors included the Baltic German von Lange s, was born on 16 March 1937 in Daugavpils (Latvia). He graduated from the Leningrad Forestry Academy in 1960 and then worked as a forestry engineer near Leningrad before joining the Komarov Botanical Institute in St Petersburg (then Leningrad) in He remained connected with the Institute for the rest of his life, progressing from PhD student to leading researcher. His candidate dissertation of 1966 concerned the Parasitic imperfect fungi of the forests of the Leningrad region, and his doctoral thesis of 1986 concerned the Coelomycetes of the USSR. He investigated conidial fungi in many regions of Russia from the north-west European sector to the Far East, giving special attention to protected areas and the republics of the former USSR. In addition he travelled widely for discussions and fieldwork, taking in Austria (2000, 2002), Turkey (2001), South Africa (2002), Sweden (2003), Korea (2003, 2004), China (2004, 2005), and Lithuania ( ). The samples he collected, also during short-term visits to the UK, Germany, Latvia, and Canada, and material from other mycologists which he identified are deposited in the LE-Fungi collection; amongst these are those of A. E. Kovalenko (countries of the Pacific basin), M. A. Bondartseva (Cuba), Yu. K. Novozhilov (Costa Rica and Puerto Rico), A. V. Alexandrova and E. S. Popov (Vietnam), and S. Stephenson (Mexico and New Zealand). Serving as curator of the fungal collections for 15 years, Vadim was active in the publication of Mycotheca Petropolitana, and he also contributed specimens to several exsiccatae including Microfungi exsiccati (Munich), Mycotheca Gracensis (Graz), and Fungi selecti exsiccati (Halle, Germany). Other specimens of his are deposited in CBS (The Netherlands), K(M)-IMI (UK), BILAS (Lithuania), and MSK (Belarus). He was active in Russian and international mycological organizations and events, acquiring friends and making scientific VOLUME 8 NO. 1 (21)

24 AWARDS AND PERSONALIA contacts in many countries. In 1994 he was able to attend IMC5 in Vancouver, where he was elected to the Executive Committee of the IMA; in 2002 the XL Congress of the Society of Phytopathologists of South Africa (Dikholo, Republic of South Africa); and in 2007 the XV Congress of European Mycologists. His scientific work resulted in more than 220 publications, including The determination of fungi of the genus Ascochyta Lib. (1977; translated into English in 2000), Imperfect fungi on trees and shrubs (1992), a summary of the cercosporoid fungi of Russia and neighboring countries (1997), and the Micromycetes volume of the Fungi of the Nizhne-Svirsky Reserve (1996). He also contributed to the ongoing Handbook of Fungi of Russia, with accounts of numerous rare and little-known genera of coelomycetes (1997) and dark-coloured hyphomycetes (2000). He also published in foreign journals in co-authorship with key world mycologists including Brian C. Sutton, T. R. Nag Raj, Uwe Braun, David W. Minter, Kevin D. Hyde, Ovidiu Constantinescu, Rafael Castañeda, and Pedro W. Crous. In total, he described three new genera, and over 70 new species, as well as making numerous new combinations. Vadim also served as editor of more than eight mycological books, and from the first issue of Micologia y Fitopatologia in 1967 he worked closely with the journal for the last 50 years, reviewing numerous submitted manuscripts. Further, at various times he was a member of the editorial boards of several international mycological journals. His endless devotion to science was widely appreciated by the scientific community, he was one of the first to be awarded the A. A. Yachevsky Medal in 2012, and had several genera and species named in his honour. He found the aesthetic pleasure of a true scientist, observing, and recording with the help of a microscope and his remarkable drawings the morphological forms of conidial fungi. When he discovered something new and interesting, the whole laboratory and colleagues in other institutions were immediately aware of this. He shared with us not only his knowledge and experience, but also his energy, Eliyathamby Punithalingam ( ) enthusiasm, and love of specimens. In recent years, when it became difficult for him to go on expeditions himself, he concentrated on material collected by colleagues in Vietnam, usually issuing detailed instructions on what substrates and where to look, how best to select them, and how to transport them. He waited with great impatience for expeditions to Vietnam to return with fresh samples of substrates (wood, dry leaves and fruits), enthusiastically and joyfully analyzing them. Vadim generously shared with us, and many other Russian students and researchers, not only his knowledge and experience, but also his energy, enthusiasm, and great love for his fungi. He will be sorely missed. Alexander Kovalenko, Yuri Novozhilov, Mark Levitin, Alina Alexandrova, and Mikhail Zhurbenko (alkov_@mail.ru; yurinovozhilov@gmail. com; mark_levitin@rambler.ru; alinaalex2011@yandex.ru; zhurb58@gmail. com) Born in Sri Lanka, Puni (as he was fondly known by his colleagues), studied at Jaffna College in Sri Lanka from which he secured a place to read for a PhD at the Imperial College University of London Field Station at Silwood Park, Ascot. He studied the Septoria diseases of chrysanthemum there from under the supervision of Ronald K. S. Wood and Brian E. Wheeler. In 1965 he joined the then Commonwealth Mycological Institute at Kew as an Assistant Mycologist, with a particular responsibility for the identification of coelomycetes that could be cultured, and which had been sent to the Institute for an opinion from mycologists and plant pathologists around the world. He was a meticulous researcher and excellent microscopist, and pioneered the use of Giemsa stain not only to visualize fungal nuclei and even chromosomes, but delicate conidial appendages that had often been overlooked or misinterpreted by others. Amongst some 190 publications, the most familiar are his monograph of plant diseases attributed to Botryodiplodia theobromae ( J. Cramer, 1980), and his Mycological Papers providing critical accounts of Ascochyta species on grasses (1979) and monocots, cryptogams and gymnosperms (1988). A superb illustrator, Puni also made many contributions to the Institute s loose-leaf Descriptions of Fungi and collaborated with others in helping them describe new coleomycetous fungi. He also had an important role in checking identification reports for the whole Institute; no small task when around identifications were made each year and all had to be prepared on manual typewriters. Following his retirement in 1995 from the now International Mycologicial Institute, which had re-located from Kew to Egham in 1993, Puni was appointed as a Research Associate in the Mycology Section of the Royal Botanic Gardens Kew a situation that enabled him to continue his research and maintain contacts with other mycologists. With a strong attachment to the Institute and its staff, he also instigated an annual reunion of past and present staff at Pembroke Lodge in Richmond Park, the next being on 14 June He died in St George s Hospital in London on 23 December 2016 after suddenly being taken ill, and was repatriated by his family to Sri Lanka for a traditional Hindu funeral. Always conscious of the troubles and violence in his home country, he made provision in his will to establish a library in his home town. I feel privileged to have been able to learn from Puni and to have had the opportunity to collaborate with him, and will always be grateful for the guidance and encouragement he gave me in my early years at the Institute after I joined it in (22) IMA FUNGUS

25 Anjali Roy ( ) On 22 January 2017, we lost Anjali Roy, a mycologist from India. Her 86 years was full of accomplishments, with contributions to the growth of science, especially the domain of her specialization, mycological studies. Born in Rajsahi, Bangladesh in preindependent India, she matriculated from Rajsahi Girls School in In an era when not many girls proceeded to higher education, she went on to graduate from Presidency College, Calcutta, with an honours degree in Botany. She endured the turmoil and transition from a colonial country to an independent nation, to complete her postgraduate studies in 1952 from Ballygunge Science College, University of Calcutta, where her doctorate was guided by S. N. Banerjee. Her zeal for study continued, leading her to the award of the degree of DSc. Her level of commitment as a young female in pursuing and sustaining a higher education trajectory, established her as an icon, even for the contemporary generations. Her search for knowledge led her to a post-doctorate position in Canada under the guidance of Mildred K. Nobles ( ) and work on the genus Coriolellus. She developed a particular interest in wood-rotting polypores, and dedicated herself to their taxonomic study based on morphology, anatomy, type of rots, culture characteristics, sexuality, and responses to chemical tests. Later, biochemical aspects attracted her attention, working with her twin sister Arati Das, a scientist at the Bose Institute in Calcutta. She went on to contribute to several emerging sectors of mycology, including as medical mycologist in the Department of Medical Mycology, School of Tropical Medicine, Calcutta. Her keenness to disseminate knowledge amongst the science community and inspire youngsters was evident in her publications, teaching, and academic responsibilities, including publishing a book for degree students with her sister. She was appointed lecturer at the University of Burdwan, West Bengal, in 1974, and in 1979 joined Visva Bharati University, Santiniketan, also in West Bengal, working there until her retirement in She mentored 10 PhD students, who benefitted from her international exposure and work experience in some of the finest laboratories of her times, such as Hubertus A. van der Aa ( ) the Royal Botanic Gardens Kew, and the Academy of Sciences in Tartu. Amongst her approximately 150 research publications, the Polyporaceae of India issued in 1996, has been an immense contribution to the field. This was coauthored with her student A. B. De, who named the polypore genus Royoporus in her honour. She inaugurated and self-funded a database of aphyllophorales fungi from India, the Indian Aphyllofungal Database (IAD), which was launched on www. fungifromindia.com in Her work earned her the appreciation and goodwill from the Indian as well as the international community of polypore scholars, and she interacted and was encouraged throughout her life-time by leading specialists of the day, including Erast Parmasto, Jose Wright, Leif Ryvarden, Robert Gilbertson, Karl-Henrik Larsen, Jim Ginns, and many more. In a period when interest in classical mycology is losing its sheen in many countries, Roy s journey of academic explorations is a gentle reminder to invest in systematic explorations of the rich fungal biota that surrounds us. For those who have known her, she will always be an inspiring figure, while for those who have not come into direct contact, her works will leave indelible impressions of appreciation and deep respect. Asit Baran De and Kiran Ramchandra Ranadive (abde.brc@gmail.com; ranadive.kiran@ gmail.com) AWARDS AND PERSONALIA In IMA Fungus 6(2): (53) (54) (2015) we paid tribute to Huub on the event of his eightieth birthday, and now we are sad to report that he passed away on 7 May An influential coelomycete expert and gifted teacher, Hubertus Antonius van der Aa was born on 5 July 1935 in Tilburg, The Netherlands. In 1965 he was appointed as mycologist at the then Centraalbureau voor Schimmelcultures (CBS) in Baarn, to become an expert in coelomycetes. Josef Adolf von Arx ( ), who had been appointed as the Director of the CBS shortly before, stimulated Huub s interest in plant pathogenic fungi. Soon Huub began his taxonomic studies of Phyllosticta, a waste-basket genus for pycnidial fungi in leaf-spots with simple hyaline conidia. In 1973 he published his thesis and first monograph of the genus Phyllosticta (van der Aa 1973). Applying his new generic concept, Huub continued to revise thousands of type specimens. He published the Revision of the species described in Phyllosticta, together with his Bulgarian colleague and friend Simeon Vanev in Huub has also been recognized for his major contributions on Phoma, in collaboration with Gerhard Boerema and co-workers of the Plant Protection Service in Wageningen. He (co-)published numerous papers on Phoma, and also contributed to the Phoma Identification Manual: differentiation of specific and infra-specific taxa in culture (Boerema et al. 2003). Many of us who followed the CBS Course of Mycology in Baarn will remember Huub, who led this annual course for many years with great passion. He had that special gift as a teacher, able to stimulate the interest for fungi in young students, impressing them with his broad knowledge of fungal taxonomy and ecology. Besides his professional focus, Huub took an interest in many subjects, like teratology of basidiomes and plant galls inflicted by fungi and insects. Huub was VOLUME 8 NO. 1 (23)

26 AWARDS AND PERSONALIA also a gifted writer and loved (old) books. He served as Treasurer of the IMA from 1983 until 1990, and received a royal decoration in 2014 for his contributions to the Baarn community and many years of service to the Dutch Mycological Society (NMV), of which he was an Honorary Member. Aa HA van der (1973) Studies in Phyllosticta I. Studies in Mycology 5: Larissa N. Vassilyeva ( ) With great sadness we note the passing of Russian mycologist Larissa N. Vassiljeva, an amazing collector and specialist in pyrenomycetes, on 23 February Larissa was born in Kursk in western Russia on 15 February From she was a student specializing in mycology at Leningrad State University. In Soviet times students were placed on a job after graduation and they could choose where they would like to have their future position. Larissa chose the Russian Far East because that region was especially interesting for mycological investigations. From August 1972 she worked in the Institute of Biology and Soil Science in Vladivostok, Siberia as a scientific researcher, and since 2002 as a principal researcher. For about ten years she studied the pyrenomycetes and loculoascomycetes of the Magadan Region, Kamchatka and Chukotka, publishing the results of these investigation in her first book Pyrenomycetes and Loculoascomycetes of the North of Russian Far-East (Vassilyeva 1987). For this work and others she received the degree of Doktor nauk (DSc) in For three years Larissa lectured on the Theory of Evolution at the Far Eastern State University, now regarded as the Far Eastern Federal University, Vladivostok. However, she decided that studying fungi was more interesting than teaching about them. Larissa had a single post-graduate student, Hai-Xia Ma, now at the Institute of Tropical Bioscience and Biotechnology in Haikou, China. Some of her major publications are listed below. She loved to collect fungi, wandering through the woods humming softly to herself, specializing in minute ascomycetes perhaps partly because she was shortsighted allowing her to see clearly the small black dots of pyrenomycetes about 500 μm diam scattered along a dead branch. She would pick up a likely substrate, pull off her thick-lensed glasses and examine the stick without a hand-lens because, basically, she had a built-in handlens for her eyes. In addition she had a sense of the biology of these fungi, and thus knew where to look for them. She was fearless in her ability to endure hot and cold weather in search of her beloved pyrenomycetes. While working in Beltsville, Maryland, during very hot humid summers, she would asked to be driven to the forest to collect and then would walk the five or so miles back to the laboratory. She served as an officer in the International Mycological Association s Committee for Asia ( ), and was a member of the Mycological Society of Japan (since 1995), the Mycological Society of America (since 1997), and Russian Academy of Sciences. We will miss the many interesting and unusual microfungal specimens that she willingly shared with others, and her sometimes unconventional and thoughtprovoking ideas about relationships among these fungi. Castlebury LA, AY Rossman, WJ Jaklitsch, LN Vassilyeva (2002) A preliminary overview of the Diaporthales based on large subunit nuclear Aa HA van der, Vanev S (2002) A Revision of the Species described in Phyllosticta. Utrecht: Centraalbureau voor Schimmelcultures. Boerema GH, de Gruyter J, Noordeloos ME, Hamers MEC (2003) Phoma Identification Manual: differentiation of specific and infra-specific taxa in culture. Wallingford: CABI Publishing. Gerard Verkley (g.verkleij@westerdijkinstitute.nl) ribosomal DNA sequences. Mycologia 94: Vassilyeva LN (1973) Die Blatterpilze und Rohrlinge (Agaricales) von Primorsky Region. Leningrad: Nauka. Vassilyeva LN (1987) [Pyrenomycetae and Loculoascomycetae in Soviet Far East.] Leningrad: Nauka. Vassilyeva LN (1988) A new treatment of the family Valsaceae. Systema Ascomycetum 7: Vassilyeva LN (1998) Plantae Non Vasculares, Fungi et Bryopsidae Orientis Extremi Rossica. Fungi. Vol. 4. Pyrenomycetidae et Loculoascomycetidae. Petropoli: Nauka. Vassilyeva L[N] (1999) Systematics in mycology. Bibliotheca Mycologica 178: Vassilyeva LN, Nazarova MM (1991) Plantae non Vasculares, Fungi et Bryopsida Orientis Extremi Sovietici. Fungi. Vol. 2. Ascomycetes, Erysiphales, Clavicipitales, Helociales [sic!]. Leningrad: Nauka. Amy Rossman, Alexey Chernyshev, and Steven Stephenson (amydianer@yahoo.com) (24) IMA FUNGUS

27 Mycelial-like 2.4 billion-year-old fossils RESEARCH NEWS Vesicle (to left) with boom-like aggregations of filaments (upper left), and branched and anastomosing filaments (centre and right) as revealed by SRXTM surface/volume renderings. Photo courtesy: Stefan Bengtson. Most fossils from the Pre-Cambrian that were claimed to perhaps be fungal have proved to be artefacts or otherwise dubious (Hawksworth 2015, Taylor et al. 2015). Some more convincing remains of mycelium-like structures have, however, now been reported from a 2.4 billion-year-old basalt from the Palaeoproterozoic of the Pre-Cambrian in South Africa (Bengston et al. 2017). The basalts were submarine and have vesicles often connected by veins evident in thin sections, and the detailed structures have been elucidated by tomographic microscopy (SRXTM) surface/volume renderings. Networks of hyphal-like threads are visualized, some with anastomoses, branches, and loops; measurements are given as 2 12 μm wide. No definite spore-like structures were discovered, but it is speculated that some bulbous protrusions of 5 10 μm diam might actually be spores. It cannot be categorically asserted or denied that these structures are fungal, but there can be little doubt that they are mycelium-like. No reference is made to septa, but none are evident in the photographs. The jury is likely to remain out for some time as to the nature of these remains, and the possibility that they could represent extinct lineages has to be borne in mind. Nevertheless, the presence of some early fungi or fungal ancestor was to be expected in Cambrian or Pre-Cambrian rocks if it is assumed that the principal fungal phyla had all become wellseparated by the Devonian. Bengston S, Rasmussen B, Ivarsson M, Muhling J, Broman C, et al. (2017) Fungus-like mycelia fossils in 2.4-billion-year-old vesicular basalt. Nature Ecology and Evolution 1: Hawksworth DL (2015) Lichenization: the origins of a fungal life-style. In: Recent Advances in Lichenology (Upreti DK, Divakar PK, Shukla V, Najpai R, eds) 1: Springer. Taylor TN, Krings M, Taylor EL (2015) Fossil Fungi. Amsterdam: Elsevier Academic Press. Secrets of the antibiotic resistant Candida auris revealed There is particular concern amongst medical mycologists of the threat to human health posed by a species of Candida resistant to whole classes of antifungals and that has been found in hospitals. This yeast, C. auris, was first described from the ear of a woman in Japan as recently as 2009, but has now been involved in invasive infections in nine countries with over 30 patients being involved in some hospital outbreaks. There is a particular problem in that it can form drug resistant biofilms on, for example, catheters. VOLUME 8 NO. 1 (25)

28 RESEARCH NEWS Biofilms formed by Candida albicans and C. auris strains as revealed by confocal scanning laser micrography, top down 3D (A-C) and side (D-F) views. Adapted from Larkin et al. (2017). Photos courtesy Mahmoud Ghannoum. A major investigation into the biology, virulence, and drug resistance of this species has now been published by Mahmoud Ghannoum s research group at the Center for Medical Mycology, Case Western Reserve University and University Hospitals Cleveland Medical Center in Cleveland, Ohio (Larkin et al. 2017). Working with 16 strains from a wide range of countries, they tested the efficacy of 11 drugs belonging to different antifungal classes. Most were ineffective although there were some differences between strains. The drugs used, however, included a novel 1,3-β-Dglucan synthase inhibitor (SCY-078) which is orally administered and completely inhibited growth of both this species and C. albicans. The drug operated by interrupting cell division, which was demonstrated by scanning electron microscopy, and also reduced the thickness of biofilms formed compared with controls. The group also explored what virulence determinants were expressed in connection with the resistance shown to fluconazole and amphotericin. Growth on different media was similar to that of C. albicans, except that no chlamydospores were formed. Further evaluation of SCY-078 for the routine treatment of C. auris infections appears to be warranted. Larkin E, Hager C, Chandra J, Mukherjee PK, Retuerto M, et al. (2017) The emerging pathogen Candida auris: growth phenotype, virulence factors, activity of antifungals, and effect of SCY-078, a novel glucan synthesis inhibitor, on growth morphology and biofilm production. Antimicrobial Agents and Chemotherapy 61 (5): e Cantharellus species concepts in Europe clarified Cantharellus cibarius. Photo: Gabriel Moreno. While the chanterelles are amongst some of the most recognizable and sought-after mushrooms in Europe and North America, agreement on species recognition has often been hard to achieve. An amazing 30 names have been applied to the European species, and the application of some has been far from clear in the absence of a Cantharellus pallens. Photo: Gabriel Moreno. modern molecular systematic approach incorporating sequences from type material. Olariaga et al. (2017) have carefully typified and/or epitypified the names proposed and carried out a new analysis using sequences from ITS2, nr DNA LSU, RPB2, and TEF- 1 genes. A set of 117 specimens was then used for DNA extractions, and a subset of 53 of these representative of the whole range of phylogenetic diversity in Europe was then studied in more detail. Just eight species could be supported in a well-structured combined tree, one of which was newly described (Cantharellus roseofagetorum). In contrast, several rather recently recognized species were relegated to synonymy (e.g. C. henrici, C. lourizanianus). Meticulous morphological studies were carried out and selected ones mapped onto the phylogenetic tree. The result was not a set of taxa that could not be recognized by field mycologists, as has been the result of studies on some other mushroom genera, but the discovery of a series of characters that could be used to separate the species without recourse to DNA sequencing. (26) IMA FUNGUS

29 The colour of the pileus when young, and the colour of the young hymenium were found to be particularly informative, and when supplemented by the coating, staining, and spore shape characteristics enabled a traditional dichotomous key to be provided. When re-circumscribed in the manner proposed here, there were also some clear geographical differences, such as the predominance of C. cibarius in northern Europe and of C. pallens in southern Europe. Further, some species were ectomycorrhizal only with deciduous trees (e.g. C. friesii) and others primarily with evergreen mediterranean oaks (e.g. C. alborufescens) or conifers (e.g. C. cibarius). Detailed descriptions and colour photographs of the accepted species are provided, along with drawings of the basidiospores. It is a pleasure to see such a work clearly prepared with field mycologists and mycophagists in mind, and all who wish to identify these fungi, whether for commercial, gastronomic, or scientific purposes, will find this paper invaluable. Olariaga I, Moreno G, Manjón JL, Salcedo I, Hofstetter V, Rodriguez D, Buyck B (2017) Cantharellus (Cantharellales, Basidiomycota) revisited in Europe through a multigene phylogeny. Fungal Diversity 83: Reservoirs of Batrachochytrium salamandrivorans infection RESEARCH NEWS Reservoirs and sources of infection of salamanders (Salamandra salamandra) by Batrachochytrium salamandrivorans. Reproduced from Fisher (2017). In addition to the devastation caused in frogs worldwide by Batrachyochytrium dendrobatis, an additional species pathogenic to salamanders, B. salamandrivorans, was described in 2013 (see IMA Fungus 4 (2): (48) (49), December 2013). This fungus is now known to have entered Europe from Asia around 2010, and is exceptionally virulent being able to kill more than 96 % of salamanders that are infected. Stegen et al. (2017) critically monitored infections in salamanders in a forest in Belgium, just 57 km from the initial outbreak site in Europe. Infections were first noted in this site in 2014, and within two years under 1 % had escaped infection and were still able to travel around the forest. In order to understand the epidemiology of this disease, Stegen et al. carried out a series of infection trials and also endeavoured to find reservoirs from which the fungus could attack. They discovered that survival of the fungus outside salamander bodies was aided by the formation of encysted zoospores, something not seen in B. dendrobatis. These spores float and are quickly picked up by not only salamanders, but also toads and waterfowl, the waterfowl being capable of spreading the disease over wide areas. DNA analysis of soils also showed that the zoospores could persist there. In addition, motile zoospores in the water are able to be carried to salamanders by newts which were also adversely affected by the disease. As noted by Fisher (2017), the combination of multiple reservoirs and an ability to persist in the environment leads to a highly infected ecosystem. It is difficult to see how such infections can be combated in the wild, and there may be a need for what he terms amphibian arks to safeguard vulnerable species. Fisher MC (2017) In peril from a perfect pathogen. Nature 544: Stegen C, Pasmans F, Schmidt BR, Rouffaer LO, Van Praet S, et al. (2017) Drivers of salamander extirpation mediated by Batrachochytrium salamandrivorans. Nature 544: Tissue types in lichen cortices Elegant microscopic methods can facilitate the elucidation of how tissues develop in complex fungal structures. Many groups of foliose lichens have well-developed cellular upper cortices, but the extent to which these parallel the cellular tissues found in plants has been uncertain. For example, are terms such as parenchyma appropriate? A true parenchyma in plants has cells subdivided from adjacent cells by cross walls adjoining older cross walls. Sanders & Rios (2017) used transmission electron microscopy (TEM) to determine just what was the case in the cortices of three foliose lichens from different families, Endocarpon pusillum (Verrucariaceae), Leptogium cyanescens (Collemataceae), and Sticta canariensis (Lobariaceae). They paid particular attention to the wall layers and found that the newer wall layers were not continuous with the older ones, the older ones tended to have amorphous electron-dense deposited in them and the newer layers increasingly welldelineated. Septal pores connecting adjacent VOLUME 8 NO. 1 (27)

30 RESEARCH NEWS Upper cortex of Sticta canariensis, chloromorph, paradermal section in TEM. Photo: W. B. Sanders and A. de los Rios. cells did, however, persist. The resultant tissue in these cases is considered comparable to true parenchyma in the unrestricted orientation of the cross walls separating the cells, and the relationship of the cells to their neighbours. The authors do, however, recognize that earlier stages in the ontogeny of the tissues involve the coalescence of filamentous hyphae and subdivision into cell-like compartments, often termed a pseudoparenchyma, in which the filamentous origins are evident in microscopic preparations. The cells in the lichen cortices studied are relatively thin-walled and rounded with no indication of a hyphal origin and so are interpreted as a parenchyma rather than a pseudoparenchyma. The lichen parenchyma is consequently homologous with plant parenchyma cells, and apomorphic (i.e. a derived character state) rather than plesiomorphic (i.e. an ancestral character state). This study shows the care that is needed to unequivocally categorize the tissue types in fungi from a developmental standpoint. It also suggests prudence in applying terms that have an implied ontogenetic meaning, and so the pragmatism of using the tissue types system (i.e textura globosa, textura intricata, etc) describing what is seen with the light microscope with no implied ontogenetic overtones. Sanders WB, de los Rios A (2017) Parenchymatous cell division characterizes the fungal cortex of some common foliose lichens. American Journal of Botany 104: Oldest fossil agaric discovered Fungi which are composed primarily of soft tissues, such as most agaric mushrooms, could be expected to only rarely be preserved in the fossil record, and that proves to be the case. An exception is where they become encased in resin, and so survive as inclusions in amber, as with the Myr-old Gerontomyces lepidotus reported on last year (see IMA Fungus 7 (2): (66), December 2016). Now a much older agaric has been discovered not in amber but in laminated limestones from Brazil from the Lower Cretaceous dating from Mya. This new mushroom is rather small, with a cap of just 10 mm diam borne on a stipe of 24 mm. It is named as Gondwanagaricites magnificus, and is clearly a member of Agaricales as gills are evident, but a family placement is not suggested in the absence of basidiospores, which were not revealed even by scanning electron microscopy. This is not just a curiosity, however, as this fossil provides a new and earlier calibration point for Agaricales for use in molecular clock phylogenies. Heads SW, Miller AN, Crane JL, Thomas MJ, Ruffatto DM, et al. (2017) The oldest fossil mushroom. PLoS One 12 (6): e Gondwanagaricites magnificus (holotype). Photo courtesy: Danielle Ruffatto. (28) IMA FUNGUS

31 Brewing Microbiology: current research, omics and microbial ecology. Edited by Nicholas A. Bokulich and Charles W. Bamforth Caister Academic Press, Norfolk. Pp. v + 331, figs. ISBN (pbk), (ebk). Price US$ 319 or 159 (pbk or ebk). Genomics is having an enormous impact on traditional as well as emerging fungal-based industries. Brewing had already became an increasingly scientific operation during the 20 th century, but genomics is now taking it to a new level of sophistication. This book aims to provide an overview of recent advances in brewing technology and their impact, particularly in the last decade. The topics covered range from physiology and handling, through genetic modification, taxonomy and evolution, to issues of contamination and spoilage. New technology has made this an exciting and developing field, and also revealed aspects that prove to be imperfectly understood. Knowledge of basic physiology has improved, for example in relation to effects of nitrogen, oxygen, and sugar levels on growth, and the complex issue of factors controlling quiescence after cropping. The considerable stresses that yeasts undergo during brewing processes are reviewed, including changes in alcohol levels, ph, temperature, carbon dioxide and oxygen, and hyperosmotic stresses. Maintaining strain quality over time is of vital importance in production to achieve a consistent product, and best-practices for propagation, storage and rejuvenation are described. Molecular phylogenetics has led to a clarification of species concepts, and the relationship between seven natural species and hybrids used in production or developed as contaminants. There is a most useful summary of how the names of 11 taxa relate to the currently accepted species (including the citation of type strains [sic!]). In the case of Saccharomyces cerevisiae, a whole-genome phylogeny of 114 top-fermenting beer strains is presented, including ones used in bread, sake, and wine production and some from nature in different regions of the world. The nomenclature and taxonomy of Brettanomyces species, also used in some beers, is also summarized. Methods of species separation are also discussed, including real-time PCR systems. A separate chapter considers the evolution of brewing yeasts in both these two genera, especially domestication and the characters associated with that which have diverged from those found in nature. In the case of traditional beers, however, inoculations often rely on back-slopping or other non-critical methods. It does, however, have to be understood that the particular strains in use in major manufacturing plants are often closely guarded by the companies because that information is commercially sensitive. As a result the available laboratory strains may not always be representative of those actually used. Also covered are the problems and potentials of genetic manipulation of brewing yeasts, and the contamination of barley and malt by a surprising variety of spoilage fungi that can lead to significant losses. Spoilage can also arise from bacteria, and omics approaches to their detection and characterization are discussed, along with the problems that can cause, whether from mycotoxins or unpleasant odours. I learnt a great deal from this volume, and there is no doubt that this will be a valuable information source not just for those involved in the brewing industry, but for those in applied mycology and food science courses. The editors are to be congratulated on putting together such an authoritative overview of brewing yeasts and their exploitation. BOOK NEWS Radical Mycology: a treatise on seeing & working with fungi. By Peter Mc- Coy Chthaeus Press, Portland, OR. Pp. xxi + 672, illustr. (16 pp. col.). ISBN (pbk). Price US$ What an incredible work! The back cover states that Radical Mycology aims to be an in-depth reference and resource manual for anyone interested in the growth of mycology as a people s science. The author has a long-standing fascination in the cultivation and collection of mushrooms, and became deeply involved in the art and activist movement in Olympia, and now lives in Portland (OR). In 2008 he started a zine when he just a few friends interested in mycology which developed into a worldwide network of contacts, Radical Mycology Congresses, and in 2014 a Radical Mycology Collective touring North America sharing knowledge and skills. This book is the summation of his life s work with fungi, bringing together all the information he has gleaned into a single reference work and with almost no contact with professional mycologists or mycological societies. He sees mycology as a neglected megascience (citing my use of the phrase), crucial to the new millennium, and groups of Radical Mycologists Without Borders touring the world informing on the importance of fungi so that they are seen as having a central role in all aspects of human life. The book starts with basic information on the nature of fungi and major groups, structure, spore liberation, hyphal growth, and form, proceeding to life-styles: mycorrhizas, endophytes, relationships with animals, biotrophs, and saprobes. Sections on ethnomycology and the role of women VOLUME 8 NO. 1 (29)

32 BOOK NEWS conclude the first part. Part II concerns collection and examination, with a welcome chapter on lichens (by Natassja Noell) which in addition to identification and chemistry also includes uses. Part III covers mycomedicinals 1 and mycophagy, including recipes for not only food and beverages but also various medicines. Part IV has a 90-page chapter on working with fungi, including cultivation and culture techniques, and a shorter one on cultivation on waste materials, composting (including the use of Trichodema), fungus farming, and incorporating fungi with different biologies into garden design. This is followed by one looking to the importance for the future, particularly in bioremediation of soil contaminants (including petrol spills), stressing the need to have a standard of care, and the range of chemical products. Part V, Integration, unconventionally, turns to the incorporation of mycelium parallels into various activities, including searching for information, navigating human situations, decision making, building connections and support networks. There are also separately authored sections on sharing fungi with children (Maya Elson) and mushroom sex life, including sexual orientation (Willoughby Arevalo). This is followed by a 44-page chapter devoted to psychoactive fungi, covering different categories of experiences, the War on Drugs in the USA, historical uses in different regions, in religions, an assessment of Gordon Wasson s investigations, and the psychedelic movement (Peter SjoXstedt-H). The last sections comprise species profiles, and appendices covering id forms, lists of endangered fungi, fungal toxins, fungal dyes and paper, healing exercises, cultivation parameters, media for cultivation, cultivation tracking forms, projects, working with psychoactive fungi, psilocin mushroom niches, facilitating events and meetings, online resources and organizations, various games, mycojokes, Latin and Greek derivations, an extensive glossary, bibliography, and endnotes to sources used in each chapter. I was impressed by the range of sources the author has been able to put together, including many from the primary literature. I must admit that I had not heard of the Radical Mycology movement before coming across this book. Some views expressed and slants placed on particular facts may surprise, or even shock, some professional mycologists, there can be no doubt about the enthusiasm for mycology that is evident on almost any page. Weighing 1.9 kg, this is very much a source book, a vademecum for a mycological movement. Consequently, some areas such as industrial mycology, food spoilage, genetics, genomics, pharmaceuticals, plant diseases, indoor fungi, mycoses, etc, are not or scarcely mentioned. It is a remarkable and impressive achievement by a citizen scientist and a few colleagues, and is indeed likely to promote the importance and knowledge of fungi amongst an audience not normally reached through conventional mycological channels. Mycology should be grateful to him. 1 Various other terms are introduced here, perhaps for the first time, including: alchemycology, mycognosis, mycoguilds, mycomemetrics, mycomimicry, mycopsychology, and mycosystematics (which has nothing to do with taxonomy as used here). Fungi: applications and management strategies. Edited by S.K. Deshmukh, J.K. Misra, J.P. Tewari, and Tamas Papp CRC Press, Boca Raton. Pp. ix ISBN Price US$ 127, This is the fourth book in the Progress in Mycological Research series, and the first to focus on applied aspects; the previous one concerned fungi from different substrates (Misra et al. 2014). The new volume deals with some aspects of fungi which have helped us to use them for human welfare (p. v). The 17 chapters included represent a choice that is necessarily somewhat eclectic, and are mainly concerned with bioprospecting for, and pharmaceutical and industrial applications of, fungal products. The topics cover searching for endophytic fungi, with one a case study in Cameroon using plants used by local people as a target, antimycobacterials, antifungals, volatile organic compounds, statins, anti-plant viruses, myconanofactories, mycoremediation, mycotoxins, nutritional values of some wild edible fungi from India, and uses of thermophiles. There is an intriguing chapter on epigenetic modification of biosynthetic pathways. Just two chapters concern management issues, fungal pathogens on rice and on soybean. The concept of a myconanofactory was a new one for me, fungi that can extract metal ions (including platinum and silver) to form minute particles on their walls which promise to have applications in medicine as well as in bioremediation. Individual mycologists will have their own favourites, but I particularly valued the chapter on statins as this was the clearest exposition of their discovery I had seen. The volume is well produced, and I was pleased to see that the so unnecessary author citations of fungal names were omitted from almost all chapters, with an unfortunate lapse in that on edible mushrooms that even has them in the abstract. The editors are to (30) IMA FUNGUS

33 be congratulated on a job well-done, and providing a further set of papers that will be of value to those teaching applied mycology courses. It will be interesting to speculate what a fourth volume in the series might contain! Misra JK, Tewari JP, Deshmukh SK, Csaba Vágvölgyi C (2014) Fungi from Different Substrates. Boca Raton: CRC Press. Bioprospecting: success, potential and constraints. Edited by Russell Paterson and Nelson Lima Springer, Cham. Pp. x + 303, illustr. (some col.). [Topics in Biodiversity and Conservation Vol. 16.] ISBN (hbk), (ebk). Price 117 (hbk), (ebk). BOOK NEWS Although not only concerned with mycology, this title is included here as it is edited by two mycologists and includes several chapters dealing with fungi. Bioprospecting for potentially exploitable pharmaceuticals from natural products reached its zenith in the 1990s, but has since been largely abandoned by major pharmaceutical companies, Big Pharma, in favour of combatorial chemistry. This has come about as a result of the increasing realization that it was both long-term and expensive, compounded by increasing legal and bureaucratical constraints consequent on the adoption of the UN Convention on Biodiversity in 1992 and subsequent Protocols. These fundamental issues are addressed in the first two chapters. Eleven chapters look at particular examples, of which four are devoted entirely or partly to fungi. These concern the role of microbial resource centres ( J Overmann and D Smith), the Iwokrama fungal/plant project (R Pingal), Brazilian fungi ( J V Braga De Souza et al.), and mine waste fungi (A A Stierle and D B Stierle). Included in Overmann & Smith s overview, is an innovative proposed business plan designed to make well-characterized strains availed to industry in a legally compliant manner; the scheme would, however, require substantial additional funding to be made available to the collections. The Iwokrama project in Guyana ran from and was funded by the European Commission; the outcomes are frankly analysed and may inform any future similar ventures. In Brazil, a variety of initiatives are underway, focussing on endophytic and soil fungi. Mine wastes have yielded fungi with several novel products of potential importance as inhibitors of enzymes causing inflammations associated with some cancers. The book is well-edited and produced, but as is an increasingly common situation, it frustratingly lacks an index. Miracle Cure: the creation of antibiotics and the birth of modern medicine. By William Rosen Viking (Penguin Random House), New York. Pp. ix + 358, illustr. ISBN (hbk), (ebk). Price US$ (hbk), US$ (ebk). This is not just yet another book on some aspect of the discovery and utilisation of penicillins, but a meticulously researched investigation with numerous notes and an extensive bibliography and one which is also enthralling to read. The author s experience as an editor and publisher at Macmillan, Simon & Schuster for almost 25 years shows. He acknowledges it as a little ironic that its completion was only possible through the products of the pharmaceutical industry as he developed a particularly aggressive cancer; he died in April 2016 without seeing the book published. While much of the subject matter will be familiar to mycologists with an interest in the history of the discovery of penicillin in particular, it also presents new insights into how Fleming s observations were transformed into a miracle drug, and the often antagonistic stances taken by some of the key players. As Fleming s accounts of his discovery vary, he favours the suspicion that the accidental mould contamination may have actually been a part of an experiment searching for a new source of the enzyme lysozyme he had discovered in 1922, and that it was the Staphylococcus that was the contaminant. The possibility that the Penicillium came from La Touche s laboratory on the floor below Fleming s room is surprising not mentioned; the study with that hypothesis (Hare 1970) is missing from the numerous sources and 16 page bibliography. There had been reports of antibacterial properties of Penicillium by earlier authors as far back as at least the observations of John Tyndall published in1876. What Fleming did was to realise that there was a phenomenon to explore although he did not explore possible curative effects even in mice or foresee the possible value to human health. His 1929 paper attracted almost no notice until about 1937 when it came to the attention of Ernst Chain and Howard Florey in Oxford; they later both claimed to have seen and realised its significance before each other. They planned to search for antibacterial compounds from diverse microorganisms, including some strains of Penicillium notatum already held in their lab. Methods to grow the fungus in VOLUME 8 NO. 1 (31)

34 BOOK NEWS bulk were devised by Norman Heatley, and Chain elucidated the chemical structure. In1940 the first successful tests on diseased mice were carried out, and in 1941 it was used to treat an Oxford policeman with amazing results. Unable to secure adequate facilities in the UK, Florey visited the USA and established links there which included Charles Thom and later Kenneth Raper. New strains were isolated by the USDA Northern Regional Laboratory; one from cantaloupe from a Peoria market giving the best yields and facilitated mass production. In 1942 interest on both sides of the Atlantic took off. Fleming s boss, Almroth Wright, published a letter in The Times claiming Fleming should have the credit and that has remained the case in the public media to this day. The author goes on to describe production and arguments over patents, involvement of pharmaceutical companies, issues of supply and legal battles over pricing, and later discoveries. Amongst the latter, particular attention is given to the discovery of streptomycin by bacteriologist Selman Wakaman (who coined the term antibiotic ) and his team, chloromycetin, and the thalidomide disaster. He also points out that it is extremely difficult to find new antibiotics, and none working by different methods have been discovered in the last 60 years. At the same time, antibiotic resistance is increasing, with some deaths attributed to resistant bacteria each year. And the major drug companies have abandoned the search for pharmaceuticals in natural products (Borris 2017). The style makes it a difficult book to put down, and it also includes many historic photographs I had not seen reproduced before. I enjoyed it very much, and unhesitatingly commend it as my top choice for vacation reading by all mycologists this summer. Borris RF (2017) Bioprospecting: an industrial perspective. In: Bioprospecting: success, potential and constraints (Paterson R, Lima N, eds): [Topics in Biodiversity and Conservation vol. 16.] Cham: Springer Nature. Hare R (1970) The Birth of Penicillin and the disarming of Microbes. London: George Allen & Unwin. Mycorrhiza function, diversity, state of the art. Edited by Ajit Varma, Ram Prasad, and Narendra Tuteja th edn. Pp. xv + 394, illustr. (some col.). Springer, Cham. ISBN (hbk), (ebk). Price (hbk), (ebk). The study of mycorrhizal fungi is a field in which there is enormous interest and novel research findings arising from numerous laboratories worldwide. Periodic syntheses are essential to provide overviews of just where particular areas of research are today, and that is accomplished here in 18 chapters involving 45 authors drawn from 11 countries. The topics covered are wideranging, and include the mobilization of micronutrients, improving the tolerance to Fusarium wilt of tomato plants through the use of arbuscular mycorrhizal (AM) fungi, interactions with bacteria, ectomycorrhizal (ECM) fungi and their roles, ECM fungi with Fagaceae in Mediterranean ecosystems, plant flavenoids in the establishment of endosymbioses, importance to tree health, importance in plant succession in glaciers and sand dunes, truffle ecology, diversity of AM fungi in symbioses, interactions with dark septate endophytes in viticulture, and mycorrhizal colonization of wetland plants. Two chapters are concerned with responses of AM fungi to climate change, and two to Piriformospora indica and its cultivation. The book is nicely produced, with some coloured figures distributed though the text, and not just collected together and tippedin as one signature. I was also pleased to see an index something sadly missing from an increasing number of titles. This is, however, very much a collection of mainly potentially stand-alone chapters on particular aspects of mycorrhizal research, rather than a comprehensive overview, as some exciting and rapidly advancing areas are scarcely mentioned, such as host specificity, genomics, and interbiont signalling mechanisms. Mycorrhizal researchers will consequently use this as a source for reviews of the selected subjects covered, rather than commend it to students as a comprehensive state of the art review. (32) IMA FUNGUS

35 MykoLibri: Die Bibliothek der Pilzbücher. Vol. 2. Addenda. By Christian Volbracht Pp. vi + 210, figs 100 (20 full page), mostly col. Christian Volbracht, Hamburg. ISBN not provided. Price 105. Christian Volbracht is a dealer in mycological books, but also a mycophile and bibliographer. He published an account of his holdings in 2006 (Volbracht 2006). He started collecting in the mid-1970s, and the 2006 volume includes information on just over 3000 titles published between 1481 and 1959, with full bibliographic details presented in 525 large-format (29.5 x 21 cm) pages, interspersed with mainly coloured copies of title pages and plates. His library has continued to swell, and this addendum volume includes an additional 600 titles published in the years up to The format is the same, and while the emphasis is on macromycetes and mycophagy, also included are some sets of plates, and even at least one work on lichenicolous fungi; works dealing exclusively with lichenized fungi are not, however, treated. The text is in German, with the short introduction given also in English and French. Unlike the original volume, the addendum comes in just one version, hard-bound with a paper cover. It is produced to the highest standards, and will be a pleasure to own and use. I have made considerable use of the original volume when needing information on particular titles, many of which are exceedingly rare, when assisting with nomenclatural and taxonomic investigations, and the two volumes should be on the shelves of all major mycological libraries, and also those of other mycophiles. Christian has provided a great service to mycology by putting this catalogue together and making it available for sale. I understand that some copies of the original volume are still available at 140, and the two can be purchased as a set for 200. Volbracht C (2006) MykoLibri: Die Bibliothek der Pilzbücher. Hamburg: C. Volbracht. BOOK NEWS The Fungi. Edited by Sarah C. Watkinson, Lynne Boddy, and Nicholas P. Money. 3 rd edn Academic Press (Elsevier), Amsterdam. Pp.xv + 449, illustrated, col. plates 27. ISBN (pbk), (ebk). Price (pbk), (ebk), (pbk+ebk). There are few authoritative and substantial texts endeavouring to cover the whole field of mycology, so it is pleasure to see a new edition of this familiar one. The first edition was in 1994, prepared by Michael Carlile and Sarah Watkinson, and for the second in 2001 they were joined by Graham Gooday. Unusually for a co-authored book, each of the authors is named as the single author for each of the 12 chapters they prepared. Also surprising is that it is 139 pages shorter than the second edition. The balance of topics, and just what to include and what to omit within them, cannot be other than eclectic when contemplating production of a single volume aimed at addressing the whole field. As the primary target audience is given as undergraduates and graduate students, the choice is necessarily influenced by the content of the courses it is expected to be used in. It is evident that the authors had courses with an experimental fungal biology emphasis to the fore, encompassing mycology from a microbiological perspective (p. xiii). In consequence, the coverage of the structural and phylogenetic diversity of the fungi is largely limited to the first two chapters by Nick Money, occupying 36 pages as opposed to 84 in the second edition. Amazingly for the 21 st century, these two chapters lack any detailed phylogenetic classification below phylum, and dismiss the whole of the ascomycetes in just under six pages (including illustrations), without even mentioning the former practice of separately naming morphs of the same species which students are likely to encounter. I was also sorry to see the 11 page Appendix on classification giving examples of species down to order in the second edition omitted. There is something of an apologia (p. xv) for this approach on the basis of the pace of change in phylogenetic research, which I find somewhat anachronistic at a time when major relationships at class and order are now rather stable. It would have been helpful to at least direct users wishing to understand classifications and check current names to those accepted in Species Fungorum; that would have been a useful exercise for the authors to have undertaken before the work went to press as some names used are not the current ones. On the positive side, the treatment of fungal ecology and interactions with other organisms has benefitted enormously from the involvement of Lynne Boddy. There is now a rather full treatment of genetic variation, mating systems, speciation, and evolution, and no less than five chapters on interactions with other organisms: mutualisms, autotrophs, humans and other animals, with other microbes, and role in VOLUME 8 NO. 1 (33)

36 BOOK NEWS ecosystem and global change. Particularly topical are the treatments of the effects of climate change and pollutants, and fungal conservation; the latter includes a table showing the use of IUCN Red List categories which had merited much more exposure to mycologists in general. I also enjoyed Sarah Watkinson s chapter on molecular ecology explaining the issues that needed to be addressed, especially in relating environmental sequences to naming fungi and the issue of cryptic speciation, but also the promise of genomics. In contrast, the coverage of industrial uses seemed rather limited for the intended audience. Just a single chapter is devoted to fungal biotechnology and occupies just 24 pages with only five references, three relating to yeasts. I would also have expected to see much more on the uses of mushrooms, mushroom cultivation, fungal products, and biocontrol, and also sections on the biodeterioration of manufactured materials, food spoilage, and fungi in the indoor environment. Each chapter ends with a Further Reading section, in most cases with general titles followed by others arranged by topic. There is also a helpful glossary and an index. Colour was not used in the last edition, so I was also pleased to see a final 28-page signature with colour versions of some of illustrations that appeared as half-tones in the text; a second best to having them in colour where they appeared, but better than having none at all. I am sure that the book will prove valuable in some fungal biology courses, but while particular chapters can be highly commended for further reading on topics in broader mycological ones, the uneven coverage will sadly limit its international appeal. (34) IMA FUNGUS

37 International and regional meetings which are entirely mycological or have a major mycological component. Only meetings supported by mycological organizations or groups are included, and not those arranged by commercial organizations th International Botanical Congress July 2017 [Nomenclature Section July 2017] Shenzhen Convention and Exhibition Center, Shenzhen, Guangdong, China IX Latin American Congress of Mycology (IXCLAM) August 2017 Miraflores, Lima, Peru Contact: almycperu@ gmail.com FORTHCOMING MEETINGS Westerdijk Institute Symposium Week: Famous Fungi 28 August 2017 Evening Symposium, open to the general public University of Utrecht, Academy Building, Domplein 29, Utrecht, The Netherlands Contact: info@westerdijkinstitute.nl Westerdijk Institute Symposium Week: 2 nd Symposium on Plant Biomass Conversion by Fungi August 2016 Westerdijk Fungal Biodiversiti Institute, Uppsalalaan 8, Utrecht, The Netherlands Contact: info@westerdijkinstitute.nl Westerdijk Institute Symposium Week: Leading Women in Fungal Biology August 2017 University of Utrecht, The Netherlands Contact: info@westerdijkinstitute.nl Westerdijk Institute Symposium Week: Cryptic Speciation in Classifications 1 September 2017 Westerdijk Fungal Biodiversiti Institute, Uppsalalaan 8, Utrecht, The Netherlands Contact: info@westerdijkinstitute.nl 9 th International Medicinal Mushroom Conference (IMMC9) September 2017 Spendid Hotel La Torre, Mondello, Palermo, Italy Contact: secretary@immc9.com X th Meeting of the European Council for the Conservation of Fungi 3 8 October 2017 Ohrid, Former Yugoslav Republic of Macedonia Contact: Mitko Karadalev Asian Mycological Congress (AMC) October 2017 Rex Hotel Saigon, Hochiminh City, Vietnam Contact: Hoang Pham; pndhoang@gail.com VOLUME 8 NO. 1 (35)

38 2018 FORTHCOMING MEETINGS 3 rd International Conference on Basic and Applied Mycology March 2018 Assiut Mycological Centre, University of Assiut, Assiut, Egypt Contact: Abdelaal H. Moubasher; ahamaumc@yahoo.com 8 th Advances Against Aspergillosis 1 3 March 2018 Lisboa Congress Centre, Lisbon, Portugal Contact: alex@hartleytaylor.co.uk AAA2018.org 20 th Congress, International Society for Human and Animal Mycology (ISHAM) 2 6 July 2018 RAI Amsterdam, Amsterdam, The Netherlands Contact: info@congresscare.com 11 th International Mycological Congress (IMC11) July 2018 San Juan, Puerto Rico Contact: Sharon A. Cantrell Rodríguez; scantrel@suagm.edu th Congress of European Mycologists September 2019 Warsaw and Białowieża Primeval Forest, Poland Contact: Julia Pawlowska; jzpawlowska@gmail.com ERRATUM In IMA Genome-F-7B (Hammerbacher et al. 2016), the culture number of the sequenced strain of Ceratocystis harringtonii should be corrected from CBS to CBS The authors regret any confusion this may have caused. Hammerbacher A, Wilken PM, Van der Nest MA, Wilson A, Naidoo K, et al. (2016) Draft genome sequence of the poplar pathogen Ceratocystis haringtonii. IMA Fungus 7: Brenda D. Wingfield (Brenda.Wingfield@up.ac.za) NOTICE IMA Fungus is compiled by David L. Hawksworth (Comparative Plant and Fungal Biology, Royal Botanic Gardens, Kew, Surrey TW93DS, UK) on behalf of the Executive Committee of the International Mycological Association. All unsigned items in the journal are by, and may be attributed to, him. Items for consideration for inclusion in all sections of the journal should be submitted to him at d.hawksworth@kew.org with a copy to the Managing Editor, Pedro W. Crous (p.crous@ westerdijkinstitute.nl). Books for consideration for coverage in the Book News section should be mailed to: IMA Fungus, Milford House, 10 The Mead, Ashtead, Surrey KT21 2LZ, UK. (36) IMA FUNGUS

39 doi: /imafungus IMA FUNGUS 8(1): 1 15 (2017) A plant pathology perspective of fungal genome sequencing Janneke Aylward 1, Emma T. Steenkamp 2, Léanne L. Dreyer 1, Francois Roets 3, Brenda D. Wingfield 4, and Michael J. Wingfield 2 1 Department of Botany and Zoology, Stellenbosch University, Private Bag X1, Matieland 7602, South Africa; corresponding author janneke@sun.ac.za 2 Department of Microbiology and Plant Pathology, University of Pretoria, Pretoria 0002, South Africa 3 Department of Conservation Ecology and Entomology, Stellenbosch University, Private Bag X1, Matieland 7602, South Africa 4 Department of Genetics, University of Pretoria, Pretoria 0002, South Africa Abstract: The majority of plant pathogens are fungi and many of these adversely affect food security. This minireview aims to provide an analysis of the plant pathogenic fungi for which genome sequences are publically available, to assess their general genome characteristics, and to consider how genomics has impacted plant pathology. A list of sequenced fungal species was assembled, the taxonomy of all species verified, and the potential reason for sequencing each of the species considered. The genomes of 1090 fungal species are currently (October 2016) in the public domain and this number is rapidly rising. Pathogenic species comprised the largest category (35.5 %) and, amongst these, plant pathogens are predominant. Of the 191 plant pathogenic fungal species with available genomes, 61.3 % cause diseases on food crops, more than half of which are staple crops. The genomes of plant pathogens are slightly larger than those of other fungal species sequenced to date and they contain fewer coding sequences in relation to their genome size. Both of these factors can be attributed to the expansion of repeat elements. Sequenced genomes of plant pathogens provide blueprints from which potential virulence factors were identified and from which genes associated with different pathogenic strategies could be predicted. Genome sequences have also made it possible to evaluate adaptability of pathogen genomes and genomic regions that experience selection pressures. Some genomic patterns, however, remain poorly understood and plant pathogen genomes alone are not sufficient to unravel complex pathogen-host interactions. Genomes, therefore, cannot replace experimental studies that can be complex and tedious. Ultimately, the most promising application lies in using fungal plant pathogen genomics to inform disease management and risk assessment strategies. This will ultimately minimize the risks of future disease outbreaks and assist in preparation for emerging pathogen outbreaks. Key words: genome size pathogen evolution pathogen lifestyle plant pathology Article info: Submitted: 26 October 2016; Accepted: 19 January 2017; Published: 9 February INTRODUCTION Sequencing of fungal genomes is being driven by various groups of scientists having different interests and needs from genomic data. Mycologists desire genome data to understand how fungi live and evolve, while industries require information on how to improve metabolic pathways or how to find new sources of natural products. The medical and plant pathology sectors need this information to understand diseases, improve diagnoses, understand how they function, and ultimately prevent or at least manage disease outbreaks (Kelman 1985). By 2007, the genomes of 42 eukaryotes were available (Cornell et al. 2007) and by 2008 the number of fungal genomes exceeded 90 (Park et al. 2008). Today, more than 3000 fungi are in completed or ongoing genome projects and the genomes of more than 900 fungal species have been released. The substantial and growing investment in determining genome sequences reflects the positive impact that this field is having on research. Our question here is what the impact has been for plant pathogenic fungi. This mini-review aims to summarise the number of available fungal plant pathogen genomes, determine their general characteristics, and consider the impact that the availability of these genomes is having on the study of plant pathology. In order to determine which fungal plant pathogens have been sequenced, we surveyed fungal species (including Microsporidia, but excluding Oomycota) listed in 11 online genome repositories (Table 1), including MycoCosm (Grigoriev et al. 2012, 2013), NCBI Genome ( the Broad Institute (www. broadinstitute.org), and the universal cataloguing database, Genomes OnLine Database (GOLD; Reddy et al. 2014). Fungal species that were found in more than one database were clustered and the current classification of all species was verified up to ordinal level using MycoBank (Robert et al. 2013) and Index Fungorum ( The most recent scientific literature was consulted where the two online reference databases were not in agreement. Synonymous names associated with each species were noted by consulting MycoBank. We used this non-redundant list to accurately determine the number of fungal species with 2017 International Mycological Association You are free to share - to copy, distribute and transmit the work, under the following conditions: Attribution: You must attribute the work in the manner specified by the author or licensor (but not in any way that suggests that they endorse you or your use of the work). Non-commercial: You may not use this work for commercial purposes. No derivative works: You may not alter, transform, or build upon this work. For any reuse or distribution, you must make clear to others the license terms of this work, which can be found at Any of the above conditions can be waived if you get permission from the copyright holder. Nothing in this license impairs or restricts the author s moral rights. VOLUME 8 NO. 1 1

40 Aylward et al. Table 1. Fungal genome resources used to populate the genome species list. Name Abbreviation Number of fungal URL genome projects Aspergillus Genome Database AspGD 4 Candida Genome Database CGD 4 EnsemblFungi 53 fungi.ensembl.org Fungal Genome Resource 4 gene.genetics.uga.edu Genomes OnLine Database GOLD 3362 genomesonline.org JGI Genome Portal: MycoCosm >1200 genome.jgi.doe.gov/fungi NCBI Genome > PomBase 1 Saccharomyces Genome Database SGD >50 The Broad Institute >100 fungal-genome-initiative The Institute of Bioinformatics and Systems Biology IBIS 20 institute/groups/fungal-microbial-genomics/ resources/index.html University of Kentucky 29 available genome sequences and, specifically, the extent to which fungal plant pathogens have been sequenced. BEYOND THE 1000 MARK The lower cost of genome sequencing, due to highthroughput technologies, has encouraged large scale genome initiatives. These include the 5000 Insect Genome Project (i5k; Robinson et al. 2011), Genome 10K (Genome 10K, Community of Scientists 2009) and 1000 Plants (www. onekp.com). These projects aim to sample species diversity by sequencing, respectively, whole genomes of insects and vertebrates and the transcriptomes of plant species. Similarly, fungal genome sequencing programmes such as the Fungal Genome Initiative (Fungal Research Community 2002, The Fungal Genome Initiative Steering Committee 2003), the Fungal Genomics Program (Grigoriev et al. 2011, Martin et al. 2011), and its extension, the 1000 Fungal Genomes (1KFG) Project (Spatafora 2011), have contributed significantly to the number of fungal genomes currently available and continue to do so. The prevalence of fungi as study organisms is evident when considering the on-going and completed genome sequencing projects. A catalogue of genome projects, GOLD (Reddy et al. 2014), began in 1997 with six genome entries (Bernal et al. 2001) and in October 2016 included 7422 eukaryote whole genome sequencing projects, of which 3515 (47.4 %) are fungal. Although the respective genome databases (Table 1) list numerous completed and on-going fungal projects, many entries do not represent different species. Of the 1459 completed fungal genome projects in GOLD, slightly more than half (ca. 775) are different species, whilst the remainder comprise additional strains of alreadysequenced species. To illustrate the extent and prevalence of sequenced genomes in the fungal kingdom, we mapped species with publically available genomes onto ordinal consensus trees (Fig. 1A C). The most recent of the fungal genome sequencing initiatives, the five-year international collaborative 1KFG Project, aims to sequence and annotate two species from each of the more than 500 known fungal families (Spatafora 2011). In three years, there has been a shift from obtaining representative genomes for all the fungal phyla (Buckley 2008) to targeting genome sequencing at the family level. By October 2016, the genomes of 1090 different fungal species were publically available (Supplementary File 1). Of this number, the 1KFG Project has released approximately 60 % of the fungal species genomes available. Although the target of 1000 sequenced fungal species has been reached, the goal of two genomes from each family is a bigger task than the number 1000 (Fig. 1). The goal of having the genome sequences for two representatives have only been achieved in 85 families in the Ascomycota, 66 in the Basidiomycota, and 11 in the remainder of the fungi. Not surprisingly, some economically and medically important families (e.g. Aspergillaceae, Clavicipitaceae, Mucoraceae, Mycosphaerellaceae, Saccharomycetaceae, Tremella-ceae, Ustilaginaceae) have many more than two representatives. Additionally, taxonomic revision and species descriptions continue to generate new fungal families and orders. In the almost ten years since the publication of the Hibbett et al. (2007) consensus tree, more than 50 fungal orders have been described, somewhat increasing the workload of the 1KFG Project. Additionally, less than 10 % of the conservative estimate of 1.5 million total fungal species are known (Hawksworth 2012) and new species descriptions continuously emerge. Therefore, the combined goals of sampling fungal biodiversity and sequencing the genomes of representative species are a continuous process. 2 IMA FUNGUS

41 Plant pathogenic fungal genomes Fig. 1. Ordinal consensus trees depicting the taxonomic (subphylum, class and order) distribution of publically available genomes for the Ascomycota (A), Basidiomycota (B) and early-diverging fungi (C). The number of sequenced genomes from each order is indicated after the order name. Where sequenced species are not classified into a family or have not been described, these are indicated as incertae sedis (inc. sed.) or unknown (unk.), respectively. The number of families with sequenced representatives out of the total number of described families is indicated in brackets. For each order, horizontal bars show the current progress of sequencing two genomes per family, indicated according to the scale bar below the figure. Dikarya consensus trees are according to Hibbett et al. (2007), while the classification of Spatafora et al. (2016) was included in the tree of early-diverging fungi. Orders described after Hibbett et al. (2007) have been added in grey (see Supplementary File 3 for references). The figures do not include unclassified fungi that have not been sequenced. VOLUME 8 NO. 1 3

42 Aylward et al. Fig. 1A. (Continued). ARE PATHOGENS PREFERENTIALLY SEQUENCED? More than 90 % of known fungal species reside in the subkingdom Dikarya (Kirk et al. 2008) comprised of the two largest phyla, Ascomycota and Basidiomycota. The large number of ascomycete and basidiomycete species for which genome sequences have been determined (Table 2) is, therefore, not an over-emphasis of these common phyla, but rather reflects the size and diversity of the Dikarya (Fig. 2). In the majority of cases, the proportion of sequenced species in the phyla of early-diverging fungi is congruent with the known species, suggesting that genome projects have not neglected them (Fig. 2). Mucoromycota has a larger proportion of sequenced species than known species due to the sequencing of several Mucoraceae species that cause human mucoromycosis. One phylum (Olpidiomycota) and one subphylum (Zoopagomycotina) of early-diverging fungi, 4 IMA FUNGUS

43 Plant pathogenic fungal genomes Fig. 1B. (Continued). VOLUME 8 NO. 1 5

44 Aylward et al. Fig. 1C. (Continued). however, do not have any sequenced representatives and do not have targeted or in progress projects listed on GOLD. Olpidiomycota was only described recently (Doweld 2013) and its members appear to be poorly known. The lack of Zoopagomycotina sequences can likely be ascribed to very few available pure cultures of this predominantly parasitic group of fungi (Spatafora et al. 2016). Within the subphyla of Ascomycota and Basidio-mycota, the proportion of sequenced species also largely corresponds to the number of known species (Fig. 3), with the major exception being Saccharomycotina (budding yeasts; Fig. 3A). The emphasis placed on this subphylum is even more pronounced when considering not only the number of species, but also the number of strains that have been sequenced. Although Pezizomycotina (filamentous fungi) is by far the most species-rich subphylum on the genome list (547 spp.), most sequenced strains are in Saccharomycotina (416), maintaining its previous status as the most sequenced subphylum in the kingdom (Cuomo & Birren 2010). Other than members of Saccharomycotina, seven highly sequenced ( 10 strains) species are listed in GOLD (Table 3). One is of industrial importance (Rhodotorula toruloides), whereas the remainder influence food security (Aspergillus flavus, Fusarium oxysporum, and Magnaporthe oryzae) or human health (Cryptococcus gattii, Coccidioides posadasii, and Trichophyton rubrum). A 2008 report by the American Academy of Microbiology (Buckley 2008) stated that fungal genome sequencing is heavily skewed in the favour of human pathogens. At that time whole-genome sequencing of eukaryotes, especially fungi, was in its infancy and the statement was based on fungal representatives. The initial high cost of genome sequencing would have favoured fungi of medical importance, but as the number of sequenced fungi grew and the cost decreased, this pattern was bound to change. We assessed whether pathogens are highly sequenced by consulting recent scientific literature (where available) on each fungal species on the genome list and categorising them according to their significance and reason for being sequenced. The largest (41.4 %) category consisted of pathogenic fungi and fungi of medical importance (Fig. 4), of which plant pathogens were the most prevalent group (49.4 %). Currently, 191 plant pathogenic species have publically available genomes (Supplementary File 2) and all belong to Dikarya. Of these, 117 are pathogens of at least one food crop and 43 affect gymnosperms, the majority of which are commercially important (Table 4). The 117 food crop species include pathogens of cereals, fruit, vegetables, and legumes. At least 60 of these species are responsible for diseases on 10 of the 15 global staple food crops (FAO 1995). 6 IMA FUNGUS

45 Plant pathogenic fungal genomes Table 2. Number of fungal species from each phylum and subphylum with at least one available genome. Phylum and subphylum Sequenced species Known species a ASCOMYCOTA 684 > Pezizomycotina 547 Saccharomycotina 123 Taphrinomycotina 11 Incertae sedis 3 BASIDIOMYCOTA 311 > Agaricomycotina 227 Pucciniomycotina 43 Ustilaginomycotina 41 BLASTOCLADIOMYCOTA 2 > 175 CHYTRIDIOMYCOTA 5 > 700 CRYPTOMYCOTA 1? MICROSPORIDIA 25 > MUCOROMYCOTA 47 Glomeromycotina 1 > 165 Mortierellomycotina 5 Mucoromycotina 41 > 325 NEOCALLIMASTIGOMYCOTA 5 > 20 ZOOPAGOMYCOTA 8 Entomophthoromycotina 4 > 275 Kickxellomycotina 4 > 260 UNKNOWN 2 Total sequenced 1090 a According to Kirk et al. (2008). Table 3. Fungal species on the Genomes OnLine Database (Bernal et al. 2001) with 10 or more completed whole-genome sequencing projects. Species Strains Phylum Subphylum Saccharomyces 166 Ascomycota Saccharomycotina cerevisiae Magnaporthe 48 Ascomycota Pezizomycotina oryzae Candida 35 Ascomycota Saccharomycotina albicans Komagataella 32 Ascomycota Saccharomycotina pastoris Saccharomyces 20 Ascomycota Saccharomycotina kudriavzevii Cryptococcus 18 Basidiomycota Agaricomycotina gattii Fusarium 17 Ascomycota Pezizomycotina oxysporum Trichophyton 12 Ascomycota Pezizomycotina rubrum Aspergillus 10 Ascomycota Pezizomycotina flavus Coccidioides 10 Ascomycota Pezizomycotina posadasii Rhodotorula 10 Basidiomycota Pucciniomycotina toruloides Saccharomyces pastorianus 10 Ascomycota Saccharomycotina Clearly, genome sequencing projects have placed an emphasis on plant pathogenic species, specifically those affecting food security or commercial forestry. In general, fungal pathogens are highly represented in the genome list. Furthermore, as the number of available genomes has increased, plant pathogens have replaced human pathogens as the predominantly sequenced category of species. Since most plant pathogens are fungi (Carris et al. 2012), the emphasis placed on fungal genome sequencing may (at least partly) be attributed to food security. For example, M. oryzae, a plant pathogen for which numerous strains have been sequenced (Table 3), is predicted to increase its distribution range and impact due to increased temperature and carbon dioxide levels (Gautam et al. 2013). The sheer number of plant species and their associated diseasecausing fungi makes this change in the focus of genome sequencing understandable. Sequencing a large number of plant pathogens that affect a range of plant species is, after all, less of a bias than sequencing many pathogenic species associated with a single species (humans). GENOME SIZE AND GENE NUMBERS IN PLANT PATHOGENIC FUNGAL GENOMES As far as we are aware, this review includes the first comprehensive list of plant pathogenic fungal genomes that have been sequenced to date. We, therefore, briefly present an overview of the genome characteristics of these species in comparison to other sequenced fungal species. We specifically looked at genome size and the numbers of genes encoded, because previous studies have revealed a link between plant pathogenicity and genome size and gene content (Duplessis et al. 2011, Ohm et al. 2012, Spanu et al. 2010). The 1090 fungal species with publically available genome sequences have haploid genome sizes ranging between two and 336 million base pairs (Mbp; Fig. 5A). The majority of these genomes fall within the Mbp range (average = 37.2, median = 33.6), consistent with the size distribution of the 1940 entries in the Fungal Genome Size Database (Kullman et al. 2005). The genome sizes of sequenced plant pathogens are only slightly, but significantly, larger compared to this norm. This difference was most apparent in the pathogenic ascomycetes for which Mann-Whitney U tests indicated the highest level of significance (Median = 38.0; U = 48131, P < 0.01). The average genome size of plant pathogenic basidiomycetes (57.3 Mb) was much larger than that of the plant pathogenic ascomycetes (39.4 Mb) and the remainder of the fungal genomes (34.8 Mb), owing to several pathogenic pucciniomycete (rust) species with genomes larger than 100 Mb. Somewhat larger genome sizes in plant pathogens are congruent with the hypothesis that they often contain more repeated elements than other species (discussed below) (Castanera et al. 2016, Ma et al. 2010). Sequenced plant pathogens also have larger genomes than human, animal and opportunistic fungal pathogens (Fig. 5B). Although sequencing has thus far sampled the genome size distribution of the majority of the fungal kingdom, species with excessively large genome sizes have been omitted. This is not necessarily VOLUME 8 NO. 1 7

46 Aylward et al. Fig. 2. Comparison between the proportion of known and sequenced fungal species in the major fungal taxonomic groups. The number of known species were obtained from Kirk et al. (2008). only due to the higher cost of sequencing large genomes, but probably also the complexity of obtaining sufficient biomaterial from a single obligately parasitic individual cultured on a live host (Barnes & Szabo 2008). Since the majority of species in Pucciniomycotina reside in the order Pucciniales of obligate plant pathogens (Kirk et al. 2008), the latter may also explain why the proportion of sequenced species in this group is slightly less than the known species (Fig. 3B). Therefore, genome-sequencing efforts so far, most likely underestimate the maximum size of plant pathogen genomes. Considering the number of predicted open reading frames (ORFs), 714 of the available genomes have publically accessible gene annotations. The sequenced fungi have, on average, ± total predicted ORFs at a density of ± ORFs per Mbp (Fig. 5C). In comparison to the other genomes, plant pathogenic fungi do not differ in the number of predicted ORFs, but they do have significantly fewer ORFs when accounting for genome size (U = 35647; P < 0.01). This trend was also observed in the animal pathogenic fungi (including entomopathogens; U = 8847; P < 0.01). Previous whole-genome studies suggest that the number of coding genes does not necessarily increase with genome size, since transposable elements and repetitive sequences proliferate in large genomes (Kidwell 2002). Fig. 3. Comparison between the proportion of known and sequenced fungal species in the subphyla of Ascomycota (A) and Basidiomycota (B). The number of known species were obtained from Kirk et al. (2008). 8 IMA FUNGUS

47 Plant pathogenic fungal genomes Fig. 4. Categories of significance identified in the 1090 sequenced fungal species. Pathogens comprise the largest category within which plant pathogens are predominant. Medically important species represent fungi that are not directly pathogenic, but cause food or environmental contamination. Interesting species are studied for their development or metabolism. Niche-specific refers to species occupying abiotic niches, whereas symbiotic species are associated with other organisms. Economically important species have a use in the economy, (e.g. culinary, biocontrol or pharmaceutical industries). Most of the parasites belong to Microsporidia. Table 4. Categories of plants affected by the 191 sequenced fungal plant pathogens. Plant pathogen categories Genomes available % Cash Crop Pathogens Food Crop Pathogens Grains Fruit Vegetables Legumes Multiple crop types Gymnosperm Pathogens Other a TOTAL a Non-gymnosperms not cultivated for food. The lower number of ORFs/Mb in the genome of plant pathogenic Ascomycetes is, therefore, consistent with their larger genome size possibly being due to repetitive elements. Additionally, some pathogens have lost genes redundant in their lifecycles (Spanu et al. 2010), which may also decrease their ORFs/Mb. This trend could, however, not be detected in the genomes of human and opportunistic pathogens (Fig. 5D). IMPACT OF GENOMES ON PLANT PATHOLOGY Ever since the advent of plant pathology, researchers have been interested in the biology of plant pathogens and how this can be translated into means for disease control. In this regard, genome data are not used in isolation, but provide context for observational and experimental data, thereby accelerating the pace of traditional research methods. For emerging pathogens, a disease may be known, but the mechanisms relating to infection biology and virulence are not necessarily understood. In these cases, a genome can provide the first glimpse of the potential effectors and toxins that are present (e.g. Ellwood et al. 2010). In some plant pathogens, genomes have resulted in a shift of conventional paradigms. Here a classic example is the discovery of entire horizontally transferrable chromosomes related to pathogenicity in Fusarium oxysporum f. sp. lycopersici (Ma et al. 2010). The primary impacts of genome sequences on plant pathology have been a better understanding of the pathogenicity, life-style and genome evolution of pathogens. Furthermore, genomes are also resources from which genetic tools can be used to mine information. Pathogenicity and life-style Secreted and cell surface proteins mediate the interaction between pathogen and host and are often the first to be characterised from plant pathogen genomes. Genome sequences enable in silico predictions of secreted virulence proteins (effectors), bypassing traditional enzyme assays or chromatography/spectrometry techniques that were ineffective at detecting less abundant effectors. For the corn smut fungus, Mycosarcosoma maydis (syn. Ustilago maydis), previous experimental studies were not able to identify the virulence factors that were eventually highlighted by interrogating the genome sequence (Kamper et al. 2006). The proteins identified in silico could then be used to experimentally determine the function of specific effectors in the infection process (Liu et al. 2015). This genome-based effector identification VOLUME 8 NO. 1 9

48 Aylward et al. Fig. 5. Genome size (A and B) and number of open reading frames (ORFs) per million base pairs (Mbp; C and D) in the plant pathogen genomes compared to the remainder of the genome list and other pathogens. Boxplots were drawn in BoxplotR (Spitzer et al. 2014) using the Tukey whisker extent. Width of the boxes is proportional to the square root of the sample size; notches show the 95 % confidence interval of the median. Opp. = opportunistic pathogens. In B and D, animal pathogens include entomopathogenic fungi and Microsporidia are excluded from other genomes, since they have small genomes with many ORFs/Mb. and subsequent phenotype determination has contributed significantly to the online Pathogen Host Interactions Database (Urban et al. 2015). Similarly, genomic prediction of fungal product biosynthesis genes have been well established (Keller et al. 2005) and gene deletion systems could subsequently be used to determine the phenotypes that they confer (Lee et al. 2005). Gene inventories of plant pathogens have also revealed proteins not previously known to be involved in pathogenicity, for example the high diversity of membrane transporters in the F. oxysporum and Pyrenochaeta lycopersici genomes strongly implicates them in the pathogenicity of these fungi (Aragona et al. 2014). Intuitively, cell wall degrading enzymes can be expected to be important in plant pathogenesis and the presence of these enzymes in plant pathogens was well established before the genomic era (Jones et al. 1972, Schneider & Collmer 2010). Genome sequences have confirmed that most plant pathogens encode an array of cell wall degrading enzymes, specifically pectinolytic enzymes in dicot pathogens (Klosterman et al. 2011, Olson et al. 2012). The diversity of cell wall degrading enzymes in a genome appears to increase with host range, as exemplified by the massive number of carbohydrate degrading enzymes in Macrophomina phaseolina that infects over 500 plant species (Islam et al. 2012). Exceptions to this perceived norm typically occur in specialised modes of pathogenesis. For example, cell wall degrading enzymes are absent from the genome of the anther smut fungus Microbotryum lychnidis-dioicae (Perlin et al. 2015). Rather than attacking plant cells, this pathogen has an array of enzymes to influence host development, enabling fungal spores to be substituted for pollen. In contrast, gene inventories suggest that necrotrophic pathogens induce apoptosis in host cells rather than breaking down their cell walls (McDonald et al. 2015). Metabolism-related enzymes in fungal genomes, therefore, have great potential to predict infection strategies and lifestyle. 10 IMA FUNGUS

49 Plant pathogenic fungal genomes Beyond the analysis of single genome sequences, comparing the genomes of ecologically different strains and species has substantial value. For example, analysis of the Rhizoctonia solani AG2-2IIIB genome would have revealed only an abundance of cell wall degrading enzymes. However, comparisons with less aggressive R. solani strains revealed that its virulence can be linked to a significant expansion of polysaccharide lyase enzymes (Wibberg et al. 2016). Similarly, comparisons of resistant and non-resistant Penicillium digitatum strains has enabled identification of mutations conferring tolerance to antifungal compounds (Marcet-Houben et al. 2012). Comparisons between the genomes of 18 dothideo-mycete species has suggested that the number of effectors encoded by these fungi are linked to the pathogenic lifestyle (Ohm et al. 2012). The greatest number of effectors was identified from necrotrophic pathogens, whereas hemibiotrophs have apparently reduced their effector arsenal to evade plant defences before they switch to necrotrophy (Ohm et al. 2012). Furthermore, multiple genome comparisons have been used to highlight specific genes under diversifying selection, revealing that evolutionary pressure on plant pathogen effector proteins drive their adaptation (Schirawski et al. 2010, Stukenbrock et al. 2011). Comparison between plant and fungal genomes have also become essential tools to tease apart pathogen and plant RNA sequences when analysing in planta transcript data (McDonald et al. 2015). The value of multiple genome comparisons has prompted projects such as the Fungal Genome Initiative and the 1KFP to focus not on sequencing single species, but groups of species useful in a comparative context (Grigoriev et al. 2013, The Fungal Genome Initiative Steering Committee 2004). The evolution of different fungal lifestyles is a fascinating topic considered by many comparative genomics studies. Some plant pathogenic fungi with different lifestyles have surprisingly similar gene contents (De Wit et al. 2012), yet unique genes mediate their host interactions. The large proportion of unique secreted effector proteins and hostspecific hydrolytic enzymes in plant pathogenic fungi implies that host association drives their adaptation and, therefore, evolution (De Wit et al. 2012, Duplessis et al. 2011, O Connell et al. 2012, Spanu et al. 2010). The effect of host association is further emphasised by the diversification of both effectors and hydrolytic enzymes in broad host range pathogens such as Colletotrichum higginsianum (O Connell et al. 2012). In contrast, host association cannot explain the loss of primary metabolism genes that led to obligate biotrophy in powdery mildew fungi (Spanu et al. 2010). Similarly, selective pressures that mediate the evolution of a hemibiotrophic strategy, where the pathogen transitions between a biotrophic and necrotrophic lifestyle, are poorly understood. Genome evolution of plant pathogens The arms race between pathogen and host (Stahl & Bishop 2000) makes pathogen adaptability, or evolu-tionary potential, particularly interesting (McDonald & Linde 2002). Reproduction and gene diversity are two of the factors that influence evolutionary potential (McDonald & Linde 2002) and these can be estimated from genome sequences. For example, an analysis of the mating type genes that govern sexual reproduction can provide insights into the mating strategy of a fungus. Heterothallic ascomycete fungi are identified by the occurrence of a single mating type in a genome (Kronstad & Staben 1997), whereas homothallic fungi contain both mating types, either in the same genome or in a dikaryotic cellular state (Wilson et al. 2015). Many fungi propagate only vegetatively or sexual reproduction is difficult to observe. In such cases, genomic analyses have been able to reveal the presence of mating type genes (e.g. Bihon et al. 2014, Marcet-Houben et al. 2012), suggesting that these species could have a cryptic sexual cycle (Bihon et al. 2014). The mating type sequence information can subsequently be used to determine the distribution of different mating types in a population (Aylward et al. 2016, Haasbroek et al. 2014). In contrast, genomes can also reveal the importance of mitotic recombination for generating new allelic combinations. For example, a whole genome survey concluded that mating type genes are completely absent from the tomato pathogen P. lycopersici and that it has an expansion of gene modules associated with heterokaryon incompatibility (Aragona et al. 2014). Adaptability to a changing environment or a resistant host can also be mediated by genome plasticity. Transposons are repetitive elements in DNA known to contribute to genome plasticity and evolution (Wöstemeyer & Kreibich 2002). The expansion of repeats in many plant pathogen genomes points to their role in diversification and adaptation (Raffaele & Kamoun 2012, Spanu et al. 2010, Thon et al. 2006) and has been directly implicated in the pathogenicity of the wheat necrotroph Pyrenophora tritici-repentis (Manning et al. 2013). Surveys of transposons across plant pathogen genomes have revealed differences in their number and activity between the essential core and dispensable supernumerary chromosomes (Ohm et al. 2012, Vanheule et al. 2016). In Fusarium poae, repeat expansion in the core chromosomes is contained, while the non-essential supernumerary chromosomes have many active transposons that invade the core chromosomes (Vanheule et al. 2016). The supernumerary chromosomes also provide opportunity for duplication and diversification of core genes, thereby facilitating adaptation. An example of such diversification and adaptation in the post-harvest spoilage fungus Penicillium digitatum is the association of DNA transposons and ABC transporters in drug resistant strains (Sun et al. 2013b). Horizontal gene transfer (HGT) may add novel ecological capabilities to the genomes of recipient species. Although not historically considered relevant to eukaryotic evolution, genome level investigations have revealed multiple HGT events in fungi, often from other kingdoms (e.g. Marcet- Houben & Gabaldón 2010, Sun et al. 2013a). Such phylogenomic studies have reported, amongst others, horizontal acquisition of genes that mediate pathogenicity (Friesen et al. 2006, Kroken et al. 2003, Slot & Rokas 2011, Thynne et al. 2015), tolerance to host defences (Marcet- Houben & Gabaldón 2010, Sun et al. 2013a), and nutrient uptake and metabolism (Soanes & Richards 2014, Sun et al. 2013a). Moreover, a comparative genomics study found evidence of HGT at chromosome level in F. oxysporum f. sp. lycopersici, as entire pathogenicity-related chromosomes could be transferred between strains (Ma et al. 2010). VOLUME 8 NO. 1 11

50 Aylward et al. Acquiring new ecological capabilities through HGT has previously played a causal role in the emergence of new pathogens and will likely do so again in future (Friesen et al. 2006, Soanes & Richards 2014, Thynne et al. 2015). Resources for genetic tools Other than facilitating whole-genome related studies, genome sequences have become ideal resources for mining genetic tools. Previously, species-specific population genetic tools such as microsatellites had to be developed painstakingly by cloning and genome walking (Barnes et al. 2001, Burgess et al. 2001). Now, any genome sequence enables rapid identification of such genetic markers (e.g. Haasbroek et al. 2014). This holds true for diagnostic markers: genome regions that unambiguously and rapidly identify a pathogen and/or differentiate between pathogens can be designed by inspecting whole genome sequences. Although development of such markers in fungi is lagging behind viral and bacterial pathogens, some examples have recently become available. A pathotype specific marker has been developed from the genome of M. oryzae f.sp. triticum (Pieck et al. 2016) and comparative genomics has detected diagnostic regions in two Calonectria species (Malapi-Wight et al. 2016) and in Pseudoperonospora cubensis (Withers et al. 2016). Continued application of fungal genomes to generate identification tools is bound to increase the efficiency of quarantine procedures (McTaggart et al. 2016). CHALLENGES Although the availability of fungal genomes has dramatically increased our knowledge and understan-ding of infection processes and genome evolution, there remains much to learn. For example, the regulatory elements in most genomes remain poorly annotated and require complex experimental methodologies for accurate identification (e.g. Shen et al. 2012). In a recent review, Schatz (2015) commented that sequencing human genomes has been one of the greatest accomplishments of the past two decades but one of the greatest pursuits for the next twenty years will be trying to understand what it all means. The same can be said for fungal genomes. The information that can be gleaned from a genome sequence is bound to increase as our understanding of these sequences grows. Genome sequences should not be seen to provide silver bullets, although they are often sold this way. They provide the blueprint of potential cellular activities, but are not sufficient to unravel the complexity of pathogen-host interactions. For example, in Fusarium oxysporum, the cell wall degrading enzymes secreted during infection of tomato displayed a clear succession (Jones et al. 1972), an ecologically relevant process that could not be deduced from a gene inventory. In combination with transcriptome data, however, genomic data has revealed how pathogens tolerate host defences (DiGuistini et al. 2011) and how hosts can resist pathogen infection (Zhu et al. 2012). Experimental work, both in vitro and in plantae, will remain essential components in studying fungal plant pathogens. The end goal of studying any host-pathogen relationship is clearly to inform disease management and control. Thus far, identifying specific molecular targets has had little impact on developing new antifungal inhibitors (Odds 2005) and integrative management strategies must, therefore, be a priority. Ultimately, the elucidated effector proteins, host targets, and the overall insights gained into the biology of pathogens must inform disease management strategies (Maloy 2005). It is also crucial that they inform risk assessment protocols governing biosecurity (McTaggart et al. 2016). It is, therefore, essential that the ecological significance of genome patterns is studied to ensure that this knowledge can be extrapolated to emerging pathogen threats. As revealed by comparative genomics, deciphering plant pathogen evolution is in many cases dependent on being able to do comparisons with species having other lifestyles. A large scale example of this is the revised classification of species previously known as Zygomycetes (Spatafora et al. 2016); an endeavour possible because of the availability of multiple genome sequences for this group. In this regard, filling in the gaps in the list of sequenced species is crucial to our understanding of relationships and pathogenesis. The challenge is, therefore, to continue sequencing apparently uninteresting or unimportant taxonomic groups along with the economically important in order to ultimately gain a holistic view. CONCLUSIONS The activities of independent research groups and several fungal sequencing initiatives (Fungal Research Community 2002, Grigoriev et al. 2011, Martin et al. 2011, Spatafora 2011, The Fungal Genome Initiative Steering Commitee 2003), have resulted in the number of publically available fungal genomes growing exponentially since 1996 when the first genome was sequenced (Goffeau et al. 1996). The taxonomic distribution of sequenced fungal genomes is currently roughly congruent with the number of species known from each phylum and subphylum. This is an important and impressive achievement in the goal of sampling biodiversity and representing the phylogenetic groups of the fungal kingdom (Fungal Research Community 2002, The Fungal Genome Initiative Steering Commitee 2003). Many of the genomes have been sequenced to sample environmental and ecological diversity. However, investment continues to be primarily focused on projects that have direct human importance. The emphasis on genomes of plant pathogenic fungi has specifically increased subsequent to the Buckley (2008) overview of sequenced fungal species. The genomes of more than fungal species are already publically available and this number is growing steadily. Fungal genomics has enabled rapid characterization of plant pathogen genomes and revealed features that allow better understanding of the biology of these species. It has also made it possible to rapidly develop tools to study pathogen biology and genetics. In a field where delayed action has profound consequences for livelihoods and food security, genome sequences provide us with essential tools to prepare for the emergence of new plant pathogens and future disease outbreaks. In this regard, the medical example provided by Bill Gates (Gates 2015) that the application of available technologies could significantly have reduced the impact of the recent Ebola epidemic also holds for plant 12 IMA FUNGUS

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54 Supplementary File 1: Ascomycota, Basidiomycota, and Earlydiverging

55 Sequenced Ascomycota species Classification Genome statistics SPECIES PHYLUM SUBPHYLUM CLASS SUBCLASS ORDER FAMILY Strain GOLD Accession GENOME Accession NCBI Project ID Significance Size (Mb) coverage # of contigs # of scaffolds # genes GC Technology Cerataphis brasiliensis yeast-like syascomycota Incertae sedis Incertae sedis Incertae sedis Incertae sedis Incertae sedis not specified Gp AOFP arthropod associated (aphid symbiont) 25,5 39, Illumina HiSeq Endocalyx cinctus Ascomycota Incertae sedis Incertae sedis Incertae sedis Incertae sedis Incertae sedis JCM 7946 Gp BCKC uncertain saprotroph 44,77 252, ,4 HiSeq 2500 Nilaparvata lugens yeast-like symbiascomycota Incertae sedis Incertae sedis Incertae sedis Incertae sedis Incertae sedis not specified Gp JRMI uncertain 26, , ; Illumina Aplosporella prunicola Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Botryosphaeriales Aplosporellaceae CBS Gp JGI probable tree pathogen 32, Aplosporella prunicola Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Botryosphaeriales Aplosporellaceae CBS Gp JGI probable tree pathogen 32,82 194, Botryosphaeria dothidea Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Botryosphaeriales Botryosphaeriaceae CBS Gp AYEP plant pathogen (broad host range of trees and shrubs) 43, Botryosphaeria dothidea Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Botryosphaeriales Botryosphaeriaceae CBS Gp AYEP plant pathogen (broad host range of trees and shrubs) 43, Diplodia pinea Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Botryosphaeriales Botryosphaeriaceae CMW39103 Gp JHUM plant pathogen (pines) 36, , ,9 Illumina MiSeq Diplodia pinea Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Botryosphaeriales Botryosphaeriaceae CMW39103 Gp JHUM plant pathogen (pines) 36, , ,9 Illumina MiSeq Diplodia scrobiculata Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Botryosphaeriales Botryosphaeriaceae CMW30223 Gp LAEG plant pathogen (conifer spp.) 34, Illumina MiSeq Diplodia scrobiculata Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Botryosphaeriales Botryosphaeriaceae CMW30223 Gp LAEG plant pathogen (conifer spp.) 34,93 15, Illumina MiSeq Diplodia seriata Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Botryosphaeriales Botryosphaeriaceae DS831 Gp LAQI plant pathogen (grapevine bot canker) 37,12 78, ,7 Illumina HiSeq Diplodia seriata Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Botryosphaeriales Botryosphaeriaceae DS831 Gp LAQI plant pathogen (grapevine bot canker) 37,12 78, ,7 Illumina HiSeq Guignardia citricarpa Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Botryosphaeriales Botryosphaeriaceae CGMCC Gp AOTE plant pathogen (citrus) 32, , ,6 454; Illumina Guignardia citricarpa Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Botryosphaeriales Botryosphaeriaceae CGMCC Gp AOTE plant pathogen (citrus) 32, , ,6 454; Illumina Macrophomina phaseolina Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Botryosphaeriales Botryosphaeriaceae MS6 Gp AHHD plant pathogen (broad host range) 48,882 66, ,3 454; Illumina Macrophomina phaseolina Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Botryosphaeriales Botryosphaeriaceae MS6 Gp AHHD plant pathogen (broad host range) 48,882 66, ,3 454; Illumina Neofusicoccum parvum Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Botryosphaeriales Botryosphaeriaceae UCRNP2 Gp AORE plant pathogen (grapevines) 42, ,8 Illumina HiSeq Neofusicoccum parvum Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Botryosphaeriales Botryosphaeriaceae UCRNP2 Gp AORE plant pathogen (grapevines) 42,59 100, ,8 Illumina HiSeq Phyllosticta capitalensis Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Botryosphaeriales Phyllostictaceae Gm33 LOEO endophyte; weak plant pathogen 32, ,6 IonTorrent Phyllosticta capitalensis Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Botryosphaeriales Phyllostictaceae Gm33 LOEO endophyte; weak plant pathogen 32,45 70, ,6 IonTorrent Phyllosticta citricarpa Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Botryosphaeriales Phyllostictaceae Gc12 Gp LOEN plant pathogen (citrus black spot) 31, ,1 IonTorrent Phyllosticta citricarpa Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Botryosphaeriales Phyllostictaceae Gc12 Gp LOEN plant pathogen (citrus black spot) 31,13 75, ,1 IonTorrent Phyllosticta citriasiana Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Botryosphaeriales Phyllostictaceae CBS Gp plant pathogen (citrus tan spot) 32,7 88, Illumina Saccharata proteae Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Botryosphaeriales Saccharataceae CBS Gp JGI endophyte; occasional plant pathogen 31,43 98, Illumina HiSeq Saccharata proteae Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Botryosphaeriales Saccharataceae CBS Gp JGI endophyte; occasional plant pathogen 31,43 98, Illumina HiSeq Polychaeton citri Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Capnodiaceae CBS Gp JGI uncertain; mold (non-pathogenic sooty) 27,21 145, Illumina HiSeq Polychaeton citri Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Capnodiaceae CBS Gp JGI uncertain; mold (non-pathogenic sooty) 27,21 145, Illumina HiSeq Cladosporium fulvum Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Cladosporiaceae not specified JGI plant pathogen (leaf mold of tomato) 61, Cladosporium fulvum Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Cladosporiaceae not specified JGI plant pathogen (leaf mold of tomato) 61, Cladosporium sphaerospermum Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Cladosporiaceae UM 843 Gp AIIA human allergen; dermatiaceous 26,89 98, ,6 Illumina Cladosporium sphaerospermum Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Cladosporiaceae UM 843 Gp AIIA human allergen; dermatiaceous 26,89 98, ,6 Illumina Dissoconium aciculare Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Dissoconiaceae CBS Gp JGI mycoparasitic (Erysiphaceae) 26,54 97, Illumina Dissoconium aciculare Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Dissoconiaceae CBS Gp JGI mycoparasitic (Erysiphaceae) 26,54 97, Illumina Pseudovirgaria hyperparasitica Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Incertae sedis CBS Gp JGI biocontrol (mycoparasite of rust) 35,44 86, Illumina Pseudovirgaria hyperparasitica Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Incertae sedis CBS Gp JGI biocontrol (mycoparasite of rust) 35,44 86, Illumina Cercospora canescens Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Mycosphaerellaceae BHU Gp ANSM plant pathogen (leaf spot of various bean crops and tomato) 33,96 12, Illumina Cercospora canescens Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Mycosphaerellaceae BHU Gp ANSM plant pathogen (leaf spot of various bean crops and tomato) 33,96 12, Illumina Cercospora zeae-maydis Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Mycosphaerellaceae SC01 Gp JGI plant pathogen (maize); toxin-producing 46,61 39, Cercospora zeae-maydis Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Mycosphaerellaceae SC01 Gp JGI plant pathogen (maize); toxin-producing 46,61 39, Lecanosticta acicola Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Mycosphaerellaceae CBS Gp AWYC plant pathogen (pine) 34, , ,7 Illumina HiSeq Lecanosticta acicola Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Mycosphaerellaceae CBS Gp AWYC plant pathogen (pine) 34, , ,7 Illumina HiSeq Mycosphaerella arachidis (CercospoAscomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Mycosphaerellaceae CALF-13A Gp LIHB plant pathogen (leaf spot on peanuts) 33,24 100, ,4 Mycosphaerella eumusae Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Mycosphaerellaceae CBS Gp LFZN plant pathogen (banana) 47, ,9 Illumina HiSeq Mycosphaerella eumusae Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Mycosphaerellaceae CBS Gp LFZN plant pathogen (banana) 47,12 165, ,9 Illumina HiSeq Mycosphaerella fijiensis Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Mycosphaerellaceae CIRAD86 Gp JGI plant pathogen (leaf spot of banana) 74,14 7, ,2 Sanger Mycosphaerella fijiensis Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Mycosphaerellaceae CIRAD86 Gp JGI plant pathogen (leaf spot of banana) 74,14 7, ,2 Sanger Mycosphaerella laricina Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Mycosphaerellaceae CBS Gp AWYE plant pathogen (larch) 25, , ,5 Illumina HiSeq Mycosphaerella laricina Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Mycosphaerellaceae CBS Gp AWYE plant pathogen (larch) 25, , ,5 Illumina HiSeq Mycosphaerella pini Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Mycosphaerellaceae NZE10 Gp AIEN plant pathogen (pine) 30,21 34, ,1 Sanger; 454; Illumina Mycosphaerella pini Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Mycosphaerellaceae NZE10 Gp AIEN plant pathogen (pine) 30,21 34, ,1 Sanger; 454; Illumina Mycosphaerella sp. Ston1 Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Mycosphaerellaceae STON1 Gp AWYF poplar-associated 27, , ,2 Illumina HiSeq Mycosphaerella sp. Ston1 Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Mycosphaerellaceae STON1 Gp AWYF poplar-associated 27, , ,2 Illumina HiSeq Passalora (Cladosporium) fulva Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Mycosphaerellaceae CBS Gp AMRR plant pathogen (tomato) 61,1 21, ,8 454 Pseudocercospora musae Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Mycosphaerellaceae CBS Gp LFZO plant pathogen (banana) 60, Illumina HiSeq Pseudocercospora musae Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Mycosphaerellaceae CBS Gp LFZO plant pathogen (banana) 60, Illumina HiSeq Pseudocercospora pini-densiflorae Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Mycosphaerellaceae CBS Gp AWYD animal pathogen (bats) 45, , ,5 Illumina HiSeq Pseudocercospora pini-densiflorae Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Mycosphaerellaceae CBS Gp AWYD animal pathogen (bats) 45, , ,5 Illumina HiSeq Sphaerulina musiva Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Mycosphaerellaceae SO2202 Gp AEFD plant pathogen (poplar) 29,35 35, ,1 454; Illumina Sphaerulina musiva Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Mycosphaerellaceae SO2202 Gp AEFD plant pathogen (poplar) 29, ,1 454; Illumina Sphaerulina populicola Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Mycosphaerellaceae P02.02b Gp AIDU plant pathogen (poplar) 33, ,2 Sphaerulina populicola Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Mycosphaerellaceae P02.02b Gp AIDU plant pathogen (poplar) 33, ,2 Zasmidium cellare Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Mycosphaerellaceae ATCC Gp JGI mold (wine cellar); uses volatile carbon sources (alcohol) 38,25 143, Zasmidium cellare Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Mycosphaerellaceae ATCC Gp JGI mold (wine cellar); uses volatile carbon sources (alcohol) 38,25 143, Zymoseptoria ardabiliae Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Mycosphaerellaceae STIR Gp AFIV plant pathogen (grasses); hemibiotroph 31,136 45, ,8 Illumina Zymoseptoria ardabiliae Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Mycosphaerellaceae STIR Gp AFIV plant pathogen (grasses); hemibiotroph 31, ,8 Illumina Zymoseptoria brevis Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Mycosphaerellaceae ZB163 Gp JYJE plant pathogen (bunchgrass) 36, ,2 Illumina HiSeq; MiSeq Zymoseptoria brevis Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Mycosphaerellaceae ZB163 Gp JYJE plant pathogen (bunchgrass) 36, ,2 Illumina HiSeq; MiSeq Zymoseptoria passerinii Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Mycosphaerellaceae P63 Gp AFIY plant pathogen (barley leaf blotch) 28,786 45, ,6 Illumina Zymoseptoria passerinii Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Mycosphaerellaceae P63 Gp AFIY plant pathogen (barley leaf blotch) 28, ,6 Illumina Zymoseptoria pseudotritici Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Mycosphaerellaceae STIR Gp AFIO plant pathogen (grasses) 33,456 45, ,4 Illumina Zymoseptoria pseudotritici Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Mycosphaerellaceae STIR Gp AFIO plant pathogen (grasses) 33, ,4 Illumina Zymoseptoria tritici Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Mycosphaerellaceae STIR04 A26b Gp AFIP plant pathogen (wheat leaf blotch) 32,641 45, ,1 Illumina Zymoseptoria tritici Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Mycosphaerellaceae STIR04 A26b Gp AFIP plant pathogen (wheat leaf blotch) 32, ,1 Illumina Piedraia hortae Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Piedraiaceae CBS Gp JGI opportunistic dermatophyte; keratinolytic 16,95 78, Illumina Piedraia hortae Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Piedraiaceae CBS Gp JGI opportunistic dermatophyte; keratinolytic 16,95 78, Illumina not publically available Acidomyces richmondensis Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Teratosphaeriaceae BFW Gp JGI acidophile 29,88 17, Acidomyces richmondensis Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Teratosphaeriaceae BFW Gp JGI acidophile 29,88 17, Baudoinia compniacensis Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Teratosphaeriaceae UAMH Gp JGI extremophile 21,88 43, ,8 454; Sanger; Illumina Baudoinia compniacensis Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Teratosphaeriaceae UAMH Gp JGI extremophile 21,88 43, ,8 454; Sanger; Illumina Hortaea acidophila Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Teratosphaeriaceae CBS v industrial importance (acid-tolerant black yeast) 20,43 100, Illumina Hortaea werneckii Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Teratosphaeriaceae EXF-2000 Gp AIJO halotolerant 51,647 70, Illumina GAIIx Hortaea werneckii Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Teratosphaeriaceae EXF-2000 Gp AIJO halotolerant 51,647 70, Illumina GAIIx Teratosphaeria nubilosa Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Teratosphaeriaceae CBS Gp plant pathogen (leaf spot of Eucalyptus) 28,44 146, Illumina HiSeq Aureobasidium melanogenum Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Dothideales Dothideaceae CBS JGI industrial importance 26,2 140, Illumina Aureobasidium melanogenum Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Dothideales Dothideaceae CBS JGI industrial importance 26,2 140, Illumina Aureobasidium namibiae Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Dothideales Dothideaceae CBS JGI uncertain 25,43 138, Illumina Aureobasidium namibiae Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Dothideales Dothideaceae CBS JGI uncertain 25,43 138, Illumina Aureobasidium pullulans Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Dothideales Dothideaceae AY4 Gp AMCU industrial importance; opportunistic pathogen; endophyte 26,7 46, Illumina GAIIx Aureobasidium pullulans Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Dothideales Dothideaceae AY4 Gp AMCU industrial importance; opportunistic pathogen; endophyte 26, Illumina GAIIx Aureobasidium subglaciale Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Dothideales Dothideaceae EXF-2481 AYYB uncertain 25,79 814, ,8 Illumina Aureobasidium subglaciale Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Dothideales Dothideaceae EXF-2481 AYYB uncertain 25, ,8 Illumina Hysterium pulicare Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Hysteriales Hysteriaceae CBS Gp AJFK saprotroph 37, ,8 Illumina GAII Hysterium pulicare Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Hysteriales Hysteriaceae CBS Gp AJFK saprotroph 37,8 75, ,8 Illumina GAII Lepidopterella palustris Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Incertae sedis Argynnaceae CBS Gp JGI saprotroph (freshwater) 45,67 133, Illumina HiSeq Lepidopterella palustris Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Incertae sedis Argynnaceae CBS Gp JGI saprotroph (freshwater) 45,67 133, Illumina HiSeq Aulographum hederae Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Incertae sedis Aulographaceae CBS Gp JGI saprotroph (plant) 31, Illumina HiSeq Aulographum hederae Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Incertae sedis Aulographaceae CBS Gp JGI saprotroph (plant) 31,98 116, Illumina HiSeq

56 Eremomyces bilateralis Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Incertae sedis Eremomycetaceae CBS Gp uncertain 26,86 91, Illumina HiSeq Cryomyces antarcticus Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Incertae sedis Incertae sedis CCFEE 534 Gp AYQD rock-inhabiting; stress-tolerant 24,32 108, ,9 Ion Torrent Cryomyces antarcticus Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Incertae sedis Incertae sedis CCFEE 534 Gp AYQD rock-inhabiting; stress-tolerant 24,32 108, ,9 Ion Torrent Peltaster fructicola Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Incertae sedis Incertae sedis LNHT1506 Gp LJAO plant pathogen (apples) 19, ,9 Peltaster fructicola Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Incertae sedis Incertae sedis LNHT1506 Gp LJAO plant pathogen (apples) 19, ,9 Lizonia empirigonia Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Incertae sedis Pseudoperisporiaceae CBS v1.0 Gp uncertain 51,52 21, PacBio Lizonia empirigonia Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Incertae sedis Pseudoperisporiaceae CBS v1.0 Gp uncertain 51,52 21, PacBio Microthyrium microscopicum Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Microthyriales Microthyriaceae CBS Gp saprotroph 37,12 72, Illumina HiSeq Microthyrium microscopicum Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Microthyriales Microthyriaceae CBS Gp saprotroph 37,12 72, Illumina HiSeq Myriangium duriaei Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Myriangiales Myriangiaceae CBS Gp JGI parasite (insect) 25,69 97, Illumina HiSeq Myriangium duriaei Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Myriangiales Myriangiaceae CBS Gp JGI parasite (insect) 25,69 97, Illumina HiSeq Cenococcum geophilum Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Mytilinidiales Gloniaceae 1,58 Gp JGI ectomycorrhizal 177, Illumina HiSeq Cenococcum geophilum Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Mytilinidiales Gloniaceae 1,58 Gp JGI ectomycorrhizal 177,57 75, Illumina HiSeq Lophium mytilinum Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Mytilinidiales Mytilinidiaceae CBS Gp JGI uncertain 43,41 74, Illumina HiSeq Lophium mytilinum Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Mytilinidiales Mytilinidiaceae CBS Gp JGI uncertain 43,41 74, Illumina HiSeq Patellaria atrata Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Patellariales Patellariaceae CBS Gp JGI saprotroph 28,69 192, Illumina HiSeq Patellaria atrata Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Patellariales Patellariaceae CBS Gp JGI saprotroph 28,69 192, Illumina HiSeq Rhizodiscina lignyota Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Patellariales Patellariaceae CBS Gp mycobiont (lichen-forming) 33,43 95, Illumina HiSeq Rhytidhysteron rufulum Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Patellariales Patellariaceae CBS Gp AJFL saprotroph (mainly); occasionaly plant pathogen 40,18 109, ,9 Illumina GAII Rhytidhysteron rufulum Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Patellariales Patellariaceae CBS Gp AJFL saprotroph (mainly); occasionaly plant pathogen 40, ,9 Illumina GAII Amniculicola lignicola Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Pleosporales Amniculicolaceae CBS Gp JGI saprotroph 49, Illumina HiSeq Amniculicola lignicola Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Pleosporales Amniculicolaceae CBS Gp JGI saprotroph 49,58 99, Illumina HiSeq Nigrograna mackinnonii Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Biatriosporaceae E5202H Gp JGVQ endophyte (root) 51, ,9 Illumina HiSeq Corynespora cassicola Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Pleosporales Corynesporascaceae UM 591 Gp JAQF plant pathogen (broad host range) 41, , ,5 Illumina HiSeq Corynespora cassicola Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Pleosporales Corynesporascaceae UM 591 Gp JAQF plant pathogen (broad host range) 41, ,5 Illumina HiSeq Cucurbitaria berberidis Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Pleosporales Cucurbitariaceae CBS Gp JGI saprotroph 32,91 145, Illumina HiSeq Cucurbitaria berberidis Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Pleosporales Cucurbitariaceae CBS Gp JGI saprotroph 32,91 145, Illumina HiSeq Delitschia confertaspora Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Pleosporales Delitschiaceae ATCC Gp JGI coprophilous 31,16 74, Illumina HiSeq Delitschia confertaspora Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Pleosporales Delitschiaceae ATCC Gp JGI coprophilous 31,16 74, Illumina HiSeq Clathrospora elynae Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Pleosporales Diademaceae CBS Gp JGI uncertain 37,39 80, Illumina HiSeq Clathrospora elynae Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Pleosporales Diademaceae CBS Gp JGI uncertain 37,39 80, Illumina HiSeq Ascochyta rabiei Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Pleosporales Didymellaceae ArDII Gp JYNV plant pathogen (blight on chickpea) 34, ,7 Illumina Didymella exigua Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Didymellaceae CBS Gp JGI saproptroph 34,39 82, Illumina Didymella exigua Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Didymellaceae CBS Gp JGI saproptroph 34,39 82, Illumina Macroventuria anomochaeta Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Didymellaceae CBS Gp JGI saprotroph 33,34 116, Illumina HiSeq Macroventuria anomochaeta Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Didymellaceae CBS Gp JGI saprotroph 33,34 116, Illumina HiSeq Phoma herbarum Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Didymellaceae JCM Gp BCGR plant pathogen (hop & hemp) 39, ,7 HiSeq 2500 Phoma herbarum Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Didymellaceae JCM Gp BCGR plant pathogen (hop & hemp) 39, ,7 HiSeq 2500 Stagonosporopsis tanaceti Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Didymellaceae Pooled DNA Gp JUDZ plant pathogen (Asteraceae) 40, ,3 Illumina Stagonosporopsis tanaceti Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Didymellaceae Pooled DNA Gp JUDZ plant pathogen (Asteraceae) 40, ,3 Illumina Karstenula rhodostoma Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Didymosphaeriaceae CBS Gp JGI saprotroph 45, Illumina HiSeq Karstenula rhodostoma Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Didymosphaeriaceae CBS Gp JGI saprotroph 45,07 121, Illumina HiSeq Verruculina enalia Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Didymosphaeriaceae CBS Gp JGI marine fungus; mangrove-associated 61,21 81, Illumina HiSeq Verruculina enalia Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Didymosphaeriaceae CBS Gp JGI marine fungus; mangrove-associated 61,21 81, Illumina HiSeq Dothidotthia symphoricarpi Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Dothidotthiaceae CBS Gp JGI saprotroph 34,43 93, Illumina HiSeq Dothidotthia symphoricarpi Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Dothidotthiaceae CBS Gp JGI saprotroph 34,43 93, Illumina HiSeq Lineolata rhizophorae Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Fenestellaceae ATCC v marine fungus 31,14 97, Illumina Aaosphaeria arxii Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Incertae sedis CBS Gp JGI uncertain 38,9 86, Illumina HiSeq Aaosphaeria arxii Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Incertae sedis CBS Gp JGI uncertain 38,9 86, Illumina HiSeq Periconia macrospinosa Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Incertae sedis DSE2036 Gp endophyte (root) 54,99 139, Trinosporium guianense Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Incertae sedis CBS Gp uncertain; environmental contaminant 24,23 140, ,00% 8062 Illumina HiSeq Lentithecium fluviatile Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Lentitheciaceae CBS Gp JGI saprotroph (freshwater) 54,69 85, Illumina HiSeq Lentithecium fluviatile Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Lentitheciaceae CBS Gp JGI saprotroph (freshwater) 54,69 85, Illumina HiSeq Leptosphaeria maculans Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Leptosphaeriaceae JN3 Gp ASM plant pathogen (Brassica crops) 45, ,3 Leptosphaeria maculans Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Leptosphaeriaceae JN3 Gp ASM plant pathogen (Brassica crops) 45, ,3 Ophiobolus disseminans Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Leptosphaeriaceae CBS Gp JGI uncertain; isolated from poisonous Mercurialis perennis 41,68 97, Illumina HiSeq Ophiobolus disseminans Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Leptosphaeriaceae CBS Gp JGI uncertain; isolated from poisonous Mercurialis perennis 41,68 97, Illumina HiSeq Plenodomus tracheiphilus Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Leptosphaeriaceae IPT5 Gp JGI plant pathogen (citrus) 34,24 98, Illumina HiSeq Plenodomus tracheiphilus Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Leptosphaeriaceae IPT5 Gp JGI plant pathogen (citrus) 34,24 98, Illumina HiSeq Lophiostoma macrostomum Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Lophiostomataceae CBS Gp JGI saprotroph 42,58 140, Illumina HiSeq Lophiostoma macrostomum Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Lophiostomataceae CBS Gp JGI saprotroph 42,58 140, Illumina HiSeq Lophiotrema nucula Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Lophiostomataceae CBS Go uncertain 48,64 138, Illumina Byssothecium circinans Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Pleosporales Massarinaceae CBS Gp JGI saprotroph 49,29 96, Illumina HiSeq Byssothecium circinans Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Pleosporales Massarinaceae CBS Gp JGI saprotroph 49,29 96, Illumina HiSeq Helminthosporium solani Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Massarinaceae B-AC-16A Gp AWWW plant pathogen (potato) 34,513 50, ,1 454; Illumina HiSeq Helminthosporium solani Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Massarinaceae B-AC-16A Gp AWWW plant pathogen (potato) 34,513 50, ,1 454; Illumina HiSeq Massarina eburnea Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Massarinaceae CBS Gp uncertain 38,24 56, Illumina HiSeq Beverwykella pulmonaria Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Pleosporales Melanommataceae JCM 9230 Gp BCHH saprotroph 45, ,2 HiSeq 2500 Beverwykella pulmonaria Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Pleosporales Melanommataceae JCM 9230 Gp BCHH saprotroph 45,97 111, ,2 HiSeq 2500 Melanomma pulvis-pyrius Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Melanommataceae CBS Gp JGI saprotroph 42,09 119, Illumina HiSeq Melanomma pulvis-pyrius Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Melanommataceae CBS Gp JGI saprotroph 42,09 119, Illumina HiSeq Bimuria novae-zelandiae Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Montagnulaceae CBS v1.0 Gp parasite; lichenicolous 78,18 62, Illumina Paraconiothyrium sporulosum Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Montagnulaceae AP3s5-JAC2a Gp manganese(ii)-oxidizing; saprotroph (soil) 38,46 118, Illumina HiSeq Parastagonospora nodorum Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Phaeosphaeriaceae SN15 Gp AAGI plant pathogen (wheat) 37,21 10, Sanger Parastagonospora nodorum Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Phaeosphaeriaceae SN15 Gp AAGI plant pathogen (wheat) 37,21 10, Sanger Phaeosphaeriaceae sp. Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Phaeosphaeriaceae PMI808 Gp uncertain 56,37 148, Setomelanomma holmii Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Phaeosphaeriaceae CBS v1.0 Gp plant pathogen (spruce needle drop) 39,18 109, Illumina Setomelanomma holmii Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Phaeosphaeriaceae CBS v1.0 Gp plant pathogen (spruce needle drop) 39,18 109, Illumina Stagonospora sp. SRC1lsM3a Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Phaeosphaeriaceae SRC1lsM3a Gp Mn(II)-oxidizing; mineral cycling 36,08 87, Trichodelitschia bisporula Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Phaeotrichaceae CBS Gp coprophilous 25,79 85, Illumina HiSeq Pleomassaria siparia Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Pleomassariaceae CBS Gp JGI saprotroph 43,18 87, Illumina HiSeq Pleomassaria siparia Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Pleomassariaceae CBS Gp JGI saprotroph 43,18 87, Illumina HiSeq Alternaria alternata Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Pleosporaceae SRC1lrK2f Gp plant pathogen (leaf spot; broad host range) 32,99 147, Illumina HiSeq Alternaria arborescens Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Pleosporaceae EGS Gp AIIC plant pathogen (tomato) 33, Illumina GA II Alternaria arborescens Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Pleosporaceae EGS Gp AIIC plant pathogen (tomato) 33,9 90, Illumina GA II Alternaria brassicicola Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Pleosporaceae ATCC Gp ACIW plant pathogen (Brassica dark leaf spot) 29,5 6, Alternaria brassicicola Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Pleosporaceae ATCC Gp ACIW plant pathogen (Brassica dark leaf spot) 29,5 6, Bipolaris maydis Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Pleosporales Pleosporaceae C5 Gp AIHU plant pathogen (wheat) 36,46 67, ,8 Illumina not publically released Bipolaris maydis Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Pleosporales Pleosporaceae C5 Gp AIHU plant pathogen (wheat) 36,46 67, ,8 Illumina Bipolaris oryzae Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Pleosporales Pleosporaceae WK-1C, ATCC Gp AMCO plant pathogen (rice) 31,36 191, ,5 Illumina Bipolaris oryzae Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Pleosporales Pleosporaceae WK-1C, ATCC Gp AMCO plant pathogen (rice) 31, ,5 Illumina Bipolaris sorokiniana Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Pleosporales Pleosporaceae ND90Pr Gp AEIN plant pathogen (cereals) 34,41 42, ,8 454; Sanger; Illumina Bipolaris sorokiniana Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Pleosporales Pleosporaceae ND90Pr Gp AEIN plant pathogen (cereals) 34,41 42, ,8 454; Sanger; Illumina Bipolaris victoriae Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Pleosporales Pleosporaceae FI3 Gp AMCY plant pathogen (blight of oats) 32, ,1 Illumina Bipolaris victoriae Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Pleosporales Pleosporaceae FI3 Gp AMCY plant pathogen (blight of oats) 32, ,1 Illumina Bipolaris zeicola Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Pleosporales Pleosporaceae 26-R-13 Gp AMCN plant pathogen (sorghum, maize and apple) 31, ,8 Illumina Bipolaris zeicola Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Pleosporales Pleosporaceae 26-R-13 Gp AMCN plant pathogen (sorghum, maize and apple) 31,27 200, ,8 Illumina Cochliobolus lunatus Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Pleosporales Pleosporaceae CX-3 Gp JGI plant pathogen (sorghum); pharmaceutical importance 31,17 129, Illumina HiSeq Cochliobolus lunatus Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Pleosporales Pleosporaceae CX-3 Gp JGI plant pathogen (sorghum); pharmaceutical importance 31,17 129, Illumina HiSeq Curvularia papendorfii Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Pleosporales Pleosporaceae UM 226 Gp JXCC opportunistic human pathogen; rare 33, ,6 Illumina HiSeq; PacBio Curvularia papendorfii Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Pleosporales Pleosporaceae UM 226 Gp JXCC opportunistic human pathogen; rare 33,38 121, ,6 Illumina HiSeq; PacBio

57 Decorospora gaudefroyi Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Pleosporales Pleosporaceae CBS marine saprotroph 30,56 152, Illumina Decorospora gaudefroyi Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Pleosporales Pleosporaceae CBS marine saprotroph 30,56 152, Illumina Pyrenochaeta lycopersici Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Pleosporaceae CRA-PAV_ER 1211 Gp ASRS plant pathogen (tomato) 50, , ,7 Illumina GAIIx Pyrenochaeta lycopersici Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Pleosporaceae CRA-PAV_ER 1211 Gp ASRS plant pathogen (tomato) 50, , ,7 Illumina GAIIx Pyrenochaeta sp. DS3sAY3a Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Pleosporaceae DS3sAY3a Gp JGI Mn(II)-oxidizing; mineral cycling 38,5 86, Illumina HiSeq Pyrenochaeta sp. DS3sAY3a Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Pleosporaceae DS3sAY3a Gp JGI Mn(II)-oxidizing; mineral cycling 38,5 86, Illumina HiSeq Pyrenochaeta sp. UM 256 Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Pleosporaceae UM 256 Gp AOUM human pathogen 35,484 27, ,4 454 Pyrenochaeta sp. UM 256 Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Pleosporaceae UM 256 Gp AOUM human pathogen 35,484 27, ,4 454 Pyrenophora seminiperda Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Pleosporaceae CCB06 Gp ATLS plant pathogen (necrotrophic; seeds of grasses/cereals) 32,539 17, ,8 454 Pyrenophora seminiperda Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Pleosporaceae CCB06 Gp ATLS plant pathogen (necrotrophic; seeds of grasses/cereals) 32,539 17, ,8 454 Pyrenophora teres Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Pleosporaceae 0-1 Gp AEEY plant pathogen (barley and some other crops) 33,583 20, ,9 Illumina Pyrenophora teres Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Pleosporaceae 0-1 Gp AEEY plant pathogen (barley and some other crops) 33,583 20, ,9 Illumina Pyrenophora tritici-repentis Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Pleosporaceae Pt-1C-BFP Gp AAXI plant pathogen (cereals and grasses; necrotrophic) 37,997 6, Sanger Pyrenophora tritici-repentis Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Pleosporaceae Pt-1C-BFP Gp AAXI plant pathogen (cereals and grasses; necrotrophic) 37,997 6, Sanger Setosphaeria turcica Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Pleosporaceae Et28A Gp AIHT plant pathogen (maize) 43,01 58, ,4 454; Sanger; Illumina Setosphaeria turcica Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Pleosporaceae Et28A Gp AIHT plant pathogen (maize) 43,01 58, ,4 454; Sanger; Illumina Stemphylium lycopersici Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Pleosporaceae CIDEFI 216 Gp LGLR plant pathogen (fruits) 35,17 77, ,7 Illumina HiSeq Stemphylium lycopersici Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Pleosporaceae CIDEFI 216 Gp LGLR plant pathogen (fruits) 35,17 77, ,7 Illumina HiSeq Shiraia sp. slf14 Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Shiraiaceae Slf14 Gp AXZN endophyte 32,067 55, Illumina HiSeq Shiraia sp. slf14 Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Shiraiaceae Slf14 Gp AXZN endophyte 32,067 55, Illumina HiSeq Sporormia fimetaria Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Sporormiaceae CBS Gp JGI coprophilous 25, Illumina HiSeq Sporormia fimetaria Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Sporormiaceae CBS Gp JGI coprophilous 25, Illumina HiSeq Westerdykella ornata Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Sporormiaceae CBS Gp JGI pharmaceutical importance (antibiotic-producing) 26,98 162, Illumina HiSeq Westerdykella ornata Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Sporormiaceae CBS Gp JGI pharmaceutical importance (antibiotic-producing) 26,98 162, Illumina HiSeq Polyplosphaeria fusca Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Tetraplosphaeriaceae CBS Gp JGI bambusicolous 37, Illumina HiSeq Polyplosphaeria fusca Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Tetraplosphaeriaceae CBS Gp JGI bambusicolous 37, Illumina HiSeq Trematosphaeria pertusa Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Trematosphaeriaceae CBS Gp JGI saprotroph (freshwater) 47,75 96, Illumina HiSeq Trematosphaeria pertusa Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Trematosphaeriaceae CBS Gp JGI saprotroph (freshwater) 47,75 96, Illumina HiSeq Pleosporales sp. UM Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales unknown UM 1110 Gp AJMS uncertain 36, , ,4 Illumina GAIIx Pleosporales sp. UM Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales unknown UM 1110 Gp AJMS uncertain 36, ,4 Illumina GAIIx Zopfia rhizophila Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Zopfiaceae CBS Gp JGI opportunistic root rot 152,78 95, Illumina HiSeq; PacBio Zopfia rhizophila Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Zopfiaceae CBS Gp JGI opportunistic root rot 152,78 95, Illumina HiSeq; PacBio Lineolata rhizophorae Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Fenestellaceae ATCC v marine fungus 31,14 97, Illumina Trypethelium eluteriae Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Trypetheliales Trypetheliaceae not specified JGI mycobiont (lichen-forming) 32,16 142, Illumina Trypethelium eluteriae Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Trypetheliales Trypetheliaceae not specified JGI mycobiont (lichen-forming) 32,16 142, Illumina Ochroconis constricta Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Venturiales Sympoventuriaceae UM 578 Gp AZYM animal pathogen; opportunistic human pathogen 34,434 87, ,1 Illumina HiSeq Ochroconis constricta Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Venturiales Sympoventuriaceae UM 578 Gp AZYM animal pathogen; opportunistic human pathogen 34, ,1 Illumina HiSeq Verruconis gallopava Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Venturiales Sympoventuriaceae CBS JYBX medical importance (black yeast) 31, Illumina Tothia fuscella Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Venturiales Venturiaceae CBS Gp uncertain 36,78 146, Illumina HiSeq Tothia fuscella Ascomycota Pezizomycotina Dothideomycetes Incertae sedis Venturiales Venturiaceae CBS Gp uncertain 36,78 146, Illumina HiSeq Venturia pyrina Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Venturiales Venturiaceae ICMP Gp JEMP plant pathogen (hemibiotrophic; pear) 41,177 40, ,6 Illumina GAIIx Venturia pyrina Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Venturiales Venturiaceae ICMP Gp JEMP plant pathogen (hemibiotrophic; pear) 41, ,6 Illumina GAIIx Hortaea acidophila Ascomycota Pezizomycotina Dothideomycetes Dothideomycetidae Capnodiales Teratosphaeriaceae CBS v industrial importance (acid-tolerant black yeast) 20,43 100, Illumina Bimuria novae-zelandiae Ascomycota Pezizomycotina Dothideomycetes Pleosporomycetidae Pleosporales Montagnulaceae CBS v parasite; lichenicolous 78,18 62, Illumina Rhizodiscina lignyota Ascomycota Pezizomycotina Dothidiomycetes Incertae sedis Patellariales Patellariaceae CBS Gp mycobiont (lichen-forming) 33,43 95, Illumina HiSeq Ascosphaera apis Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Ascosphaerales Ascosphaeraceae USDA-ARSEF 7405 Gp AARE animal pathogen (honey bees) 21, Sanger Ascosphaera apis Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Ascosphaerales Ascosphaeraceae USDA-ARSEF 7405 Gp AARE animal pathogen (honey bees) 21,5 4, Sanger Cyphellophora europaea Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Cyphellophoraceae CBS AOBU human pathogen (superficial infection); dermatiaceous 28, , Illumina Cyphellophora europaea Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Cyphellophoraceae CBS AOBU human pathogen (superficial infection); dermatiaceous 28, Illumina Capronia coronata Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Herpotrichiellaceae CBS Gp AMWN ; Broad saprotroph; possible beetle associate; black yeast 25,8 90, ,8 Capronia coronata Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Herpotrichiellaceae CBS Gp AMWN ; Broad saprotroph; possible beetle associate; black yeast 25, ,8 Capronia epimyces Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Herpotrichiellaceae CBS Gp ; Broad AMGY uncertain; yeast (black) 28,9 83, ,4 Capronia epimyces Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Herpotrichiellaceae CBS Gp ; Broad AMGY uncertain; yeast (black) 28, ,4 Capronia semiimmersa Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Herpotrichiellaceae CBS27337 Go JYCC medical importance (black yeast) 31, Illumina Cladophialophora bantiana Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Herpotrichiellaceae CBS Go JYBT medical importance (black yeast) 36, Illumina Cladophialophora carrionii Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Herpotrichiellaceae CBS Gp AOFF human pathogen (chromoblastomycosis ); dermatiaceous 28,99 80, ,3 Illumina Cladophialophora carrionii Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Herpotrichiellaceae CBS Gp AOFF human pathogen (chromoblastomycosis ); dermatiaceous 28,99 80, ,3 Illumina Cladophialophora immunda Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Herpotrichiellaceae CBS Gp JSEJ industrial importance (black yeast; bioremediation; degrades polyaromatic hydrocarbons) 41, ,8 IonTorrent Cladophialophora immunda Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Herpotrichiellaceae CBS Gp JSEJ industrial importance (black yeast; bioremediation; degrades polyaromatic hydrocarbons) 41,58 318, ,8 IonTorrent Cladophialophora psammophila Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Herpotrichiellaceae CBS Gp AMGX industrial importance (bioremediation) 39,42 133, ,6 Illumina Cladophialophora psammophila Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Herpotrichiellaceae CBS Gp AMGX industrial importance (bioremediation) 39,42 133, ,6 Illumina Cladophialophora yegresii Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Herpotrichiellaceae CBS Gp AMGW endophyte (cactus); dermatiaceous; potential human pathogen 27,89 132, Illumina Cladophialophora yegresii Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Herpotrichiellaceae CBS Gp AMGW endophyte (cactus); dermatiaceous; potential human pathogen 27,89 132, Illumina Coniosporium apollinis Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Herpotrichiellaceae CBS Gp AJKL rock-inhabiting black yeast 28,64 128, ,1 Illumina Coniosporium apollinis Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Herpotrichiellaceae CBS Gp AJKL rock-inhabiting black yeast 28, ,1 Illumina Exophiala alcalophila Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Herpotrichiellaceae JCM 1751 Gp BCHY black yeast; soil-associated 29, ,1 HiSeq 2500 Exophiala alcalophila Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Herpotrichiellaceae JCM 1751 Gp BCHY black yeast; soil-associated 29, ,1 HiSeq 2500 Exophiala aquamarina Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Herpotrichiellaceae CBS Gp AMGV animal pathogen (marine) 41,566 79, Illumina Exophiala aquamarina Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Herpotrichiellaceae CBS Gp AMGV animal pathogen (marine) 41, Illumina Exophiala calicioides Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Herpotrichiellaceae JCM 6030 Gp BCHZ insect-associated (bark beetles) 36, ,8 HiSeq 2500 Exophiala calicioides Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Herpotrichiellaceae JCM 6030 Gp BCHZ insect-associated (bark beetles) 36,37 153, ,8 HiSeq 2500 Exophiala dermatitidis Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Herpotrichiellaceae NIH 8656 Gp AFPA human pathogen (rare); thermophilic 26,37 151, ,5 Illumina not publically released Exophiala dermatitidis Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Herpotrichiellaceae NIH 8656 Gp AFPA human pathogen (rare); thermophilic 26,37 151, ,5 Illumina Exophiala mesophila Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Herpotrichiellaceae CBS Gp JSEI industrual importance (bioremediation; black yeast) 30, ,4 IonTorrent Exophiala mesophila Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Herpotrichiellaceae CBS Gp JSEI industrual importance (bioremediation; black yeast) 30,38 393, ,4 IonTorrent Exophiala oligosperma Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Herpotrichiellaceae CBS72588 JYCA medical importance (black yeast) 38, Illumina Exophiala sideris Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Herpotrichiellaceae CBS JYBR medical importance (black yeast) 29, Illumina Exophiala spinifera Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Herpotrichiellaceae JCM Gp BCHD human pathogen (cutaneous) 32, ,7 HiSeq 2500 Exophiala spinifera Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Herpotrichiellaceae JCM Gp BCHD human pathogen (cutaneous) 32, ,7 HiSeq 2500 Exophiala xenobiotica Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Herpotrichiellaceae CBS Gp JTCI opportunistic human pathogen; bioremdiation 32, ,5 IonTorrent Exophiala xenobiotica Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Herpotrichiellaceae CBS Gp JTCI opportunistic human pathogen; bioremdiation 32,27 354, ,5 IonTorrent Fonsecaea erecta Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Herpotrichiellaceae CBS Gp LVYI medical importance (black yeast) 34, Illumina; Ion Torrent Fonsecaea monophora Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Herpotrichiellaceae CBS Gp LVKK medical importance (black yeast) 35,23 90, ,2 Illumina; Ion Torrent Fonsecaea multimorphosa Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Herpotrichiellaceae CBS Go JYBV medical importance (black yeast) 33, Illumina Fonsecaea nubica Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Herpotrichiellaceae CBS Gp LVCJ medical importance (black yeast) 33, ,5 Illumina; Ion Torrent Fonsecaea pedrosoi Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Herpotrichiellaceae CBS Go JYBS medical importance (black yeast) 34, Illumina Herpotrichiellaceae sp. UM238 Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Herpotrichiellaceae UM238 Gp AMYF opportunistic human pathogen 28, , ,8 Illumina GaII Herpotrichiellaceae sp. UM238 Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Herpotrichiellaceae UM238 Gp AMYF opportunistic human pathogen 28, ,8 Illumina GaII Phialophora attae Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Herpotrichiellaceae CBS Gp LFJN insect-associated (ants) 30, ,6 IonTorrent Phialophora attae Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Herpotrichiellaceae CBS Gp LFJN insect-associated (ants) 30, ,6 IonTorrent Rhinocladiella mackenziei Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Chaetothyriales Herpotrichiellaceae CBS Gp JYBU human pathogen (neurotropic; cerebral phaeohyphomycosis) 32, Aspergillus acidus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae ATCC Gp JGI saprotroph 37,47 65, GS-FLX Aspergillus acidus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae ATCC Gp JGI saprotroph 37,47 65, GS-FLX Aspergillus aculeatus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae ATCC Gp JGI industrial importance 35, ; Sanger Aspergillus aculeatus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae ATCC Gp JGI industrial importance 35, ; Sanger Aspergillus awamori Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae JCM Gp BCGT industrial importance (fermentation; starch degradation) 36,06 209, ,8 Illumina Aspergillus brasiliensis Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae CBS Gp JGI industrial importance 35,81 156, ; Illumina Aspergillus brasiliensis Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae CBS Gp JGI industrial importance 35,81 156, ; Illumina Aspergillus calidoustus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae ACAL_DNA_1 Gp CDMC human pathogen 41, ,1 Aspergillus calidoustus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae ACAL_DNA_1 Gp CDMC human pathogen 41, ,1 Aspergillus carbonarius Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae ITEM 5010 Gp JGI industrial importance 36,3 17, ; Sanger

58 Aspergillus carbonarius Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae ITEM 5010 Gp JGI industrial importance 36,3 17, ; Sanger Aspergillus chevalieri Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae JCM Gp BCIE opportunistic human pathogen 28, HiSeq 2500 Aspergillus chevalieri Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae JCM Gp BCIE opportunistic human pathogen 28,01 313, HiSeq 2500 Aspergillus clavatus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae NRRL 1 Gp AAKD soil- and manure-associated; occasional pathogen 27,9 11, Sanger Aspergillus clavatus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae NRRL 1 Gp AAKD soil- and manure-associated; occasional pathogen 27,9 11, Sanger Aspergillus flavus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae NRRL 3357 Gp AAIH post-harvest decay 36, Sanger Originally submitted as A. delicata Aspergillus flavus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae NRRL 3357 Gp AAIH post-harvest decay 36,9 5, Sanger Aspergillus fumigatus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae A1163 (CEA10) Gp ABDB opportunistic human pathogen; saprotroph 29,2 10,00 55 Sanger Aspergillus fumigatus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae A1163 (CEA10) Gp ABDB opportunistic human pathogen; saprotroph 29,2 10,00 55 Sanger Aspergillus glaucus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae CBS Gp JGI saprotroph 27,99 175, GS-FLX Aspergillus glaucus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae CBS Gp JGI saprotroph 27,99 175, GS-FLX Aspergillus kawachii Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae IFO 4308 Gp BACL industrial importance 37,1 17, GS-FLX Aspergillus kawachii Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae IFO 4308 Gp BACL industrial importance 37,1 17, GS-FLX Aspergillus lentulus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae IFM Gp BCLY opportunistic human pathogen 30, ,5 PacBio RSII Aspergillus lentulus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae IFM Gp BCLY opportunistic human pathogen 30, ,5 PacBio RSII Aspergillus luchuensis Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae RIB 2604 Gp BCWF industrial importance 34, ,9 ABI 3730 Aspergillus luchuensis Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae RIB 2604 Gp BCWF industrial importance 34, ,9 ABI 3730 Aspergillus nidulans Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae FGSC A4 Gp ; Gp000JGI 130 human pathogen 30,48 1, Sanger Aspergillus nidulans Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae FGSC A4 Gp ; Gp000JGI 130 human pathogen 30,48 1, Sanger Aspergillus niger Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae CBS Gp ; Gp001JGI industrial importance 34,85 8, Aspergillus niger Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae CBS Gp ; Gp001JGI industrial importance 34,85 8, Aspergillus nomius Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae NRRL13137 Gp JNOM human pathogen (aspergillosis) 36, ,9 IonTorrent Aspergillus nomius Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae NRRL13137 Gp JNOM human pathogen (aspergillosis) 36,14 42, ,9 IonTorrent Aspergillus ochraceoroseus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae SRRC1432 Gp JYKN medical importance (mycotoxin-producing) 24, ,2 IonTorrent Aspergillus ochraceoroseus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae SRRC1432 Gp JYKN medical importance (mycotoxin-producing) 24,27 39, ,2 IonTorrent Aspergillus oryzae Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae AS Gp AKXN industrial importance 36,3 7, Illumina GAII Aspergillus oryzae Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae AS Gp AKXN industrial importance 36,3 7, Illumina GAII Aspergillus parasiticus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae SU-1 Gp JMUG medical importance (aflatoxin-producing) 40 50, Illumina HiSeq Aspergillus parasiticus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae SU-1 Gp JMUG medical importance (aflatoxin-producing) 40 50, Illumina HiSeq Aspergillus rambellii Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae SRRC1468 Gp JZBS medical importance (mycotoxin-producing) 26, ,8 IonTorrent Aspergillus rambellii Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae SRRC1468 Gp JZBS medical importance (mycotoxin-producing) 26,44 89, ,8 IonTorrent Aspergillus ruber Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae CBS Gp JGI halotolerant 26,21 91, ; Illumina Aspergillus ruber Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae CBS Gp JGI halotolerant 26,21 91, ; Illumina Aspergillus sojae Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae NBRC 4239 Gp BACA traditional use 39,7 14, Aspergillus sojae Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae NBRC 4239 Gp BACA traditional use 39,7 14, Aspergillus sp. Z5 Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae Z5 Gp LDZW marine fungus; secondary metabolite biosynthesis 33, ,5 Illumina HiSeq Aspergillus sp. Z5 Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae Z5 Gp LDZW marine fungus; secondary metabolite biosynthesis 33,81 100, ,5 Illumina HiSeq Aspergillus sydowii Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae CBS Gp JGI animal pathogen (sea fan corals) 34,38 95, GS-FLX Aspergillus sydowii Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae CBS Gp JGI animal pathogen (sea fan corals) 34,38 95, GS-FLX Aspergillus terreus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae NIH2624 Gp JGI soil-associated; cosmopolitan 29,33 11, Sanger Aspergillus terreus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae NIH2624 Gp JGI soil-associated; cosmopolitan 29,33 11, Sanger Aspergillus tubingensis Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae CBS Gp JGI industrial importance 35,15 125, ; Illumina Aspergillus tubingensis Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae CBS Gp JGI industrial importance 35,15 125, ; Illumina Aspergillus udagawae Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae IFM Gp BBXM animal pathogen; aspergillosis 32, ,6 HiSeq 1500 Aspergillus udagawae Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae IFM Gp BBXM animal pathogen; aspergillosis 32, ,6 HiSeq 1500 Aspergillus ustus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae Gp JOMC opportunistic human pathogen 38, Illumina HiSeq Aspergillus ustus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae Gp JOMC opportunistic human pathogen 38,35 100, Illumina HiSeq Aspergillus versicolor Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae CBS Gp JGI industrial importance 33,13 178, GS-FLX Aspergillus versicolor Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae CBS Gp JGI industrial importance 33,13 178, GS-FLX Aspergillus wentii Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae DTO 134E9 Gp JGI industrial importance 31,35 95, Illumina Aspergillus wentii Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae DTO 134E9 Gp JGI industrial importance 31,35 95, Illumina Aspergillus westerdijkiae Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae CBS Gp LKBE medical importance (mycotoxin-producing) 36, ,2 Illumina MiSeq Aspergillus westerdijkiae Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae CBS Gp LKBE medical importance (mycotoxin-producing) 36,07 142, ,2 Illumina MiSeq Aspergillus zonatus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae CBS Gp JGI industrial importance 28,92 84, Illumina Aspergillus zonatus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae CBS Gp JGI industrial importance 28,92 84, Illumina Basipetospora chlamydospora Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae JCM Gp BCHP uncertain 33, ,7 HiSeq 2500 Basipetospora chlamydospora Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae JCM Gp BCHP uncertain 33,49 162, ,7 HiSeq 2500 Monascus purpureus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae NRRL 1596 Gp JGI industrial importance (toxin producing) 23,44 102, Illumina HiSeq Monascus purpureus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae NRRL 1596 Gp JGI industrial importance (toxin producing) 23,44 102, Illumina HiSeq Monascus ruber Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae NRRL 1597 Gp JGI industrial importance 24,8 123, Illumina HiSeq Monascus ruber Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae NRRL 1597 Gp JGI industrial importance 24,8 123, Illumina HiSeq Penicillium aurantiogriseum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae NRRL Gp ALJY endophyte 31, , ,5 Illumina GAIIx Penicillium aurantiogriseum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae NRRL Gp ALJY endophyte 31, , ,5 Illumina GAIIx Penicillium biforme Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae FM169 Gp CBXO pharmaceutical importance (rugulovasine production) 34, ,1 Penicillium biforme Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae FM169 Gp CBXO pharmaceutical importance (rugulovasine production) 34, ,1 Penicillium bilaiae Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae ATCC Gp JGI P-solubilizing 37, Illumina HiSeq Penicillium bilaiae Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae ATCC Gp JGI P-solubilizing 37,54 101, Illumina HiSeq Penicillium brasilianum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae PMG11 Gp CDHK endophyte 35, ,5 Penicillium brasilianum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae PMG11 Gp CDHK endophyte 35, ,5 Penicillium brevicompactum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae AgRF18 Gp JGI industrial importance (metabolite production); water-damage contaminant 31,68 98, Illumina Penicillium brevicompactum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae AgRF18 Gp JGI industrial importance (metabolite production); water-damage contaminant 31,68 98, Illumina Penicillium camemberti Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae FM 013 Gp CBVV industrial importance (cheese-making) 35, ,2 Penicillium camemberti Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae FM 013 Gp CBVV industrial importance (cheese-making) 35, ,2 Penicillium canescens Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae ATCC Gp JGI calcium phosphate solubilizer 33, Illumina Penicillium canescens Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae ATCC Gp JGI calcium phosphate solubilizer 33,26 97, Illumina Penicillium capsulatum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae CBS Gp JPLR industrial importance (paper production); opportunitsic human pathogen 34, ,1 Illumina; PacBio Penicillium capsulatum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae CBS Gp JPLR industrial importance (paper production); opportunitsic human pathogen 34,34 231, ,1 Illumina; PacBio Penicillium carneum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae LCP05634 Gp CBXS saprotroph; medical importance (mycotoxin-producing) 25, ,1 Penicillium carneum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae LCP05634 Gp CBXS saprotroph; medical importance (mycotoxin-producing) 25, ,1 Penicillium chrysogenum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae IB 08/921 Gp JPDR saprotroph 31, ,9 Illumina HiSeq Penicillium chrysogenum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae IB 08/921 Gp JPDR saprotroph 31, ,9 Illumina HiSeq Penicillium citrinum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae DSM 1997 Gp LKUP saprotroph; medical importance (mycotoxin-producing) 31, ,9 Illumina MiSeq Penicillium citrinum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae DSM 1997 Gp LKUP saprotroph; medical importance (mycotoxin-producing) 31,53 64, ,9 Illumina MiSeq Penicillium digitatum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae Pd1 Gp AKCU post-harvest decay 26,082 24, ,9 454 Penicillium digitatum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae Pd1 Gp AKCU post-harvest decay 26,082 24, ,9 454 Penicillium expansum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae d1 Gp JQFY post-harvest decay 32, ,5 Illumina HiSeq Penicillium expansum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae d1 Gp JQFY post-harvest decay 32,1 605, ,5 Illumina HiSeq Penicillium fellutanum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae ATCC Gp JGI calcium phosphate solubilizer 30, Illumina HiSeq Penicillium fellutanum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae ATCC Gp JGI calcium phosphate solubilizer 30,2 121, Illumina HiSeq Penicillium freii Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae DAOM Gp LLXE psychrophile 33, ,4 Illumina HiSeq Penicillium freii Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae DAOM Gp LLXE psychrophile 33,55 79, ,4 Illumina HiSeq Penicillium fuscoglaucum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae FM041 Gp CBXP contaminant (cheese) 36, ,7 Penicillium fuscoglaucum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae FM041 Gp CBXP contaminant (cheese) 36, ,7 Penicillium glabrum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae DAOM Gp JGI post-harvest decay 33, Illumina HiSeq Penicillium glabrum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae DAOM Gp JGI post-harvest decay 33,16 96, Illumina HiSeq Penicillium griseofulvum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae PG3 Gp LHQR post-harvest decay 29, ,3 Illumina MiSeq Penicillium griseofulvum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae PG3 Gp LHQR post-harvest decay 29,14 270, ,3 Illumina MiSeq Penicillium italicum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae PHI-1 Gp JQGA post-harvest decay 30, ,2 Illumina HiSeq Penicillium italicum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae PHI-1 Gp JQGA post-harvest decay 30,16 421, ,2 Illumina HiSeq Penicillium janthinellum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae ATCC Gp JGI endophyte 35,15 99, Penicillium janthinellum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae ATCC Gp JGI endophyte 35,15 99,

59 Penicillium lanosocoeruleum v1.0 Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae ATCC Gp JGI pharmaceutical importance (antibiotic- and mycotoxin-producing) 29, Illumina Penicillium lanosocoeruleum v1.0 Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae ATCC Gp JGI pharmaceutical importance (antibiotic- and mycotoxin-producing) 29,05 113, Illumina Penicillium nalgiovense Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae FM193 Gp CBXQ industrial importance (fermentation; metabolite production) 31, ,5 Penicillium nalgiovense Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae FM193 Gp CBXQ industrial importance (fermentation; metabolite production) 31, ,5 Penicillium nordicum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae BFE487 Gp JNNR contaminant (food; toxigenic) 30,421 20, ,7 Illumina HiSeq Penicillium nordicum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae BFE487 Gp JNNR contaminant (food; toxigenic) 30,421 20, ,7 Illumina HiSeq Penicillium oxalicum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae AGIH lignocellulolytic 30, Penicillium oxalicum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae AGIH lignocellulolytic 30,18 28, Penicillium paneum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae FM227 Gp CBXN contaminant (cereal; toxigenic) 26, Penicillium paneum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae FM227 Gp CBXN contaminant (cereal; toxigenic) 26, Penicillium paxilli Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae ATCC Gp AOTG pharmaceutical importance (production of paxelline and antimicrobial com 34,8 150, ,9 Illumina MiSeq Penicillium paxilli Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae ATCC Gp AOTG pharmaceutical importance (production of paxelline and antimicrobial com 34,8 150, ,9 Illumina MiSeq Penicillium raistrickii Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae ATCC Gp JGI calcium phosphate solubilizer 31,44 95, Penicillium raistrickii Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae ATCC Gp JGI calcium phosphate solubilizer 31,44 95, Penicillium roqueforti Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae FM164 Gp CBMR industrial importance (cheese-making) 29, ,7 Penicillium roqueforti Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae FM164 Gp CBMR industrial importance (cheese-making) 29, ,7 Penicillium rubens Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae Wisconsin Gp JGI industrial importance (penicillin production) 32,22 9, Penicillium rubens Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae Wisconsin Gp JGI industrial importance (penicillin production) 32,22 9, Penicillium solitum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae Gp JYNM post-harvest decay 35,23 7,00 47,1 Penicillium verrucosum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae BFE808 Gp LAKW medical importance (ochratoxin-producing) 31, ,1 Illumina MiSeq Penicillium verrucosum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae BFE808 Gp LAKW medical importance (ochratoxin-producing) 31,15 60, ,1 Illumina MiSeq Xeromyces bisporus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae FRR 0525 Gp CCCX xerophile 22, ,4 Xeromyces bisporus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Aspergillaceae FRR 0525 Gp CCCX xerophile 22, ,4 Byssochlamys spectabilis Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Thermoascaceae No. 5 Gp BAUL industrial importance (formaldehyde resistant) 29,76 53, ,6 Illumina Byssochlamys spectabilis Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Thermoascaceae No. 5 Gp BAUL industrial importance (formaldehyde resistant) 29,76 53, ,6 Illumina Thermoascus aurantiacus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Thermoascaceae ATCC Gp JGI industrial importance (thermophile) 28,49 95, Illumina Thermoascus aurantiacus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Thermoascaceae ATCC Gp JGI industrial importance (thermophile) 28,49 95, Illumina Thermoascus crustaceus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Thermoascaceae JCM Gp BCIC thermophile 31, ,2 HiSeq 2500 Thermoascus crustaceus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Thermoascaceae JCM Gp BCIC thermophile 31,64 195, ,2 HiSeq 2500 Neosartorya fischeri Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Trichocomaceae NRRL 181 Gp AAKE opportunistic human pathogen 32,55 1, Sanger Neosartorya fischeri Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Trichocomaceae NRRL 181 Gp AAKE opportunistic human pathogen 32,55 1, Sanger Rasamsonia emersonii Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Trichocomaceae CBS Gp LASV thermophile 28,25 14, ,6 Sanger; 454 Rasamsonia emersonii Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Trichocomaceae CBS Gp LASV thermophile 28,25 14, ,6 Sanger; 454 Talaromyces aculeatus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Trichocomaceae ATCC Gp JGI iron phosphate solubilizer 37,27 91, Illumina Talaromyces aculeatus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Trichocomaceae ATCC Gp JGI iron phosphate solubilizer 37,27 91, Illumina Talaromyces cellulolyticus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Trichocomaceae Y-94 Gp BBPS industrial importance (cellulase production) 36, ,6 454 Talaromyces cellulolyticus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Trichocomaceae Y-94 Gp BBPS industrial importance (cellulase production) 36,4 19, ,6 454 Talaromyces islandicus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Trichocomaceae PIS Gp CVMT industrial importance (biopolymer degradation, mycotoxins) 34, ,3 Talaromyces islandicus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Trichocomaceae PIS Gp CVMT industrial importance (biopolymer degradation, mycotoxins) 34, ,3 Talaromyces leycettanus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Trichocomaceae CBS Gp JSYV industrial importance (thermophilic enzzymes) 25, ,7 Illumina HiSeq Talaromyces leycettanus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Trichocomaceae CBS Gp JSYV industrial importance (thermophilic enzzymes) 25,95 180, ,7 Illumina HiSeq Talaromyces marneffei Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Trichocomaceae ATCC Gp ABAR opportunistic pathogen (dimorphic; penicilliosis) 28,64 8, Sanger Talaromyces marneffei Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Trichocomaceae ATCC Gp ABAR opportunistic pathogen (dimorphic; penicilliosis) 28,64 8, Sanger Talaromyces pinophilus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Trichocomaceae 1-95 Gp LSFK industrial importance (enzyme production) 36, ,8 Illumina HiSeq Talaromyces pinophilus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Trichocomaceae 1-95 Gp LSFK industrial importance (enzyme production) 36,41 150, ,8 Illumina HiSeq Talaromyces purpureogenus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Trichocomaceae MYA-38 Gp LIAB medical importance (mycotoxin-producing) 38, ,7 Illumina HiSeq; PacBio Talaromyces purpureogenus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Trichocomaceae MYA-38 Gp LIAB medical importance (mycotoxin-producing) 38,85 213, ,7 Illumina HiSeq; PacBio Talaromyces stipitatus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Trichocomaceae ATCC Gp ABAS saprotroph 35,69 8, Sanger Talaromyces stipitatus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Trichocomaceae ATCC Gp ABAS saprotroph 35,69 8, Sanger Talaromyces verruculosus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Trichocomaceae TS63-9 Gp LHCL industrial importance (cellulase production) 37, ,2 Illumina HiSeq Talaromyces verruculosus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Trichocomaceae TS63-9 Gp LHCL industrial importance (cellulase production) 37,63 100, ,2 Illumina HiSeq Thermomyces lanuginosus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Trichocomaceae SSBP Gp ANHP industrial importance (thermophile; xylanase producer) 19,156 48, ,2 454; Illumina GAII Thermomyces lanuginosus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Eurotiales Trichocomaceae SSBP Gp ANHP industrial importance (thermophile; xylanase producer) 19,156 48, ,2 454; Illumina GAII Ajellomyces dermatitidis Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Ajellomycetaceae SLH14081 Gp ACBU human pathogen (blastomycosis) 75,4 8, Sanger Ajellomyces dermatitidis Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Ajellomycetaceae SLH14081 Gp ACBU human pathogen (blastomycosis) 75,4 8, Sanger Emmonsia crescens Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Ajellomycetaceae UAMH 3008 Gp LCZI occasional human pathogen (dimorphic) 30,7 163, ,2 Emmonsia parva Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Ajellomycetaceae UAMH 139 Gp LDEV occasional animal pathogen (dimorphic) , ,4 Histoplasma capsulatum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Ajellomycetaceae NAm1 Gp AMXR human pathogen (histoplasmosis) 32, ,1 Illumina HiSeq Histoplasma capsulatum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Ajellomycetaceae NAm1 Gp AMXR human pathogen (histoplasmosis) 32,99 7, ,1 Illumina HiSeq Paracoccidioides brasiliensis Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Ajellomycetaceae Pb03 Gp ABHV human pathogen (paracoccidioidomycosis) 29,06 8, ,3 Paracoccidioides brasiliensis Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Ajellomycetaceae Pb03 Gp ABHV human pathogen (paracoccidioidomycosis) 29,06 8, ,3 Paracoccidioides lutzii Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Ajellomycetaceae Gp ABKH human pathogen (paracoccidioidomycosis) 32,93 148, ,8 Illumina Arthroderma benhamiae Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Arthrodermataceae CBS Gp ABSU human pathogen (dermatophytosis) 22, Arthroderma benhamiae Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Arthrodermataceae CBS Gp ABSU human pathogen (dermatophytosis) 22,2 8, Microsporum canis Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Arthrodermataceae CBS Gp ABVF human pathogen 23,24 7, Sanger Microsporum canis Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Arthrodermataceae CBS Gp ABVF human pathogen 23,24 7, Sanger Microsporum gypseum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Arthrodermataceae CBS Gp ABQE occasional human pathogen; dermatophyte 23,2682 9, ,5 Sanger Microsporum gypseum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Arthrodermataceae CBS Gp ABQE occasional human pathogen; dermatophyte 23,2682 9, ,5 Sanger Trichophyton equinum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Arthrodermataceae CBS Gp ABWI animal pathogen (horse); dermatophyte 24,158 4, ,4 Sanger Trichophyton equinum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Arthrodermataceae CBS Gp ABWI animal pathogen (horse); dermatophyte 24,158 4, ,4 Sanger Trichophyton interdigitale Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Arthrodermataceae MR816 Gp AOKY human pathogen (dermatophyte ) 22,469 56, Illumina Trichophyton interdigitale Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Arthrodermataceae MR816 Gp AOKY human pathogen (dermatophyte ) 22,469 56, Illumina Trichophyton rubrum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Arthrodermataceae CBS Gp ACPH human pathogen (dermatophyte ) 22,5 8, ,3 Sanger Trichophyton rubrum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Arthrodermataceae CBS Gp ACPH human pathogen (dermatophyte ) 22,5 8, ,3 Sanger Trichophyton soudanense Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Arthrodermataceae CBS Gp AOKW human pathogen (tinea capitis) 22,85 75, ,5 Illumina Trichophyton tonsurans Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Arthrodermataceae CBS Gp ACPI human pathogen (dermatophyte; ringworm of scalp) 22,988 5, ,1 Sanger Trichophyton tonsurans Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Arthrodermataceae CBS Gp ACPI human pathogen (dermatophyte; ringworm of scalp) 22,988 5, ,1 Sanger Trichophyton verrucosum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Arthrodermataceae HKI 0517 Gp ACYE animal pathogen (cattle, hoerse); dermatophyte 22, ; Sanger Trichophyton verrucosum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Arthrodermataceae HKI 0517 Gp ACYE animal pathogen (cattle, hoerse); dermatophyte 22,54 3, ; Sanger Trichophyton violaceum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Arthrodermataceae CMCC(F)T3l Gp LHPN human pathogen (dermatophyte) 23, Illumina HiSeq; PacBio Trichophyton violaceum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Arthrodermataceae CMCC(F)T3l Gp LHPN human pathogen (dermatophyte) 23,37 250, Illumina HiSeq; PacBio Gymnascella aurantiaca Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Gymnoascaceae NRRL 5967 Gp JGI saprotroph 25,35 143, Illumina HiSeq Gymnascella aurantiaca Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Gymnoascaceae NRRL 5967 Gp JGI saprotroph 25,35 143, Illumina HiSeq Gymnascella citrina Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Gymnoascaceae NRRL 5970 Gp JGI saprotroph 25,16 146, Illumina HiSeq Gymnascella citrina Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Gymnoascaceae NRRL 5970 Gp JGI saprotroph 25,16 146, Illumina HiSeq Lacazia loboi Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Incertae sedis LDI48194 Gp LBNG human pathogen (blastomycosis) IonTorrent Lacazia loboi Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Incertae sedis LDI48194 Gp LBNG human pathogen (blastomycosis) 14 80, IonTorrent Amauroascus mutatus Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Onygenaceae UAMH 3576 Gp LJPJ uncertain; comparsion with patogenic Onygenales 30,38 50, ,9 Illumina Amauroascus niger Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Onygenaceae UAMH 3544 Gp LJPK uncertain; comparsion with patogenic Onygenales 36,72 50, ,7 Illumina Byssoonygena ceratinophila Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Onygenaceae UAMH 5669 Gp LJPH keratinophilic 27,45 50, ,2 Illumina Chrysosporium lucknowense Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Onygenaceae VKM F-3500-D Gp industrial importance 38,5 12, ,6 454; Sanger Chrysosporium lucknowense Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Onygenaceae VKM F-3500-D Gp industrial importance 38,5 12, ,6 454; Sanger Chrysosporium queenslandicum Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Onygenaceae CBS Gp LJPI industrial importance (antimicorbial compunds); comparsion with pathogenic Onygenales 32,34 50, ,6 Illumina Coccidioides immitis Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Onygenaceae RMSCC 2394 Gp AATX human pathogen (Coccidiomycosis) 28,9 8, Sanger Coccidioides immitis Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Onygenaceae RMSCC 2394 Gp AATX human pathogen (Coccidiomycosis) 28,9 8, Sanger Coccidioides posadasii Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Onygenaceae RMSCC 2133 Gp ABFM human pathogen (Coccidiomycosis/valley fever) 27,9 6, ,6 Sanger Coccidioides posadasii Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Onygenaceae RMSCC 2133 Gp ABFM human pathogen (Coccidiomycosis/valley fever) 27,9 6, ,6 Sanger Onygena corvina Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Onygenaceae CBS Gp JWPT saprotroph 21, ,2 Illumina Onygena corvina Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Onygenaceae CBS Gp JWPT saprotroph 21,71 370, ,2 Illumina Uncinocarpus reesii Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Onygenaceae UAMH 1704 Gp JGI saprotroph (soil); closely related to Coccidioides 22,33 5, Sanger Uncinocarpus reesii Ascomycota Pezizomycotina Eurotiomycetes Eurotiomycetidae Onygenales Onygenaceae UAMH 1704 Gp JGI saprotroph (soil); closely related to Coccidioides 22,33 5, Sanger

60 Phaeomoniella chlamydospora Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Phaeomoniellales Incertae sedis UCRPC4 Gp LCWF plant pathogen (grapevine trunk disease) 27,51 107, Illumina HiSeq Phaeomoniella chlamydospora Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Phaeomoniellales Incertae sedis UCRPC4 Gp LCWF plant pathogen (grapevine trunk disease) 27,51 107, Illumina HiSeq Endocarpon pusillum Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Verrucariales Verrucariaceae Z07020 Gp APWS mycobiont (lichen-forming) 37,5 78, ; Illumina GAIIx Endocarpon pusillum Ascomycota Pezizomycotina Eurotiomycetes Chaetothyriomycetidae Verrucariales Verrucariaceae Z07020 Gp APWS mycobiont (lichen-forming) 37,5 78, ; Illumina GAIIx Symbiotaphrina buchneri Ascomycota Pezizomycotina Incertae sedis Incertae sedis Incertae sedis Incertae sedis JCM 9740 Gp BCIG insect-associated (beetle endosymbiont) 23, ,6 HiSeq 2500 Symbiotaphrina buchneri Ascomycota Pezizomycotina Incertae sedis Incertae sedis Incertae sedis Incertae sedis JCM 9740 Gp BCIG insect-associated (beetle endosymbiont) 23,71 251, ,6 HiSeq 2500 Symbiotaphrina kochii Ascomycota Pezizomycotina Incertae sedis Incertae sedis Incertae sedis Incertae sedis CBS Gp insect-associated (beetle endosymbiont) 24, ; Illumina Symbiotaphrina kochii Ascomycota Pezizomycotina Incertae sedis Incertae sedis Incertae sedis Incertae sedis CBS Gp insect-associated (beetle endosymbiont) 24,83 69, ; Illumina Cladonia grayi Ascomycota Pezizomycotina Lecanoromycetes Lecanoromycetidae Lecanorales Cladoniaceae Cgr/DA2myc/ss JGI mycobiont (lichen-forming) 39, ; Illumina Cladonia grayi Ascomycota Pezizomycotina Lecanoromycetes Lecanoromycetidae Lecanorales Cladoniaceae Cgr/DA2myc/ss JGI mycobiont (lichen-forming) 39, ; Illumina Cladonia macilenta Ascomycota Pezizomycotina Lecanoromycetes Lecanoromycetidae Lecanorales Cladoniaceae KoLRI Gp AUPP mycobiont (lichen-forming) 37,12 540, ,8 Illumina Cladonia macilenta Ascomycota Pezizomycotina Lecanoromycetes Lecanoromycetidae Lecanorales Cladoniaceae KoLRI Gp AUPP mycobiont (lichen-forming) 37,12 540, ,8 Illumina Cladonia metacorallifera Ascomycota Pezizomycotina Lecanoromycetes Lecanoromycetidae Lecanorales Cladoniaceae KoLRI Gp AXCT mycobiont (lichen-forming) 36, , Illumina Cladonia metacorallifera Ascomycota Pezizomycotina Lecanoromycetes Lecanoromycetidae Lecanorales Cladoniaceae KoLRI Gp AXCT mycobiont (lichen-forming) 36, , Illumina Usnea florida Ascomycota Pezizomycotina Lecanoromycetes Lecanoromycetidae Lecanorales Parmeliaceae ATCC18376 v mycobiont (lichen-forming) 44,32 87, Illumina Usnea florida Ascomycota Pezizomycotina Lecanoromycetes Lecanoromycetidae Lecanorales Parmeliaceae ATCC18376 v mycobiont (lichen-forming) 44,32 87, Illumina Gyalolechia flavorubescens Ascomycota Pezizomycotina Lecanoromycetes Lecanoromycetidae Teloschistales Teloschistaceae KoLRI Gp AUPK mycobiont (lichen-forming) 34, , Illumina HiSeq Gyalolechia flavorubescens Ascomycota Pezizomycotina Lecanoromycetes Lecanoromycetidae Teloschistales Teloschistaceae KoLRI Gp AUPK mycobiont (lichen-forming) 34, , Illumina HiSeq Xanthoria parietina Ascomycota Pezizomycotina Lecanoromycetes Lecanoromycetidae Teloschistales Teloschistaceae 46-1 Gp JGI mycobiont (lichen-forming) 31,9 43, Xanthoria parietina Ascomycota Pezizomycotina Lecanoromycetes Lecanoromycetidae Teloschistales Teloschistaceae 46-1 Gp JGI mycobiont (lichen-forming) 31,9 43, Lasallia pustulata Ascomycota Pezizomycotina Lecanoromycetes Incertae sedis Umbilicariales Umbilicariaceae Sardinia_ Gp JYIL mycobiont (lichen-forming) 39, ,2 PacBio; Illumina HiSeq Lasallia pustulata Ascomycota Pezizomycotina Lecanoromycetes Incertae sedis Umbilicariales Umbilicariaceae Sardinia_ Gp JYIL mycobiont (lichen-forming) 39,23 90, ,2 PacBio; Illumina HiSeq Umbilicaria muehlenbergii Ascomycota Pezizomycotina Lecanoromycetes Incertae sedis Umbilicariales Umbilicariaceae Gp JFDN mycobiont (lichen-forming); genetically tractable 34, , ,1 Illumina HiSeq Umbilicaria muehlenbergii Ascomycota Pezizomycotina Lecanoromycetes Incertae sedis Umbilicariales Umbilicariaceae Gp JFDN mycobiont (lichen-forming); genetically tractable 34, , ,1 Illumina HiSeq Blumeria graminis Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Erysiphales Erysiphaceae A6 Gp AOLT plant pathogen (mildew on grasses and cereals) 47,46 70, ,4 454; Illumina HiSeq Blumeria graminis Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Erysiphales Erysiphaceae A6 Gp AOLT plant pathogen (mildew on grasses and cereals) 47,46 70, ,4 454; Illumina HiSeq Erysiphe necator Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Erysiphales Erysiphaceae branching Gp JNUS plant pathogen ( powdery mildew of grape) 50, Illumina HiSeq Erysiphe necator Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Erysiphales Erysiphaceae branching Gp JNUS plant pathogen ( powdery mildew of grape) 50,66 29, Illumina HiSeq Erysiphe pisi Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Erysiphales Erysiphaceae not specified Gp CACN plant pathogen (powdery mildew of pea) 41 8, ,2 454 Erysiphe pisi Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Erysiphales Erysiphaceae not specified Gp CACN plant pathogen (powdery mildew of pea) 41 8, ,2 454 Marssonina brunnea Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Dermateaceae MB_m1 Gp AFXC plant pathogen (poplar leaf spot) 51,949 34, ,9 454; Sanger Marssonina brunnea Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Dermateaceae MB_m1 Gp AFXC plant pathogen (poplar leaf spot) 51,949 34, ,9 454; Sanger Ascocoryne sarcoides Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Helotiaceae NRRL Gp AIAA pharmaceutical importance (antibiotic-producing); saprotroph 34, Ascocoryne sarcoides Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Helotiaceae NRRL Gp AIAA pharmaceutical importance (antibiotic-producing); saprotroph 34,2 250, Glarea lozoyensis Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Helotiaceae Gp AGUE pharmaceutical importance (produces antifugal compounds) 38,718 34, Illumina HiSeq Glarea lozoyensis Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Helotiaceae Gp AGUE pharmaceutical importance (produces antifugal compounds) 38,718 34, Illumina HiSeq Hymenoscyphus fraxineus Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Helotiaceae CBS Gp LLCC plant pathogen (ash dieback) 51, ,7 Illumina HiSeq Hymenoscyphus fraxineus Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Helotiaceae CBS Gp LLCC plant pathogen (ash dieback) 51,52 90, ,7 Illumina HiSeq Hymenoscyphus fructigenus Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Helotiaceae CBS Gp LKUV saprotroph 61, ,1 Illumina HiSeq Hymenoscyphus fructigenus Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Helotiaceae CBS Gp LKUV saprotroph 61,12 400, ,1 Illumina HiSeq Hymenoscyphus infarciens Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Helotiaceae CBS Gp LLCB saprotroph 68, ,6 Illumina HiSeq Hymenoscyphus infarciens Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Helotiaceae CBS Gp LLCB saprotroph 68,15 250, ,6 Illumina HiSeq Hymenoscyphus laetus Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Helotiaceae CBS Gp LLCA saprotroph 36, ,9 Illumina HiSeq not publically available Hymenoscyphus laetus Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Helotiaceae CBS Gp LLCA saprotroph 36,47 140,00 51,9 Illumina HiSeq Hymenoscyphus repandus Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Helotiaceae CBS Gp LLCE saprotroph 42, ,6 Illumina HiSeq Hymenoscyphus repandus Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Helotiaceae CBS Gp LLCE saprotroph 42,81 50, ,6 Illumina HiSeq Hymenoscyphus salicellus Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Helotiaceae CBS Gp LLCD saprotroph 57, ,4 Illumina HiSeq Hymenoscyphus salicellus Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Helotiaceae CBS Gp LLCD saprotroph 57,95 30,00 44,4 Illumina HiSeq Hymenoscyphus scutula Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Helotiaceae CBS Gp LKTO saprotroph 62, ,1 Illumina HiSeq Hymenoscyphus scutula Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Helotiaceae CBS Gp LKTO saprotroph 62,23 100, ,1 Illumina HiSeq Meliniomyces bicolor Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Helotiaceae E Gp JGI mycorrhizal 82,38 190, Illumina HiSeq Meliniomyces bicolor Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Helotiaceae E Gp JGI mycorrhizal 82,38 190, Illumina HiSeq Meliniomyces variabilis Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Helotiaceae F Gp JGI mycorrhizal 55,86 108, Illumina HiSeq Meliniomyces variabilis Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Helotiaceae F Gp JGI mycorrhizal 55,86 108, Illumina HiSeq Rhizoscyphus ericae Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Helotiaceae UAMH 7357 Gp mycorrhizal (ericoid) 57,41 120, Illumina HiSeq Calycina herbarum Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Hyaloscyphaceae CBS Gp LLEY uncertain saprotroph 69, ,7 Illumina HiSeq Calycina herbarum Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Hyaloscyphaceae CBS Gp LLEY uncertain saprotroph 69,31 300, ,7 Illumina HiSeq Acephala macrosclerotiorum Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Incertae sedis EW76-UTF0540 v ectomycorrhizal 73,68 141, Illumina Acephala macrosclerotiorum Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Incertae sedis EW76-UTF0540 v ectomycorrhizal 73,68 141, Illumina Cadophora sp. DSE1049 Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Incertae sedis DSE1049 Gp JGI endophyte (grasslands) 70,46 79, Illumina HiSeq Cadophora sp. DSE1049 Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Incertae sedis DSE1049 Gp JGI endophyte (grasslands) 70,46 79, Illumina HiSeq Cairneyiella heathii Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Incertae sedis VPRI Gp AYLM mycorrhizal 50,69 20, ,69 Chalara longipes Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Incertae sedis BDJ Gp JGI saprotroph 52,43 104, Illumina Chalara longipes Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Incertae sedis BDJ Gp JGI saprotroph 52,43 104, Illumina Leptodontium sp. Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Incertae sedis PMI_412 v1.0 Gp endophyte; root 73,78 140, Illumina HiSeq Leptodontium sp. Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Incertae sedis PMI_412 v1.0 Gp endophyte; root 73,78 140, Illumina HiSeq Loramyces juncicola Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Loramycetaceae v saprotroph; marine 42,68 86, Illumina Loramyces juncicola Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Loramycetaceae v saprotroph; marine 42,68 86, Illumina Oidiodendron maius Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Myxotrichaceae Zn Gp JGI endomycorrhizal; metal-tolerant 46,43 28, ; Illumina Oidiodendron maius Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Myxotrichaceae Zn Gp JGI endomycorrhizal; metal-tolerant 46,43 28, ; Illumina Rutstroemia firma Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Rutstroemiaceae CBS v saprotroph 44,27 81, Illumina Rutstroemia sydowiana Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Rutstroemiaceae CBS Gp JWJB saprotroph 51, ,1 Illumina MiSeq Rutstroemia sydowiana Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Rutstroemiaceae CBS Gp JWJB saprotroph 51,99 101, ,1 Illumina MiSeq Botrytis cinerea (Botryotinia fuckeliana) Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Sclerotiniaceae B05.10 Gp AAID plant pathogen (grapevines) 41, Sanger Botrytis cinerea (Botryotinia fuckeliana) Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Sclerotiniaceae B05.10 Gp AAID plant pathogen (grapevines) 41,2 65, Sanger Botrytis paeoniae Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Sclerotiniaceae AR05 Gp LBGX plant pathogen (peony; Botrytis blight) 44, ,1 IonTorrent Botrytis paeoniae Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Sclerotiniaceae AR05 Gp LBGX plant pathogen (peony; Botrytis blight) 44,24 236, ,1 IonTorrent Ciborinia camelliae Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Sclerotiniaceae ICMP Gp LGKQ plant pathogen (ornamental camellia cultivars) 40, ,5 Illumina MiSeq Ciborinia camelliae Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Sclerotiniaceae ICMP Gp LGKQ plant pathogen (ornamental camellia cultivars) 40,73 142, ,5 Illumina MiSeq Rutstroemia (Sclerotinia) echinophilaascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Sclerotiniaceae CBS JWJA saprotroph 40, ,1 Rutstroemia (Sclerotinia) echinophilaascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Sclerotiniaceae CBS JWJA saprotroph 40, ,1 Sclerotinia borealis Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Sclerotiniaceae F-4157 Gp AYSA plant pathogen (cereals; sychrophilic) 39,45 23, ,9 454 Sclerotinia borealis Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Sclerotiniaceae F-4157 Gp AYSA plant pathogen (cereals; sychrophilic) 39,45 23, ,9 454 Sclerotinia homoeocarpa Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Sclerotiniaceae ShHRS10 Gp LNGN plant pathogen (turfgrass) 42, ,9 Illumina HiSeq; PacBio Sclerotinia homoeocarpa Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Sclerotiniaceae ShHRS10 Gp LNGN plant pathogen (turfgrass) 42,27 50, ,9 Illumina HiSeq; PacBio Sclerotinia sclerotiorum Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Sclerotiniaceae ATCC Gp AAGT plant pathogen (broadest host range known) 38,532 8, ,8 Sanger Sclerotinia sclerotiorum Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Sclerotiniaceae ATCC Gp AAGT plant pathogen (broadest host range known) 38,532 8, ,8 Sanger Phialocephala scopiformis Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Vibrisseaceae CBS Gp LKNI endophyte 48, ,6 Illumina HiSeq Phialocephala scopiformis Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Vibrisseaceae CBS Gp LKNI endophyte 48, ,6 Illumina HiSeq Amorphotheca resinae Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Incertae sedis Amorphothecaceae DAOM Gp JGI industrial importance 28,63 42, GS-FLX-Titanium; Illumina HiSeq Amorphotheca resinae Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Incertae sedis Amorphothecaceae DAOM Gp JGI industrial importance 28,63 42, GS-FLX-Titanium; Illumina HiSeq Pseudogymnoascus destructans Ascomycota Pezizomycotina Leotiomycetes Incertae sedis Incertae sedis Pseudeurotiaceae M1379 Gp AYKP animal pathogen (white nose syndrome in bats); psychrophile 30, , ,1 Illumina HiSeq Pseudogymnoascus destructans Ascomycota Pezizomycotina Leotiomycetes Incertae sedis Incertae sedis Pseudeurotiaceae M1379 Gp AYKP animal pathogen (white nose syndrome in bats); psychrophile 30, , ,1 Illumina HiSeq Pseudogymnoascus pannorum Ascomycota Pezizomycotina Leotiomycetes Incertae sedis Incertae sedis Pseudeurotiaceae M1372 Gp AYKR human pathogen (geomycosis); soil-dwelling psychrophile 29, , ,5 Illumina HiSeq Pseudogymnoascus pannorum Ascomycota Pezizomycotina Leotiomycetes Incertae sedis Incertae sedis Pseudeurotiaceae M1372 Gp AYKR human pathogen (geomycosis); soil-dwelling psychrophile 29, , ,5 Illumina HiSeq Pseudogymnoascus sp. BL549 Ascomycota Pezizomycotina Leotiomycetes Incertae sedis Incertae sedis Pseudeurotiaceae BL549 Gp LNAR soil-associated 25,86 26, ,1 Illumina Rutstroemia firma Ascomycota Pezizomycotina Leotiomycetes Leotiomycetidae Helotiales Rutstroemiaceae CBS v saprotroph 44,27 81, Illumina Arthrobotrys oligospora Ascomycota Pezizomycotina Orbiliomycetes Orbiliomycetidae Orbiliales Orbiliaceae ATCC Gp ADOT biocontrol (nematophagous) 40, Sanger; 454 Arthrobotrys oligospora Ascomycota Pezizomycotina Orbiliomycetes Orbiliomycetidae Orbiliales Orbiliaceae ATCC Gp ADOT biocontrol (nematophagous) 40,1 33, Sanger; 454 Dactylellina haptotyla Ascomycota Pezizomycotina Orbiliomycetes Orbiliomycetidae Orbiliales Orbiliaceae CBS Gp AQGS biocontrol (nematophagous) 39,531 28, ,3 454 Dactylellina haptotyla Ascomycota Pezizomycotina Orbiliomycetes Orbiliomycetidae Orbiliales Orbiliaceae CBS Gp AQGS biocontrol (nematophagous) 39,531 28, ,3 454

61 Drechslerella stenobrocha Ascomycota Pezizomycotina Orbiliomycetes Orbiliomycetidae Orbiliales Orbiliaceae YNWS Gp biocontrol (nematophagous) 29,02 80, Illumina Drechslerella stenobrocha Ascomycota Pezizomycotina Orbiliomycetes Orbiliomycetidae Orbiliales Orbiliaceae YNWS Gp biocontrol (nematophagous) 29,02 80, Illumina Ascobolus immersus Ascomycota Pezizomycotina Pezizomycetes Pezizomycetidae Pezizales Ascobolaceae RN42 Gp JGI coprophilous 59,53 86, Illumina HiSeq Ascobolus immersus Ascomycota Pezizomycotina Pezizomycetes Pezizomycetidae Pezizales Ascobolaceae RN42 Gp JGI coprophilous 59,53 86, Illumina HiSeq Ascodesmis nigricans Ascomycota Pezizomycotina Pezizomycetes Pezizomycetidae Pezizales Ascodesmidaceae CBS Gp coprophilous 27,39 96, Illumina Caloscypha fulgens Ascomycota Pezizomycotina Pezizomycetes Pezizomycetidae Pezizales Caloscyphaceae ATCC v plant pathogen (seed rot of conifers) 44, Illumina Caloscypha fulgens Ascomycota Pezizomycotina Pezizomycetes Pezizomycetidae Pezizales Caloscyphaceae ATCC v plant pathogen (seed rot of conifers) 44, Illumina Gyromitra esculenta Ascomycota Pezizomycotina Pezizomycetes Pezizomycetidae Pezizales Discinaceae CBS v saprotroph 45,05 83, Illumina Gyromitra esculenta Ascomycota Pezizomycotina Pezizomycetes Pezizomycetidae Pezizales Discinaceae CBS v saprotroph 45,05 83, Illumina Morchella conica Ascomycota Pezizomycotina Pezizomycetes Pezizomycetidae Pezizales Morchellaceae CCBAS932 Gp JGI mycorrhizal; saprotroph 48,21 67, Illumina HiSeq Morchella conica Ascomycota Pezizomycotina Pezizomycetes Pezizomycetidae Pezizales Morchellaceae CCBAS932 Gp JGI mycorrhizal; saprotroph 48,21 67, Illumina HiSeq Morchella importuna Ascomycota Pezizomycotina Pezizomycetes Pezizomycetidae Pezizales Morchellaceae SCYDJ1-A1 v saprotroph; edible 48,8 83, Illumina Morchella importuna Ascomycota Pezizomycotina Pezizomycetes Pezizomycetidae Pezizales Morchellaceae SCYDJ1-A1 v saprotroph; edible 48,8 83, Illumina Kalaharituber pfeilii Ascomycota Pezizomycotina Pezizomycetes Pezizomycetidae Pezizales Pezizaceae F3 v mycorrhizal; edible desert truffle 78, PacBio Kalaharituber pfeilii Ascomycota Pezizomycotina Pezizomycetes Pezizomycetidae Pezizales Pezizaceae F3 v mycorrhizal; edible desert truffle 78, PacBio Terfezia boudieri Ascomycota Pezizomycotina Pezizomycetes Pezizomycetidae Pezizales Pezizaceae S1 Gp JGI mycorrhizal 63,23 104, ; Illumina; PacBio Terfezia boudieri Ascomycota Pezizomycotina Pezizomycetes Pezizomycetidae Pezizales Pezizaceae S1 Gp JGI mycorrhizal 63,23 104, ; Illumina; PacBio Pyronema confluens Ascomycota Pezizomycotina Pezizomycetes Pezizomycetidae Pezizales Pyronemataceae CBS Gp CATG saprotroph (soil) 50, ; Illumina HiSeq Pyronema confluens Ascomycota Pezizomycotina Pezizomycetes Pezizomycetidae Pezizales Pyronemataceae CBS Gp CATG saprotroph (soil) 50, ; Illumina HiSeq Ramularia collo-cygni Ascomycota Pezizomycotina Pezizomycetes Pezizomycetidae Pezizales Pyronemataceae Gs CZLF plant pathogen (barley) 30,3 90, ,4 Ramularia collo-cygni Ascomycota Pezizomycotina Pezizomycetes Pezizomycetidae Pezizales Pyronemataceae Gs CZLF plant pathogen (barley) 30,3 90, ,4 Wilcoxina mikolae Ascomycota Pezizomycotina Pezizomycetes Pezizomycetidae Pezizales Pyronemataceae CBS Gp JGI ectomycorrhizal 117, Illumina HiSeq; PacBio Wilcoxina mikolae Ascomycota Pezizomycotina Pezizomycetes Pezizomycetidae Pezizales Pyronemataceae CBS Gp JGI ectomycorrhizal 117,29 95, Illumina HiSeq; PacBio Sarcoscypha coccinea Ascomycota Pezizomycotina Pezizomycetes Pezizomycetidae Pezizales Sarcoscyphaceae ATCC v saprotroph 39,09 95, Illumina Sarcoscypha coccinea Ascomycota Pezizomycotina Pezizomycetes Pezizomycetidae Pezizales Sarcoscyphaceae ATCC v saprotroph 39,09 95, Illumina Plectania melastoma Ascomycota Pezizomycotina Pezizomycetes Pezizomycetidae Pezizales Sarcosomataceae CBS v saprotroph 92,51 72, Illumina Plectania melastoma Ascomycota Pezizomycotina Pezizomycetes Pezizomycetidae Pezizales Sarcosomataceae CBS v saprotroph 92,51 72, Illumina Choiromyces venosus Ascomycota Pezizomycotina Pezizomycetes Pezizomycetidae Pezizales Tuberaceae Gp JGI ectomycorrhizal 126,04 89, Illumina; PacBio Choiromyces venosus Ascomycota Pezizomycotina Pezizomycetes Pezizomycetidae Pezizales Tuberaceae Gp JGI ectomycorrhizal 126,04 89, Illumina; PacBio Tuber borchii Ascomycota Pezizomycotina Pezizomycetes Pezizomycetidae Pezizales Tuberaceae Tbo3840 Go ectomycorrhizal 97,18 97, PacBio Tuber melanosporum Ascomycota Pezizomycotina Pezizomycetes Pezizomycetidae Pezizales Tuberaceae Mel28 Gp CABJ ectomycorrhizal (edible delicacy) 124,945 10, Sanger Tuber melanosporum Ascomycota Pezizomycotina Pezizomycetes Pezizomycetidae Pezizales Tuberaceae Mel28 Gp CABJ ectomycorrhizal (edible delicacy) 124,945 10, Sanger Microdochium bolleyi Ascomycota Pezizomycotina Sordariomycetes Xylariomycetidae Amphisphaeriales Incertae sedis J235TASD1 Gp LSSP endophyte 38,84 136, ,1 HiSeq 2500 Pestalotiopsis fici Ascomycota Pezizomycotina Sordariomycetes Xylariomycetidae Amphisphaeriales Pestalotiopsidaceae W106-1 Gp ARNU endophyte 51,91 80, ; Illumina HiSeq Pestalotiopsis sp. JCM 9685 Ascomycota Pezizomycotina Sordariomycetes Xylariomycetidae Amphisphaeriales Pestalotiopsidaceae JCM 9685 Gp BCGF endophyte (Taxus brevifolia) 48,23 235, ,6 Illumina Thozetella sp. PMI_491 Ascomycota Pezizomycotina Sordariomycetes Sordariomycetidae Chaetosphaeriales Chaetosphaeriaceae PMI_491 Gp JGI endophyte 72,86 112, Illumina HiSeq Thozetella sp. PMI_491 Ascomycota Pezizomycotina Sordariomycetes Sordariomycetidae Chaetosphaeriales Chaetosphaeriaceae PMI_491 Gp JGI endophyte 72,86 112, Illumina HiSeq Coniochaeta ligniaria Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Coniochaetales Coniochaetaceae NRRL Gp JGI industrial importance 42,38 94, Illumina HiSeq Coniochaeta ligniaria Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Coniochaetales Coniochaetaceae NRRL Gp JGI industrial importance 42,38 94, Illumina HiSeq Coniochaeta sp. PMI_546 Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Coniochaetales Coniochaetaceae PMI_546 Gp JGI endophyte (root); saproptroph 33, Illumina HiSeq Coniochaeta sp. PMI_546 Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Coniochaetales Coniochaetaceae PMI_546 Gp JGI endophyte (root); saproptroph 33,51 102, Illumina HiSeq Chrysoporthe austroafricana Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Diaporthales Cryphonectriaceae CMW 2113 Gp JYIP plant pathogen (eucalyptus, Tibouchina, Syzygium) 44, ,3 Illumina MiSeq; IonTorrent Chrysoporthe austroafricana Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Diaporthales Cryphonectriaceae CMW 2113 Gp JYIP plant pathogen (eucalyptus, Tibouchina, Syzygium) 44,66 40, ,3 Illumina MiSeq; IonTorrent Chrysoporthe cubensis Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Diaporthales Cryphonectriaceae CMW Gp LJCY plant pathogen (eucalyptus, Tibouchina, Syzygium) 42, ,1 Illumina MiSeq Chrysoporthe cubensis Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Diaporthales Cryphonectriaceae CMW Gp LJCY plant pathogen (eucalyptus, Tibouchina, Syzygium) 42,62 40, ,1 Illumina MiSeq Chrysoporthe deuterocubensis Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Diaporthales Cryphonectriaceae CMW 8650 Gp LJDD plant pathogen (eucalyptus, Tibouchina, Syzygium) 43, ,3 Illumina MiSeq Chrysoporthe deuterocubensis Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Diaporthales Cryphonectriaceae CMW 8650 Gp LJDD plant pathogen (eucalyptus, Tibouchina, Syzygium) 43,97 45, ,3 Illumina MiSeq Cryphonectria parasitica Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Diaporthales Cryphonectriaceae EP155 Gp JGI plant pathogen (chestnut blight) 43,9 8, Cryphonectria parasitica Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Diaporthales Cryphonectriaceae EP155 Gp JGI plant pathogen (chestnut blight) 43,9 8, Diaporthe ampelina Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Diaporthales Diaporthaceae UCDDA912 Gp LCUC endophyte 47, ,5 Illumina HiSeq Diaporthe ampelina Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Diaporthales Diaporthaceae UCDDA912 Gp LCUC endophyte 47,33 50, ,5 Illumina HiSeq Diaporthe aspalathi Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Diaporthales Diaporthaceae DPM Gp LJJS plant pathogen (soybean stem canker) 55, ,6 Illumina HiSeq Diaporthe aspalathi Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Diaporthales Diaporthaceae DPM Gp LJJS plant pathogen (soybean stem canker) 55,03 100, ,6 Illumina HiSeq Diaporthe longicolla Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Diaporthales Diaporthaceae MS10-6 Gp AYRD plant pathogen (soybean) 62,26 97, Illumina HiSeq Diaporthe longicolla Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Diaporthales Diaporthaceae MS10-6 Gp AYRD plant pathogen (soybean) 62,26 97, Illumina HiSeq Ophiognomonia clavigignenti-juglanascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Diaporthales Gnomoniaceae pooled sample Gp AEGN plant pathogen (butternut) 15,906 2, ,8 454 Ophiognomonia clavigignenti-juglanascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Diaporthales Gnomoniaceae pooled sample Gp AEGN plant pathogen (butternut) 15,906 2, ,8 454 Lollipopaia minuta Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Diaporthales Incertae sedis CBS v saprotroph tropical 37, Illumina Lollipopaia minuta Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Diaporthales Incertae sedis CBS v saprotroph tropical 37,42 138, Illumina Melanconium sp. 1 NRRL Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Diaporthales Melanconidaceae NRRL Gp JGI plant pathogen (probable maize pathogen) 58,52 92, Illumina Melanconium sp. 1 NRRL Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Diaporthales Melanconidaceae NRRL Gp JGI plant pathogen (probable maize pathogen) 58,52 92, Illumina Valsa mali Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Diaporthales Valsaceae SXYL134 Gp JUIZ plant pathogen (apple & pear) 35, ,3 Illumina GAIIx Valsa mali Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Diaporthales Valsaceae SXYL134 Gp JUIZ plant pathogen (apple & pear) 35,73 180, ,3 Illumina GAIIx Acremonium chrysogenum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Glomerallales Plectosphaerellaceae ATCC Gp JPKY industrial importance; pharamceutical importance 28, Illumina HiSeq Acremonium furcatum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Glomerallales Plectosphaerellaceae JCM 9210 Gp BCIA biocontrol (mycoparasite) 40, ,7 HiSeq 2500 Colletotrichum godetiae Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Glomerelalles Glomerellaceae CBS v insect-associated 51, PacBio Colletotrichum lupini Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Glomerelalles Glomerellaceae CBS v plant pathogen (Lupinus flowering plants) 58, PacBio Colletotrichum caudatum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Glomerellales Glomerellaceae CBS v plant pathogen (warm-season grasses) 44, Illumina Colletotrichum caudatum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Glomerellales Glomerellaceae CBS v plant pathogen (warm-season grasses) 44,2 143, Illumina Colletotrichum falcatum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Glomerellales Glomerellaceae Cf671 Gp LPVI plant pathogen (sugar-cane red rot) 48, ,6 Colletotrichum falcatum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Glomerellales Glomerellaceae Cf671 Gp LPVI plant pathogen (sugar-cane red rot) 48,19 150, ,6 Colletotrichum fioriniae Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Glomerellales Glomerellaceae MH 18 Gp JGI plant pathogen (anthracnose in various fruit) ,5 Illumina HiSeq Colletotrichum fioriniae Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Glomerellales Glomerellaceae MH 18 Gp JGI plant pathogen (anthracnose in various fruit) ,5 Illumina HiSeq Colletotrichum gloeosporioides Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Glomerellales Glomerellaceae Nara gc5 Gp ANPB plant pathogen (disease and anthracnose on a range of fuit and vegetables) 55,6 37, ,6 Illumina HiSeq Colletotrichum gloeosporioides Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Glomerellales Glomerellaceae Nara gc5 Gp ANPB plant pathogen (disease and anthracnose on a range of fuit and vegetables) 55,6 37, ,6 Illumina HiSeq Colletotrichum godetiae Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Glomerellales Glomerellaceae CBS v insect-associated 51,65 235, PacBio Colletotrichum graminicola Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Glomerellales Glomerellaceae M1.001 Gp ACOD plant pathogen (anthracnose in cereals) 51,64 9, ,1 454; Sanger Colletotrichum graminicola Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Glomerellales Glomerellaceae M1.001 Gp ACOD plant pathogen (anthracnose in cereals) 51,64 9, ,1 454; Sanger Colletotrichum higginsianum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Glomerellales Glomerellaceae IMI Gp CACQ plant pathogen (anthracnose in Brassicaceae) 44, ,1 Colletotrichum higginsianum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Glomerellales Glomerellaceae IMI Gp CACQ plant pathogen (anthracnose in Brassicaceae) 44, ,1 Colletotrichum incanum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Glomerellales Glomerellaceae MAFF Gp LFIW plant pathogen (broad host range; including soybean) 53,6 230, Colletotrichum lupini Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Glomerellales Glomerellaceae CBS v plant pathogen (Lupinus flowering plants) 58,76 97, PacBio Colletotrichum orbiculare Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Glomerellales Glomerellaceae MAFF Gp AMCV plant pathogen (melons and cucumber) 90 55, ,6 454; Illumina GAIIx Colletotrichum orbiculare Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Glomerellales Glomerellaceae MAFF Gp AMCV plant pathogen (melons and cucumber) 90 55, ,6 454; Illumina GAIIx Colletotrichum phormii Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Glomerellales Glomerellaceae CBS Go plant pathogen (anthracnose on flax) 51,67 97, PacBio Colletotrichum salicis Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Glomerellales Glomerellaceae CBS Gp JFFI plant pathogen (black canker of willow) 48,37 40, ,8 Colletotrichum simmondsii Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Glomerellales Glomerellaceae CBS Gp JFBX plant pathogen (safflower oil crop) 50,47 39, ,7 Illumina Colletotrichum simmondsii Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Glomerellales Glomerellaceae CBS Gp JFBX plant pathogen (safflower oil crop) 50,47 39, ,7 Illumina Colletotrichum sublineola Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Glomerellales Glomerellaceae TX430BB Gp JMSE plant pathogen (anthracnose in wild rice and sorghum) 46,75 92, ,3 Illumina GAII Colletotrichum sublineola Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Glomerellales Glomerellaceae TX430BB Gp JMSE plant pathogen (anthracnose in wild rice and sorghum) 46,75 92, ,3 Illumina GAII Colletotrichum tofieldiae Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Glomerellales Glomerellaceae CBS Gp LFHQ endophyte (root) 52,96 300, Illumina Colletotrichum zoysiae Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Glomerellales Glomerellaceae MAFF plant pathogen (warm-season grasses) 46,53 102, Illumina Acremonium chrysogenum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Glomerellales Plectosphaerellaceae ATCC Gp JPKY industrial importance; pharamceutical importance 28,5 137, Illumina HiSeq Acremonium furcatum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Glomerellales Plectosphaerellaceae JCM 9210 Gp BCIA biocontrol (mycoparasite) 40,32 281, ,7 HiSeq 2500 Sodiomyces alkalinus Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Glomerellales Plectosphaerellaceae F11 Gp JGI alkaliphile 43,45 113, Illumina Sodiomyces alkalinus Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Glomerellales Plectosphaerellaceae F11 Gp JGI alkaliphile 43,45 113, Illumina Verticillium alfalfae Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Glomerellales Plectosphaerellaceae VaMs.102 Gp ABPE plant pathogen (broad host range); wilt disease 32,83 4, Sanger Verticillium alfalfae Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Glomerellales Plectosphaerellaceae VaMs.102 Gp ABPE plant pathogen (broad host range); wilt disease 32,83 4, Sanger Verticillium dahliae Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Glomerellales Plectosphaerellaceae VdLs.17 Gp ABJE plant pathogen (broad host range); wilt disease 33,9 7, Verticillium dahliae Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Glomerellales Plectosphaerellaceae VdLs.17 Gp ABJE plant pathogen (broad host range); wilt disease 33,9 7, Verticillium longisporum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Glomerellales Plectosphaerellaceae VL1 Gp CVQH plant pathogen (canola) 99, ,9

62 Verticillium longisporum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Glomerellales Plectosphaerellaceae VL1 Gp CVQH plant pathogen (canola) 99, ,9 Verticillium tricorpus Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Glomerellales Plectosphaerellaceae MUCL9297 Gp JPET opportunistic plant pathogen (lettuce); saprotroph 36,06 65, ,3 Illumina HiSeq; PacBio Verticillium tricorpus Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Glomerellales Plectosphaerellaceae MUCL9297 Gp JPET opportunistic plant pathogen (lettuce); saprotroph 36,06 65, ,3 Illumina HiSeq; PacBio Clonostachys rosea Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Bionectriaceae CBS Gp JGI endophyte; saprotroph; parasite; biocontrol 52, Illumina HiSeq Clonostachys rosea Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Bionectriaceae CBS Gp JGI endophyte; saprotroph; parasite; biocontrol 52,44 59, Illumina HiSeq Gliomastix tumulicola Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Bionectriaceae JCM Gp BCHX uncertain 42, ,8 HiSeq 2500 Gliomastix tumulicola Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Bionectriaceae JCM Gp BCHX uncertain 42,39 295, ,8 HiSeq 2500 Stanjemonium grisellum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Bionectriaceae CBS Gp soil-associated , Illumina Aciculosporium take Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae MAFF Gp University of Kentucky plant pathogen (bamboo parasite) 58,836 18, ,1 454 GS FLX Titanium Aciculosporium take Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae MAFF Gp University of Kentucky plant pathogen (bamboo parasite) 58,836 18, ,1 454 GS FLX Titanium Aschersonia aleyrodis Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae RCEF 2490 Gp AZGY entomopathogen 30,87 76, ,87 Illumina HiSeq Atkinsonella hypoxylon Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae B4728 Gp JFHB parasite (systemic grass) 35,6 31, Atkinsonella hypoxylon Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae B4728 Gp JFHB parasite (systemic grass) 35,6 31, Atkinsonella texensis Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae B6155 Gp LBNC epiphyte 29,37 63, ,8 Illumina MiSeq Atkinsonella texensis Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae B6155 Gp LBNC epiphyte 29,37 63, ,8 Illumina MiSeq Balansia obtecta Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae B249 Gp JFZS parasite (systemic grass) 30,1 21, ,5 Balansia obtecta Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae B249 Gp JFZS parasite (systemic grass) 30,1 21, ,5 Claviceps fusiformis Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae PRL 1980 Gp AFRA plant pathogen (pearl millet) 52,585 34, ,9 454 Claviceps fusiformis Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae PRL 1980 Gp AFRA plant pathogen (pearl millet) 52,585 34, ,9 454 Claviceps paspali Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae 7990 Gp AFRC medical importance (ergot fungus; mycotoxin-producing) 28,97 54, ,5 454 Claviceps paspali Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae 7990 Gp AFRC medical importance (ergot fungus; mycotoxin-producing) 28,97 54, ,5 454 Claviceps purpurea Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae 20,1 Gp CAGA plant pathogen (cereals) 32, ,6 Claviceps purpurea Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae 20,1 Gp CAGA plant pathogen (cereals) 32, ,6 Epichloe (Neotyphodium) gansuensascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae E7080 AFRE endophyte (grass) 39,62 41, ,4 454 Epichloe amarillans Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae E57 Gp AFRF endophyte; plant pathogen 38,06 27, ,3 454 Epichloe amarillans Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae E57 Gp AFRF endophyte; plant pathogen 38,06 27, ,3 454 Epichloe baconii Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae ATCC Gp JFGY endophyte; plant pathogen 38,08 18, ,6 454 Epichloe baconii Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae ATCC Gp JFGY endophyte; plant pathogen 38,08 18, ,6 454 Epichloe brachyelytri Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae E4804 Gp AFRB endophyte; plant pathogen 44,23 24, ,2 454 Epichloe brachyelytri Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae E4804 Gp AFRB endophyte; plant pathogen 44,23 24, ,2 454 Epichloe bromicola Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae ATCC Gp JFHA endophyte; plant pathogen 28,57 6, ,3 454 Epichloe bromicola Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae ATCC Gp JFHA endophyte; plant pathogen 28,57 6, ,3 454 Epichloe elymi Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae E56 Gp AMDJ endophyte 32,335 31, ,6 454; Ion Torrent Epichloe elymi Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae E56 Gp AMDJ endophyte 32,335 31, ,6 454; Ion Torrent Epichloe festucae Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae FI1 Gp AFRX endophyte 35,105 27, ,1 454; Sanger Epichloe festucae Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae FI1 Gp AFRX endophyte 35,105 27, ,1 454; Sanger Epichloe glyceriae Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae E277 Gp AFRG endophyte; plant pathogen 46,72 27, Epichloe glyceriae Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae E277 Gp AFRG endophyte; plant pathogen 46,72 27, Epichloe mollis Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae AL9924 Gp JFGW endophyte; plant pathogen 36,169 28, Epichloe mollis Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae AL9924 Gp JFGW endophyte; plant pathogen 36,169 28, Epichloe sylvatica Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae GR10156 Gp LCTT endophyte; plant pathogen (woodland grass sp.) 36, ,8 Illumina MiSeq Epichloe sylvatica Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae GR10156 Gp LCTT endophyte; plant pathogen (woodland grass sp.) 36,1 160, ,8 Illumina MiSeq Epichloe typhina Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae E5819 Gp AFSE plant pathogen (Poaceae) 34,185 18, ,5 454 Epichloe typhina Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae E5819 Gp AFSE plant pathogen (Poaceae) 34,185 18, ,5 454 Epichloe uncinata Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae e167 Gp LELE endophyte (grasses) 53,34 131, ,6 Illumina MiSeq Epichloe uncinata Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae e167 Gp LELE endophyte (grasses) 53,34 131, ,6 Illumina MiSeq Hypocrella siamensis Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae MTCC Gp JMQE entomopathogenic 28,856 90, ,1 Illumina HiSeq Hypocrella siamensis Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae MTCC Gp JMQE entomopathogen 28,856 90, ,1 Illumina HiSeq Metacordyceps chlamydosporia Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae 123 Gp AOSW biocontrol (nematophagous) 42, , ,7 Illumina HiSeq Metacordyceps chlamydosporia Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae 123 Gp AOSW biocontrol (nematophagous) 42, , ,7 Illumina HiSeq Metarhizium acridum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae CQMa102 Gp ADNI entomopathogenic ; biocontrol 39, , ,9 Illumina Metarhizium acridum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae CQMa102 Gp ADNI entomopathogen ; biocontrol 39, , ,9 Illumina Metarhizium album Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae ARSEF 1941 Gp AZHE entomopathogen 30,45 117, ,45 Illumina HiSeq Metarhizium anisopliae Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae ARSEF 23 Gp ADNJ entomopathogenic ; biocontrol 39, , ,5 Illumina Metarhizium anisopliae Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae ARSEF 23 Gp ADNJ entomopathogen ; biocontrol 39, , ,5 Illumina Metarhizium brunneum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae ARSEF 3297 Gp AZNG biocontrol (entomopathogen) 37,07 80, ,5 Illumina HiSeq Metarhizium guizhouense Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae ARSEF 977 Gp AZNH biocontrol (entomopathogen) 43,47 95, ,6 Illumina HiSeq Metarhizium majus Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae ARSEF 297 Gp AZNE biocontrol (entomopathogen) 42,02 71, Illumina HiSeq Metarhizium rileyi Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae RCEF 4871 Gp AZHC entomopathogen 32,01 107, ,3 Illumina HiSeq Metarhizium robertsii Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae ARSEF 2575 Gp ELW parasite (arthropod) 40,317 25, ,8 454 Metarhizium robertsii Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae ARSEF 2575 Gp ELW parasite (arthropod) 40,317 25, ,8 454 Neotyphodium aotearoae Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae ATCC MYA-1229 Gp JFGX endophyte (grass) 34,423 26, ,6 454 Neotyphodium aotearoae Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae ATCC MYA-1229 Gp JFGX endophyte (grass) 34,423 26, ,6 454 Neotyphodium gansuense Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae E7080 Gp AFRE endophyte (grass); medical importance (mycotoxin-producing) 39,616 41, ,4 454 Neotyphodium gansuense Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae E7080 Gp AFRE endophyte (grass); medical importance (mycotoxin-producing) 39,616 41, ,4 454 Paecilomyces hepiali Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae FENG Gp LNDK pharmaceutical importance (anticancer properties) 34, ,9 Illumina HiSeq Paecilomyces hepiali Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae FENG Gp LNDK pharmaceutical importance (anticancer properties) 34,68 100, ,9 Illumina HiSeq Periglandula ipomoeae Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae IasaF13 Gp AFRD pharmaceutical importance (ergoline-alkaloid-producing) 35, ,6 454 Periglandula ipomoeae Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae IasaF13 Gp AFRD pharmaceutical importance (ergoline-alkaloid-producing) 35, ,6 454 Tolypocladium inflatum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae NRRL 8044 Gp AOHE entomopathogen 30,217 82, ; Illumina Tolypocladium inflatum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae NRRL 8044 Gp AOHE entomopathogen 30,217 82, ; Illumina Torrubiella hemipterigena Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae BCC 1449 Gp CDHN entomopathogen 28, ,6 Torrubiella hemipterigena Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae BCC 1449 Gp CDHN entomopathogen 28, ,6 Villosiclava virens Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae UV-8b Gp JHTR plant pathogen; (false smut of rice) 39, , ,9 454; Illumina HiSeq Villosiclava virens Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Clavicipitaceae UV-8b Gp JHTR plant pathogen; (false smut of rice) 39, , ,9 454; Illumina HiSeq Beauveria bassiana Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Cordycipitaceae ARSEF 2860 Gp ADAH entomopathogenic 33,69 28, ,5 454 Beauveria bassiana Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Cordycipitaceae ARSEF 2860 Gp ADAH entomopathogen 33,69 28, ,5 454 Beauveria sp. YA-2014 Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Cordycipitaceae YA-2014 MTCC8017 Gp JMNB insect-associated 36,58 66, ,1 Illumina HiSeq Beauveria sp. YA-2014 Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Cordycipitaceae YA-2014 MTCC8017 Gp JMNB insect-associated 36,58 66, ,1 Illumina HiSeq Cordyceps brongniartii Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Cordycipitaceae RCEF 3172 Gp AZHA entomopathogen 32,52 73, ,5 Illumina HiSeq Cordyceps cicadae Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Cordycipitaceae BA-001 Gp AEIW pharmaceutical importance; traditional use 32,52 28, ,7 454 Cordyceps confragosa Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Cordycipitaceae RCEF 1005 Gp AZHF entomopathogen 35,59 35, ,1 Illumina HiSeq Cordyceps militaris Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Cordycipitaceae CM01 Gp AEVU entomopathogenic 32,26 100, ,4 454; Illumina Cordyceps militaris Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Cordycipitaceae CM01 Gp AEVU entomopathogen 32,26 100, ,4 454; Illumina Isaria farinosa Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Cordycipitaceae MTCC 4114 Gp JMNC entomopathogenic 34,157 60, ,2 Illumina HiSeq Isaria farinosa Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Cordycipitaceae MTCC 4114 Gp JMNC entomopathogen 34,157 60, ,2 Illumina HiSeq Isaria fumosorosea Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Cordycipitaceae ARSEF 2679 Gp AZHB entomopathogen 33,49 86, ,5 Illumina HiSeq Trichoderma asperellum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Hypocreaceae CBS Gp JGI biocontrol (mycoparasite); saprotroph to biotroph 37, ; Illumina Trichoderma asperellum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Hypocreaceae CBS Gp JGI biocontrol (mycoparasite); saprotroph to biotroph 37,46 130, ; Illumina Trichoderma atroviride Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Hypocreaceae IMI Gp ABDG biocontrol (mycoparasite) 36,1 8, ,7 Sanger Trichoderma atroviride Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Hypocreaceae IMI Gp ABDG biocontrol (mycoparasite) 36,1 8, ,7 Sanger Trichoderma citrinoviride Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Hypocreaceae TUCIM opportunistic human pathogen 33,22 63, Illumina Trichoderma citrinoviride Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Hypocreaceae TUCIM opportunistic human pathogen 33,22 63, Illumina Trichoderma hamatum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Hypocreaceae GD12 Gp ANCB biocontrol (mycoparasite) 38,176 40, ,5 Illumina HiSeq Trichoderma hamatum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Hypocreaceae GD12 Gp ANCB biocontrol (mycoparasite) 38,176 40, ,5 Illumina HiSeq Trichoderma harzianum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Hypocreaceae CBS Gp JGI biocontrol (mycoparasite) 40, Illumina Trichoderma harzianum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Hypocreaceae CBS Gp JGI biocontrol (mycoparasite) 40,98 120, Illumina Trichoderma longibrachiatum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Hypocreaceae ATCC Gp JGI biocontrol (plant pathogenic soil nematodes) 32,24 104, Illumina Trichoderma longibrachiatum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Hypocreaceae ATCC Gp JGI biocontrol (plant pathogenic soil nematodes) 32,24 104, Illumina Trichoderma parareesei Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Hypocreaceae CBS Gp LFMI biocontrol (mycoparasite) 32, ,7 Illumina Trichoderma parareesei Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Hypocreaceae CBS Gp LFMI biocontrol (mycoparasite) 32,1 28, ,7 Illumina

63 Trichoderma pseudokoningii Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Hypocreaceae SMF2 Gp ANBJ biocontrol (nematode parasite) 31,743 69, ; Illumina HiSeq Trichoderma pseudokoningii Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Hypocreaceae SMF2 Gp ANBJ biocontrol (nematode parasite) 31,743 69, ; Illumina HiSeq Trichoderma reesei Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Hypocreaceae QM6a Gp AAIL industrial importance (cellulases; hemicellulases) 33,3 9, ,8 Sanger Trichoderma reesei Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Hypocreaceae QM6a Gp AAIL industrial importance (cellulases; hemicellulases) 33,3 9, ,8 Sanger Trichoderma virens Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Hypocreaceae Gv29-8 Gp ABDF biocontrol 39 8, ,2 Sanger Trichoderma virens Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Hypocreaceae Gv29-8 Gp ABDF biocontrol 39 8, ,2 Sanger Acremonium (Sarocladium) strictumascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Incertae sedis DS1bioAY4a Gp endophyte (grasses); mycoparasite 35,79 120, Illumina Sarocladium oryzae Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Incertae sedis Saro-13 LOPT plant pathogen (rice sheath rot) 32,78 82, ,5 Illumina Stachybotrys chartarum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Incertae sedis IBT Gp ASEQ medical importance (stachybotryotoxicosis; mycotoxin-producing-satratox 36,48 192, ,3 Illumina HiSeq Stachybotrys chartarum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Incertae sedis IBT Gp ASEQ medical importance (stachybotryotoxicosis; mycotoxin-producing-satratox 36,48 192, ,3 Illumina HiSeq Stachybotrys chlorohalonata Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Incertae sedis IBT Gp APWP medical importance (atranone-producing) 34,39 196, ,4 Illumina HiSeq Stachybotrys chlorohalonata Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Incertae sedis IBT Gp APWP medical importance (atranone-producing) 34,39 196, ,4 Illumina HiSeq Ustilaginoidea virens Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Incertae sedis IPU010 Gp BBTG plant pathogen (rice) 33, ,9 Illumina MiSeq; PacBio Ustilaginoidea virens Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Incertae sedis IPU010 Gp BBTG plant pathogen (rice) 33, ,9 Illumina MiSeq; PacBio Dactylonectria macrodidyma Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Nectriaceae JAC Gp JYGD plant pathogen (grapevine, avocado, and olive) Illumina MiSeq Dactylonectria macrodidyma Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Nectriaceae JAC Gp JYGD plant pathogen (grapevine, avocado, and olive) 58 74, Illumina MiSeq Fusarium acuminatum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Nectriaceae CS5907 Gp CBMG plant pathogen (cereals) 43, Illumina HiSeq Fusarium acuminatum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Nectriaceae CS5907 Gp CBMG plant pathogen (cereals) 43, Illumina HiSeq Fusarium avenaceum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Nectriaceae FaLH03 Gp JQGD plant pathogen (generalist, including grain crops) 43, Illumina HiSeq Fusarium avenaceum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Nectriaceae FaLH03 Gp JQGD plant pathogen (generalist, including grain crops) 43,17 459, Illumina HiSeq Fusarium circinatum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Nectriaceae FSP 34 Gp AYJV plant pathogen (pitch canker of pines) 44,138 11, ,3 454 Fusarium circinatum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Nectriaceae FSP 34 Gp AYJV plant pathogen (pitch canker of pines) 44,138 11, ,3 454 Fusarium equiseti Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Nectriaceae CS3069 Gp CBMI plant pathogen (members of the Leguminoseae and some cereals) 38, Illumina HiSeq Fusarium equiseti Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Nectriaceae CS3069 Gp CBMI plant pathogen (members of the Leguminoseae and some cereals) 38, Illumina HiSeq Fusarium fujikuroi Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Nectriaceae B14 Gp ANFV plant pathogen (rice) 43,81 75, ,3 Illumina HiSeq Fusarium fujikuroi Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Nectriaceae B14 Gp ANFV plant pathogen (rice) 43,81 75, ,3 Illumina HiSeq Fusarium graminearum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Nectriaceae CS3005 Gp JATU plant pathogen (wheat and barley) 36,66 50, ,3 Illumina HiSeq Fusarium graminearum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Nectriaceae CS3005 Gp JATU plant pathogen (wheat and barley) 36,66 50, ,3 Illumina HiSeq Fusarium langsethiae Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Nectriaceae Fl Gp JXCE medical importance (mycotoxin-producing) 37, ,3 Illumina GAIIx; 454 Fusarium langsethiae Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Nectriaceae Fl Gp JXCE medical importance (mycotoxin-producing) 37,54 500, ,3 Illumina GAIIx; 454 Fusarium nygamai Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Nectriaceae MRC8546 Gp LBNR plant pathogen (rice) 51, ,5 Illumina HiSeq Fusarium nygamai Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Nectriaceae MRC8546 Gp LBNR plant pathogen (rice) 51,62 333, ,5 Illumina HiSeq Fusarium oxysporum f. sp. conglutiascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Nectriaceae NRRL Gp AGNF plant pathogen (broad host range) 53,57 124, ,4 Illumina Fusarium oxysporum f. sp. conglutiascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Nectriaceae NRRL Gp AGNF plant pathogen (broad host range) 53,57 124, ,4 Illumina Fusarium pseudograminearum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Nectriaceae CS3427 Gp CBMD plant pathogen (wheat) 36, ,8 Illumina HiSeq Fusarium pseudograminearum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Nectriaceae CS3427 Gp CBMD plant pathogen (wheat) 36, ,8 Illumina HiSeq Fusarium sambucinum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Nectriaceae F-4 Gp LSRD plant pathogen (potato dry rot); mycotoxin-producing 37, ,8 454 Fusarium sambucinum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Nectriaceae F-4 Gp LSRD plant pathogen (potato dry rot); mycotoxin-producing 37,78 18, ,8 454 Fusarium sp. JS1030 Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Nectriaceae JS1030 Gp JWIW uncertain 53, ,7 Illumina HiSeq; MiSeq Fusarium sp. JS1030 Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Nectriaceae JS1030 Gp JWIW uncertain 53, , ,7 Illumina HiSeq; MiSeq Fusarium sp. JS626 Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Nectriaceae JS626 Gp JWIV uncertain 42, ,2 Illumina HiSeq; MiSeq Fusarium sp. JS626 Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Nectriaceae JS626 Gp JWIV uncertain 42, , ,2 Illumina HiSeq; MiSeq Fusarium temperatum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Nectriaceae CMWF389 Gp LJGR plant pathogen (maize); opportunistic human pathogen 45,46 414, Illumina HiSeq Fusarium verticillioides Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Nectriaceae FGSC 7600 Gp AAIM plant pathogen (maize) 41,8851 8, ,7 Sanger Fusarium verticillioides Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Nectriaceae FGSC 7600 Gp AAIM plant pathogen (maize) 41,8851 8, ,7 Sanger Fusarium virguliforme Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Nectriaceae Mont-1 Gp AEYB plant pathogen (soybean) 50,448 20, ,4 454 Fusarium virguliforme Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Nectriaceae Mont-1 Gp AEYB plant pathogen (soybean) 50,448 20, ,4 454 Ilyonectria europaea Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Nectriaceae PMI-82 Gp JGI root-associated 63,66 153, Illumina HiSeq Ilyonectria europaea Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Nectriaceae PMI-82 Gp JGI root-associated 63,66 153, Illumina HiSeq Nectria haematococca Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Nectriaceae MPVI Gp JGI plant pathogen (broad host range) ,8 Nectria haematococca Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Nectriaceae MPVI Gp JGI plant pathogen (broad host range) ,8 Neonectria ditissima Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Nectriaceae RS324p Gp LDPL plant pathogen (apple canker) 44, ,8 Illumina HiSeq Neonectria ditissima Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Nectriaceae RS324p Gp LDPL plant pathogen (apple canker) 44,95 200, ,8 Illumina HiSeq Niesslia exilis Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Niessliaceae CBS Gp saprotroph 35,38 85, Illumina HiSeq Valetoniellopsis laxa Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Niessliaceae CBS Gp uncertain; monotypic genus 22,13 95, Illumina HiSeq Drechmeria coniospora Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Ophiocordycipitaceae ATCC Gp JYHR biocontrol (nematode endoparasite) 31, ,9 Illumina MiSeq; SOLiD Drechmeria coniospora Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Ophiocordycipitaceae ATCC Gp JYHR biocontrol (nematode endoparasite) 31,89 100, ,9 Illumina MiSeq; SOLiD Hirsutella minnesotensis Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Ophiocordycipitaceae 3608 Gp JPUM biocontrol (nematode pathogen) 51,11 128, ,1 Illumina HiSeq Hirsutella thompsonii Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Ophiocordycipitaceae MTCC3556 Gp APKB arthropod pathogen (acarine ) 34, , ,6 Illumina HiSeq Hirsutella thompsonii Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Ophiocordycipitaceae MTCC3556 Gp APKB arthropod pathogen (acarine ) 34, , ,6 Illumina HiSeq Ophiocordyceps polyrhachis-furcataascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Ophiocordycipitaceae BCC Gp LKCN uncertain 43,16 37, ,8 454; Illumina Ophiocordyceps sinensis Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Ophiocordycipitaceae CO18 Gp ANOV traditional use 78, , ,2 454; Illumina HiSeq Ophiocordyceps sinensis Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Ophiocordycipitaceae CO18 Gp ANOV traditional use 78, , ,2 454; Illumina HiSeq Ophiocordyceps unilateralis Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Ophiocordycipitaceae SC16a Gp LAZP entomopathogen 25, Illumina MiSeq Ophiocordyceps unilateralis Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Ophiocordycipitaceae SC16a Gp LAZP entomopathogen 25,49 120, Illumina MiSeq Purpureocillium lilacinum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Ophiocordycipitaceae TERIBC 1 Gp LOFA saprotroph 38, ,9 Illumina HiSeq Purpureocillium lilacinum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Ophiocordycipitaceae TERIBC 1 Gp LOFA saprotroph 38,82 200, ,9 Illumina HiSeq Tolypocladium (Elaphocordyceps) ophioglossoides Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Ophiocordycipitaceae CBS Gp LFRF parasite 31,25 76, Illumina HiSeq Tolypocladium sp. Salcha MEA-2 Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Ophiocordycipitaceae MEA-2 Gp JPIJ uncertain 30, ,9 PacBio Tolypocladium sp. Salcha MEA-2 Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Ophiocordycipitaceae MEA-2 Gp JPIJ uncertain 30,83 42, ,9 PacBio Tolypocladium sp. Sup5 PDA-1 Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Ophiocordycipitaceae PDA-1 Gp JPHH uncertain 37, PacBio; Illumina HiSeq; Oxford Nanopore Tolypocladium sp. Sup5 PDA-1 Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Ophiocordycipitaceae PDA-1 Gp JPHH uncertain 37,87 469, PacBio; Illumina HiSeq; Oxford Nanopore Myrothecium inundatum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Stachybotriaceae CBS Gp saprotroph; biocontrol; industrial importance 39,21 95, Illumina HiSeq Stachybotrys echinata Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Stachybotriaceae JCM Gp BCHF medical importance (mycotoxin-producing) 43, ,8 HiSeq 2500 Stachybotrys echinata Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Hypocreales Stachybotriaceae JCM Gp BCHF medical importance (mycotoxin-producing) 43,68 215, ,8 HiSeq 2500 Albophoma yamanashiensis Ascomycota Pezizomycotina Sordariomycetes Incertae sedis Incertae sedis JCM Gp BCKH uncertain 31, ,7 HiSeq 2500 Albophoma yamanashiensis Ascomycota Pezizomycotina Sordariomycetes Incertae sedis Incertae sedis Incertae sedis JCM Gp BCKH uncertain 31,39 325, ,7 HiSeq 2500 Didymobotryum rigidum Ascomycota Pezizomycotina Sordariomycetes Incertae sedis Incertae sedis JCM 8837 Gp BCKI uncertain 41, ,7 HiSeq 2500 Didymobotryum rigidum Ascomycota Pezizomycotina Sordariomycetes Incertae sedis Incertae sedis Incertae sedis JCM 8837 Gp BCKI uncertain 41,73 204, ,7 HiSeq 2500 Plectosphaerella cucumerina Ascomycota Pezizomycotina Sordariomycetes Sordariomycetidae Incertae sedis Plectosphaerellaceae DS2psM2a2 v plant pathogen (blight of cucurbits); nematophagous 36,84 86, Illumina Plectosphaerella cucumerina Ascomycota Pezizomycotina Sordariomycetes Sordariomycetidae Incertae sedis Plectosphaerellaceae DS2psM2a2 v plant pathogen (blight of cucurbits); nematophagous 36,84 86, Illumina Torpedospora radiata Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Incertae sedis Torpedosporaceae JK5252C Gp marine fungus 33,2 93, Illumina HiSeq Torpedospora radiata Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Incertae sedis Torpedosporaceae JK5252C Gp marine fungus 33,2 93, Illumina HiSeq Lindra thalassiae Ascomycota Pezizomycotina Sordariomycetes Lulworthiomycetidae Lulworthiales Lulworthiaceae JK4322 Gp probable plant pathogen (turtle grass) 31,92 145, Illumina HiSeq Gaeumannomyces graminis Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Magnaporthales Magnaporthaceae R3-111a-1 Gp ADBI plant pathogen (root rot of cereals) 43,76 25, ,8 454 Gaeumannomyces graminis Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Magnaporthales Magnaporthaceae R3-111a-1 Gp ADBI plant pathogen (root rot of cereals) 43,76 25, ,8 454 Harpophora oryzae Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Magnaporthales Magnaporthaceae R5-6-1 Gp JNVV endophyte 50, , ,5 Illumina HiSeq Harpophora oryzae Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Magnaporthales Magnaporthaceae R5-6-1 Gp JNVV endophyte 50, , ,5 Illumina HiSeq Magnaporthe oryzae Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Magnaporthales Magnaporthaceae KJ201 Gp ANSL plant pathogen (rice) 45, , ,6 Illumina GAIIx Magnaporthe oryzae Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Magnaporthales Magnaporthaceae KJ201 Gp ANSL plant pathogen (rice) 45, , ,6 Illumina GAIIx Magnaporthiopsis poae Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Magnaporthales Magnaporthaceae ATCC Gp ADBL plant pathogen (turfgrass) 39, , Magnaporthiopsis poae Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Magnaporthales Magnaporthaceae ATCC Gp ADBL plant pathogen (turfgrass) 39, , Melanospora tiffanyae Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Melanosporales Ceratostomataceae F1KG0001 v biocontrol (mycoparasite) 28,36 136, Illumina Melanospora tiffanyae Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Melanosporales Ceratostomataceae F1KG0001 v biocontrol (mycoparasite) 28,36 136, Illumina Ceratocystis adiposa Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Microascales Ceratocystidaceae CBS Gp LXGU plant pathogen (black root in sugar cane) 28,34 93, Illumina Ceratocystis albifundus Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Microascales Ceratocystidaceae CMW17620 Gp JSSU plant pathogen (woody & herbaceaous) 26, ,6 Illumina GAIIx Ceratocystis albifundus Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Microascales Ceratocystidaceae CMW17620 Gp JSSU plant pathogen (woody & herbaceaous) 26,88 24, ,6 Illumina GAIIx Ceratocystis eucalypticola Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Microascales Ceratocystidaceae CMW9998 Gp LJOA probable plant pathogen (eucalyptus) 30, ,1 Illumina HiSeq 2000 Ceratocystis eucalypticola Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Microascales Ceratocystidaceae CMW9998 Gp LJOA probable plant pathogen (eucalyptus) 30,72 80, ,1 Illumina HiSeq 2000 Ceratocystis fimbriata Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Microascales Ceratocystidaceae C1421 Gp APWK plant pathogen (sweet potato) 29,4 27, , Ceratocystis fimbriata Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Microascales Ceratocystidaceae C1421 Gp APWK plant pathogen (sweet potato) 29,4 27, ,06 454

64 Ceratocystis manginecans Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Microascales Ceratocystidaceae CMW17570 Gp JJRZ plant pathogen (mango) 31,7 22, Illumina Ceratocystis manginecans Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Microascales Ceratocystidaceae CMW17570 Gp JJRZ plant pathogen (mango) 31,7 22, Illumina Ceratocystis platani Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Microascales Ceratocystidaceae CFO Gp LBBL plant pathogen (trees) 29, ,2 Illumina HiSeq Ceratocystis platani Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Microascales Ceratocystidaceae CFO Gp LBBL plant pathogen (trees) 29,18 655, ,2 Illumina HiSeq Chalaropsis thielavioides Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Microascales Ceratocystidaceae JCM 1933 Gp BCGU plant pathogen (Fabaceae sp.) 29, ,1 HiSeq 2500 Chalaropsis thielavioides Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Microascales Ceratocystidaceae JCM 1933 Gp BCGU plant pathogen (Fabaceae sp.) 29,33 387, ,1 HiSeq 2500 Endoconidiophora laricicola Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Microascales Ceratocystidaceae CBS Gp LXGT insect-associated 32,69 93, ,4 Illumina Huntiella moniliformis Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Microascales Ceratocystidaceae CBS Gp JMSH saprotroph 25,4 38, ,9 Illumina GAIIx Huntiella moniliformis Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Microascales Ceratocystidaceae CBS Gp JMSH saprotroph 25,4 38, ,9 Illumina GAIIx Huntiella omanensis Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Microascales Ceratocystidaceae CMW Gp JSUI plant pathogen (weak mango pathogen) 31, ,7 Illumina Huntiella omanensis Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Microascales Ceratocystidaceae CMW Gp JSUI plant pathogen (weak mango pathogen) 31,13 9, ,7 Illumina Huntiella savannae Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Microascales Ceratocystidaceae CBS Gp LCZG saproptroph 28, ,3 Illumina HiSeq Huntiella savannae Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Microascales Ceratocystidaceae CBS Gp LCZG saproptroph 28,54 22, ,3 Illumina HiSeq Thielaviopsis musarum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Microascales Ceratocystidaceae CMW1546 Gp LKBB post harvest pathogen (banana) 28, ,2 Illumina GAIIx Thielaviopsis musarum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Microascales Ceratocystidaceae CMW1546 Gp LKBB post harvest pathogen (banana) 28,42 95, ,2 Illumina GAIIx Thielaviopsis paradoxa Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Microascales Ceratocystidaceae JCM 6020 Gp BCHJ plant pathogen (palm) 29, HiSeq 2500 Thielaviopsis paradoxa Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Microascales Ceratocystidaceae JCM 6020 Gp BCHJ plant pathogen (palm) 29,63 345, HiSeq 2500 Thielaviopsis punctulata Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Microascales Ceratocystidaceae CR-DP1 Gp LAEV plant pathogen (date palm) 28, ,3 Thielaviopsis punctulata Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Microascales Ceratocystidaceae CR-DP1 Gp LAEV plant pathogen (date palm) 28,12 75, ,3 Knoxdaviesia capensis Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Microascales Gondwanamycetaceae Gs LNGK saprotroph (plant-associated) 35,54 188, ,8 HiSeq 2500 Knoxdaviesia capensis Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Microascales Gondwanamycetaceae Gs LNGK saprotroph (plant-associated) 35,54 188, ,8 HiSeq 2500 Knoxdaviesia proteae Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Microascales Gondwanamycetaceae SB2.3 Gp LNGL saprotroph (plant-associated) 35, ,7 HiSeq 2500 Knoxdaviesia proteae Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Microascales Gondwanamycetaceae SB2.3 Gp LNGL saprotroph (plant-associated) 35, , ,7 HiSeq 2500 Corollospora maritima Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Microascales Halosphaeriaceae CBS v marine saprotroph 36,97 144, Illumina Corollospora maritima Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Microascales Halosphaeriaceae CBS v marine saprotroph 36,97 144, Illumina Microascus trigonosporus Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Microascales Microascaceae CBS Gp JGI human pathogen 36, Illumina Microascus trigonosporus Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Microascales Microascaceae CBS Gp JGI human pathogen 36,1 104, Illumina Scedosporium apiospermum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Microascales Microascaceae IHEM Gp JOWA human pathogen (infects lungs, especially in cystic fibrosis patients) 41, , ,4 Illumina HiSeq; PacBio Scedosporium apiospermum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Microascales Microascaceae IHEM Gp JOWA human pathogen (infects lungs, especially in cystic fibrosis patients) 41, , ,4 Illumina HiSeq; PacBio Scedosporium aurantiacum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Microascales Microascaceae WM Gp JUDQ human pathogen 39, ,2 Illumina HiSeq Scedosporium aurantiacum Ascomycota Pezizomycotina Sordariomycetes Hypocreomycetidae Microascales Microascaceae WM Gp JUDQ human pathogen 39,89 162, ,2 Illumina HiSeq Graphilbum fragrans Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Ophiostomatales Ophiostomataceae CBS Gp LLKO insect-associated (conifer-infesting beetles) 34, ,7 Illumina HiSeq Graphilbum fragrans Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Ophiostomatales Ophiostomataceae CBS Gp LLKO insect-associated (conifer-infesting beetles) 34,27 64, ,7 Illumina HiSeq Grosmannia clavigera Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Ophiostomatales Ophiostomataceae kw1407 Gp ACXQ plant pathogen; blue stain 29,79 50, ,4 Sanger; 454; Illumina Grosmannia clavigera Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Ophiostomatales Ophiostomataceae kw1407 Gp ACXQ plant pathogen; blue stain 29,79 50, ,4 Sanger; 454; Illumina Leptographium lundbergii Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Ophiostomatales Ophiostomataceae CBS Gp LDEF insect-associated (beetles) 26, ,9 Illumina HiSeq Leptographium lundbergii Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Ophiostomatales Ophiostomataceae CBS Gp LDEF insect-associated (beetles) 26,54 70, ,9 Illumina HiSeq Leptographium procerum Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Ophiostomatales Ophiostomataceae CMW34542 Gp JRUC plant pathogen (pine) 28, ,8 Illumina MiSeq Leptographium procerum Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Ophiostomatales Ophiostomataceae CMW34542 Gp JRUC plant pathogen (pine) 28,57 32, ,8 Illumina MiSeq Ophiostoma novo-ulmi Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Ophiostomatales Ophiostomataceae H327 Gp AMZD plant pathogen (Dutch Elm Disease) 31,855 61, ,1 454 Ophiostoma novo-ulmi Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Ophiostomatales Ophiostomataceae H327 Gp AMZD plant pathogen (Dutch Elm Disease) 31,855 61, ,1 454 Ophiostoma piceae Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Ophiostomatales Ophiostomataceae UAMH Gp AQHS saprotroph; wood-staining 32, ,4 454; Illumina HiSeq Ophiostoma piceae Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Ophiostomatales Ophiostomataceae UAMH Gp AQHS saprotroph; wood-staining 32,84 735, ,4 454; Illumina HiSeq Sporothrix brasiliensis Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Ophiostomatales Ophiostomataceae 5110 Gp AWTV human and animal pathogen (sporotrichosis) 33,21 20, Sporothrix globosa Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Ophiostomatales Ophiostomataceae CBS Gp LVYW human and animal pathogen (sporotrichosis) 33,47 146, ,3 Illumina Sporothrix insectorum Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Ophiostomatales Ophiostomataceae RCEF 264 Gp AZHD entomopathogen 34, , Illumina HiSeq Sporothrix pallida Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Ophiostomatales Ophiostomataceae SPA8 Gp JNEX saprotroph (environmental) 39,881 20, Ion Torrent Sporothrix pallida Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Ophiostomatales Ophiostomataceae SPA8 Gp JNEX saprotroph (environmental) 39,881 20, Ion Torrent Sporothrix schenckii Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Ophiostomatales Ophiostomataceae ATCC Gp AWEQ human and animal pathogen (sporotrichosis) 32,228 56, ,1 Illumina Sporothrix schenckii Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Ophiostomatales Ophiostomataceae ATCC Gp AWEQ human and animal pathogen (sporotrichosis) 32,228 56, ,1 Illumina Chaetomium globosum Ascomycota Pezizomycotina Sordariomycetes Sordariomycetidae Sordariales Chaetomiaceae CBS Gp AAFU human pathogen and allergen (mycosis; mycotoxin-producing) 34,88 7, ,6 Sanger Chaetomium globosum Ascomycota Pezizomycotina Sordariomycetes Sordariomycetidae Sordariales Chaetomiaceae CBS Gp AAFU human pathogen and allergen (mycosis; mycotoxin-producing) 34,88 7, ,6 Sanger Chaetomium thermophilum Ascomycota Pezizomycotina Sordariomycetes Sordariomycetidae Sordariales Chaetomiaceae DSM 1495 Gp ADUW thermophile 28,32 24, ,6 454 Chaetomium thermophilum Ascomycota Pezizomycotina Sordariomycetes Sordariomycetidae Sordariales Chaetomiaceae DSM 1495 Gp ADUW thermophile 28,32 24, ,6 454 Myceliophthora heterothallica Ascomycota Pezizomycotina Sordariomycetes Sordariomycetidae Sordariales Chaetomiaceae CBS v1.0 Gp thermophile 35,36 132, Illumina HiSeq Myceliophthora heterothallica Ascomycota Pezizomycotina Sordariomycetes Sordariomycetidae Sordariales Chaetomiaceae CBS v1.0 Gp thermophile 35,36 132, Illumina HiSeq Myceliophthora thermophila Ascomycota Pezizomycotina Sordariomycetes Sordariomycetidae Sordariales Chaetomiaceae ATCC Gp GCA_ thermophile 38, Myceliophthora thermophila Ascomycota Pezizomycotina Sordariomycetes Sordariomycetidae Sordariales Chaetomiaceae ATCC Gp GCA_ thermophile 38, Thielavia antarctica Ascomycota Pezizomycotina Sordariomycetes Sordariomycetidae Sordariales Chaetomiaceae CBS Gp JGI industrial importance 40,66 88, Illumina Thielavia antarctica Ascomycota Pezizomycotina Sordariomycetes Sordariomycetidae Sordariales Chaetomiaceae CBS Gp JGI industrial importance 40,66 88, Illumina Thielavia appendiculata Ascomycota Pezizomycotina Sordariomycetes Sordariomycetidae Sordariales Chaetomiaceae CBS Gp JGI industrial importance 32,74 95, Illumina HiSeq Thielavia appendiculata Ascomycota Pezizomycotina Sordariomycetes Sordariomycetidae Sordariales Chaetomiaceae CBS Gp JGI industrial importance 32,74 95, Illumina HiSeq Thielavia arenaria Ascomycota Pezizomycotina Sordariomycetes Sordariomycetidae Sordariales Chaetomiaceae CBS Gp JGI industrial importance 30,99 95, Illumina HiSeq Thielavia arenaria Ascomycota Pezizomycotina Sordariomycetes Sordariomycetidae Sordariales Chaetomiaceae CBS Gp JGI industrial importance 30,99 95, Illumina HiSeq Thielavia hyrcaniae Ascomycota Pezizomycotina Sordariomycetes Sordariomycetidae Sordariales Chaetomiaceae CBS Gp JGI industrial importance 31,18 88, Illumina Thielavia hyrcaniae Ascomycota Pezizomycotina Sordariomycetes Sordariomycetidae Sordariales Chaetomiaceae CBS Gp JGI industrial importance 31,18 88, Illumina Thielavia terrestris Ascomycota Pezizomycotina Sordariomycetes Sordariomycetidae Sordariales Chaetomiaceae NRRL 8126 Gp GCA_ industrial importance (thermophile, acidophile) 36, Sanger Thielavia terrestris Ascomycota Pezizomycotina Sordariomycetes Sordariomycetidae Sordariales Chaetomiaceae NRRL 8126 Gp GCA_ industrial importance (thermophile, acidophile) 36, Sanger Madurella mycetomatis Ascomycota Pezizomycotina Sordariomycetes Sordariomycetidae Sordariales Incertae sedis mm55 Gp LCTW human pathogen (eumycetoma) 36,7 43, ,8 PacBio; Illumina; 454 Podospora anserina Ascomycota Pezizomycotina Sordariomycetes Sordariomycetidae Sordariales Lasiosphaeriaceae S mat+ Gp GCA_ saprotroph (dung-inhabiting) 35,01 10, Sanger Neurospora africana Ascomycota Pezizomycotina Sordariomycetes Sordariomycetidae Sordariales Sordariaceae FGSC 1740 Gp CAPO model organism 36, ,9 Neurospora crassa Ascomycota Pezizomycotina Sordariomycetes Sordariomycetidae Sordariales Sordariaceae OR74A Gp AABX model organism 41,102 20, Sanger; 454 Neurospora discreta Ascomycota Pezizomycotina Sordariomycetes Sordariomycetidae Sordariales Sordariaceae FGSC 8579 Gp model organism 37,3 8, Illumina Neurospora pannonica Ascomycota Pezizomycotina Sordariomycetes Sordariomycetidae Sordariales Sordariaceae FGSC 7221 Gp CAPQ model organism 38, ,8 Illumina Neurospora sublineolata Ascomycota Pezizomycotina Sordariomycetes Sordariomycetidae Sordariales Sordariaceae FGSC 5508 Gp CAPP model organism 35, ,3 Illumina Neurospora terricola Ascomycota Pezizomycotina Sordariomycetes Sordariomycetidae Sordariales Sordariaceae FGSC 1889 Gp CAPR model organism 39, ,2 Illumina Neurospora tetrasperma Ascomycota Pezizomycotina Sordariomycetes Sordariomycetidae Sordariales Sordariaceae FGSC 2509 Gp AFCY model organism 39,123 47, ,4 454; Sanger Sordaria macrospora Ascomycota Pezizomycotina Sordariomycetes Sordariomycetidae Sordariales Sordariaceae k-hell Gp CABT model organism 38, ,5 Phaeoacremonium aleophilum Ascomycota Pezizomycotina Sordariomycetes Diaportheomycetidae Togniniales Togniniaceae UCRPA7 Gp AORD plant pathogen (grapevine trunk disease) 47,47 100, ,7 Illumina HiSeq Apiospora montagnei Ascomycota Pezizomycotina Sordariomycetes Xylariomycetidae Xylariales Apiosporaceae NRRL Gp JGI cellulolytic 47,67 95, Illumina Anthostoma avocetta Ascomycota Pezizomycotina Sordariomycetes Xylariomycetidae Xylariales Diatrypaceae NRRL 3190 Gp JGI pharmaceutical importance (antibiotic producing-heptelidic acid) 56,23 117, Ilumina Eutypa lata Ascomycota Pezizomycotina Sordariomycetes Xylariomycetidae Xylariales Diatrypaceae UCREL1 Gp AORF plant pathogen; grapevine dieback , ,6 Illumina HiSeq Daldinia eschscholtzii Ascomycota Pezizomycotina Sordariomycetes Xylariomycetidae Xylariales Xylariaceae UM1020 Gp AIID saproptroph 35, , ,8 Illumina GAIIx Hypoxylon sp. CI-4A Ascomycota Pezizomycotina Sordariomycetes Xylariomycetidae Xylariales Xylariaceae CI-4A Gp JGI endophyte 37,7 114, Illumina Hypoxylon sp. CO27-5 Ascomycota Pezizomycotina Sordariomycetes Xylariomycetidae Xylariales Xylariaceae CO27-5 Gp JGI endophyte 46,59 138, ; Illumina Hypoxylon sp. E7406B Ascomycota Pezizomycotina Sordariomycetes Xylariomycetidae Xylariales Xylariaceae E7406B Gp JYCQ uncertain 39,9 450, ,5 Illumina HiSeq Hypoxylon sp. EC38 Ascomycota Pezizomycotina Sordariomycetes Xylariomycetidae Xylariales Xylariaceae EC38 Gp endophyte 47,72 116, Illumina; PacBio Rosellinia necatrix Ascomycota Pezizomycotina Sordariomycetes Xylariomycetidae Xylariales Xylariaceae W97 Gp BBSO plant pathogen (fruits) 45,03 20, ,9 454 Xylaria hypoxylon Ascomycota Pezizomycotina Sordariomycetes Xylariomycetidae Xylariales Xylariaceae OSC Go saprotroph (white rot) 42,85 148, Illumina Xylaria sp. JS573 Ascomycota Pezizomycotina Sordariomycetes Xylariomycetidae Xylariales Xylariaceae JS573 Gp JWIU saprotroph 40, , ,6 Illumina HiSeq; MiSeq Xylona heveae Ascomycota Pezizomycotina Xylonomycetes Incertae sedis Xylonomycetales Xylonomycetaceae TC161 Gp JGI endophyte (sapwood) 24,34 124, Illumina HiSeq Alloascoidea hylecoeti Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Alloascoideaceae JCM 7604 Gp BCKZ insect-associated 24,82 139, ,1 HiSeq 2500 Ascoidea asiatica Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Ascoideaceae JCM 7603 Gp BCKQ insect-associated 20,31 184, ,8 Illumina Ascoidea rubescens Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Ascoideaceae DSM 1968 Gp JGI insect-vectored 17,5 140, Illumina HiSeq Babjeviella inositovora Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Debaryomycetaceae NRRL Y Gp JGI grows on myo-inositol 15,22 80, ; Illumina Candida sojae Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Debaryomycetaceae GF41 Gp LMTL insect-associated 11,75 48, ,4 Illumina Debaryomyces fabryi Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Debaryomycetaceae CBS 789 Gp LMYN probable human pathogen 11,7 1000, ,6 IonTorrent Debaryomyces hansenii Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Debaryomycetaceae CBS767 Gp GCA_ industrial importance 12,18 9, ,4 Sanger Meyerozyma caribbica Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Debaryomycetaceae MG20W Gp BADS biocontrol (mycoparasite) 10,61 21, ,8 454 Meyerozyma guilliermondii Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Debaryomycetaceae ATCC 6260 Gp AAFM uncertain; yeast (nonpathogenic) 10,609 12, Millerozyma acaciae Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Debaryomycetaceae JCM Gp BCKO metabolically interesting (toxin immunity) 11,07 248, ,4 Illumina

65 Millerozyma farinosa Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Debaryomycetaceae CBS 7064 Gp GCA_ halotolerant; osmotolerant 21,459 9, ,33 Sanger Priceomyces haplophilus Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Debaryomycetaceae JCM 1635 Gp BCIF insect-associated (bark beetles) 10,52 273, HiSeq 2500 Scheffersomyces lignosus Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Debaryomycetaceae JCM 9837 Gp BCGS fermentation (xylose) 16,59 171, ,2 HiSeq 2500 Scheffersomyces stipitis Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Debaryomycetaceae CBS 6054 Gp AAVQ fermentation 15, Spathaspora arborariae Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Debaryomycetaceae UFMG-19.1A Gp AYLH industrial importance 12,87 23, ,9 454 Spathaspora passalidarum Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Debaryomycetaceae NRRL Y Gp AEIK industrial importance (biofuel production-xylose fermentation) 13, ; Sanger Wickerhamia fluorescens Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Debaryomycetaceae JCM 1821 Gp BCGE saprotroph 13,18 239, ,3 HiSeq 2500 Geotrichum candidum Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Dipodascaceae 3C Gp JMRO plant pathogen (citrus) 41,38 31, , Saprochaete clavata Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Dipodascaceae CNRMA Gp CBXB human pathogen 17, ,8 Sporopachydermia quercuum Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Dipodascaceae JCM 9486 Gp BCGN endophyte 16,41 182, ,2 Illumina HiSeq Yarrowia deformans Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Dipodascaceae JCM 1694 Gp BCIW industrial importance (Alkane-using) 20,88 131, HiSeq 2500 Yarrowia keelungensis Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Dipodascaceae JCM Gp BCJD Industrial importance; oil-degrading 21,82 203, HiSeq 2500 Yarrowia lipolytica Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Dipodascaceae CLIB122 Gp GCA_ industrial importance (Alkane-using) 20,5 10, Sanger Yarrowia sp. JCM Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Dipodascaceae JCM Gp BCKF uncertain 21,76 217, HiSeq 2500 Yarrowia sp. JCM Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Dipodascaceae JCM Gp BCLX uncertain 21,9 251, ,8 Illumina Yarrowia sp. JCM Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Dipodascaceae JCM Gp BCKG uncertain 21,75 230, ,9 HiSeq 2500 Ashbya aceri Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Eremotheciaceae FD-2008 Gp GCA_ insect-associated 8, Sanger Eremothecium (Ashbya) gossypii Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Eremotheciaceae ATCC Gp GCA_ plant pathogen (insect-associated; cotton) 9,1 4, Sanger Eremothecium coryli Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Eremotheciaceae CBS 5749 Gp AZAH plant pathogen (soybean) 9,09 18, ,6 Illumina HiSeq Eremothecium cymbalariae Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Eremotheciaceae DBVPG#7215 Gp GCA_ plant pathogen (stigmatomycosis; fruit rot; crops) 9,66 40, ,3 Ambrosiozyma kashinagacola Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Incertae sedis JCM Gp BCGA insect-associated 12,32 227, ,1 HiSeq 2500 Ambrosiozyma monospora Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Incertae sedis JCM 7599 Gp BCIP industrial importance; ethanol production 24,98 116, ,2 HiSeq 2500 Candida albicans Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Incertae sedis SC5314 Gp AACQ opportunistic human pathogen 27, ,4 Candida apicola Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Incertae sedis NRRL Y Gp LBNK industrial importance (sophorolipid producing) 9,77 211, ,2 Illumina GAIIx Candida arabinofermentans Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Incertae sedis NRRL YB-2248 Gp JGI industrial importance 13,23 74, ; Illumina Candida auris Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Incertae sedis 6684 Gp LGST human pathogen (multridrug healthcare infections) 12,5 179, ,1 Illumina HiSeq; MiSeq Candida boidinii Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Incertae sedis GF002 Gp LMZO industrial importance (methylotrophic) 19,11 144, ,4 Illumina HiSeq Candida bracarensis Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Incertae sedis CBS Gp CAPU opportunistic human pathogen 12,2 51 Candida carpophila Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Incertae sedis JCM 9396 Gp BCGK probable human pathogen 10,24 273, ,4 HiSeq 2500 Candida caseinolytica Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Incertae sedis NRRL Y Gp JGI plant pathogen (possible; associated with rot in cacti) 9,18 47, ; Illumina Candida castellii Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Incertae sedis CBS 4332 Gp CAPW uncertain 10,2 101 Candida dubliniensis Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Incertae sedis CD36 Gp ASM2694v opportunistic human pathogen 14,62 11, Sanger Candida ethanolica Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Incertae sedis M2 Gp ANNA industrial importance 23,58 85, ,4 Illumina HiSeq Candida glabrata Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Incertae sedis CBS 138 Gp ASM254v human pathogen 12,3 8, Sanger Candida homilentoma Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Incertae sedis JCM 1507 Gp BCGB insect-associated (Bostrychid beetles) 12,18 223, ,4 HiSeq 2500 Candida intermedia Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Incertae sedis JCM 1607 Gp BCGD opportunistic human pathogen (candidiasis) 13,03 218, ,2 HiSeq 2500 Candida maltosa Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Incertae sedis Xu316 Gp AOGT industrial importance 12,8 130, ,2 Illumina HiSeq Candida nivariensis Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Incertae sedis CBS 9983 Gp CAPV human pathogen; drug-resistant 11, Candida orthopsilosis Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Incertae sedis AY2 Gp AMDC human pathogen 12,65 67, ,6 Illumina GAIIx Candida parapsilosis Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Incertae sedis CDC317 Gp CABE human pathogen (sepsis) 13,08 9, ,7 Sanger Candida sorboxylosa Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Incertae sedis JCM 1536 Gp BCGC industrial importance (fermentative yeast) 10,71 247, ,2 HiSeq 2500 Candida succiphila Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Incertae sedis JCM 9445 Gp BCGL industrial importance (methanol-assimilating) 12,15 234, ,9 HiSeq 2500 Candida tanzawaensis Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Incertae sedis NRRL Y Gp JGI insect-associated 13,14 74, ; Illumina Candida tenuis Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Incertae sedis ATCC Gp JGI industrial importance 10,7 26, ; Sanger Candida tropicalis Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Incertae sedis MYA-3404 Gp AAFN human pathogen (candidiasis); industrial importance (biodiesel production 14,6 10, Sanger Candida versatilis Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Incertae sedis JCM 5958 Gp BCJV industrial importance (salt-tolerant) 9,31 441, ,9 HiSeq 2500 Lodderomyces elongisporus Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Incertae sedis NRRL YB-4239 Gp AAPO possible human pathogen 15,547 8, Nadsonia fulvescens Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Incertae sedis DSM 6958 Gp JGI uncertain; yeast (bipolar budding) 13,75 72, ; Illumina Pachysolen tannophilus Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Incertae sedis NRRL Y-2460 Gp CAHV Fermentation; osmotolerant 12,6 90, ,8 454; Illumina; Sanger Starmerella bombicola Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Incertae sedis JCM 9596 Gp BCGO industrial importance (Sophorolipids, Glycolipid Biosurfactant production) 9,56 299, ,6 HiSeq 2500 Zygoascus hellenicus Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Incertae sedis Y-7136 v human pathogen (fungemia) 12,17 98, Illumina Lipomyces starkeyi Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Lipomycetaceae NRRL Y11557 Gp JGI metabolically interesting 21,27 47, ; Illumina; Sanger Clavispora lusitaniae Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Metschnikowiaceae ATCC Gp AAFT human pathogen 12,1 9, ,5 Sanger Metschnikowia bicuspidata Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Metschnikowiaceae Baker2002 Gp JGI parasite (brine shrimp); hypersaline 16,06 16, ; Illumina MiSeq Metschnikowia fructicola Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Metschnikowiaceae 277 Gp biocontrol (spoilage control) 24, , ,5 Illumina HiSeq Cyberlindnera fabianii Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Phaffomycetaceae JCM 3601 Gp BCGI industrial importance (waste water treatment; alcoholic fermentation) 12,3 223, ,2 HiSeq 2500 Cyberlindnera jadinii Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Phaffomycetaceae NBRC 0988 Gp BAEL industrial importance 14,27 20, ; Sanger Brettanomyces (Dekkera) bruxellensis Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Pichiaceae CBS 2499 Gp AHMD industrial importance 13,37 127, ,3 454; Illumina Brettanomyces anomalus Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Pichiaceae YV396 Gp LCTY industrial importance (fermentative yeast) 12,88 100, ,9 Illumina HiSeq Hyphopichia burtonii Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Pichiaceae NRRL Y-1933 Gp spoilage yeast 12,4 50, ; Illumina Nakazawaea peltata Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Pichiaceae JCM 9829 Gp BCGQ fermentation 11,65 239, ,5 HiSeq 2500 Ogataea methanolica Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Pichiaceae JCM Gp BCKW industrial importance (methylotrophic ) 15,1 248, ,2 HiSeq 2500 Ogataea polymorpha Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Pichiaceae BY4329 Gp BBNV industrial importance (methylotrophic ) 8,89 20, ,7 454 GS FLX; GAIIx Pichia kudriavzevii Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Pichiaceae NBRC1279 Gp BBOI spoilage yeast 10,18 45, ,5 454 Pichia membranifaciens Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Pichiaceae NRRL Y-2026 Gp JGI spoilage yeast; contaminant 11,58 38, Eremothecium sinecaudum Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Saccharomycetaceae ATCC Gp industrial importance (oxylipin synthesis) 8, ,13 Kazachstania africana Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Saccharomycetaceae CBS 2517 Gp GCA_ uncertain 11, ,4 Kazachstania naganishii Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Saccharomycetaceae CBS 8797 Gp GCA_ saprotroph (soil) 10, ,1 Kluyveromyces aestuarii Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Saccharomycetaceae ATCC Gp AEAS marine yeast 9,91 14, ,3 454 Kluyveromyces dobzhanskii Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Saccharomycetaceae CBS 2104 Gp CCBQ insect-associated (Drosophila) 10, ,3 PacBio RS II; Ion Torrent Kluyveromyces lactis Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Saccharomycetaceae NRRL Y-1140 Gp GCA_ industrial importance 10,729 11, Sanger Kluyveromyces marxianus Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Saccharomycetaceae DMKU Gp AKFM industrial importance (thermotolerant) 10, ,1 Illumina Kluyveromyces wickerhamii Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Saccharomycetaceae UCD Gp AEAV gut associate (Drosophila) 9,8 12, ,9 454 Kuraishia capsulata Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Saccharomycetaceae CBS 1993 Gp CBUD nitrate assimilating 11, ,6 Lachancea kluyveri Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Saccharomycetaceae NRRL Y Gp AACE uncertain; yeast (budding) 11,37 0, ,6 Sanger Lachancea lanzarotensis Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Saccharomycetaceae CBS Gp CDLU industrial importance (grape fermentation) 11, ,3 Illumina MiSeq Lachancea thermotolerans Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Saccharomycetaceae CBS 6340 Gp ASM industrial importance 10,392 12, Sanger Lachancea waltii Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Saccharomycetaceae NCYC 2644 Gp AADM uncertain; yeast (budding) 10,912 8, ,3 Sanger Nakaseomyces bacillisporus Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Saccharomycetaceae CBS 7720 Gp CAPX fermentation 10, Nakaseomyces delphensis Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Saccharomycetaceae CBS 2170 Gp CAPT fermentation 10, Naumovozyma castellii Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Saccharomycetaceae NRRL Y Gp AACF uncertain; yeast (budding) 11,242 3, ,86 Sanger Naumovozyma dairenensis Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Saccharomycetaceae CBS 421 Gp uncertain; yeast (budding) 13, Ogataea angusta Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Saccharomycetaceae ATCC MYA-335 Gp JGI industrial importance (thermotolerant) 8, ; Sanger Ogataea parapolymorpha Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Saccharomycetaceae DL-1 Gp AEOI industrial importance 8,874 49, ,9 454 Pichia pastoris (Komagataella phaffii) Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Saccharomycetaceae GS115 Gp ASM industrial importance (methylotroph); model organism 9,216 20, Saccharomyces arboricola Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Saccharomycetaceae H-6 Gp ALIE uncertain; yeast (budding); closely related to S. cerevisae 11,619 50, ,7 454; SOLiD Saccharomyces bayanus Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Saccharomycetaceae MCYC 623 Gp AACA fermentation (wine- and cider-making) 11,477 6, ,2 Sanger Saccharomyces boulardii Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Saccharomycetaceae EDRL Gp ATCS pharmaceutical importance (probiotic) 11,482 50, ,2 454 Saccharomyces cerevisiae Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Saccharomycetaceae S288c Gp GCA_ fermentation 12, Sanger Saccharomyces eubayanus Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Saccharomycetaceae CBS12357 Gp BBYU industrial importance (fermentation; brewing) 11,65 360, Illumina Saccharomyces kudriavzevii Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Saccharomycetaceae IFO 1802 Gp AACI uncertain; yeast (budding) 11,189 3, ,4 Sanger Saccharomyces mikatae Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Saccharomycetaceae IFO 1815 Gp AABZ uncertain; yeast (budding) 11,47 5, Sanger Saccharomyces paradoxus Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Saccharomycetaceae NRRL Y Gp AABY fermentation (wine-making) 11,872 7, ,7 Sanger Saccharomyces pastorianus Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Saccharomycetaceae CBS 1513 Gp AZCJ fermentation (lager beer) 19,367 18, ,3 454 Saccharomyces uvarum Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Saccharomycetaceae A9 Gp JNVO Fermentation (Bottom-fermenting wine yeast) 11,595 30, ,1 ion Torrent Tetrapisispora blattae Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Saccharomycetaceae CBS 6284 Gp GCA_ gut symbiont (cockroach ) 14, ,79 Tetrapisispora phaffii Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Saccharomycetaceae CBS 4417 Gp GCA_ biocontrol (spoilage control); killer yeast 12, Torulaspora delbrueckii Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Saccharomycetaceae CBS 1146 Gp GCA_ industrial importance (biotechnological interest) 9, Vanderwaltozyma polyspora Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Saccharomycetaceae DSM Gp AAZN uncertain; yeast; close relative of S. cerevisiae 14,674 7, Sanger Zygosaccharomyces bailii Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Saccharomycetaceae ISA1307 Gp CBTC spoilage yeast; acetic acid tolerant 12, ,5 Zygosaccharomyces rouxii Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Saccharomycetaceae CBS 732 Gp GCA_ food spoilage yeast (Halotolerant and osmotolerant) 9,764 11, Sanger

66 Hanseniaspora uvarum Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Saccharomycodaceae 34-9 Gp JPPO saprotroph 8,1 234, ,6 Illumina HiSeq Hanseniaspora valbyensis Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Saccharomycodaceae NRRL Y-1626 Gp JGI traditional use 11,46 53, ; Illumina Hanseniaspora vineae Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Saccharomycodaceae T02/19AF Gp JFAV industrial importance 11, , ,4 Illumina GAIIx Arthroascus fermentans Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Saccharomycopsidaceae v fermenting yeast 14,37 89, Illumina Saccharomycopsis malanga Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Saccharomycopsidaceae JCM 7620 Gp BCGJ uncertain 16,72 190, ,8 HiSeq 2500 Sugiyamaella (Candida) lignohabitaascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Trichomonascaceae CBS industrial importance (biorefinery) 15, ,9 Sugiyamaella americana Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Trichomonascaceae NRRL YB-2067 v insect-associated (beetles) 16,48 100, Illumina Sympodiomyces attinorum Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Trichomonascaceae NRRL Y v insect-associated (ants) 14,02 89, Illumina Trichomonascus petasosporus Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Trichomonascaceae NRRL YB-2093 Gp saprotrophic yeast 14,46 91, Illumina Wickerhamiella domercqiae Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Trichomonascaceae JCM 9478 Gp BCGM pharmaceutical importance (anticancer sophorolipid producing) 8,47 344, ,4 HiSeq 2500 Wickerhamomyces anomalus Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Wickerhamomycetaceae NRRL Y-366 Gp AEGI industrial importance (wine yeast); spoilage; biocontrol (mycotoxic compou 14,15 97, ,1 454; Illumina Wickerhamomyces ciferrii Ascomycota Saccharomycotina Saccharomycetes Saccharomycetidae Saccharomycetales Wickerhamomycetaceae NRRL Y-1031 Gp CAIF pharmaceutical importance (sphingolipid biosynthesis pathway) 15, ,4 Pneumocystis jirovecii (carinii) Ascomycota Taphrinomycotina Pneumocystidomycetes Pneumocystidomycetidae Pneumocystidales Pneumocystidaceae SE8 Gp CAKM parasite (obligate); colonizes lungs (pneumonia) 8,15 675, ,4 454; Illumina Pneumocystis murina Ascomycota Taphrinomycotina Pneumocystidomycetes Pneumocystidomycetidae Pneumocystidales Pneumocystidaceae B123 Gp AFWA parasite (obligate); colonizes lungs (pneumonia) 7, , ,9 454; Illumina Schizosaccharomyces cryophilus Ascomycota Taphrinomycotina Schizosaccharomycetes Schizosaccharomycetidae Schizosaccharomycetales Schizosaccharomycetaceae OY26 Gp ACQJ uncertain; yeast (fission) 11,56 51, ,7 454 Schizosaccharomyces japonicus Ascomycota Taphrinomycotina Schizosaccharomycetes Schizosaccharomycetidae Schizosaccharomycetales Schizosaccharomycetaceae yfs275 Gp AATM model organism; yeast (fission) 11,73 9, Sanger Schizosaccharomyces octosporus Ascomycota Taphrinomycotina Schizosaccharomycetes Schizosaccharomycetidae Schizosaccharomycetales Schizosaccharomycetaceae yfs286 Gp ABHY uncertain; yeast (fission) 11,63 39, ,5 Sanger Schizosaccharomyces pombe Ascomycota Taphrinomycotina Schizosaccharomycetes Schizosaccharomycetidae Schizosaccharomycetales Schizosaccharomycetaceae 972h- Gp GCA_ model organism 12, Saitoella complicata Ascomycota Taphrinomycotina Taphrinomycetes Taphrinomycetidae Taphrinales Protomycetaceae NRRL Y Gp BACD saprotroph 14,14 52, ,5 454; Illumina Taphrina deformans Ascomycota Taphrinomycotina Taphrinomycetes Taphrinomycetidae Taphrinales Taphrinaceae PYCC 5710 Gp CAHR plant pathogen (peach) 13, ,5 Taphrina flavorubra Ascomycota Taphrinomycotina Taphrinomycetes Taphrinomycetidae Taphrinales Taphrinaceae JCM Gp BAVW plant pathogen (Prunus fruit) 15,73 200, ,6 Illumina HiSeq Taphrina populina Ascomycota Taphrinomycotina Taphrinomycetes Taphrinomycetidae Taphrinales Taphrinaceae JCM Gp BAVX plant pathogen (cottonwood) , ,4 Illumina HiSeq Taphrina wiesneri Ascomycota Taphrinomycotina Taphrinomycetes Taphrinomycetidae Taphrinales Taphrinaceae JCM Gp BAVU plant pathogen (cherry trees) 13,1 200, ,1 Illumina HiSeq

67 Sequenced Basidiomycota species Classification Genome statistics SPECIES PHYLUM SUBPHYLUM CLASS SUBCLASS ORDER FAMILY Strain GOLD Accession GENOME Accession NCBI Project ID Significance Size (Mb) coverage # of contigs # of scaffolds # genes GC Technology Agaricus bisporus Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Agaricaceae JB137-s8 Gp JGI edible; saprotroph 30, ; Illumina Leucoagaricus gongylophorus Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Agaricaceae Ac12 Gp ANIS insect symbiont 101, ,1 454 Leucoagaricus sp. SymC.cos Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Agaricaceae SymC.cos Gp LSHD insect-associated (ants) 119, ,4 Illumina HiSeq Macrolepiota fuliginosa Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Agaricaceae MF-IS2 Gp JGI saprotroph 46,4 165, Illumina HiSeq Amanita brunnescens Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Amanitaceae BX004 Gp JNHV mycorrhizal 57, Illumina HiSeq Amanita inopinata Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Amanitaceae Kibby_2008Gp JNHW saprotroph 22, ,9 Illumina HiSeq Amanita jacksonii Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Amanitaceae TRTC168611Gp AYNK ectomycorrhizal 30, Amanita muscaria var. guessowbasidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Amanitaceae BX008 Gp JNHX ectomycorrhizal 67, ,5 Illumina HiSeq Amanita polypyramis Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Amanitaceae BW_CC Gp JNHY ectomycorrhizal 23, ,3 Amanita thiersii Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Amanitaceae SKay4041 Gp JGI saprotroph 33,69 135, Bolbitius vitellinus Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Bolbitiaceae SZMC-NL-19Go saprotroph 51, PacBio Clavaria fumosa Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Clavariaceae KM Gp CVRD saprotroph 70, ,7 Illumina HiSeq Cortinarius glaucopus Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Cortinariaceae AT Gp JGI ectomycorrhizal 63,45 107, Illumina HiSeq Hebeloma cylindrosporum Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Cortinariaceae h7 Gp JGI mycorrhizal 38, Illumina; PacBio; 454; Sanger Crepidotus sp. BD-2015 Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Crepidotaceae MCA163 Gp CVRE uncertain 70, ,3 IonTorrent Crepidotus variabilis Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Crepidotaceae CBS v saprotroph 38,58 111, Illumina Fistulina hepatica Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Fistulinaceae ATCC 64428Gp JYFI saprotroph; wood-decay 33,85 137, ,2 Illumina; PacBio Laccaria amethystina Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Hydnangiaceae LaAM-08-1 Gp JGI ectomycorrhizal 52,2 156, ; Illumina HiSeq Laccaria bicolor Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Hydnangiaceae S238N-H82 Gp JGI ectomycorrhizal 60,71 9, Sanger Hypsizygus marmoreus Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Lyophyllaceae Gp LUEZ saprotroph; edible 41,63 287, ,7 Illumina HiSeq; MiSeq Termitomyces sp. J132 Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Lyophyllaceae J132 Gp JDCH insect-cultivated fungus 67, ,3 Illumina HiSeq Marasmius fiardii Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Marasmiaceae PR-910 Gp uncertain 59,45 96, Illumina HiSeq Moniliophthora perniciosa Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Marasmiaceae FA553 Gp ABRE plant pathogen (cacao) 26, Sanger Moniliophthora roreri Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Marasmiaceae MCA 2997 Gp AWSO plant pathogen (pod rot of cacao) 52,204 15, ,9 454; Illumina Mycena chlorophos Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Mycenaceae Gp BAYG bioluminescent saprotroph 43, ,6 Illumina Mycena galopus Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Mycenaceae ATCC-62051Go saprtotroph 211,36 80, PacBio Panellus stipticus Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Mycenaceae LUM Gp saprotroph 53,17 112, Illumina Crucibulum laeve Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Nidulariaceae CBS v saprotroph 44,73 106, Illumina Cyathus striatus Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Nidulariaceae AH Go saptrotroph; bird's nest fungus 91, Illumina; PacBio Lentinula edodes Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Omphalotaceae L-54 Gp edible 40,2 11, ; SOLiD Omphalotus olearius Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Omphalotaceae VT Gp AHIW bioluminescent mushroom 28,15 85, ,4 Illumina Rhodocollybia butyracea Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Omphalotaceae AH v saprotroph 96, PacBio Armillaria gallica Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Physalacriaceae Ar21-2 Gp JGI saprotroph; pathogen (root rot; opportunistic tree p 85,34 41, Illumina HiSeq Armillaria ostoyae Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Physalacriaceae 28-4 v plant pathogen; conifer root rot (parasitic or saprop 58,01 67, PacBio Cylindrobasidium torrendii Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Physalacriaceae FP15055 ss-gp JNEL saproptroph 31,57 134, Illumina HiSeq Flammulina velutipes Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Physalacriaceae W23 Gp AQHU model organism 35,64 37, ,8 454 Guyanagaster necrorhiza Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Physalacriaceae MCA 3950 Gp saprotroph 53,69 81, Illumina HiSeq Oudemansiella mucida Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Physalacriaceae CBS Go saptrotroph; white-rot 61,73 131, PacBio Pleurotus eryngii Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Pleurotaceae ATCC v bioremediation; edible 44,61 96, Pleurotus ostreatus Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Pleurotaceae PC15 Gp AYUK saprotrophic (white rot) 34,342 8, ,9 Sanger Pluteus cervinus Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Pluteaceae NL-1719 v saprotroph 52,19 138, Illumina Volvariella volvacea Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Pluteaceae PYd21 Gp ANCH edible mushroom 36,626 90, ,8 Illumina GAII Coprinellus micaceus Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Psathyrellaceae FP vgp saprotroph 77,39 74, PacBio Coprinellus pellucidus Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Psathyrellaceae TEP2b coprophilous 41,56 167, Illumina Coprinopsis cinerea Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Psathyrellaceae okayama7# Gp AACS model organism 36, Sanger Coprinopsis marcescibilis Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Psathyrellaceae CBS Gp coprophilous 38,91 94, Illumina Pterula gracilis Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Pterulaceae CBS v developmentally interesting 34,77 88, Illumina Schizophyllum commune Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Schizophyllaceae H4-8 Gp ADMJ saprotroph; opportunistic human pathogen 38,67 8, ,5 Sanger Galerina marginata Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Strophariaceae CBS Gp AYUM saprotroph (white rot); toxic 59,42 34, ; Illumina Gymnopilus chrysopellus Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Strophariaceae PR-1187 Gp JGI saprotroph 47,22 142, Illumina HiSeq Gymnopilus junonius Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Strophariaceae AH Go saptrotroph; white-rot; hallucinogenic 59,46 101, Illumina; PacBio Hypholoma sublateritium Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Strophariaceae FD-334 SS-4Gp industrial importance 48, ; Illumina Baeospora myosura Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Tricholomataceae KM Gp CEMG saprotroph 61, ,5 Gymnopus androsaceus Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Tricholomataceae JB14 Gp JGI saprotrophic (white rot); important in carbon cycle 89, Illumina Gymnopus luxurians Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Tricholomataceae FD-317 M1 Gp JGI saprotroph 66,28 29, ; Illumina Lepista nuda Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Tricholomataceae CBS v saprotroph; edible 43,49 93, Illumina Macrocystidia cucumis Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Tricholomataceae KM Gp CEMH saprotroph 46, ,8 Tricholoma matsutake Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Agaricales Tricholomataceae 945 Gp ectomycorrhizal (edible) 175,76 100, Illumina HiSeq; PacBio Anomoporia bombycina Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Amylocorticales Amylocorticiaceae ATCC Go uncertain 44,48 126, Illumina Plicaturopsis crispa Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Amylocorticales Amylocorticiaceae FD-325 SS-3 Gp JGI saprotrophic (white rot) 34,5 33, ; Illumina Athelia rolfsii Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Atheliales Atheliaceae 1010 Gp JZWR plant pathogen (blight of vegetables) 59, ,4 IonTorrent; PGM Fibulorhizoctonia psychrophilabasidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Atheliales Atheliaceae CBS Gp psychrophile; spoilage 95,13 30, Illumina HiSeq Piloderma croceum Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Atheliales Atheliaceae F 1598 Gp ectomycorrhizal 59,33 102, Illumina Auricularia subglabra Basidiomycota Agaricomycotina Agaricomycetes Auriculariomycetidae Auriculariales Auriculariaceae SS-5 Gp JGI saprotrophic (white rot) 74,92 46, ,6 Sanger; 454; Illumina Exidia glandulosa Basidiomycota Agaricomycotina Agaricomycetes Auriculariomycetidae Auriculariales Auriculariaceae HHB12029 Gp JGI saproptroph 78,17 83, Illumina HiSeq Aporpium (Elmerina) caryae Basidiomycota Agaricomycotina Agaricomycetes Auriculariomycetidae Auriculariales Incertae sedis L Gp lignocellulotic cosmopolitan fungus 39,53 26, Illuminal; PacBio Boletus edulis Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Boletales Boletaceae BED1 Gp JGI mycorrhizal 46, Illumina Xerocomus badius Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Boletales Boletaceae v ectomycorrhizal 38,39 166, Illumina Phlebopus portentosus Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Boletales Boletinellaceae PP33 Gp JROP ectomycorrhizal; edible 30, Illumina HiSeq Coniophora puteana Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Boletales Coniophoraceae RWD Gp SS2 AEIT saprotroph (brown rot) 42,97 49, ,4 454; Illumina Leucogyrophana mollusca Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Boletales Hygrophoropsidaceae KUC A-06 Gp JGI saprotroph 35,19 110, Illumina HiSeq Gyrodon lividus Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Boletales Paxillaceae BX Gp JGI ectomycorrhizal 43,05 113, Illumina HiSeq Hydnomerulius pinastri Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Boletales Paxillaceae MD-312 Gp JGI saprotroph (brown rot) 38,28 41, ; Illumina HiSeq Paxillus ammoniavirescens Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Boletales Paxillaceae Pou09.2 v ectomycorrhizal 35,76 176, Illumina Paxillus involutus Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Boletales Paxillaceae ATCC Gp ectomycorrhizal 58,3 36, ; Illumina Paxillus rubicundulus Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Boletales Paxillaceae Ve08.2h10 Gp ectomycorrhizal 53,01 133, Illumina Rhizopogon salebrosus Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Boletales Rhizopogonaceae TDB-379 Gp mycorrhizal; model organism 82,29 120, Illumina HiSeq Rhizopogon vinicolor Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Boletales Rhizopogonaceae AM-OR Gp ectomycorrhizal 36,1 153, Illumina Pisolithus microcarpus Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Boletales Sclerodermataceae 441 Gp JGI mycorrhizal 53,03 87, Illumina HiSeq; 454; Sanger Pisolithus tinctorius Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Boletales Sclerodermataceae Marx 270 Gp JGI Mycorrhizal 71, Illumina Scleroderma citrinum Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Boletales Sclerodermataceae Foug A Gp ectomycorrhizal 56,14 80, Illumina Serpula lacrymans Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Boletales Serpulaceae S7.9 Gp AEQB saprotroph (brown rot) 42, Sanger Suillus americanus Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Boletales Suillaceae EM31 v mycorrhizal 50, Illumina Suillus brevipes Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Boletales Suillaceae Sb2 Gp JGI ectomycorrhizal 51,71 101, Illumina HiSeq Suillus decipiens Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Boletales Suillaceae EM49 v mycorrhizal; edible 62,78 91, Illumina Suillus granulatus Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Boletales Suillaceae EM37 v mycorrhizal 42,34 80, Illumina Suillus hirtellus Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Boletales Suillaceae EM16 v ectomycorrhizal 49,94 109, Illumina Suillus luteus Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Boletales Suillaceae UH-Slu-Lm8-n1 Gp JGI ectomycorrhizal 37,01 93, ; Illumina HiSeq Botryobasidium botryosum Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Cantharellales Botryobasidiaceae FD-172 SS1 Gp JGI saprotroph 46,67 31, ; Illumina Cantharellus anzutake Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Cantharellales Cantharellaceae C23 Gp ectomycorrhizal 120, Illumina HiSeq Rhizoctonia solani Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Cantharellales Ceratobasidiaceae AG-1 IB Gp CAOJ plant pathogen (broad host range) 47, Clavulina sp. PMI_390 Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Cantharellales Clavulinaceae PMI_390 Gp JGI ectomycorrhizal 41,64 131, Illumina HiSeq Hydnum rufescens Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Cantharellales Hydnaceae UP504 v ectomycorrhizal; edible 67,14 102, Illumina Tulasnella calospora Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Cantharellales Tulasnellaceae AL13/4D Gp JGI mycorrhizal 62, Illumina HiSeq

68 Dendrothele bispora Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Corticiales Corticiaceae CBS Gp JGI saproptroph 130, Illumina HiSeq Limonomyces culmigenus Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Corticiales Corticiaceae CBS plant pathogen (turfgrass) 36, ,8 Punctularia strigosozonata Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Corticiales Corticiaceae HHB Gp AEGM saprotrophic (white rot) 34,17 45, ,2 Sanger; 454; Illumina Sphaerobolus stellatus Basidiomycota Agaricomycotina Agaricomycetes Phallomycetidae Geastrales Geastraceae SS14 Gp saprotroph 176,37 57, Illumina Sclerogaster hysterangioides Basidiomycota Agaricomycotina Agaricomycetes Phallomycetidae Geastrales Sclerogastraceae SCL2.BST uncertain 124, Illumina; PacBio Gloeophyllum trabeum Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Gloeophyllales Gloeophyllaceae ATCC 11539Gp AFVP saprotroph (brown rot) 37,18 54, ,2 Sanger; 454; Illumina Heliocybe sulcata Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Gloeophyllales Gloeophyllaceae OMC1185 v saprotroph (brown rot) 31,95 82, Neolentinus lepideus Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Gloeophyllales Gloeophyllaceae HHB14362 sgp JGI saprotroph (brown rot) 35,64 92, Illumina HiSeq Gautieria morchelliformis Basidiomycota Agaricomycotina Agaricomycetes Phallomycetidae Gomphales Gomphaceae GMNE.BST v uncertain, hypogeal 122, PacBio Ramaria acris Basidiomycota Agaricomycotina Agaricomycetes Phallomycetidae Gomphales Gomphaceae UT TGp JGI ectomycorrhizal 105,46 78, Illumina Fomitiporia mediterranea Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Hymenochaetales Hymenochaetaceae MF3/22 Gp AEJJ saprotrophic (white rot) 63,35 39, ,8 Sanger; 454; Illumina Onnia scaura Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Hymenochaetales Hymenochaetaceae P-53A Gp saprotroph; wood-decomposinbg 37,1 88, Illumina HiSeq Phellinus noxius Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Hymenochaetales Hymenochaetaceae OVT-YTM/9Gp AYOR plant pathogen (broad host range, typically trees) 31,259 35, ,6 Ion Torrent Porodaedalea chrysoloma Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Hymenochaetales Hymenochaetaceae FP Go plant pathogen (conifer parasite; white pocket rot) 44,69 50, Illumina; PacBio Porodaedalea niemelaei Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Hymenochaetales Hymenochaetaceae PN IPGo plant pathogen (conifer white rot) 53, PacBio Sanghuangporus baumii Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Hymenochaetales Hymenochaetaceae 821 Gp LNZH pharmaceutical importance (medicinal mushroom) 31,64 186, ,6 Illumina Rickenella fibula Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Hymenochaetales Rickenellaceae HBK v saprotroph, probable 59,35 91, PacBio Schizopora paradoxa Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Hymenochaetales Schizoporaceae KUC8140 Gp JGI wood-inhabiting 44, Illumina HiSeq Jaapia argillacea Basidiomycota Agaricomycotina Agaricomycetes Agaricomycetidae Jaapiales Jaapiaceae MUCL 3360Gp AYUL saprotroph 45,05 20, ,8 454; Illumina Mutinus elegans Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Phallales Phallaceae ME.BST v saprotroph 52,78 85, Illumina Taiwanofungus camphoratus Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Polyporales Coriolaceae S27 Gp JNBV pharmaceutical importance (traditional medicine) 32,16 878, ,6 Illumina GAIIx; 454 Trametes cinnabarina Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Polyporales Coriolaceae DSM 3022 Gp JSYY saprotrophic (white rot) 31,62 180, ,6 Illumina HiSeq Trametes hirsuta Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Polyporales Coriolaceae 072 Gp LIYB saprotrophic (white rot) 33,62 101, ,8 Illumina HiSeq; MiSeq Trametes sp. AH28-2 Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Polyporales Coriolaceae AH28-2 Gp LJJJ saprotrophic (white rot); industrial importance (lacc 38,9 100, ,8 Illumina HiSeq Antrodia sinuosa Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Polyporales Fomitopsidaceae LB1 Gp JGI saprotroph 30,17 128, Illumina HiSeq Daedalea quercina Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Polyporales Fomitopsidaceae L Gp JGI saproptroph 32,74 144, Illumina HiSeq Fomitopsis pinicola Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Polyporales Fomitopsidaceae FP SSGp AEHC saprotroph (brown rot) 41,61 85, ,8 Illumina; PacBio Laetiporus sulphureus Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Polyporales Fomitopsidaceae Gp JGI saprotroph 39,92 85, Illumina HiSeq Postia placenta Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Polyporales Fomitopsidaceae MAD 698 Gp ABWF saprotroph (brown rot) 90,9 7, Sanger Ganoderma lucidum Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Polyporales Ganodermataceae BCRC 37180Gp AGAX traditional use 43, ,1 454; Illumina; Sanger Ganoderma sp SS1 Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Polyporales Ganodermataceae SS1 Gp JGI saprotroph 39,52 33, Sanger; 454; Illumina Trametopsis cervina Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Polyporales Hapalopilaceae CIRM-BRFM 1824 v biocontrol (mycoparasite) 29, Illumina Hydnopolyporus fimbriatus Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Polyporales Meripilaceae CBS Go saprotroph 34,57 92, Illumina Bjerkandera adusta Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Polyporales Meruliaceae HHB-12826Gp JGI saprotrophic (white rot) 42,73 47, ; Illumina Ceriporiopsis (Gelatoporia) subbasidiomycota Agaricomycotina Agaricomycetes Incertae sedis Polyporales Meruliaceae B Gp JGI saprotrophic (white rot) 39 56, ; Sanger Obba rivulosa Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Polyporales Meruliaceae 3A-2 Gp saprotrophic (white rot) 34, Illumina Phlebia brevispora Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Polyporales Meruliaceae HHB-7030 SGp JGI saprotrophic (white rot) 49,96 42, ; Illumina not publically available Phanerochaete carnosa Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Polyporales Phanerochaetaceae HHB Gp AEHB saprotrophic (white rot) 46,29 58, ,1 Sanger; 454; Illumina Phanerochaete chrysosporiumbasidiomycota Agaricomycotina Agaricomycetes Incertae sedis Polyporales Phanerochaetaceae RP-78 Gp AADS saprotrophic (white rot) 35,15 10, Sanger Phlebiopsis gigantea Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Polyporales Phanerochaetaceae 11061_1 CRGp JGI saprotrophic (white rot) 30,14 144, Illumina Cerrena unicolor Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Polyporales Polyporaceae 303 Gp JGI saprotrophic (white rot) , Illumina Dichomitus squalens Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Polyporales Polyporaceae LYAD-421 SSGp AEID saproptroph 42,75 50, ,6 454; Illumina Fibroporia radiculosa Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Polyporales Polyporaceae TFFH 294 Gp CAGC saprotroph (brown rot) 28, ,2 Illumina GAII Leiotrametes sp. BRFM 1775 Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Polyporales Polyporaceae BRFM 1775Gp lignocellulolytic 34,52 142, Illumina Lentinus polychrous Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Polyporales Polyporaceae BCC7694 Gp JSYW saprotroph; edible 38,05 180, Lentinus tigrinus Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Polyporales Polyporaceae ALCF2SS1-7Gp saprotroph 39,76 103, Illumina HiSeq Lignosus rhinocerotis Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Polyporales Polyporaceae TM02 Gp AXZM pharmaceutical importance (traditional medicine) 34, ,7 Illumina HiSeq Polyporus arcularius Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Polyporales Polyporaceae HHB13444 Gp JGI saprotrophic (white rot) 43,45 118, Illumina HiSeq Polyporus brumalis Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Polyporales Polyporaceae BRFM 1820 v saprotroph 45,72 94, Illumina Pycnoporus cinnabarinus Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Polyporales Polyporaceae BRFM 137 CCBP saprotrophic (white rot) 33, ,6 Pycnoporus coccineus Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Polyporales Polyporaceae CIRM1662 Gp JGI saprotrophic (white rot) 32,1 128, Illumina HiSeq Pycnoporus sanguineus Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Polyporales Polyporaceae BRFM 1264 JGI saprotrophic (white rot) 36, Trametes cingulata Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Polyporales Polyporaceae BRFM 1805 v saprotrophic (white rot) , Illumina Trametes elegans Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Polyporales Polyporaceae CIRM1663 Gp JGI saprotrophic (white rot) 33,08 86, Illumina Trametes ljubarskyi Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Polyporales Polyporaceae CIRM1659 Gp JGI saprotrophic (white rot) 34 86, Illumina HiSeq Trametes versicolor Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Polyporales Polyporaceae FP SGp AEJI saprotrophic (white rot) 44, ,7 454 Trichaptum abietinum Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Polyporales Polyporaceae L ss-1gp JGI saprotrophic (white rot) 40,61 125, Illumina HiSeq Wolfiporia cocos Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Polyporales Polyporaceae MD-104 SS10Gp AEHD saprotroph (brown rot); traditional use 50,48 40, Sanger; 454; Illumina Xenasmatella vaga Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Polyporales Xenasmataceae CBS Go saprotroph 64,17 91, Illumina Artomyces pyxidatus Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Russulales Auriscalpiaceae HHB10654 Gp JGI saprotroph 37,99 90, Illumina HiSeq Auriscalpium vulgare Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Russulales Auriscalpiaceae FP SpGp saprotroph 39,65 122, Illumina HiSeq Lentinellus vulpinus Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Russulales Auriscalpiaceae AHS73672-sp v saprotroph (white rot) 34,71 94, Heterobasidion annosum Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Russulales Bondarzewiaceae Gp AOSL plant pathogen (conifers) 31,009 66, ,5 Illumina Heterobasidion irregulare Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Russulales Bondarzewiaceae TC 32-1 Gp AEOJ plant pathogen (conifers and hardwoods) 33,649 8, ,2 Sanger Dentipellis sp. Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Russulales Hericiaceae KUC8613 v bioremediation; white rot 36,71 88, Illumina Scytinostroma sp. Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Russulales Lachnocladiaceae KUC9335 Go uncertain 60,64 N/A PacBio Vararia minispora Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Russulales Lachnocladiaceae EC-137 Gp saprotroph 36,81 149, Illumina HiSeq Peniophora aff. cinerea. Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Russulales Peniophoraceae CBS Gp JGI saprotroph 46,03 141, Illumina HiSeq Peniophora sp. Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Russulales Peniophoraceae CONTA v uncertain 48,44 85, Illumina Lactarius quietus Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Russulales Russulaceae SC ectomycorrhizal 115,9 97, PacBio Stereum hirsutum Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Russulales Stereaceae FP SS1Gp AEGX saprotrophic (white rot) 46,51 44, ,3 Sanger; 454; Illumina Piriformospora indica Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Sebacinales Sebacinaceae DSM Gp CAFZ endophyte 24,98 22, ,7 454 Sebacina vermifera Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Sebacinales Sebacinaceae MAFF Gp JGI mycorrhizal (orchid) 38,09 117, ; Illumina Thelephora ganbajun Basidiomycota Agaricomycotina Agaricomycetes Incertae sedis Thelephorales Thelephoraceae P2 v ectomycorrhizal ; edible 36,34 176, Illumina Sistotremastrum niveocremeumbasidiomycota Agaricomycotina Agaricomycetes Incertae sedis Trechisporales Hydnodontaceae HHB9708 ss-1 Gp JGI saprotroph (brown rot) 33,86 143, Illumina Cantharellus anzutake Basidiomycota Agaricomycotina Dacrymycetes Incertae sedis Dacrymycetales Cerinomycetaceae C mycorrhizal;edible 120, PacBio Cerinomyces ceraceus Basidiomycota Agaricomycotina Dacrymycetes Incertae sedis Dacrymycetales Cerinomycetaceae ATCC v saprotroph (brown rot) 33,2 91, Illumina Calocera cornea Basidiomycota Agaricomycotina Dacrymycetes Incertae sedis Dacrymycetales Dacrymycetaceae HHB12733 Gp JGI saprotroph (brown rot) 33,24 85, Illumina HiSeq Calocera viscosa Basidiomycota Agaricomycotina Dacrymycetes Incertae sedis Dacrymycetales Dacrymycetaceae TUFC12733Gp JGI saprotrophic (white rot) 29,1 81, Illumina HiSeq Dacryopinax sp. DJM-731 SS1 Basidiomycotates Agaricomycotina Dacrymycetes Incertae sedis Dacrymycetales Dacrymycetaceae DJM-731 SS1 Gp AEUS saprotroph (brown rot) 29,5 43, ,2 Sanger; 454; Illumina Mrakia blollopis Basidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Cystofilobasidiales Mrakiaceae SK-4 Gp BBPU cryophilic 30, ,7 PacBio Mrakia frigida Basidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Cystofilobasidiales Mrakiaceae JCM 7857 Gp BCJW biocontrol (toxibn production ); psychrotolerant 28, HiSeq 2500 Phaffia (Xanthophyllomyces) dendrorhous Basidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Cystofilobasidiales Mrakiaceae JCM 9681 Gp BCJZ industrial importance (carotenoid) 19, ,3 HiSeq 2500 Phaffia rhodozyma Basidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Cystofilobasidiales Mrakiaceae CBS 7918 Gp LSVH industrial importance (carotenoid biosynthesis) 18,76 10, ,2 Illumina GAIIx Filobasidium wieringae Basidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Filobasidiales Filobasidiaceae JCM Gp BCIV saprotroph, probable 19,78 277, ,9 HiSeq 2500 Naganishia (Cryptococcus) vishniacii Basidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Filobasidiales Filobasidiaceae ANT Gp JGI psychrophile 19,69 146, Illumina HiSeq Naganishia albida (Cryptococcus Basidiomycota albidus) Agaricomycotina Tremellomycetes Tremellomycetidae Filobasidiales Filobasidiaceae NRRL Y-1402Gp LKPZ opportunistic human pathogen 24,81 43, ,8 Solicoccozyma (Cryptococcus) Basidiomycota phenolicus Agaricomycotina Tremellomycetes Tremellomycetidae Filobasidiales Piskurozymaceae JCM Gp BCIS saprotroph 22,26 278, ,9 HiSeq 2500 Solicoccozyma (Cryptococcus) Basidiomycota terricola Agaricomycotina Tremellomycetes Tremellomycetidae Filobasidiales Piskurozymaceae JCM Gp BCLD industrial importance (oleaginous) 24, ,4 HiSeq 2500 Holtermannia corniformis Basidiomycota Agaricomycotina Tremellomycetes Incertae sedis Holtermanniales Incertae sedis JCM 1743 Gp BCIJ uncertain 20, ,2 HiSeq 2500 Holtermanniella nyarrowii Basidiomycota Agaricomycotina Tremellomycetes Incertae sedis Holtermanniales Incertae sedis JCM Gp BCIT psychrophile 17,74 309, ,7 HiSeq 2500 Bulleromyces albus Basidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Tremellales Bulleraceae JCM 2954 Gp BCIX uncertain 19,42 271, ,3 HiSeq 2500 Sirobasidium intermedium Basidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Tremellales Bulleraceae CBS7805 v saprotroph 21,96 92, Illumina Dioszegia aurantiaca Basidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Tremellales Bulleribasidiaceae JCM 2956 Gp BCKN uncertain; plant-associated 19,34 188, ,6 Illumina Dioszegia crocea Basidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Tremellales Bulleribasidiaceae JCM 2961 Gp BCKK uncertain; plant-associated 20, ,2 Illumina Dioszegia cryoxerica Basidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Tremellales Bulleribasidiaceae ANT Gp JGI psychrophile 39,52 98, Illumina HiSeq Cryptococcus amylolentus Basidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Tremellales Cryptococcaceae CBS 6039 Gp AWGJ saprotroph 20,25 117, ,4 Illumina

69 Cryptococcus depauperatus Basidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Tremellales Cryptococcaceae CBS 7841 Gp AWGK parasite (mycoparasite on Lecanicillium lecanii) 15, ,7 Illumina Cryptococcus gattii Basidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Tremellales Cryptococcaceae R265 Gp AAFP human pathogen 17, ,87 Sanger Cryptococcus neoformans Basidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Tremellales Cryptococcaceae JEC 21 Gp GCA_ human pathogen 19,05 12, Sanger Kwoniella (Cryptococcus) best Basidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Tremellales Cryptococcaceae CBS Gp ASCK saprotroph 24, ,3 Illumina Kwoniella (Cryptococcus) deje Basidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Tremellales Cryptococcaceae CBS Gp ASCJ saprotroph 23, ,4 Illumina Kwoniella heveanensis Basidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Tremellales Cryptococcaceae BCC8398 Gp ASQB saprotroph 25, ,9 Illumina Kwoniella mangrovensis Basidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Tremellales Cryptococcaceae CBS 8507 Gp ASQE saprotroph (marine) 22, ,9 Illumina Kwoniella pini (Cryptococcus pbasidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Tremellales Cryptococcaceae CBS Gp ASCL saprotroph 20, ,2 Illumina Fellomyces penicillatus Basidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Tremellales Cuniculitremaceae Phaff54-35 v uncertain 21,05 97, Illumina Kockovaella imperatae Basidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Tremellales Cuniculitremaceae NRRL Y-179Go uncertain; yeast 17,47 428, PacBio Phaeotremella (Cryptococcus) Basidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Tremellales Phaeotremellaceae JCM 9039 Gp BCHT saprotroph 20, ,2 HiSeq 2500 Papiliotrema (Cryptococcus) flbasidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Tremellales Rhynchogastremataceae NRRL Y-503Gp CAUG biocontrol (of fusarium head blight) 22, ,5 Papiliotrema (Cryptococcus) labasidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Tremellales Rhynchogastremataceae RY1 Gp JDSR occasional human pathogen 19, ,1 Illumina MiSeq Fibulobasidium inconspicuum Basidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Tremellales Sirobasidiaceae Phaff89-39 v saprotroph, probable 20,29 91, Illumina Tremella fuciformis Basidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Tremellales Tremellaceae Tr26 Gp LBGW biocontrol (mycoparasite) 23, Illumina HiSeq Tremella mesenterica Basidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Tremellales Tremellaceae DSM 1558 Gp AFVY industrial importance; white rot 28,6 7, ,7 Sanger Phaeotremella (Cryptococcus) Basidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Trichosporonales Phaeotremellaceae JCM Gp BCHU saprotroph 22, ,4 Illumina Apiotrichum (Trichosporon) brbasidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Trichosporonales Trichosporonaceae JCM 1599 Gp BCJI saprotrophic (cabbage) 23, ,5 HiSeq 2500 Apiotrichum (Trichosporon) dobasidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Trichosporonales Trichosporonaceae JCM 9580 Gp BCFW human allergen 24, Illumina Apiotrichum (Trichosporon) gabasidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Trichosporonales Trichosporonaceae JCM 9941 Gp BCJN saprotrophic (cabbage) 24, HiSeq 2500 Apiotrichum (Trichosporon) grbasidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Trichosporonales Trichosporonaceae JCM Gp BCJO saprotrophic (soil) 24, HiSeq 2500 Apiotrichum (Trichosporon) labasidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Trichosporonales Trichosporonaceae JCM 2947 Gp BCKV soil or animal-associated 30, ,6 Illumina Apiotrichum (Trichosporon) mbasidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Trichosporonales Trichosporonaceae JCM 9937 Gp BCFV opportunistic human pathogen 24, HiSeq 2500 Apiotrichum (Trichosporon) vebasidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Trichosporonales Trichosporonaceae JCM Gp BCKJ alkane-assimilating 31, ,6 Illumina Cryptococcus sp. JCM Basidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Trichosporonales Trichosporonaceae JCM Gp BCLC industrial importance (oleaginous) 28, ,5 HiSeq 2500 Cutaneotrichosporon (CryptocBasidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Trichosporonales Trichosporonaceae JCM 1532 Gp BCJH industrial importance (oleaginous) 18, ,9 HiSeq 2500 Trichosporon asahii Basidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Trichosporonales Trichosporonaceae CBS 8904 Gp AMBO human pathogen 25,299 12, ,5 454; Illumina Trichosporon chiarellii Basidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Trichosporonales Trichosporonaceae MYA-4694 v insect-associated (ants) 24,81 86, Illumina Trichosporon cutaneum Basidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Trichosporonales Trichosporonaceae B3 Gp LRUG industrial importance (lipid production) 38, ,6 Illumina Trichosporon guehoae Basidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Trichosporonales Trichosporonaceae JCM Gp BCJX saprotrophic (soil) 33, ,2 HiSeq 2500 Trichosporon oleaginosus Basidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Trichosporonales Trichosporonaceae IBC0246 Gp JGI industrial importance (lipid producer); biotechnolog 19,84 115, Illumina HiSeq Trichosporon porosum Basidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Trichosporonales Trichosporonaceae JCM 1458 Gp BCJG saprotrophic (soil) 25, HiSeq 2500 Vanrija humicola Basidiomycota Agaricomycotina Tremellomycetes Tremellomycetidae Trichosporonales Trichosporonaceae JCM 1457 Gp BCJF saprotroph 22, ,7 HiSeq 2500 Basidioascus undulatus Basidiomycota Agaricomycotina Wallemiomycetes Incertae sedis Geminibasidiales Geminibasidiaceae DAOM 2419Gp JTLS xerotolerant 32, Wallemia ichthyophaga Basidiomycota Agaricomycotina Wallemiomycetes Incertae sedis Wallemiales Wallemiaceae EXF-994 Gp APLC halophile 9, ,3 Illumina HiSeq Wallemia sebi Basidiomycota Agaricomycotina Wallemiomycetes Incertae sedis Wallemiales Wallemiaceae CBS Gp AFQX xerophile 9, ; Illumina Agaricostilbum hyphaenes Basidiomycota Pucciniomycotina Agaricostilbomycetes Agaricostilbomycetidae Agaricostilbales Agaricostilbaceae ATCC MYA-Gp JGI uncertain (associated with palms) 17,89 197, Illumina HiSeq Chionosphaera apobasidialis Basidiomycota Pucciniomycotina Agaricostilbomycetes Agaricostilbomycetidae Agaricostilbales Chionosphaeraceae v parasite (lichens) 21,83 94, Illumina Chionosphaera cuniculicola Basidiomycota Pucciniomycotina Agaricostilbomycetes Agaricostilbomycetidae Agaricostilbales Chionosphaeraceae CBS10063 v insect-associated (conifer-inhabiting bark beetles) 14,16 89, Illumina Cystobasidiopsis (SporobolomBasidiomycota Pucciniomycotina Agaricostilbomycetes Agaricostilbomycetidae Agaricostilbales Chionosphaeraceae JCM 7595 Gp BCIO uncertain 22,18 241, ,2 Illumina Atractiellales sp. 95 Basidiomycota Pucciniomycotina Atractiellomycetes Incertae sedis Atractiellales unknown PMI 95 Gp JGI endophyte (root) 51,47 66, Illumina HiSeq Classicula fluitans Basidiomycota Pucciniomycotina Classiculomycetes Incertae sedis Classiculales Classiculaceae ATCC 64713Gp JGI aquatic 52,67 82, Illumina HiSeq Cystobasidium pallidum Basidiomycota Pucciniomycotina Cystobasidiomycetes Incertae sedis Cystobasidiales Cystobasidiaceae JCM 3780 Gp BCIL uncertain 21, ,7 HiSeq 2500 Erythrobasidium hasegawianubasidiomycota Pucciniomycotina Cystobasidiomycetes Incertae sedis Erythrobasidiales Erythrobasidiaceae ATCC 9536 (Gp uncertain 40,69 155, Illumina HiSeq Erythrobasidium yunnanense Basidiomycota Pucciniomycotina Cystobasidiomycetes Incertae sedis Erythrobasidiales Erythrobasidiaceae JCM Gp BCJC uncertain 20, HiSeq 2500 Naohidea sebacea Basidiomycota Pucciniomycotina Cystobasidiomycetes Incertae sedis Naohideales Naohideaceae CBS 8477 (PGp biocontrol (mycoparasite) 20,43 87, Illumina HiSeq Heterogastridium pycnidioideubasidiomycota Pucciniomycotina Microbotryomycetes Incertae sedis Heterogastridiales Heterogastridiaceae ATCC MYA-Gp parasite 17,58 109, Illumina; PacBio; 454; Sanger Hyalopycnis blepharistoma Basidiomycota Pucciniomycotina Microbotryomycetes Incertae sedis Heterogastridiales Heterogastridiaceae ATCC v saprotroph, probable 17,93 102, Illumina Meredithblackwellia eburnea Basidiomycota Pucciniomycotina Microbotryomycetes Incertae sedis Kriegeriales Kriegeriaceae MCA 4105 vgp uncertain; yeast (budding) 30,68 112, Leucosporidiella creatinivora Basidiomycota Pucciniomycotina Microbotryomycetes Incertae sedis Leucosporidiales Leucosporidiaceae JCM Gp BCKL aquatic saprotroph 27,97 190, ,7 Illumina Microbotryum lychnidis-dioicabasidiomycota Pucciniomycotina Microbotryomycetes Incertae sedis Microbotryales Microbotryaceae p1a1 LamolGp CDLV plant pathogen (anther smut of campion flowers) 3,47 3, ,5 Microbotryum violaceum Basidiomycota Pucciniomycotina Microbotryomycetes Incertae sedis Microbotryales Microbotryaceae p1a1 LamolGp AEIJ plant pathogen (anther smut of Caryophyllaceae); s 26, ,4 454 Rhodosporidium toruloides Basidiomycota Pucciniomycotina Microbotryomycetes Incertae sedis Sporidiobolales Sporidiobolaceae MTCC 457 Gp AJMJ industrial importance (lipid producer) 20,06 121, Illumina GAIIx Rhodotorula glutinis Basidiomycota Pucciniomycotina Microbotryomycetes Incertae sedis Sporidiobolales Sporidiobolaceae ATCC 20409Gp AEVR industrial importance (lipid producer) 20, ,9 Sanger; 454; Illumina Rhodotorula graminis Basidiomycota Pucciniomycotina Microbotryomycetes Incertae sedis Sporidiobolales Sporidiobolaceae WP1 Gp JGI endophyte 21,01 8, ; Sanger Rhodotorula minuta Basidiomycota Pucciniomycotina Microbotryomycetes Incertae sedis Sporidiobolales Sporidiobolaceae MCA 4210 Gp JGI occasional human pathogen 20,82 94, Illumina HiSeq Rhodotorula mucilaginosa Basidiomycota Pucciniomycotina Microbotryomycetes Incertae sedis Sporidiobolales Sporidiobolaceae C2.5t1 Gp JWTJ human pathogen (fungemia) 19, ,5 Rhodotorula sp. JG-1b Basidiomycota Pucciniomycotina Microbotryomycetes Incertae sedis Sporidiobolales Sporidiobolaceae JG-1b Gp LQXB Opportunistic Pathogen 19,4 133, ,6 Illumina HiSeq Rhodotorula toruloides Basidiomycota Pucciniomycotina Microbotryomycetes Incertae sedis Sporidiobolales Sporidiobolaceae JCM Gp BCIY industrial importance (oleaginous) 20, HiSeq 2500 Sporidiobolus salmonicolor Basidiomycota Pucciniomycotina Microbotryomycetes Incertae sedis Sporidiobolales Sporidiobolaceae SPOSA68321S011 Gp CENE opportunistic human pathogen 50, ,3 Illumina HiSeq Sporobolomyces linderae Basidiomycota Pucciniomycotina Microbotryomycetes Incertae sedis Sporidiobolales Sporidiobolaceae CBS 7893 (P93) Gp JGI uncertain; only known strain in existence 18,24 93, Illumina HiSeq Sporobolomyces roseus Basidiomycota Pucciniomycotina Microbotryomycetes Incertae sedis Sporidiobolales Sporidiobolaceae IAM Gp JGI free-living; non-pathogenic 21,2 8, Illumina Mixia osmundae Basidiomycota Pucciniomycotina Mixiomycetes Incertae sedis Mixiales Mixiaceae IAM Gp JGI plant pathogen (fern parasite) 13,63 150, ,5 454; Illumina Cronartium comandrae Basidiomycota Pucciniomycotina Pucciniomycetes Incertae sedis Pucciniales Cronartiaceae C4 Gp AUZW plant pathogen; rust fungus (pine) 68, ,7 Illumina HiSeq Cronartium quercuum Basidiomycota Pucciniomycotina Pucciniomycetes Incertae sedis Pucciniales Cronartiaceae G11 Gp JGI plant pathogen; rust fungus (fusiform rust of pine) 76,57 70, Illumina Cronartium ribicola Basidiomycota Pucciniomycotina Pucciniomycetes Incertae sedis Pucciniales Cronartiaceae 11-2 Gp AWVX plant pathogen; rust fungus (pine) 94, ,3 Illumina HiSeq Endocronartium harknessii Basidiomycota Pucciniomycotina Pucciniomycetes Incertae sedis Pucciniales Cronartiaceae PhW48OC Gp AXDO plant pathogen; rust fungus (pine-pine gall rust) 56, ,9 Illumina HiSeq Melampsora allii-populina Basidiomycota Pucciniomycotina Pucciniomycetes Incertae sedis Pucciniales Melampsoraceae 12AY07 v1.0gp plant pathogen; rust fungus (poplar) 335, PacBio Melampsora larici-populina Basidiomycota Pucciniomycotina Pucciniomycetes Incertae sedis Pucciniales Melampsoraceae 98AG31_300Gp AECX plant pathogen (poplar); rust fungus 101,1 6, Sanger Melampsora pinitorqua Basidiomycota Pucciniomycotina Pucciniomycetes Incertae sedis Pucciniales Melampsoraceae Mpini7 Gp AUYS plant pathogen (pine twisting rust); rust fungus 34, , ,00% 44,9 Illumina HiSeq Puccinia arachidis Basidiomycota Pucciniomycotina Pucciniomycetes Incertae sedis Pucciniales Pucciniaceae MRf11 Gp LBLN plant pathogen (peanut) 87, ,4 IonTorrent not publically available Puccinia graminis Basidiomycota Pucciniomycotina Pucciniomycetes Incertae sedis Pucciniales Pucciniaceae CRL Gp AAWC plant pathogen; rust fungus (cereals) 88,724 7, ,3 Sanger Puccinia psidii Basidiomycota Pucciniomycotina Pucciniomycetes Incertae sedis Pucciniales Pucciniaceae Aus_3 Gp LKHF plant pathogen; rust fungus (guava) 129, ,6 Illumina HiSeq Puccinia sorghi Basidiomycota Pucciniomycotina Pucciniomycetes Incertae sedis Pucciniales Pucciniaceae Rojas Gp LAVV plant pathogen (maize); rust 99, ,8 Illumina HiSeq Puccinia striiformis Basidiomycota Pucciniomycotina Pucciniomycetes Incertae sedis Pucciniales Pucciniaceae PST-08/21 Gp AORS plant pathogen (wheat); rust fungus 56, ,2 Illumina HiSeq Puccinia triticina Basidiomycota Pucciniomycotina Pucciniomycetes Incertae sedis Pucciniales Pucciniaceae 1-1 BBBD Gp ADAS plant pathogen (wheat, barley, rye); rust fungus 162,948 16, ; Sanger Uromyces viciae-fabae Basidiomycota Pucciniomycotina Pucciniomycetes Incertae sedis Pucciniales Pucciniaceae I2 Gp JNCO plant pathogen (beans); rust 215, ,7 Illumina HiSeq Septobasidium sp. Basidiomycota Pucciniomycotina Pucciniomycetes Incertae sedis Septobasidiales Septobasidiaceae PNB30-8B v endophyte 27,51 80, Illumina Tritirachium sp. CBS Basidiomycota Pucciniomycotina Tritirachiomycetes Incertae sedis Tritirachiales Tritirachiaceae CBS Gp JGI uncertain; mold 18,89 86, Illumina Ceraceosorus bombacis Basidiomycota Ustilaginomycotina Exobasidiomycetes Exobasidiomycetidae Ceraceosorales Ceraceosoraceae Cbombacis1Gp CCYA plant pathogen (lumber tree); smut 26, ,9 Ceraceosorus sp. Basidiomycota Ustilaginomycotina Exobasidiomycetes Exobasidiomycetidae Ceraceosorales Ceraceosoraceae MCA 4658 v1.0 Gp plant pathogen (lumber tree); smut 24, Illumina Meira miltonrushii Basidiomycota Ustilaginomycotina Exobasidiomycetes Exobasidiomycetidae Exobasidiales Brachybasidiaceae MCA 3882 Gp mite-associate 17,38 99, Illumina HiSeq Meira nashicola Basidiomycota Ustilaginomycotina Exobasidiomycetes Exobasidiomycetidae Exobasidiales Brachybasidiaceae JCM Gp BCJU probable plant pathogen (pear) 18, HiSeq 2500 Acaromyces ingoldii Basidiomycota Ustilaginomycotina Exobasidiomycetes Exobasidiomycetidae Exobasidiales Cryptobasidiaceae MCA 4198 v biocontrol (toxigenic on mite spp.) 19,33 91, Illumina Exobasidium vaccinii Basidiomycota Ustilaginomycotina Exobasidiomycetes Exobasidiomycetidae Exobasidiales Exobasidiaceae MPITM Gp JGI plant pathogen (endoparasitic) 16,99 96, Illumina HiSeq Tilletiaria anomala Basidiomycota Ustilaginomycotina Exobasidiomycetes Exobasidiomycetidae Georgefischeriales Tilletiariaceae UBC 951 Gp JMSN uncertain 18,7 25, Golubevia (Tilletiopsis) pallescens Basidiomycota Ustilaginomycotina Exobasidiomycetes Exobasidiomycetidae Golubeviales Golubeviaceae JCM 5230 Gp BCHO biocontrol (of podwery mildew) 27, ,2 HiSeq 2500 Tilletiopsis washingtonensis Basidiomycota Ustilaginomycotina Exobasidiomycetes Exobasidiomycetidae Incertae sedis Incertae sedis MCA 4816 Gp uncertain; yeast 18,76 84, Illumina HiSeq Tilletia caries Basidiomycota Ustilaginomycotina Exobasidiomycetes Exobasidiomycetidae Tilletiales Tilletiaceae DAOM Gp LWDD plant pathogen (bunt of wheat) 29, ,1 Illumina Tilletia controversa Basidiomycota Ustilaginomycotina Exobasidiomycetes Exobasidiomycetidae Tilletiales Tilletiaceae DAOM Gp LWDE plant pathogen (dwarf bunt of wheat) 28, ,6 Illumina Tilletia horrida Basidiomycota Ustilaginomycotina Exobasidiomycetes Exobasidiomycetidae Tilletiales Tilletiaceae QB-1 Gp LAXH plant pathogen (rice); smut 20, ,8 Illumina HiSeq Tilletia indica Basidiomycota Ustilaginomycotina Exobasidiomycetes Exobasidiomycetidae Tilletiales Tilletiaceae DAOM Gp LWDF plant pathogen (Karnal bunt on wheat an triticale) 30, ,6 Illumina Tilletia walkeri Basidiomycota Ustilaginomycotina Exobasidiomycetes Exobasidiomycetidae Tilletiales Tilletiaceae DAOM Gp LWDG plant pathogen (Ryegrass bunt) 24,34 58, ,8 Illumina Malassezia caprae Basidiomycota Ustilaginomycotina Malasseziomycetes Incertae sedis Malasseziales Malasseziaceae CBS Gp LFFV opportunistic animal pathogen 7, ,8 Illumina HiSeq Malassezia cuniculi Basidiomycota Ustilaginomycotina Malasseziomycetes Incertae sedis Malasseziales Malasseziaceae CBS Gp LFFW skin-associated; lipophilic 7, Illumina HiSeq Malassezia dermatis Basidiomycota Ustilaginomycotina Malasseziomycetes Incertae sedis Malasseziales Malasseziaceae JCM Gp BCKX animal pathogen (dermatitis) 7,55 486, HiSeq 2500 Malassezia equina Basidiomycota Ustilaginomycotina Malasseziomycetes Incertae sedis Malasseziales Malasseziaceae CBS 9969 Gp LFFY opportunistic animal pathogen 7,66 244, Illumina HiSeq

70 Malassezia furfur Basidiomycota Ustilaginomycotina Malasseziomycetes Incertae sedis Malasseziales Malasseziaceae CBS 1878 Gp LFGL skin-associated; lipophilic 7, ,1 Illumina HiSeq Malassezia globosa Basidiomycota Ustilaginomycotina Malasseziomycetes Incertae sedis Malasseziales Malasseziaceae CBS 7966 Gp AAYY human pathogen 8, Sanger Malassezia japonica Basidiomycota Ustilaginomycotina Malasseziomycetes Incertae sedis Malasseziales Malasseziaceae JCM Gp BCKY skin-associated; lipophilic 8,36 466, ,3 HiSeq 2500 Malassezia nana Basidiomycota Ustilaginomycotina Malasseziomycetes Incertae sedis Malasseziales Malasseziaceae JCM Gp BCLA opportunistic animal pathogen 7, HiSeq 2500 Malassezia obtusa Basidiomycota Ustilaginomycotina Malasseziomycetes Incertae sedis Malasseziales Malasseziaceae CBS 7876 Gp LFGC skin-associated; lipophilic 7, ,2 Illumina HiSeq Malassezia pachydermatis Basidiomycota Ustilaginomycotina Malasseziomycetes Incertae sedis Malasseziales Malasseziaceae CBS 1879 Gp LGAV animal pathogen (dermatitis) 8, ,1 Illumina HiSeq Malassezia restricta Basidiomycota Ustilaginomycotina Malasseziomycetes Incertae sedis Malasseziales Malasseziaceae CBS 7877 Gp AAXK human pathogen 4, ,3 Sanger Malassezia slooffiae Basidiomycota Ustilaginomycotina Malasseziomycetes Incertae sedis Malasseziales Malasseziaceae CBS 7956 Gp LFGK opportunistic animal pathogen 8, ,9 Illumina HiSeq Malassezia sympodialis Basidiomycota Ustilaginomycotina Malasseziomycetes Incertae sedis Malasseziales Malasseziaceae ATCC 42132Gp CANK opportunistic human and animal pathogen (cutabeo 7,67 130, ,1 454; Illumina HiSeq Malassezia yamatoensis Basidiomycota Ustilaginomycotina Malasseziomycetes Incertae sedis Malasseziales Malasseziaceae MY9725 Gp LFCX human pathogen (dermatitis) 8,11 235, ,7 Illumina HiSeq Ustilaginomycotina sp. SA 807 Basidiomycota Ustilaginomycotina unknown unknown unknown unknown SA 807 Gp uncertain; plant-associated 27,32 140, Illumina HiSeq Melanotaenium endogenum Basidiomycota Ustilaginomycotina Ustilaginomycetes Ustilaginomycetidae Urocystidales Melanotaeniaceae CBS v plant pathogen (bedstraw); smut 19,17 97, Illumina Kalmanozyma (Pseudozyma) bbasidiomycota Ustilaginomycotina Ustilaginomycetes Ustilaginomycetidae Ustilaginales Ustilaginaceae GHG001 Gp AWXO industrial importance 17, ,1 Illumina HiSeq Moesziomyces (Pseudozyma) abasidiomycota Ustilaginomycotina Ustilaginomycetes Ustilaginomycetidae Ustilaginales Ustilaginaceae T-34 Gp BAFG industrial importance 18, ,1 454 Moesziomyces (Pseudozyma) abasidiomycota Ustilaginomycotina Ustilaginomycetes Ustilaginomycetidae Ustilaginales Ustilaginaceae DSM70725 Gp AWNI industrial importance 17, ,9 Illumina HiSeq Moesziomyces antarcticus Basidiomycota Ustilaginomycotina Ustilaginomycetes Ustilaginomycetidae Ustilaginales Ustilaginaceae JCM Gp BBIZ industrial importance (psychrophili lipid and lipase p 18, ,9 Illumina MiSeq Pseudozyma flocculosa Basidiomycota Ustilaginomycotina Ustilaginomycetes Ustilaginomycetidae Ustilaginales Ustilaginaceae DAOM 1969Gp AOUS biocontrol (powdery mildew) 23, ,3 454 Pseudozyma hubeiensis Basidiomycota Ustilaginomycotina Ustilaginomycetes Ustilaginomycetidae Ustilaginales Ustilaginaceae SY62 Gp BAOW industrial importance (biosurfactant producing) 18, ,5 Illumina HiSeq Sporisorium reilianum Basidiomycota Ustilaginomycotina Ustilaginomycetes Ustilaginomycetidae Ustilaginales Ustilaginaceae SRZ2 Gp GCA_ plant pathogen (maize); smut 18, , Sporisorium scitamineum Basidiomycota Ustilaginomycotina Ustilaginomycetes Ustilaginomycetidae Ustilaginales Ustilaginaceae Gp JFOL plant pathogen (sugarcane); smut 19, ,64 HiSeq 2500 Ustilago esculenta Basidiomycota Ustilaginomycotina Ustilaginomycetes Ustilaginomycetidae Ustilaginales Ustilaginaceae MMT Gp JTLW plant pathogen (wild rice); smut 20, ,4 Illumina HiSeq Ustilago hordei Basidiomycota Ustilaginomycotina Ustilaginomycetes Ustilaginomycetidae Ustilaginales Ustilaginaceae Uh Gp CAGI plant pathogen (barley); smut 21, ,9 Ustilago maydis Basidiomycota Ustilaginomycotina Ustilaginomycetes Ustilaginomycetidae Ustilaginales Ustilaginaceae 521 Gp AACP plant pathogen (maize and teosinte); smut 19, Sanger

71 Sequenced early-diverging fungal species Classification Genome statistics SPECIES PHYLUM SUBPHYLUM CLASS SUBCLASS ORDER FAMILY Strain GOLD Accession GENOME Accession NCBI Project ID Significance Size (Mb) coverage # of contigs # of scaffolds # genes GC Technology Allomyces macrogynus Blastocladiomycota none Blastocladiomycetes none Blastocladiales Blastocladiaceae ATCC Gp ACDU model organism (sexual cycle); cosmopolitan 57 11, Catenaria anguillulae Blastocladiomycota none Blastocladiomycetes none Blastocladiales Catenariaceae PL171 Gp JGI biocontrol (nematode parasite) 41,34 93, Illumina; PacBio Globomyces pollinis-pini Chytridiomycota none Chytridiomycetes Chytridiomycetidae Rhizophydiales Globomycetaceae Arg68 Go aquatic saprtotroph 21,65 163, Illumina Batrachochytrium dendrobatidis Chytridiomycota none Chytridiomycetes Chytridiomycetidae Rhizophydiales unknown JEL423 Gp AATT animal pathogen 23,89 7, ,3 Sanger Homoloaphlyctis polyrhiza Chytridiomycota none Chytridiomycetes Chytridiomycetidae Rhizophydiales unknown JEL 142 Gp AFSM saprotroph 26,11 11, ,7 454 Spizellomyces punctatus Chytridiomycota none Chytridiomycetes Chytridiomycetidae Spizellomycetales Spizellomycetaceae DAOM BR117 Gp ACOE model organism 23,906 9, Sanger Gonapodya prolifera Chytridiomycota none Monoblepharidomycetes Monoblepharidales Gonapodyaceae JEL478 Gp JGI aqautic 48, Illumina Rozella allomycis Cryptomycota none none none none unknown CSF55 Gp JGI parasite (water moulds) 11,86 264, ; Illumina HiSeq Antonospora locustae Microsporidia Nosematidae HM-2013 Gp biocontrol; parasite (grasshoppers) 6,07 7, Illumina Edhazardia aedis Microsporidia Amblyosporidae USNM Gp AFBI parasite 46,6 20, ,5 454 Spraguea lophii Microsporidia Spraguidae 42_110 Gp ATCN parasite (teleost fish) 4,98 70, ,4 Illumina GAii Enterocytozoon bieneusi Microsporidia Enterocytozoonidae H348 Gp ABGB parasite 3, Nosema antheraeae Microsporidia Nosematidae YY Gp Silkworm pathogen data parasite (silkworm) 6, Sanger; Illumina Nosema apis Microsporidia Nosematidae BRL01 Gp ANPH parasite (honeybee) 8, ,8 454 Nosema bombycis Microsporidia Nosematidae CQ1 Gp ACJZ parasite (silkworm) 15, Sanger; Illumina Nosema ceranae Microsporidia Nosematidae BRL01 Gp ACOL parasite (honeybee) 7,86 24, ,3 454 Vittaforma corneae Microsporidia Nosematidae ATCC Gp AEYK opportunistic pathogen 3, ,5 454 Encephalitozoon bieneusi Microsporidia Encephalitozoonidae H348 ABGB parasite 3, ,7 454; Illumina Encephalitozoon cuniculi Microsporidia Encephalitozoonidae GB-M1 Gp GCA_ parasite 2, Encephalitozoon hellem Microsporidia Encephalitozoonidae ATCC Gp GCA_ parasite 2,25 53, Encephalitozoon intestinalis Microsporidia Encephalitozoonidae ATCC Gp GCA_ parasite 2, Sanger Encephalitozoon romaleae Microsporidia Encephalitozoonidae SJ-2008 Gp GCA_ parasite 2,19 300, ,4 Anncaliia algerae Microsporidia Tubulinosematidae PRA109 Gp AOMV parasite (aquatic) 17, Illumina Nematocida parisii Microsporidia Incertae sedis ERTm1 Gp JGI parasite (C.elegans) 4,07 97, ,4 454 Pseudoloma neurophilia Microsporidia Glugeidae Gp LGUB parasite (zebrafish) 5,25 138, Illumina Trachipleistophora hominis Microsporidia Pleistophoridae Gp ANCC parasite 8,498 32, ,1 454; SOLiD Vavraia culicis Microsporidia Pleistophoridae floridensis Gp AEUG parasite (mosquito) 6, ,7 454 Hamiltosporidium tvaerminnensis Microsporidia Dubosqiidae OER-3-3 Gp ACSZ parasite 13, ,5 Nematocida displodere Microsporidia None JUm2807 Gp LTDL parasite (Caenorhabditis elegans) 3,1 2500, ,1 Illumina Nematocida ironsii Microsporidia None ERTm5 Gp LTDK parasite (Caenorhabditis elegans) 4,4 101, ,5 Illumina Nematocida sp. 1 Microsporidia None ERTm2 Gp AERB parasite (Caenorhabditis elegans) 4,7 65, ,4 454 Ordospora colligata Microsporidia Ordosporidae OC4 Gp JOKQ parasite (Daphnia) 2,29 627, ,5 Illumina HiSeq Mitosporidium daphniae Microsporidia unknown UGP3 Gp JMKJ parasite 5, Illumina HiSeq Rhizophagus irregularis Mucoromycota Glomeromycotina Glomeromycetes none Glomerales Glomeraceae DAOM Gp AUPC mycorrhizal (arbuscular) 91, ,5 Sanger; 454; Illumina Lobosporangium transversale Mucoromycota Mortierellomycotina none none Mortierellales Mortierellaceae NRRL 3116 v industrial importance (lipid biosynthesis) 42,77 195, PacBio Mortierella alpina Mucoromycota Mortierellomycotina none none Mortierellales Mortierellaceae ATCC Gp ADAG industrial importance 38,042 31, ; Illumina; Sanger Mortierella elongata Mucoromycota Mortierellomycotina none none Mortierellales Mortierellaceae AG-77 Gp JGI endophyte; saprotroph 49,96 112, Illumina HiSeq Mortierella multidivaricata Mucoromycota Mortierellomycotina none none Mortierellales Mortierellaceae RSA 2152 T Go saprotroph; soil; garlic-odour 37,67 184, PacBio Mortierella verticillata Mucoromycota Mortierellomycotina none none Mortierellales Mortierellaceae NRRL 6337 Gp AEVJ animal pathogen 41, ,9 Illumina; Sanger Backusella circina Mucoromycota Mucoromycotina none none Mucorales Backusellaceae FSU 941 Gp JGI thermophile 48,65 100, Illumina HiSeq Blakeslea trispora Mucoromycota Mucoromycotina none none Mucorales Choanephoraceae NRRL 2456 v industrial importance (beta-carotene); plant pathogen 37, PacBio Absidia (Chlamydoabsidia) padenii Mucoromycota Mucoromycotina none none Mucorales Cunninghamellaceae NRRL 2977 Go uncertain 34,33 139, PacBio Cunninghamella bertholletiae Mucoromycota Mucoromycotina none none Mucorales Cunninghamellaceae B7461 Gp JNEL human pathogen (mucoromycosis) 31,1 82, ,8 Illumina HiSeq not publically available Cunninghamella elegans Mucoromycota Mucoromycotina none none Mucorales Cunninghamellaceae B9769 Gp JNDR degrades xenobiotics, some herbicides and fungicides 31,74 115, ,5 Illumina HiSeq Gongronella butleri Mucoromycota Mucoromycotina none none Mucorales Cunninghamellaceae CBS Go industrial importance (chitosan production) 33,01 150, PacBio Hesseltinella vesiculosa Mucoromycota Mucoromycotina none none Mucorales Cunninghamellaceae NRRL 3301 Gp uncertain 25,86 157, Illumina Lichtheimia corymbifera Mucoromycota Mucoromycotina none none Mucorales Lichtheimiaceae CBS Gp CBTN human pathogen (mucoromycosis) 33, ,9 HiSeq 2500; 454 Lichtheimia hyalospora Mucoromycota Mucoromycotina none none Mucorales Lichtheimiaceae FSU Gp JGI industrial importance (thermotolerant) 33,28 140, Illumina not publically available Lichtheimia ramosa Mucoromycota Mucoromycotina none none Mucorales Lichtheimiaceae :FSU:6197 Gp opportunistic human pathogen (mucoromycosis) 30, ,1 HiSeq 2500; 454 Phascolomyces articulosus Mucoromycota Mucoromycotina none none Mucorales Lichtheimiaceae RSA 2281 Go coprophilous 47,61 121, PacBio Rhizomucor miehei Mucoromycota Mucoromycotina none none Mucorales Lichtheimiaceae CAU432 Gp AGBC industrial importance (thermophile) 27, ,3 454; Illumina HiSeq Rhizomucor variabilis Mucoromycota Mucoromycotina none none Mucorales Lichtheimiaceae B7584 Gp JNES human pathogen 33,037 59, ,8 Illumina HiSeq Thermomucor indicae-seudaticae Mucoromycota Mucoromycotina none none Mucorales Lichtheimiaceae HACC 243 Gp JSYX industrial importance (dairy industry) 29,57 180, ,8 Illumina HiSeq Zychaea mexicana Mucoromycota Mucoromycotina none none Mucorales Lichtheimiaceae RSA 1403 Go coprophilous 50, PacBio Actinomucor elegans Mucoromycota Mucoromycotina none none Mucorales Mucoraceae JCM Gp BCHK human pathogen; mucormycosis 34,17 317, ,9 HiSeq 2500 Apophysomyces elegans Mucoromycota Mucoromycotina none none Mucorales Mucoraceae B7760 Gp JNDQ human pathogen (mucoromycosis); thermotolerant 38,5 53, Illumina HiSeq Apophysomyces trapeziformis Mucoromycota Mucoromycotina none none Mucorales Mucoraceae B9324 Gp JNDP human pathogen (mucoromycosis) 35,8 102, Illumina HiSeq Cokeromyces recurvatus Mucoromycota Mucoromycotina none none Mucorales Mucoraceae B5483 Gp JNEH human pathogen (mucoromycosis) 29,34 63, ,4 Mucor (Kirkomyces) cordense Mucoromycota Mucoromycotina none none Mucorales Mucoraceae RSA 1222 Go uncertain 40,95 157, PacBio Mucor ambiguus Mucoromycota Mucoromycotina none none Mucorales Mucoraceae NBRC 6742 Gp BBKB industrial importance (novel carbonyl reductase) 40,74 200, ,7 Illumina MiSeq Mucor circinelloides Mucoromycota Mucoromycotina none none Mucorales Mucoraceae 1006PhL Gp AOCY opportunistic pathogen; model organism 36,348 45, ,5 Illumina Mucor heterogamus Mucoromycota Mucoromycotina none none Mucorales Mucoraceae NRRL 1489 Go soil-associated; homothallic 54,49 285, PacBio Mucor indicus Mucoromycota Mucoromycotina none none Mucorales Mucoraceae B7402 Gp JNEK human pathogen (mucoromycosis) 39,797 55, ,8 Illumina HiSeq Mucor irregularis Mucoromycota Mucoromycotina none none Mucorales Mucoraceae B50 Gp AZYI human pathogen (mucoromycosis) 33, , ,9 Illumina Mucor racemosus Mucoromycota Mucoromycotina none none Mucorales Mucoraceae B9645 Gp JNEI human pathogen (mucoromycosis) 65,553 49, ,5 Illumina HiSeq Mucor ramosissimus Mucoromycota Mucoromycotina none none Mucorales Mucoraceae Gp JNEF human pathogen (mucoromycosis) 42,9 73, ,3 Illumina HiSeq Mucor velutinosus Mucoromycota Mucoromycotina none none Mucorales Mucoraceae B5328 Gp JNDK human pathogen (mucoromycosis) 35, , ,5 Illumina HiSeq Parasitella parasitica Mucoromycota Mucoromycotina none none Mucorales Mucoraceae CBS Gp CCXP biocontrol (mycoparasite) 44, Illumina HiSeq Mycotypha africana Mucoromycota Mucoromycotina none none Mucorales Mycotyphaceae NRRL 2978 Go soil-associated; dimorphic 29,2 610, PacBio Phycomyces blakesleeanus Mucoromycota Mucoromycotina none none Mucorales Phycomycetaceae NRRL1555 Gp JGI model organism 53,9 7, Sanger Rhizopus delemar Mucoromycota Mucoromycotina none none Mucorales Rhizopodaceae RA Gp AACW opportunistic human pathogen 46, ,6 Sanger Rhizopus microsporus Mucoromycota Mucoromycotina none none Mucorales Rhizopodaceae CCTCC M Gp ANKS traditional use 45,74 97, ,9 Illumina Rhizopus oryzae Mucoromycota Mucoromycotina none none Mucorales Rhizopodaceae NRRL Gp JNDU opportunistic human pathogen (mucormycosis); traditional fermentation 43,351 86, ,4 Illumina HiSeq Rhizopus stolonifer Mucoromycota Mucoromycotina none none Mucorales Rhizopodaceae B9770 Gp JNDS opportunistic human pathogen (mucoromycosis); black bread mold; 38,025saprotroph 42, ,5 Illumina HiSeq Saksenaea oblongispora Mucoromycota Mucoromycotina none none Mucorales Saksenaeaceae B3353 Gp JNEV human pathogen (mucoromycosis) 40,75 72, ,9 Illumina HiSeq Saksenaea vasiformis Mucoromycota Mucoromycotina none none Mucorales Saksenaeaceae B4078 Gp JNDT human pathogen (mucoromycosis) 42,502 88, ,7 Illumina HiSeq Syncephalastrum monosporum Mucoromycota Mucoromycotina none none Mucorales Syncephalastraceae B8922 Gp JNEN human pathogen (mucoromycosis) 29,565 75, ,3 Illumina HiSeq Syncephalastrum racemosum Mucoromycota Mucoromycotina none none Mucorales Syncephalastraceae B6101 Gp JNDN human pathogen (mucoromycosis) 29,568 76, ,2 Illumina HiSeq Umbelopsis isabellina Mucoromycota Mucoromycotina none none Mucorales Umbelopsidaceae NBRC 7884 Gp BAVE oleaginous 22, SOLiD Umbelopsis ramanniana Mucoromycota Mucoromycotina none none Mucorales Umbelopsidaceae AG Gp JGI saprotroph (soil) 23, Illumina HiSeq Anaeromyces robustus Neocallimastigomycota Incertae sedis Neocallimastigomycetes none Neocallimastigales Neocallimastigaceae S gut-associated; sheep 71, PacBio Neocallimastix californiae Neocallimastigomycota Incertae sedis Neocallimastigomycetes none Neocallimastigales Neocallimastigaceae G gut-associated; goat 193, PacBio Orpinomyces sp. C1A Neocallimastigomycota Incertae sedis Neocallimastigomycetes none Neocallimastigales Neocallimastigaceae C1A Gp ASRE anaerobic gut fungus 100,954 94, Illumina; PacBio Piromyces finnis Neocallimastigomycota Incertae sedis Neocallimastigomycetes none Neocallimastigales Neocallimastigaceae v gut-associated; horse 56,46?? PacBio Piromyces sp. E2 Neocallimastigomycota Incertae sedis Neocallimastigomycetes none Neocallimastigales Neocallimastigaceae ATCC anaerobic gut fungus 71, fungal sp. EF0021 unknown unknown unknown unknown unknown unknown EF0021 Gp AIET endophyte (Taxus ) 44, ,1 454 fungal sp. No unknown unknown unknown unknown unknown unknown No Gp BBSH industrial importance (lipopeptidolactone and,3-beta-glucan synthase 21,72 inhibitor 135production) ; Sanger Basidiobolus meristosporus Zoopagomycota Entomophthoromycotina Basidiobolomycetes none Basidiobolales Basidiobolaceae B9252 Gp JNEO human pathogen (mucoromycosis) 103,87 22, ,9 Illumina HiSeq Conidiobolus coronatus Zoopagomycota Entomophthoromycotina Entomophthoromycetes none Entomophthorales Ancylistaceae NRRL Gp JGI human and animal pathogen (conidiobolomycosis) 39,9 16, Conidiobolus incongruus Zoopagomycota Entomophthoromycotina Entomophthoromycetes none Entomophthorales Ancylistaceae B7586 Gp JNEM human and animal pathogen (conidiobolomycosis) 93,89 75, ,4 Illumina HiSeq Conidiobolus thromboides Zoopagomycota Entomophthoromycotina Entomophthoromycetes none Entomophthorales Ancylistaceae FSU 785 Gp biocontrol (entomopathogen) 24,64 127, Illumina HiSeq Coemansia reversa Zoopagomycota Kickxellomycotina none none Kickxellales Kickxellaceae NRRL 1564 Gp JGI arthropod-associated 21,84 38,

72 Linderina pennispora Zoopagomycota Kickxellomycotina none none Kickxellales Kickxellaceae ATCC v1.0 Gp saprotroph (soil) 26, PacBio Martensiomyces pterosporus Zoopagomycota Kickxellomycotina none none Kickxellales Kickxellaceae CBS Gp saprotroph 19,82 115, Illumina HiSeq Ramicandelaber brevisporus Zoopagomycota Kickxellomycotina none none Kickxellales Kickxellaceae CBS Gp JGI saprotroph (soil) 25,53 168, Illumina HiSeq

73 Supplementary File 2: Cash crops, Food crops, and Gymnosperms

74 CASH CROP PATHOGENS Classification Genome SPECIES Significance PHYLUM Accession Ashbya gossypii plant pathogen (insect-associated; cotton) Ascomycota Gp Ceratocystis adiposa plant pathogen (black root in sugar cane) Ascomycota Gp Colletotrichum falcatum plant pathogen (sugar-cane red rot) Ascomycota Gp Colletotrichum phormii plant pathogen (anthracnose on flax) Ascomycota Go Colletotrichum simmondsii plant pathogen (safflower oil crop) Ascomycota Gp Phoma herbarum plant pathogen (hop & hemp) Ascomycota Gp Sporisorium scitamineum plant pathogen (sugarcane); smut Basidiomycota Gp Verticillium longisporum plant pathogen (canola) Ascomycota Gp

75 FOOD CROP PATHOGENS Classification Genome SPECIES Significance Staple crop? PHYLUM Accession Grains Bipolaris maydis plant pathogen (wheat) x Ascomycota Gp Bipolaris oryzae plant pathogen (rice) x Ascomycota Gp Bipolaris sorokiniana plant pathogen (cereals) x Ascomycota Gp Bipolaris victoriae plant pathogen (blight of oats) x Ascomycota Gp Bipolaris zeicola plant pathogen (sorghum, maize and apple) x Ascomycota Gp Blumeria graminis plant pathogen (mildew on grasses and cereals) x Ascomycota Gp Cercospora zeae-maydis plant pathogen (maize); toxin-producing x Ascomycota Gp Claviceps fusiformis plant pathogen (pearl millet) x Ascomycota Gp Claviceps purpurea plant pathogen (cereals) x Ascomycota Gp Cochliobolus lunatus plant pathogen (sorghum); pharmaceutical importance x Ascomycota Gp Colletotrichum graminicola plant pathogen (anthracnose in cereals) x Ascomycota Gp Colletotrichum sublineola plant pathogen (anthracnose in wild rice and sorghum) x Ascomycota Gp Fusarium acuminatum plant pathogen (cereals) x Ascomycota Gp Fusarium avenaceum plant pathogen (generalist, including grain crops) x Ascomycota Gp Fusarium equiseti plant pathogen (members of the Leguminoseae and some cereals) x Ascomycota Gp Fusarium fujikuroi plant pathogen (rice) x Ascomycota Gp Fusarium graminearum plant pathogen (wheat and barley) x Ascomycota Gp Fusarium nygamai plant pathogen (rice) x Ascomycota Gp Fusarium pseudograminearum plant pathogen (wheat) x Ascomycota Gp Fusarium temperatum plant pathogen (maize); opportunistic human pathogen x Ascomycota Gp Fusarium verticillioides plant pathogen (maize) x Ascomycota Gp Gaeumannomyces graminis plant pathogen (root rot of cereals) x Ascomycota Gp Magnaporthe oryzae plant pathogen (rice) x Ascomycota Gp Melanconium sp. 1 NRRL plant pathogen (probable maize pathogen) x Ascomycota Gp Parastagonospora nodorum plant pathogen (wheat) x Ascomycota Gp Puccinia graminis plant pathogen; rust fungus (cereals) x Basidiomycota Gp Puccinia sorghi plant pathogen (maize); rust x Basidiomycota Gp Puccinia striiformis plant pathogen (wheat); rust fungus x Basidiomycota Gp Puccinia triticina plant pathogen (wheat, barley, rye); rust fungus x Basidiomycota Gp Pyrenophora seminiperda plant pathogen (necrotrophic; seeds of grasses/cereals) x Ascomycota Gp Pyrenophora teres plant pathogen (barley and some other crops) x Ascomycota Gp Pyrenophora tritici-repentis plant pathogen (cereals and grasses; necrotrophic) x Ascomycota Gp Ramularia collo-cygni plant pathogen (barley) x Ascomycota Gs Sarocladium oryzae plant pathogen (rice sheath rot) x Ascomycota Sclerotinia borealis plant pathogen (cereals; sychrophilic) x Ascomycota Gp Setosphaeria turcica plant pathogen (maize) x Ascomycota Gp Sporisorium reilianum plant pathogen (maize); smut x Basidiomycota Gp Tilletia caries plant pathogen (bunt of wheat) x Basidiomycota Gp Tilletia controversa plant pathogen (dwarf bunt of wheat) x Basidiomycota Gp Tilletia horrida plant pathogen (rice); smut x Basidiomycota Gp Tilletia indica plant pathogen (Karnal bunt on wheat an triticale) x Basidiomycota Gp Ustilaginoidea virens plant pathogen (rice) x Ascomycota Gp Ustilago esculenta plant pathogen (wild rice); smut x Basidiomycota Gp Ustilago hordei plant pathogen (barley); smut x Basidiomycota Gp Ustilago maydis plant pathogen (maize and teosinte); smut x Basidiomycota Gp Villosiclava virens plant pathogen; (false smut of rice) x Ascomycota Gp Zymoseptoria passerinii plant pathogen (barley leaf blotch) x Ascomycota Gp Zymoseptoria tritici plant pathogen (wheat leaf blotch) x Ascomycota Gp Fruit Alternaria arborescens plant pathogen (tomato) Ascomycota Gp Botryotinia fuckeliana plant pathogen (grapevines) Ascomycota Gp Ceratocystis manginecans plant pathogen (mango) Ascomycota Gp Cladosporium fulvum plant pathogen (leaf mold) Ascomycota Colletotrichum fioriniae plant pathogen (anthracnose in crops and wild plants) Ascomycota Gp Dactylonectria macrodidyma plant pathogen (grapevine, avocado, and olive) Ascomycota Gp Diplodia seriata plant pathogen (grapevine bot canker) Ascomycota Gp Eremothecium cymbalariae plant pathogen (stigmatomycosis; fruit rot; crops) Ascomycota Gp Erysiphe necator plant pathogen ( powdery mildew of grape) Ascomycota Gp Eutypa lata plant pathogen; grapevine dieback Ascomycota Gp Geotrichum candidum plant pathogen (citrus) Ascomycota Gp Guignardia citricarpa plant pathogen (citrus) Ascomycota Gp Huntiella omanensis plant pathogen (weak mango pathogen) Ascomycota Gp Meira nashicola probable plant pathogen (pear) Basidiomycota Gp Moniliophthora perniciosa plant pathogen (cacao) Basidiomycota Gp Mycosphaerella eumusae plant pathogen (banana) Ascomycota Gp Mycosphaerella fijiensis plant pathogen (leaf spot of banana) Ascomycota Gp Neofusicoccum parvum plant pathogen (grapevines) Ascomycota Gp Neonectria ditissima plant pathogen (apple canker) Ascomycota Gp Passalora (Cladosporium) fulva plant pathogen (tomato) Ascomycota Gp Peltaster fructicola plant pathogen (apples) Ascomycota Gp Phaeoacremonium aleophilum plant pathogen (grapevine trunk disease) Ascomycota Gp Phaeomoniella chlamydospora plant pathogen (grapevine trunk disease) Ascomycota Gp Phyllosticta citriasiana plant pathogen (citrus tan spot) Ascomycota Gp Phyllosticta citricarpa plant pathogen (citrus black spot) Ascomycota Gp

76 Plenodomus tracheiphilus plant pathogen (citrus) Ascomycota Gp Pseudocercospora musae plant pathogen (banana) Ascomycota Gp Puccinia psidii plant pathogen; rust fungus (guava) Basidiomycota Gp Pyrenochaeta lycopersici plant pathogen (tomato) Ascomycota Gp Rosellinia necatrix plant pathogen (fruits) Ascomycota Gp Stemphylium lycopersici plant pathogen (fruits) Ascomycota Gp Taphrina deformans plant pathogen (peach) Ascomycota Gp Taphrina flavorubra plant pathogen (Prunus fruit) Ascomycota Gp Taphrina wiesneri plant pathogen (cherry trees) Ascomycota Gp Thielaviopsis punctulata plant pathogen (date palm) Ascomycota Gp Valsa mali plant pathogen (apple & pear) Ascomycota Gp Venturia pyrina plant pathogen (hemibiotrophic; pear) Ascomycota Gp Vegetables Alternaria brassicicola plant pathogen (Brassica dark leaf spot) Ascomycota Gp Athelia rolfsii plant pathogen (blight of vegetables) Basidiomycota Gp Ceratocystis fimbriata plant pathogen (sweet potato) x Ascomycota Gp Colletotrichum higginsianum plant pathogen (anthracnose in Brassicaceae) Ascomycota Gp Fusarium sambucinum plant pathogen (potato dry rot); mycotoxin-producing x Ascomycota Gp Helminthosporium solani plant pathogen (potato) x Ascomycota Gp Leptosphaeria maculans plant pathogen (Brassica crops) Ascomycota Gp Ophiognomonia clavigignenti-juglanplant pathogen (butternut) Ascomycota Gp Plectosphaerella cucumerina plant pathogen (blight of cucurbits); nematophagous Ascomycota Sclerotinia sclerotiorum plant pathogen (broadest host range known) Ascomycota Gp Legumes Ascochyta rabiei plant pathogen (blight on chickpea) Ascomycota Gp Colletotrichum incanum plant pathogen (broad host range; including soybean) Ascomycota Gp Diaporthe aspalathi plant pathogen (soybean stem canker) x Ascomycota Gp Diaporthe longicolla plant pathogen (soybean) x Ascomycota Gp Eremothecium coryli plant pathogen (soybean) x Ascomycota Gp Erysiphe pisi plant pathogen (powdery mildew of pea) Ascomycota Gp Fusarium virguliforme plant pathogen (soybean) x Ascomycota Gp Moniliophthora roreri plant pathogen (pod rot of cacao) Basidiomycota Gp Mycosphaerella arachidis (Cercospoplant pathogen (leaf spot on peanuts) Ascomycota Gp Puccinia arachidis plant pathogen (peanut) Basidiomycota Gp Uromyces viciae-fabae plant pathogen (beans); rust Basidiomycota Gp Multiple food crop types Alternaria alternata plant pathogen (leaf spot) Ascomycota Gp Cercospora canescens plant pathogen (leaf spot of various bean crops and tomato) Ascomycota Gp Colletotrichum gloeosporioides plant pathogen (disease and anthracnose on a range of fuit and vegetables) Ascomycota Gp Colletotrichum orbiculare plant pathogen (melons and cucumber) Ascomycota Gp Corynespora cassicola plant pathogen (broad host range) x Ascomycota Gp Fusarium oxysporum plant pathogen (broad host range) Ascomycota Gp Macrophomina phaseolina plant pathogen (broad host range) x Ascomycota Gp Nectria haematococca plant pathogen (broad host range) x Ascomycota Gp Rhizoctonia solani plant pathogen (broad host range) x Basidiomycota Gp Verticillium alfalfae plant pathogen (broad host range); wilt disease Ascomycota Gp Verticillium dahliae plant pathogen (broad host range); wilt disease Ascomycota Gp

77 GYMNOSPERM PATHOGENS Classification Genome SPECIES Significance PHYLUM Accession Aplosporella prunicola probable tree pathogen Ascomycota Gp Armillaria ostoyae plant pathogen; conifer root rot (parasitic or saprophytic) Basidiomycota Botryosphaeria dothidea plant pathogen (broad host range of trees and shrubs) Ascomycota Gp Caloscypha fulgens plant pathogen (seed rot of conifers) Ascomycota Ceraceosorus bombacis plant pathogen (lumber tree); smut Basidiomycota Gp Ceraceosorus sp. plant pathogen (lumber tree); smut Basidiomycota Gp Ceratocystis albifundus plant pathogen (woody & herbaceaous) Ascomycota Gp Ceratocystis eucalypticola probable plant pathogen (eucalyptus) Ascomycota Gp Ceratocystis platani plant pathogen (trees) Ascomycota Gp Chrysoporthe austroafricana plant pathogen (eucalyptus, Tibouchina, Syzygium) Ascomycota Gp Chrysoporthe cubensis plant pathogen (eucalyptus, Tibouchina, Syzygium) Ascomycota Gp Chrysoporthe deuterocubensis plant pathogen (eucalyptus, Tibouchina, Syzygium) Ascomycota Gp Colletotrichum salicis plant pathogen (black canker of willow) Ascomycota Gp Cronartium comandrae plant pathogen; rust fungus (pine) Basidiomycota Gp Cronartium quercuum plant pathogen; rust fungus (fusiform rust of pine) Basidiomycota Gp Cronartium ribicola plant pathogen; rust fungus (pine) Basidiomycota Gp Cryphonectria parasitica plant pathogen (chestnut blight) Ascomycota Gp Diplodia pinea plant pathogen (pines) Ascomycota Gp Diplodia scrobiculata plant pathogen (conifer spp.) Ascomycota Gp Endocronartium harknessii plant pathogen; rust fungus (pine-pine gall rust) Basidiomycota Gp Fusarium circinatum plant pathogen (pitch canker of pines) Ascomycota Gp Grosmannia clavigera plant pathogen; blue stain Ascomycota Gp Heterobasidion annosum plant pathogen (conifers) Basidiomycota Gp Heterobasidion irregulare plant pathogen (conifers and hardwoods) Basidiomycota Gp Hymenoscyphus fraxineus plant pathogen (ash dieback) Ascomycota Gp Lecanosticta acicola plant pathogen (pine) Ascomycota Gp Leptographium procerum plant pathogen (pine) Ascomycota Gp Marssonina brunnea plant pathogen (poplar leaf spot) Ascomycota Gp Melampsora allii-populina plant pathogen; rust fungus (poplar) Basidiomycota Gp Melampsora larici-populina plant pathogen (poplar); rust fungus Basidiomycota Gp Melampsora pinitorqua plant pathogen (pine twisting rust); rust fungus Basidiomycota Gp Mycosphaerella laricina plant pathogen (larch) Ascomycota Gp Mycosphaerella pini plant pathogen (pine) Ascomycota Gp Ophiostoma novo-ulmi plant pathogen (Dutch Elm Disease) Ascomycota Gp Phellinus noxius plant pathogen (broad host range, typically trees) Basidiomycota Gp Porodaedalea chrysoloma plant pathogen (conifer parasite; white pocket rot) Basidiomycota Go Porodaedalea niemelaei plant pathogen (conifer white rot) Basidiomycota Go Setomelanomma holmii plant pathogen (spruce needle drop) Ascomycota Gp Sphaerulina musiva plant pathogen (poplar) Ascomycota Gp Sphaerulina populicola plant pathogen (poplar) Ascomycota Gp Taphrina populina plant pathogen (cottonwood) Ascomycota Gp Teratosphaeria nubilosa plant pathogen (leaf spot of Eucalyptus) Ascomycota Gp Thielaviopsis paradoxa plant pathogen (palm) Ascomycota Gp

78 Supplementary File 3

79 Phylogenetic placement of fungal orders described after Hibbett et al. (2007). ASCOMYCOTA (Lumbsch & Huhndorf, 2010) Abrothallales: (Pérez-Ortega et al., 2014) Acrospermales, Dyfrolomycetales, Monoblastiales, Lichenotheliales, Strigulales: (Wijayawardene et al., 2014) Amphisphaeriales: (Senanayake et al., 2015) Archaeorhizomycetes: (Rosling et al., 2011) Ascosphaerales & Arachnomycetales: (Kirk et al., 2008) Asterinales: (Hongsanan et al., 2014) Asterotexiales: (Guatimosim et al., 2014) Celotheliales: (Gueidan et al., 2014) Dothideomycete families: (Schoch et al., 2009a; Wijayawardene et al., 2014) Eremithallales: (Lucking et al., 2008) Geoglossomycetes & Geoglossales: (Schoch et al., 2009b) Lecanoromycete families: (Miadlikowska et al., 2014) Lecanoromycete orders (Arctomiales, Caliciales, Hymeneliales, Sarrameanales, Trapeliales): (Miadlikowska et al., 2014) Lecideales: (Schmull et al., 2011) Leotiomycete families: (Wang et al., 2006) Mytilinidiales: (Boehm et al., 2009) Natipusillales: (Hongsanan et al., 2014) Phaeomoniellales: (Chen et al., 2015) Saccharomycotina families: (Kurtzman, 2011) Sordariomycete families and orders (Amplistromatales, Annulatascales, Cordanales, Falcocladiales, Glomerellales, Jobellisiales, Koralionastetales, Magnaporthales, Pisorisporiales, Savoryellales, Togniniales): (Maharachchikumbura et al., 2015) Taxonomic confusion exists at almost all taxonomic levels within the Leotiomycetes: (Johnston et al., 2014) Trapeliales & Sarrameanales: (Hodkinson & Lendemer, 2011) Trypetheliales: (Hyde et al., 2013) Tubeufiales: (Boonmee et al., 2014) Umbilicariales: (Miadlikowska et al., 2014) Valsariales: (Jaklitsch et al., 2015) Venturiales: (Wijayawardene et al., 2014; Zhang et al., 2011) Xylonomycetes: (Gazis et al., 2012)

80 BASIDIOMYCOTA Amylocorticiales & Jaapiales: (Binder et al., 2010) Jaapials is sister to Agaricomycetidae Ceraceosorales: (Wang et al., 2015) Geminibasidiales: (Nguyen et al., 2013) Geminibasidiomycetes: (Nguyen et al., 2015) Golubeviales: (Wang et al., 2015) Holtermanniales: (Liu et al., 2015) Kriegeriales: (Toome et al., 2013) Lepidostromatales: (Hodkinson et al., 2014) Moniliellales: (Wang et al., 2014) Robbauerales: (Wang et al., 2015) Trichosporonales: (Liu et al., 2015) Tritirachiomycetes: (Schell et al., 2011) Wallemiomycetes at base of Agaricomycotina: (Nguyen et al., 2013) EARLY-DIVERGING FUNGI Basidiobolales (Gryganskyi et al., 2012) Cladochytriales (Mozley-Standridge et al., 2009) Lobulomycetales (Simmons et al., 2009) Mortierellomycotina (Hoffmann et al., 2011) Olpidiomycota: Index Fungorum no. 42, Effectively published 27/12/ :02:48 (ISSN ), Nomenclatural novelties: Alexander B. Doweld; Publication Name: Index Fungorum 42: Dec Rhizophlyctidales (Letcher et al., 2008)

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84 doi: /imafungus IMA FUNGUS 8(1): (2017) Functional diversity in Dichomitus squalens monokaryons Sara Casado López 1,2, Bart Theelen 1, Serena Manserra 1, Tedros Yonatan Issak 1,2, Johanna Rytioja 3, Miia R. Mäkelä 1,3, and Ronald P. de Vries 1,2,3 1 Westerdijk Fungal Biodiversity Institute, Uppsalalaan 8, 3584 CT Utrecht, The Netherlands 2 Fungal Molecular Physiology, Utrecht University, Uppsalalaan 8, 3584 CT Utrecht, The Netherlands; corresponding author r.devries@ westerdijkinstitute.nl 3 Department of Food and Environmental Sciences, Division of Microbiology and Biotechnology, Viikki Biocenter 1, University of Helsinki, Finland Abstract: Dichomitus squalens is a white-rot fungus that colonizes and grows mainly on softwood and is commonly found in the northern parts of Europe, North America, and Asia. We analyzed the genetic and physiological diversity of eight D. squalens monokaryons derived from a single dikaryon. In addition, an unrelated dikaryon and a newly established dikaryon from two of the studied monokaryons were included. Both growth and lignocellulose acting enzyme profiles were highly variable between the studied monokaryotic and dikaryotic strains, demonstrating a high level of diversity within the species. Key words: AFLP carbon utilization monokaryon sexual reproduction white-rot Article info: Submitted: 23 January 2017; Accepted: 24 February 2017; Published: 8 March INTRODUCTION Basidiomycete fungi are essential in forest ecology, due to their ability to degrade wood (Eriksson et al. 1990). Wood degrading basidiomycetes have traditionally been divided in two subgroups, white-rot and brown-rot fungi, according to their method of wood decay, although the existence of intermediate species has been suggested (Riley et al. 2014). White-rot fungi are unique in possessing an array of extracellular lignin-modifying enzymes and are therefore able to completely mineralize recalcitrant aromatic lignin polymers (Mäkelä et al. 2014). In addition, they typically harbor a full repertoire of genes encoding enzymes that are targeted for depolymerization of different plant cell wall polysaccharides (Rytioja et al. 2014). Most basidiomycetes have a sexual reproductive cycle. When suitable substrate and growth conditions are present, haploid (n) spores germinate and a primary mycelium is formed. The primary mycelium can fuse with another primary mycelium of a compatible mating type and form a dikaryotic (n + n, unfused nuclei from different parents) secondary mycelium. The secondary mycelium produces basidiomata where fusion of two haploid nuclei occurs in basidia to give diploid nuclei (2n). After that, meiosis follows and each basidium bears four haploid basidiospores. Therefore, the offspring of two different dikaryons can produce a dikaryon with a different genetic make-up than either parent, which can result in different physiological abilities of the offspring. There are different mating compatibility systems in fungi. In agaricomycetes, mating is regulated by either a bipolar or a tetrapolar system. Tetrapolar behavior consists of two unlinked genetic complexes, A and B, with four potential mating interactions between spores from one basidiome. In the bipolar case, only compatible or incompatible interactions are found (Raudaskoski & Kothe 2010). In both cases, when hyphae from two compatible mating types find each other and make contact, fusion of the mycelia usually occurs through clamp connections and a dikaryon is formed (Krings et al. 2011, Kim et al. 2014). In this study, we aimed to dissect the variations in genetic profile, growth, and enzyme production in different monokaryotic strains derived from a dikaryon of the whiterot basidiomycete Dichomitus squalens. This species is commonly found in North America and northern regions of Europe and Asia (Andrews & Gill 1943), and it usually grows on softwood, but is also able to grow and degrade hardwood (Blanchette et al. 1987, Fackler et al. 2007). Dichomitus squalens has a tetrapolar mating system, and in a previous study 20 monokaryotic progeny derived from a single wildtype dikaryon of D. squalens (FBCC312, CBS ) were isolated and their growth rate, cellulose and lignin degrading ability, and isoenzyme patterns were compared (Pham et al. 1990). We performed a more detailed analysis of these strains by studying their genetic variation using amplified fragment length polymorphisms (AFLP), and their ability to grow on different carbon sources and to produce extracellular lignocellulolytic enzymes. In addition, we generated a new dikaryon from two of the monokaryons and compared that to the original parent. The results of this study provide insight into the functional diversity that exists within a single basidiomycete species in nature and how this can evolve in fungi that contain a sexual cycle International Mycological Association You are free to share - to copy, distribute and transmit the work, under the following conditions: Attribution: You must attribute the work in the manner specified by the author or licensor (but not in any way that suggests that they endorse you or your use of the work). Non-commercial: You may not use this work for commercial purposes. No derivative works: You may not alter, transform, or build upon this work. For any reuse or distribution, you must make clear to others the license terms of this work, which can be found at Any of the above conditions can be waived if you get permission from the copyright holder. Nothing in this license impairs or restricts the author s moral rights. VOLUME 8 NO. 1 17

85 Casado López et al. Table 1. Mating type assigned to Dichomitus squalens strains described in Pham et al. (1990) related to their corresponding incompatibility group. Strain no. Mating type Strain no. Incompatibility groupᵃ (in CBS and/or FBCC collection) (Pham et al. 1990) (Pham et al. 1990) CBS AB DS 1 I CBS DS 19 CBS ab DS 3 II CBS DS 4 CBS DS 6 CBS DS 7 CBS DS 10 CBS DS 15 CBS DS 16 CBS DS 2 Ab CBS DS 8 III CBS DS 9 CBS DS 11 CBS DS 14 CBS DS 17 CBS DS 18 CBS DS 20 CBS ab DS 5 IV CBS DS 12 CBS DS 13 FBCC312 (CBS ) CBS (cross of CBS and CBS ) FBCC184 AaBb AaBb AaBb MATERIALS AND METHODS Fungal strains and cultivations Dichomitus squalens strains obtained from the CBS collection (Westerdijk Fungal Biodiversity Institute, Utrecht, The Netherlands) and the Fungal Biotechnology Culture Collection (FBCC, Department of Food and Environmental Sciences, University of Helsinki) were used in this study (Table 1) and maintained on 2 % (w/v) malt extract 1.5 % (w/v) agar (MEA) plates. Liquid pre-cultures were performed in 250 ml Erlenmeyer flasks with 50 ml low-nitrogen asparagine-succinate medium (LN-AS) minimal medium (Hatakka & Uusi-Rauva 1983), ph 4.5, amended with 0.05 % glycerol as a carbon source. The medium was inoculated with 5 agar plugs (0.5 cm diam) from MEA plates that were covered with fresh fungal mycelium and incubated for 7 d at 28 C stationary. From the precultures, 4 ml of homogenized mycelium (Mäkelä et al. 2002) was transferred to 250 ml Erlenmeyer flasks containing 100 ml LN-AS with 2 % powdered wheat bran or Norway spruce (Picea abies) sawdust as carbon source. These cultures were incubated for 16 d at 28 C under agitation (120 rpm). All cultures were performed in duplicate. Culture medium samples were harvested after 9 and 16 d of growth, centrifuged at 4 C to remove residual biomass and stored at -20 C as 2 ml aliquots. The ability of D. squalens strains to grow on 37 different carbon sources was tested on plates containing LN-AS agar (1.5 % agar-agar (w/v)) amended with: (1) monomeric and oligomeric sugars: D-glucose, D-fructose, D-galactose, D-mannose, D-ribose, D-xylose, L-arabinose, L-rhamnose, D-galacturonic acid, D-glucuronic acid, cellobiose, maltose, lactose, raffinose, or sucrose; (2) polymeric substrates: arabinogalactan, beechwood xylan, birchwood xylan, arabic gum, guar gum, soluble starch, apple pectin, citrus pectin, inulin, calcium lignin, casein, cellulose, or sodium lignin; and (3) powdered complex substrates: wheat bran, sugar beet pulp, citrus pulp, soybean hulls, rice bran, cotton seed pulp, alfalfa meal, or spruce sawdust. All monomeric and oligomeric carbon sources were added to a final concentration of 25 mm, while pure polymeric and complex substrates were added to a final concentration of 1 % and 3 %, respectively. 18 IMA FUNGUS

86 Functional diversity in Dichomitus squalens monokaryons Table 2. Substrates assayed for polysaccharide degrading enzymatic activities, corresponding enzymes and their abbreviation, and suppliers of the substrates. Substrate Enzyme Abbreviation Supplier 4-Nitrophenyl α-l-arabinofuranoside α-l-arabinofuranosidase ABF Sigma 4-Nitrophenyl α-d-galactopyranoside α-1,4-d-galactosidase AGL Sigma 4-Nitrophenyl β-d-glucopyranoside β-1,4-d-glucosidase BGL Sigma 4-Nitrophenyl β-d-xylopyranoside β-xylosidase BXL Sigma 4-Nitrophenyl β-d-cellobioside Cellobiohydrolase CBH Sigma 4-Nitrophenyl α-d-glucopyranoside α-1,4-d-glucosidase AGD Koch-light 4-Nitrophenyl β-d-galactopyranoside β-1,4-d-galactosidase LAC Sigma 4-Nitrophenyl β-d-mannopyranoside β-mannosidase MND Sigma 4-Nitrophenyl maltoside Glucoamylase GLA Acros The ph of the medium was adjusted to 4.5. Monosaccharides and cellobiose were filter-sterilized and added to the medium after autoclaving, while the other substrates were added before autoclaving. LN-AS agar plates without any carbon source were used as control. The strains were inoculated by placing mycelium-covered agar plug (0.5 mm diam) obtained from the edge of 1-wk-old MEA plates in the center of the plates. Incubation time was established for 3 d at 28 C according to the growth of the first strain reaching the edge of the plate. After this period, pictures were taken and the diameter of the fungal colony was measured for every plate in duplicate. To obtain dikaryons, two agar plugs (0.5 mm diam) covered by mycelia of monokaryons of compatible mating type, CBS (Ab) and CBS (ab), were placed on a MEA plate at 3 cm distance. After 3 d of growth at 28 C, the mycelia of the different strains made contact, and successful crosses were confirmed by microscopy (Zeiss Axioplan Microscope) by staining a sample of the common mycelium with lactophenol Cotton blue (Leck 1999). Enzymatic activity assays Liquid pre-cultures and transfer conditions were performed according to Rytioja et al. (2017), but in this study 2 g of powdered wheat bran and spruce sawdust were used for 100 ml LN-AS cultures, respectively. The cultures were performed as two biological replicates and sampled after 9 and 16 d. All p-nitrophenol (pnp) assays (Table 2) were performed in triplicate in 96-well plates using a FLUORstar OPTIMA microplate reader (BMG). For pnp assays, 0.1 % of the corresponding substrate was used in 50 mm sodium acetate buffer (ph 5.0). Reaction mixtures for α-larabinofuranosidase (ABF), α-1,4-d-galactosidase (AGL), β-1,4-d-glucosidase (BGL) and β-1,4-d-galactosidase (LAC) activity were incubated for 1 h, while reaction mixtures for β-xylosidase (BXL), cellobiohydrolase (CBH) and β-mannosidase (MND) activity were incubated for 4 h. All reactions were incubated at 28 C and were stopped by adding 100 μl 0.25 M sodium carbonate. A pnp standard curve was generated to calculate the concentration of the released pnp. The activities are expressed in nmol/min/ml. Laccase and manganese peroxidase (MnP) activities were measured by following the oxidation of 2,2-azino-bis (3-ethylbenzothiazoline-6-sulfonate) (ABTS); ϵ 420 = M 1 cm 1 ; Sigma) at 440 nm using a Biochrom Libra S22 UV/ Vis Spectrophotometer. Laccase activity was determined in 50 mm sodium malonate buffer (ph 4.5) for 30 s at 25 C (Eggert et al. 1996). After this first measurement 0.5 mm manganese sulfate and 0.1 mm hydrogen peroxide were added to the same reaction to start the MnP reaction, and change of absorbance was followed again for 30 s (Hofrichter & Fritsche 1997, Hofrichter et al. 1998). Three technical replicates were performed on biological duplicates and these were averaged in the graphs. SDS-PAGE Protein profiles were obtained from the same culture liquids that were used to measure enzyme activities. Sixteen µl of each culture liquid sample was fractionated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with 10 % (wt/vol) acrylamide/30 % bisacrylamide gels and a molecular weight marker (Bio-Rad unstained marker) was used to identify the molecular mass of the protein bands. The gels were stained using the Silver staining method (Chevallet et al. 2006) and documented using the HP scanner 4400c. AFLP To describe how related the D. squalens strains are at the genetic level, they were clustered in a phylogenetic tree based on amplified fragment length polymorphisms VOLUME 8 NO. 1 19

87 Casado López et al. Fig. 1. Clamp connections of the Dichomitus squalens compatible monokaryons (CBS and CBS ) crossing to form the dikaryon (CBS ) as visualized by lactophenol cotton blue staining. Clamp connections are indicated by arrows (A D). (AFLPs). DNA isolation from mycelium grown on MEA plates was performed with a Microbial DNA isolation kit (MO BIO Laboratories, Carlsbad, USA) according to the manufacturer s recommendations. The experimental procedure was adapted from Illnait-Zaragozi et al. (2012). To each AFLP-reaction, 0.2 μl of Orange DNA size standard (MCLAB, DSMO-101) was added and samples were processed on a 3730xl DNA Analyzer (Applied Biosystems). Raw data was normalized and further analyzed with Bionumerics (v. 4.61, Applied Maths, Sint-Martens-Latem, Belgium) using the Pearson correlation coefficient and UPGMA clustering method. RESULTS Growth profiling and mating type determination Most Dichtomitus squalens strains used in this study have previously been described (Pham et al. 1990), but their genotypes and phenotypes had not been analyzed in detail. The mating type of the strains was verified by crossing them with each other, and resulted with strains of mating type AB, ab, Ab and ab fitting in the previously suggested groups I, II, III and IV, respectively (Table 1). All strains from the study of Pham et al. (1990) were initially tested by growth on glucose, fructose, xylose, galacturonic acid, cellobiose, beechwood xylan, guar gum, apple pectin, and citrus pectin (data not shown). Based on phenotypic differences that suggested different carbon source utilization abilities and compatibility of mating types, a selection of eight monokaryotic strains was made that included representatives of all mating types (AB, Ab, ab, and ab). In addition, the parental dikaryotic strain (FBCC312) of the monokaryons, and a dikaryon FBCC184 which was a separate natural isolate, were included. Finally, another dikaryon, D. squalens CBS , was formed by successfully crossing the compatible monokaryons CBS and CBS (Fig. 1), and was included in the study. Growth profiling of all the studied monokaryotic and dikaryotic strains was performed on 37 carbon sources to reveal the phenotypic differences between them (Supplementary Material Fig. 1). Monokaryon CBS (AB) produced a larger colony diameter than the other monokaryons and dikaryons on all monosaccharides, but not on complex substrates, where both the parental strain FBCC312 and monokaryon CBS (AB) showed a more similar pattern of good growth. On some substrates, such as alfalfa meal, the best growth was observed for the dikaryon FBCC312 (Supplementary Material Fig. 1). Monokaryons CBS (ab), CBS (Ab) and CBS (ab) also showed relatively good growth in most of the substrates compared to the other monokaryotic strains. Conversely, CBS (AB), CBS (ab) and CBS (Ab) showed poor growth in all substrates. The new dikaryon formed in this study (CBS ) had a similar growth profile to the 20 IMA FUNGUS

88 Functional diversity in Dichomitus squalens monokaryons Fig. 2. Clustering of AFLP patterns of Dichomitus squalens isolates. Similarity of the patterns is given in percentage. monokaryons from which it was generated (CBS and CBS ). The parental dikaryon (FBCC312) grew better on most of the carbon sources than the newly formed dikaryon (CBS ) (Supplementary Fig. 1). These two dikaryons (FBCC312 and CBS ) both grew overall better than the non-related dikaryon FBCC184. AFLP analysis reveals high genetic variation between the monokaryotic strains AFLP analysis was conducted to analyze the relationship and genetic variability between the Dichomitus squalens monokaryotic strains and also between the three dikaryons (Fig. 2). Two batches of a single dikaryon from two culture collections, FBCC312 and CBS , clustered together confirming that they are in fact the same strain, which is the parent of the progeny. Monokaryons CBS , CBS , CBS , and CBS , which showed good growth on most carbon sources (Supplementary Fig. 1) were clustered close to each other and to the parental dikaryon FBCC312. Monokaryons that showed poor growth were clustered more distantly to the other monokaryons and the dikaryon FBCC312. The dikaryon FBCC184 positioned further away from the monokaryons and their parental dikaryon (FBCC312), which reflects its origin as it is a separate isolate of this fungal species. Monokaryotic progeny demonstrated high diversity in their enzyme activity profile Since Dichomitus squalens uses wood as its main carbon source in nature, several enzyme activities related to plant biomass degradation were measured in the cultures of the strains grown on spruce sawdust or powdered wheat bran containing liquid medium. The conditions were chosen to compare the behavior of the fungus growing in wheat bran, which is rich in easily degradable polysaccharides, to spruce, which in addition to polysaccharides contains a high amount of lignin and is the natural substrate of D. squalens. These two substrates were also compared in detail in a recent study of FBCC312 (Rytioja et al. 2017) and conditions were chosen to mimic those of the previous study. In spruce medium, most of the enzyme activities related to polysaccharide degradation increased from 9 to 16 d of cultivation (Fig. 3B, D, H, J). The only exceptions were AGL (Fig. 3F), for which the monokaryons CBS and CBS and dikaryon FBCC312 presented higher activity at the earlier time point, and MND (Fig. 3N), for which monokaryon CBS and dikaryon FBCC312 showed higher activity at the earlier time point. In contrast, more variation in the production of the hydrolytic polysaccharide active enzyme activities between the strains was observed during growth on wheat bran. Interestingly, the variation in activity was larger for some enzymes (e.g. ABF, LAC; Figs 3C D and 3K L) than for others (e.g. BGL, CBH, Fig 3A B and 3G H), suggesting a different fine tuning of the regulation in the different strains. Comparing both media, in wheat bran all the activities measured were much higher (from 4 to 10 times higher depending on the enzyme) than in spruce sawdust, including the ligninolytic enzymes (laccase and MnP). The activity patterns of the ligninolytic activities differed from those of the polysaccharide active enzymes (Fig. 3). On both wheat bran and spruce sawdust amended cultures, the laccase and MnP activities were higher at the earlier time point for most of the strains (Fig. 3O R). However, a more notable difference was detected on wheat bran cultures, where much higher activity levels of these two enzymes were produced by the dikaryons than the monokaryons, reaching three-fold higher in some cases, for example dikaryon CBS at the early time point compared to monokaryons CBS , and (Fig. 3P, R). The activities detected for the newly made dikaryon (CBS ) differed from those of FBCC312, which was especially evident for AGL where CBS had significantly higher activity in wheat bran medium at both time points (Fig. 3E F). Also in spruce sawdust it demonstrated a much higher increase of activity over time compared to FBCC312 for which AGL activity decreased. For laccase and MnP, the difference was more evident in Norway spruce sawdust where CBS showed higher activity at the earlier time point (Fig. 3P R). This indicates that the two monokaryons derived from FBCC312 did not re-combine to a strain with the same capabilities as FBCC312. Differences were observed in the activity level as well as the relative activity between the two time points studied. Finally, there was generally higher ligninolytic activity for five of the monokaryons (except CBS , CBS and CBS ) and the three dikaryons at the early time point compared to polysaccharide active enzymes. VOLUME 8 NO. 1 21

89 Casado López et al. Fig. 3. Extracellular enzyme activities detected in the culture liquids of the Dichomitus squalens strains after 9 and 16 days of growth. BGL in wheat bran (A) and Norway spruce (B); ABF in wheat bran (C) and Norway spruce (D); AGL in wheat bran (E) and Norway spruce (F); CBH in wheat bran (G) and Norway spruce (H); BXL in wheat bran (I) and Norway spruce (J); LAC in wheat bran (K) and Norway spruce (L); MND in wheat bran (M) and Norway spruce (N); laccase in wheat bran (O) and Norway spruce (P); MnP in wheat bran (Q) and Norway spruce (R). Vertical bars represent standard deviation of at least four replicate measurements. DISCUSSION Fungi with a sexual reproductive cycle have the possibility to alter their genetic information through recombination during meiosis (Kothe 1996). Therefore, dikaryotic species, such as Dichomitus squalens, can produce monokaryotic offspring with diverse genetic combinations. Strain improvement by cycles of basidiome production and crosses, without the 22 IMA FUNGUS

90 Functional diversity in Dichomitus squalens monokaryons Fig. 3. (Continued). need for mutagenesis, is a feasible approach to improve the ability of white-rot fungi to degrade lignin (Wyatt & Broda 1995). In this study, we explored this variation by analyzing the genetic variation, carbon source usage profile and enzyme activity profile of eight monokaryotic strains derived from a single dikaryon of D. squalens and compared these to the parental dikaryon, a new dikaryon generated from two of these monokaryons, and an unrelated dikaryon that was isolated in nature. Regarding the growth profile, D. squalens monokaryon CBS (AB) seems to have inherited an improved system to use mono- and oligosaccharides compared to all the other tested strains, including the dikaryons, which is especially evident for the disaccharide cellobiose. This observation is in agreement with a previous study on another white-rot species, Trametes versicolor, which demonstrated that a monokaryon presented a better growth rate than its parental dikaryon on glucose-soy agar and hardwood kraft pulp (Addleman & Archibald 1993). However, the monokaryotic strains of the white-rot fungus Pleurotus ostreatus have been reported to grow markedly slower than their dikaryotic parental strains (Eichlerová & Homolka 1999). No correlation between the mating type, appearance of the colony, and the production of ligninolytic enzyme activity, was detected between the P. ostreatus monokaryons (Eichlerová & Homolka 1999). This is in line with our observations, since D. squalens strains did not have an apparent relation between the improved ability for carbon source utilization and mating type genes since the strains showing better growth on different carbon sources belong to all four mating types: CBS (AB), CBS (ab), CBS (Ab), and CBS (ab). These strains clustered close to each other in the AFLP analysis and to the parental strain (FBCC312), suggesting that they inherited common genetic abilities that VOLUME 8 NO. 1 23

91 Casado López et al. cause the improved growth. Moreover, monokaryon CBS (AB) showed poor growth in all carbon sources and, to a lesser extent this was also observed for monokaryons CBS (ab) and CBS (Ab), indicating that poor growth is also not linked to the mating type. These strains also clustered more distantly from the parental strain in the AFLP analysis. Despite the improved ability to utilize monoand oligosaccharides by the monokaryon CBS (AB), its abilities to grow on complex substrates were similar to the parental strain. The newly made D. squalens dikaryon (CBS ) showed a different enzyme activity profile than its parental dikaryon FBCC312. Previously, significantly improved properties, including enzyme activity, growth rate, and chemical conversion abilities, compared to the parental dikaryon have been reported from white-rot fungal dikaryons derived from compatible monokaryons (Eichlerová & Homolka 1999). In addition to lignin-active enzymes, white-rot fungi also possess a full arsenal of enzymes that decompose the other wood polymers (Nagy et al. 2017). This was also evident from our data, since during growth in liquid media with wheat bran (low lignin content) the D. squalens strains produced much higher levels of polysaccharide-related enzymes compared to liquid medium with spruce sawdust (high lignin content). The observation confirms a previous report that lignin can be a barrier for the utilization of the polysaccharides, and that initially the energy of the fungus is directed towards enzymes necessary to degrade the lignin to enable access to polysaccharides (Rytioja et al. 2017). Partial degradation of easily accessible hemicelluloses likely provides the energy for lignin degradation (Blanchette 1984), while production of polysaccharide degrading enzymes increases gradually when these polysaccharides become more accessible by removal of lignin (Pandey & Singh 2014). This has also been observed for other basidiomycete white-rot fungi (Patyshakuliyeva et al. 2015, Kuuskeri et al. 2016). The results of our study also show that white-rot fungi in both monokaryotic and dikaryotic forms use a similar strategy for plant biomass degradation. Our results also suggest a highly diverse ability of the mono- and dikaryotic strains of D. squalens to degrade plant biomass related substrates. This is in line with results from monokaryotic strains of the white-rot fungi Pycnoporus cinnabarinus (Herpoël et al. 2000) and P. sanguineus (Lomascolo et al. 2002) producing higher laccase activity, and P. ostreatus (Eichlerová et al. 2002) and Trametes hirsuta (Li et al. 2012) producing higher laccase and MnP activity than the parental dikaryon. This not only has implications for the efficiency of different strains of a fungal species in biomass decomposition in natural biotopes, but also indicates the importance of strain selection for biotechnological applications (Eichlerová et al. 2003). This has been shown in previous studies by demonstrating that, for example, isolates of the white-rot fungus Ceriporiopsis subvermispora were superior to those of Phanerochaete chrysosporium for biopulping (Blanchette et al. 1992). In fact, isolation of basidiosporederived monokaryotic strains has been an efficient method to obtain new white-rot fungal strains with high variation in the production of enzymes involved in lignin modification and degradation (Eichlerová-Voláková & Homolka 1997, Santoyo et al. 2008). In future studies, we plan to further dissect the mechanisms underlying these differences by analyzing genomes, transcriptomes, and proteomes, of selected D. squalens strains. This may reveal whether the observed differences are mainly at the level of genome content, for example differences in plant cell wall active enzyme encoding gene repertoire, or at the post-genomic or regulatory level. The number of genome-sequenced white-rot fungi has increased significantly in the last few years (Rytioja et al. 2014, Floudas et al. 2015, Nagy et al. 2016, 2017) and in several, but not all, cases monokaryotic strains have been sequenced. However, the variation between two genomesequenced monokaryotic strains of Pleurotus ostreatus that were produced by dedikaryotization of the parental strain was exemplified in their transcriptome analysis. The functionally annotated genes were categorized into groups of unknown functions, glycosyl hydrolases, and redox enzymes, and the relative importance of these groups was shown to differ between the monokaryotic strains (Alfaro et al. 2016). 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Journal of Chemical Technology & Biotechnology 87: Lomascolo A, Cayol J-L, Roche M, Guo L, Robert J-L, et al. (2002) Molecular clustering of Pycnoporus strains from various geographic origins and isolation of monokaryotic strains for laccase hyperproduction. Mycological Research 106: Mäkelä MR, Galkin S, Hatakka A, Lundell TK (2002) Production of organic acids and oxalate decarboxylase in lignin-degrading white rot fungi. Enzyme and Microbial Technology 30: Mäkelä MR, Hildén KS, de Vries RP (2014) Degradation and modification of plant biomass by fungi. In: The Mycota. Vol. 13. Fungal Genomics (Nowrousian M ed.): nd edn. Berlin: Springer-Verlag. Nagy LG, Riley R, Bergmann PJ, Krizsán K, Martin FM, et al. (2017) Genetic bases of fungal white rot wood decay predicted by phylogenomic analysis of correlated gene-phenotype evolution. Molecular Biology and Evolution 34: Nagy LG, Riley R, Tritt A, Adam C, Daum C, et al. (2016) Comparative genomics of early-diverging mushroom-forming fungi provides insights into the origins of lignocellulose decay capabilities. Molecular Biology and Evolution 33: Pandey VK, Singh MP (2014) Biodegradation of wheat straw by Pleurotus ostreatus. Cellular and Molecular Biology 60: Patyshakuliyeva A, Post H, Zhou M, Jurak E, Heck AJR, et al. (2015) Uncovering the abilities of Agaricus bisporus to degrade plant biomass throughout its life cycle. Environmental Microbiology 17: Pham TTT, Maaroufi A, Odier E (1990) Inheritance of celluloseand lignin- degrading ability as well as endoglucanase isozyme pattern in Dichomitus squalens. Applied Microbiology and Biotechnology 33: Raudaskoski M, Kothe E (2010) Basidiomycete mating type genes and pheromone signaling. Eukaryotic Cell 9: Riley R, Salamov AA, Brown DW, Nagy LG, Floudas D, et al. (2014) Extensive sampling of basidiomycete genomes demonstrates inadequacy of the white-rot/brown-rot paradigm for wood decay fungi. Proceedings of the National Academy of Sciences, USA 27: Rytioja J, Hildén K, Di Falco M, Zhou M, Aguilar-Pontes MV, et al. (2017) The molecular response of the white-rot fungus Dichomitus squalens to wood and non-woody biomass as examined by transcriptome and exoproteome analyses. Environmental Microbiology. DOI: / Rytioja J, Hildén KS, Yuzon J, Hatakka A, de Vries RP et al. (2014) Plant-polysaccharide-degrading enzymes from basidiomycetes. Microbiology and Molecular Biology Reviews 78: Santoyo F, González AE, Terrón MC, Ramírez L, Pisabarro AG (2008) Quantitative linkage mapping of lignin-degrading enzymatic activities in Pleurotus ostreatus. Enzyme and Microbial Technology 43: Wyatt AM, Broda P (1995) Informed strain improvement for lignin degradation by Phanerochaete chrysosporium. Microbiology 141: VOLUME 8 NO. 1 25

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94 doi: /imafungus IMA FUNGUS 8(1): (2017) New species of Tulasnella associated with terrestrial orchids in Australia Celeste C. Linde 1, Tom W. May 2, Ryan D. Phillips 1, Monica Ruibal 1, Leon M. Smith 1, and Rod Peakall 1 1 Ecology and Evolution, Research School of Biology, The Australian National University, Canberra, ACT 2601, Australia; corresponding author celeste.linde@anu.edu.au 2 Royal Botanic Gardens Victoria, Birdwood Ave, South Yarra VIC 3141, Australia Abstract: Recent studies using sequence data from eight sequence loci and coalescent-based species delimitation methods have revealed several species-level lineages of Tulasnella associated with the orchid genera Arthrochilus, Caleana, Chiloglottis, and Drakaea in Australia. Here we formally describe three of those species, Tulasnella prima, T. secunda, and T. warcupii spp. nov., as well as an additional Tulasnella species associated with Chiloglottis growing in Sphagnum, T. sphagneti sp. nov. Species were identified by phylogenetic analyses of the ITS with up to 1.3 % sequence divergence within taxa and a minimum of 7.6 % intraspecific divergence. These new Tulasnella (Tulasnellaceae, Cantharellales) species are currently only known from orchid hosts, with each fungal species showing a strong relationship with an orchid genus. In this study, T. prima and T. sphagneti associate with Chiloglottis, while T. secunda associates with Drakaea and Caleana, and T. warcupii associates with Arthrochilus oreophilus. Key words: host specificity ITS sequencing orchid mycorrhizas species delimitation Article info: Submitted: 23 November 2016; Accepted: 6 February 2017; Published: 10 March INTRODUCTION Tulasnella is a cosmopolitan saprotrophic fungal genus that often forms a mycorrhizal relationship with orchids. There are approximately 90 species epithets in Tulasnella ( fungorum.org) with Kirk et al. (2008) indicating that there are approximately 50 accepted species in the genus. Asexual morphs of Tulasnella were formerly referred to in Epulorhiza. In earlier studies on the genus, Warcup and Talbot (Warcup & Talbot 1967, Warcup 1971, 1981) were able to induce formation of basidia and basidiospores from some Australian orchid-derived cultures by placing a casing of soil over cultures on agar. However, the spore-producing tissues were often slow to form and diffuse. Indeed, sporophores could only be detected by examination under a dissecting microscope. In some cases, such as in T. calospora (Warcup & Talbot 1967), only spores were visible above the casing soil surface. Unfortunately, subsequent studies on Tulasnella have not been able to generate basidiospore formation (Suarez et al. 2006, Cruz et al. 2011). For example, Ma et al. (2003) noted that despite repeated attempts, none of the epulorhiza-like Rhizoctonia isolates produced hymenia or basidiospores on [various media] after two months. Although Warcup and Talbot (Warcup & Talbot 1967, Warcup 1971, Warcup & Talbot 1980) utilized morphological characters of the sporophores (such as the size and shape of basidiospores) for taxonomic treatments of Tulasnella from orchids, recent studies on the group have mostly designated operational taxonomic units (OTUs) based solely on phylogenetic analysis of DNA sequence data. Indeed, numerous molecular OTUs have been designated amongst Tulasnella associated with orchids (e.g. Smith et al. 2010, Jacquemyn et al. 2011, 2012, Pandey M et al. 2013, Cruz et al. 2014, Oja et al. 2015) or liverworts (Kottke et al. 2003, Bidartondo & Duckett 2010), without formally naming the species. Formal naming of the species is preferred and essential to prevent confusion of taxonomic units discovered in separate studies (Hibbett & Taylor 2013). Molecular OTUs within Tulasnella have been designated by two methods. First, application of a sequence divergence threshold for a barcode DNA region such as the ITS; with thresholds ranging from 3 5 % (Suarez et al. 2006, Cruz et al. 2014, Jacquemyn et al. 2014, 2015). Second, application of a multi-gene concordance analysis utilizing coalescent theory that explicitly incorporates gene tree conflicts into a model of phylogenetic history for the populations or species concerned (Yang & Rannala 2010, Fujita et al. 2012) and utilizing a number of independent DNA loci (Linde et al. 2014). The second approach is more rigorous for delimiting species (Taylor et al. 2000) and the similarity within and between species delimited with coalescence can be used to calibrate the cut-off threshold used in the first method. A study of Tulasnella isolates from Australian terrestrial orchids (Orchidaceae, tribe Diurideae, subtribe Drakaeinae) in the genera Arthrochilus, Chiloglottis, Drakaea, and Paracaleana (Linde et al. 2014), using eight loci analysed by a variety of methods (including phylogenies of individual loci, Bayesian coalescent based species delimitation, and population structure analysis) revealed five phylogenetic species: one associated with Chiloglottis, one with Drakaea and Paracaleana, and three with Arthrochilus (among which one was known from one isolate and another from two isolates). Analysis of the ITS alone recovered the same five 2017 International Mycological Association You are free to share - to copy, distribute and transmit the work, under the following conditions: Attribution: You must attribute the work in the manner specified by the author or licensor (but not in any way that suggests that they endorse you or your use of the work). 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95 Linde et al. phylogenetic species as well-separated and well-supported clades, revealing congruence between the widely used ITS region and the more extensive multi-locus analysis (Linde et al. 2014). The phylogenetic species were not formally named in Linde et al. (2014). Many of the orchid species associated with Tulasnella are rare or endangered (Hopper & Brown 2006), and the association between orchid and fungus has been and continues to be the subject of much research in Australia (Smith et al. 2010) and elsewhere (McCormick & Jacquemyn 2014). For Tulasnella associated with orchids identification by use of sequences is now the norm, rather than using cultural characters or features of the sporophore. It is therefore appropriate to supply formal names to three of the phylogenetic species (each known from more than two strains) already characterised on sequence data by Linde et al. (2014), along with a further phylogenetic species isolated from Chiloglottis associated with Sphagnum. After assessing information on Tulasnella from Australia, to determine if prior names exist for phylogenetic lineages, we describe four new species of Tulasnella here: T. prima and T. sphagneti spp. nov. from Chiloglottis, T. secunda sp. nov. from Drakaea and Caleana (inclusive of Paracaleana), and T. warcupii sp. nov. from Arthrochilus oreophilus. MATERIALS AND METHODS Fungal collections Taxonomy of the orchid genera, which are all members of the subtribe Drakaeinae, follows Miller & Clements (2014), who accepted the genera Arthrochilus, Caleana (inclusive of Paracaleana), Chiloglottis (inclusive of Simpliglottis), and Drakaea. Tulasnella mycorrhizal associations as identified from previous studies on associations with Australian terrestrial orchids in Arthrochilus, Caleana (as Paracaleana), Chiloglottis and Drakaea (Roche et al. 2010, Phillips et al. 2011, Linde et al. 2014, Phillips et al. 2014), were investigated. Additionally, we treat a Tulasnella isolated from Chiloglottis aff. valida and C. turfosa growing in Sphagnum hummocks within the Kosciuszko National Park, New South Wales (Table 1). Some Chiloglottis orchids growing in Sphagnum were not in flower at the time of collection, and are thus referred to as Chiloglottis sp. However, based on previous studies the Chiloglottis species involved are either C. aff. valida, C. valida, or C. turfosa (Peakall et al. 2010, Peakall & Whitehead 2014). Literature on Tulasnella from Australia was reviewed, and this literature along with GenBank and culture collection databases: CBS (CBS-KNAW Fungal Biodiversity Centre culture collection) and MAFF (culture collection, Ministry of Agriculture, Forestry and Fisheries, Tsukuba, Ibaraki, Japan, searched via NIAS [National Institute of Agro- Environmental Sciences] Genebank - go.jp/databases-micro_search_en.ph) were searched for isolates of Tulasnella from Australia (Tables 2 and 3). Fungal isolation Isolations were made within 7 d of the field collection of the plant tissue using a modified version of the protocol of Roche et al. (2010). We used two types of isolation media to grow mycorrhizal isolates: Fungal Isolation Media (FIM; Clements & Ellyard 1979) and 3MN+A-Z, which is a Melin-Norkrans medium (low CN MMN) (Wright et al. 2010) modified with 15g/L agar and human vitamin and mineral supplements (Centrum Balanced Formula, Wyeth Consumer Healthcare, Baulkham Hills, NSW, Australia) instead of thiamine. One Centrum tablet was dissolved in 100 ml water, filter sterilised, and 10 ml added per litre of 3 MN medium (post autoclaving). Peloton-rich tissues (collars) of orchids were washed several times with sterile distilled water after which the tissue was macerated in sterile distilled water to release pelotons, which were plated onto agar plates containing antibiotics (FIM + tetracycline 25 mg/ml, and 3MN+A-Z + streptomycin 50 mg/ml). Germinating pelotons were transferred to either FIM or 3MN+A-Z media after 3 10 d. The medium chosen depended on which the pelotons germinated. After 3 4 wk all colonies were hyphal-tipped and subcultured to ensure colonies consisted of a single genotype. Cultures were stored at 5 C on FIM or 3MN+A-Z agar slants covered with mineral oil. Voucher specimens of the fungi, as dried-down liquid cultures, are lodged at the National Herbarium of Victoria (MEL) and ex-type cultures are stored in the culture collection of the Department of Agriculture, Victoria (VPRI). DNA extraction, sequencing and phylogenetic analysis Small agar blocks cut from colony edges of isolates were briefly homogenised in 2 ml screw-cap tubes containing sterilise distilled water and glass beads. The blocks were homogenised in a FP120 (Thermo Scientific, Milford, MA) homogenizer for 5 s at 5.5 m/s. Petri dishes containing either half strength FIM or 3MN+A-Z broths were inoculated with the homogenised agar blocks and incubated at room temperature (approximately 23 C) in the dark. Mycelium was harvested, stored at -4 C, and lyophilized prior to DNA extraction. DNA extractions of the lyophilized-mycelium were performed using Qiagen DNeasy Plant Mini Kit or DNeasy 96 Plant Kit according to the manufacturer s instructions (Amersham Biosciences, Hilden, Germany). As previously noted, in a comprehensive evaluation of eight nuclear and mitochondrial loci, Linde et al. (2014) sequenced Tulasnella isolates from orchids in the genera Arthrochilus, Caleana (as Paracaleana), Chiloglottis, and Drakaea. That study showed that within Tulasnella a single locus, ITS (nucr ITS), revealed congruent species delimitation and phylogenetic outcomes. Therefore, for the phylogenetic analysis of additional Tulasnella isolates from Chiloglottis, we only employed ITS. ITS sequences were amplified with the primers ITS1 and ITS4 (White et al. 1990) following methods described in Roche et al. (2010) for the PCR reaction, thermal cycling, purification of PCR and extension products. Products were sequenced bi-directionally with ABI PRISM BigDye Terminator v. 3.1 sequencing kit (Applied Biosystems, Foster City, CA) on an ABI-3130 automated sequencer. Sequences were edited using the program Sequencher v. 4.7 (GeneCodes, Ann Arbor, MI to correct for base read ambiguities. Our sequences were aligned with the most similar sequences available from GenBank ( ncbi.nlm.nih.gov). Alignments were performed in Geneious v. 8 ( Kearse et al. 2012) using 28 IMA FUNGUS

96 New orchid associated Tulasnella species Table 1. Tulasnella isolates examined in this study. Identity Isolate no. Type Host GPS, if known Origin Habitat Collectors* GenBank accession no. Tulasnella sphagneti (CLM541) (CLM583) Holotype Chiloglottis aff. valida Chiloglottis turfosa Chiloglottis sp _1 Chiloglottis sp _2 Chiloglottis sp _1 Chiloglottis turfosa 13102_2 Chiloglottis turfosa Chiloglottis sp Chiloglottis sp. Tulasnella prima CLM159 Holotype Chiloglottis trilabra II.2 Chiloglottis seminuda CLM306 Chiloglottis formicifera CLM308 Chiloglottis formicifera CLM309 Chiloglottis formicifera CLM310.1 CLM310.2 Chiloglottis aff. jeanesii Chiloglottis aff. jeanesii CLM316.1 Chiloglottis seminuda CLM316.2 Chiloglottis seminuda CLM346 Chiloglottis reflexa CLM361 Chiloglottis diphylla S, E S, E S, E S, E S, E S, E S, E S, E S, E S, E S, E S, E S, E S, E S, E S, E S, E S, E S, E S, E Kosciuszko NP, NSW Kosciuszko NP, NSW Kosciuszko NP, NSW Kosciuszko NP, NSW Kosciuszko NP, NSW Kosciuszko NP, NSW Kosciuszko NP, NSW Kosciuszko NP, NSW Kosciuszko NP, NSW Alpine Sphagnum hammocks Alpine Sphagnum hammocks Alpine Sphagnum hammocks Alpine Sphagnum hammocks Alpine Sphagnum hammocks Alpine Sphagnum hammocks Alpine Sphagnum hammocks Alpine Sphagnum hammocks Alpine Sphagnum hammocks Reference YT KY This study YT KY This study YT KY This study YT KY This study YT KY This study YT KY This study YT KY This study YT KY This study YT KY This study Blue Mountains, NSW Eucalyptus woodland RP KF Exeter, NSW Eucalyptus woodland RP HM Upper Kangaroo Valley, NSW Upper Kangaroo Valley, NSW Kanangra Boyd NP, NSW Tallaganda State Forest, NSW Tallaganda State Forest, NSW Eucalyptus woodland RP KF Eucalyptus woodland RP KF Eucalyptus woodland RP KF Eucalyptus woodland RP HM Eucalyptus woodland RP HM Exeter, NSW Eucalyptus woodland RP HM Exeter, NSW Eucalyptus woodland RP HM Mt Wilson, NSW Eucalyptus woodland RP HM Bilpin, NSW Eucalyptus woodland RP HM (Roche et al. 2010) (Roche et al. 2010) (Roche et al. 2010) (Roche et al. 2010) (Roche et al. 2010) (Roche et al. 2010) (Roche et al. 2010) (Roche et al. 2010) (Roche et al. 2010) (Roche et al. 2010) (Roche et al. 2010) VOLUME 8 NO. 1 29

97 Linde et al. Table 1. (Continued). Identity Isolate no. Type Host GPS, if known Origin Habitat Collectors* GenBank accession no. CLM366 CLM371 CLM372 CLM377 CLM380.1 CLM380.2 CLM381.1 CLM381.2 CLM388 CLM389 CLM390 CLM391 Chiloglottis trapeziformis Chiloglottis trapeziformis Chiloglottis trapeziformis Chiloglottis aff. jeanesii Chiloglottis aff. jeanesii Chiloglottis aff. jeanesii Chiloglottis aff. jeanesii Chiloglottis aff. jeanesii Chiloglottis aff. jeanesii Chiloglottis aff. jeanesii Chiloglottis aff. jeanesii Chiloglottis aff. jeanesii CLM393 Chiloglottis valida CLM395 Chiloglottis valida CLM405 SRBG01.II.3 CLM407 CLM416 SRBG03.I.8 Chiloglottis trapeziformis Chiloglottis trapeziformis Chiloglottis trapeziformis Chiloglottis trapeziformis Chiloglottis trapeziformis S, E S, E S, E S, E S, E S, E S, E S, E S, E S, E S, E S, E S, E S, E S, E S, E S, E S, E S, E Black Mountain, ACT Eucalyptus woodland CCL HM Black Mountain, ACT Eucalyptus woodland CCL HM Black Mountain, ACT Eucalyptus woodland CCL HM Kanangra Boyd NP, NSW Kanangra Boyd NP, NSW Kanangra Boyd NP, NSW Kanangra Boyd NP, NSW Kanangra Boyd NP, NSW Kanangra Boyd NP, NSW Kanangra Boyd NP, NSW Kanangra Boyd NP, NSW Kanangra Boyd NP, NSW Tallaganda State Forest, NSW Kanangra Boyd NP, NSW Eucalyptus woodland RP HM Eucalyptus woodland RP HM Eucalyptus woodland RP HM Eucalyptus woodland RP HM Eucalyptus woodland RP HM Eucalyptus woodland RP HM Eucalyptus woodland RP HM Eucalyptus woodland RP HM Eucalyptus woodland RP HM Eucalyptus woodland RP HM Eucalyptus woodland RP HM ANBG, ACT Eucalyptus woodland SR HM ANBG, ACT Eucalyptus woodland SR HM ANBG, ACT Eucalyptus woodland SR HM Black Mountain, ACT Eucalyptus woodland SR HM ANBG, ACT Eucalyptus woodland SR HM Reference (Roche et al. 2010) (Roche et al. 2010) (Roche et al. 2010) (Roche et al. 2010) (Roche et al. 2010) (Roche et al. 2010) (Roche et al. 2010) (Roche et al. 2010) (Roche et al. 2010) (Roche et al. 2010) (Roche et al. 2010) (Roche et al. 2010) (Roche et al. 2010) (Roche et al. 2010) (Roche et al. 2010) (Roche et al. 2010) (Roche et al. 2010) (Roche et al. 2010) (Roche et al. 2010) 30 IMA FUNGUS

98 New orchid associated Tulasnella species Table 1. (Continued). Identity Isolate no. Type Host GPS, if known Origin Habitat Collectors* GenBank accession no. Reference SRBG03.III.12 SRBG03.IV.5 Chiloglottis trapeziformis Chiloglottis trapeziformis CLM068 Chiloglottis diphylla S, E S, E S, E ANBG, ACT Eucalyptus woodland SR HM ANBG, ACT Eucalyptus woodland SR HM Blue Mountains, NSW Eucalyptus woodland RP KF (Roche et al. 2010) (Roche et al. 2010) (Roche et al. 2010) Tulasnella secunda CLM009 Holotype Drakaea elastica NA Karnup, WA Kunzea woodland RDP KF (Linde et al. 2014) S CLM222 Paracaleana minor Black Mountain, ACT Eucalyptus woodland CCL KF E (Linde et al. 2014) CLM251 Drakaea concolor NA Mt Gregory, WA Sandplain heath RDP KF (Linde et al. 2014) CLM253 Drakaea confluens NA Lake Gnartaminny, Jarrah forest RDP KF WA (Linde et al. 2014) CLM255 Drakaea livida NA Walpole WA Jarrah forest RDP KF (Linde et al. 2014) CLM257 Drakaea glyptodon NA Moore River, WA Banksia woodland RDP KF (Linde et al. 2014) CLM258 Drakaea glyptodon NA Margaret River, WA Eucalyptus woodland RDP KF (Linde et al. 2014) CLM259 Drakaea glyptodon NA Ruabon, WA Kunzea woodland RDP KF (Linde et al. 2014) CLM260 Drakaea elastica NA Nambeelup, WA Kunzea woodland RDP KF (Linde et al. 2014) CLM261 Drakaea gracilis NA Westdale, WA Eucalyptus woodland RDP KF (Linde et al. 2014) CLM266 Drakaea confluens NA Lake Gnartaminny, Jarrah forest RDP KF WA (Linde et al. 2014) CLM267 Paracaleana NA Talbot West, WA Eucalyptus woodland RDP KF hortiorum (Linde et al. 2014) CLM268 Paracaleana NA Mt Gregory, WA Sandplain heath RDP KF terminalis (Linde et al. 2014) CLM272 Paracaleana lyonsii NA Eurardy, WA Sandplain heath RDP KF (Linde et al. 2014) CLM273 Drakaea livida NA Canning Mills, WA Jarrah forest RDP KF (Linde et al. 2014) CLM274 Paracaleana triens NA Talbot West, WA Kunzea woodland RDP KF (Linde et al. 2014) CLM276 Drakaea isolata NA Lake Chinocup, WA Mallee woodland RDP KF (Linde et al. 2014) CLM277 Drakaea gracilis NA Westdale, WA Eucalyptus woodland RDP KF (Linde et al. 2014) I3DLIsl6 Drakaea livida NA Walpole, WA Jarrah forest RDP HQ B9DGDen1 Drakaea glyptodon NA Denbarker WA Jarrah forest RDP HQ JS4 Drakaea glyptodon NA Southern WA JX (Phillips et al. 2011) (Phillips et al. 2011) (Sommer et al. 2012) VOLUME 8 NO. 1 31

99 Linde et al. Table 1. (Continued). Identity Isolate no. Type Host GPS, if known Origin Habitat Collectors* GenBank accession no. Reference Tulasnella warcupii CLM027 Holotype CLM007 CLM022 CLM028 CLM091 CLM092 Unassigned CLM084 Unassigned CLM085 Unassigned CLM031 Arthrochilus oreophilus Arthrochilus oreophilus Arthrochilus oreophilus Arthrochilus oreophilus Arthrochilus oreophilus Arthrochilus oreophilus Arthrochilus oreophilus Arthrochilus oreophilus Arthrochilus oreophilus S, E S, E S, E S, E S, E S, E E S E S E S Unassigned BB0002_2_A Terrestrial orchid Australia ANBG = Australian National Botanic Gardens; NP = National Park. *CCL = Celeste Linde; YT = Yann Triponez; RP = Rod Peakall; RDP = Ryan Phillips; DG = Don Gomez. Atherton Tablelands, Qld Atherton Tablelands, Qld Atherton Tablelands, Qld Atherton Tablelands, Qld Atherton Tablelands, Qld Atherton Tablelands, Qld Atherton Tablelands, Qld Atherton Tablelands, Qld Atherton Tablelands, Qld Eucalyptus woodland DG KF Eucalyptus woodland DG KF Eucalyptus woodland DG KF Eucalyptus woodland DG KF Eucalyptus woodland DG KF Eucalyptus woodland DG KF Eucalyptus woodland DG KF Eucalyptus woodland DG KF Eucalyptus woodland DG KF Howard,C.G. and Clements,M.A. (Linde et al. 2014) (Linde et al. 2014) (Linde et al. 2014) (Linde et al. 2014) (Linde et al. 2014) (Linde et al. 2014) (Linde et al. 2014) (Linde et al. 2014) (Linde et al. 2014) (Linde et al. 2014) JN Unpublished 32 IMA FUNGUS

100 New orchid associated Tulasnella species Table 2. Isolates of Tulasnella (some as Tulasnellaceae or Epulorhiza) from Australian orchids, additional to those analysed from orchid hosts in Drakaeinae by Roche et al. (2010) and Linde et al. (2014). All isolates collected by Warcup currently present in culture collections are shown, along with all isolates from which sequences have been obtained. Orchid hosts in Drakaeinae are in bold. Note that AY derived from isolate PN1 from Pterosylis nutans is given in GenBank as asexual morph: Epulorhiza repens, but Bougoure et al. (2005) considered the isolate was most likely a Thanatephorus (and it is therefore omitted below). Tulasnella species T. asymmetrica Thelymitra nuda T. asymmetrica Thelymitra luteocilium Host Original isolate Reference for original isolate Culture collection strain JH Warcup 0267 (Warcup 1973) MAFF AFTOL ID 1678 JH Warcup 085 (Warcup & Talbot 1967) MAFF (ex-type) Sequence ITS* DQ DQ Sequence LSU Reference for sequence Garnica & Weiss (unpub.) (Suarez et al. 2006) KC [clones c001 c006] (Cruz et al. 2014) T. asymmetrica Thelymitra epipactoides JH Warcup 0302 (Warcup 1973) MAFF DQ (Suarez et al. 2006) KC ,51,52,56 [clones c001 c005, c009] (Cruz et al. 2014) T. asymmetrica Thelymitra epipactoides JH Warcup 0591 (Warcup 1973) NIAES 5809 (Bidartondo et al. 2003) MAFF P = NIAES 5809 NIAES 5809 DQ KC , KC152350, KC , [clones c001, c003, c005, c008 c010] (Suarez et al. 2006) (Cruz et al. 2014) T. calospora JH Warcup 07 (Warcup & MAFF no sequences Acianthus Talbot 1967) exsertus T. calospora Diuris JH Warcup 0388 (Warcup 1973) MAFF no sequences maculata T. calospora Thelymitra JH Warcup 0584 (Warcup 1973) MAFF no sequences aristata T. calospora Thelymitra sp. JH Warcup 0638 (Warcup 1973) MAFF no sequences T. calospora host not specified T. calospora Caladenia reticulata T. irregularis Dendrobium dicuphum T. calospora Microtis parviflora Tulasnella sp. [presume from orchid] Tulasnella sp. [presume from orchid] Epulorhiza possibly [in GenBank as Fungi ] Acianthus pusillus JH Warcup 0689 (Warcup 1973) MAFF no sequences JH Warcup 062 CBS AY (Taylor et al. 2003) JH Warcup 0632 TM1 (Perkins et al. 1995) CBS [ex type] = JCM 9996 AY AF AY (Kristiansen et al. 2001) (Taylor et al. 2003) (Bougoure et al. 2005) JT Otero 306 DQ Otero (unpub.) JT Otero 307 DQ Otero (unpub.) AP2 AY (Bougoure et al. 2005) VOLUME 8 NO. 1 33

101 Linde et al. Table 2. (Continued). Tulasnella species Epulorhiza sp. Epulorhiza sp. Epulorhiza sp. Host Original isolate Reference for original isolate Diuris corymbosa Prasophyllum giganteum Pyrorchis nigricans Culture collection strain Sequence ITS* Kings_Park_ EF Dm01 Kings_Park_ EF Pg01 7 isolates EF , Epulorhiza sp. Disa bracteata 11 isolates EF , 79 83, 85 Tulasnella sp. terrestrial orchid T. calospora Diuris magnifica Sequence LSU Reference for sequence (Bonnardeaux et al. 2007) (Bonnardeaux et al. 2007) (Bonnardeaux et al. 2007) (Bonnardeaux et al. 2007) BB0002_2_A JN Howard & Clements (unpub.) DR88 KT Davis et al. (unpub) T. calospora Disa bracteata DR28 KT Davis et al. (unpub) T. calospora Microtis media DR126 KT Davis et al. (unpub) Tulasnellaceae sp. RP-2011 Tulasnellaceae sp. 1 3 Tulasnellaceae sp. 4 5 Tulasnellaceae sp. 6 7 Tulasnellaceae sp. 8 9 Tulasnellaceae sp. 10 Tulasnellaceae sp. 11 Tulasnellaceae sp. 12 Tulasnellaceae sp. 13 Tulasnellaceae sp. 14 Tulasnellaceae sp. 16 Tulasnella sp. [as Uncultured mycorrhizal fungus ] Tulasnella sp. [as Uncultured mycorrhizal fungus ] Drakaea elastica, D. glyptodon, D. livida, D. micrantha, D. thynniphila Thelymitra macrophylla 50 isolates HQ (Phillips et al. 2011) JP15, JP44, JP49 JX (Sommer et al. 2012) Disa bracteata JP24, JP26 JX (Sommer et al. 2012) Pyrorchis nigricans Diuris magnifica JP33, JP37 JX (Sommer et al. 2012) JP40, JP60 JX (Sommer et al. 2012) Microtis sp. JP63 JX (Sommer et al. 2012) Drakaea glyptodon Spiculaea ciliata Lyperanthus serratus Microtis capularis JS4 JX (Sommer et al. 2012) JS43 JX (Sommer et al. 2012) JS64 JX (Sommer et al. 2012) JS66, JS68 JX (Sommer et al. 2012) Microtis media JS163 JX (Sommer et al. 2012) Diuris fragrantissima 30 isolates DQ ,86-95 DQ ,84-85 Diuris punctata 7 isolates DQ DQ790763,65, 72, 77,79,82,98 (Smith et al. 2010) (Smith et al. 2010) Tulasnella sp. [as Uncultured mycorrhizal fungus ] Diuris punctata var. daltonii 2 isolates DQ790804,08 DQ790769,73 (Smith et al. 2010) 34 IMA FUNGUS

102 New orchid associated Tulasnella species Table 2. (Continued). Tulasnella species Tulasnella sp. [as Uncultured mycorrhizal fungus ] Tulasnella sp. [as Uncultured mycorrhizal fungus ] Host Original isolate Reference for original isolate Diuris dendrobioides Diuris chryeopsis *some sequences are ITS plus partial LSU. Culture collection strain Sequence ITS* Sequence LSU Reference for sequence 1 isolate DQ DQ (Smith et al. 2010)) 3 isolates DQ790796,99 DQ790761,64, DQ (Smith et al. 2010) Table 3. Tulasnella species isolated from Australian orchids by JH Warcup and PHB Talbot and other studies. Sporophore morphology was from sporophores (i.e. the perfect state ) initiated from cultures. Tulasnella species in bold were newly described by Warcup and Talbot. Orchid genera in Drakaeinae are given in bold. References do not include publications where original isolates of Warcup were later sequenced (as cited in Table 2). Unpublished observations derive from sequences in GenBank as detailed in Table 2. Tulasnella Orchid genera Identification method Comments References species T. allantospora Chiloglottis, Corybas sporophore morphology (Warcup & Talbot 1971, Warcup 1981) T. asymmetrica Chiloglottis, Cryptostylis, Dendrobium, Thelymitra T. calospora Acianthus, Caladenia, Corybas, Cymbidium, Dendrobium, Diuris, Eriochilus, Lyperanthus, Microtis, Orthoceras, Thelymitra sporophore morphology also as Tulasnella sp., isolate 086 (Warcup & Talbot 1967); see Warcup & Talbot (1971) (Warcup & Talbot 1967, 1971, Warcup 1973, 1981) sporophore morphology (Warcup & Talbot 1967, Warcup 1971, 1973, 1981, 1990) Disa, Diuris, Microtis sequence (Bougoure et al. 2005) T. cruciata Acianthus, Chiloglottis, Thelymitra sporophore morphology (Warcup & Talbot 1971, Warcup 1973, 1981, 1990) T. deliquescens Acianthus, Microtis culture morphology (Perkins et al. 1995) [as Epulorhiza repens] T. irregularis Dendrobium sporophore morphology also as Tulasnella sp. 1, isolate 0632 (Warcup 1973); see Warcup & Talbot (1980) (Warcup & Talbot 1980, Warcup 1981) T. violea Drakaea culture morphology (Warcup 1981) Thelymitra sporophore morphology (Warcup & Talbot 1971, Warcup 1973, 1990) Tulasnella sp. (some as Epulorhiza sp. or Tulasnellaceae sp.) Arthrochilus, Caladenia, Caleana, Calochilus, Chiloglottis, Corybas, Cryptostylis, Cymbidium, Dendrobium, Dipodium, Drakaea, Microtis, Thelymitra Acianthus, Caleana (as Paracaleana), Disa, Diuris, Drakaea, Lyperanthus, Microtis, Prasophyllum, Pyrorchis, Spiculaea, Thelymitra culture morphology culturally seem Tulasnella, perfect states not seen Warcup 1973, 1981, 1990, Perkins & McGee 1995, Perkins et al. 1995) sequence (Bougoure et al. 2005, Bonnardeaux et al. 2007, Smith et al. 2010, Phillips et al. 2011, Sommer et al. 2012) VOLUME 8 NO. 1 35

103 Linde et al. ClustalW followed by manual adjustments to optimise indel locations. Sequences for phylogenetic analysis included representatives of species-level clades in one of the two main subclades of phylogenetic group IV of the phylogeny of Tulasnella constructed by Cruz et al. (2011). This subclade contains Tulasnella sp. ECU 6 and T. eichleriana. To these sequences were added a selection of previously sequenced isolates from Australian orchids (Table 1) representing the phylogenetic breadth of the OTUs identified by Linde et al. (2014) along with new sequences from Chiloglottis associated with Sphagnum (Table 1). BLAST matches were carried out for representative sequences of putative OTUs from Australian orchids from our analysis to recover related sequences in GenBank. Phylogenies were estimated using Bayesian inference with MrBayes v (Ronquist & Huelsenbeck 2003) and maximum likelihood (ML) analysis through the RAxML Blackbox (Stamatakis et al. 2008). Support for the nodes was assessed with Bayesian Posterior Probabilities (BPP) in MrBayes and for ML trees using 1000 pseudoreplicates of nonparametric bootstrapping. A GTR+G substitution model was used for all analyses as other models are nested within these. Trees were visualised using FigTree v ( figtree/) and rooted to Tulasnella eichleriana sequences. Trees include identical sequences from different isolates; however the identical sequences were removed when nodes support was assessed. Pairwise sequence divergence of the ITS sequences within and among lineages were estimated with the Kimura-2-parameter distances with gap deletion in MEGA5 (Tamura et al. 2011). RESULTS Tulasnella species from Australian orchids In placing formal names on phylogenetic species of Tulasnella, it is necessary to consider any names from previous work on the genus. Essentially, type specimens (that anchor names) need to be placed into the phylogenetic framework. However, given the lack of diagnostic morphological characters for recently isolated strains, a significant issue is whether types or suitable reference material exists and if sequence data are available for that material. Three species of Tulasnella have been described from Australian orchids: T. cruciata, originally from Acianthus and Dendrobium; T. irregularis from Dendrobium; and T. asymmetrica originally from Thelymitra. May et al. (2003) collated records of Tulasnella from all sources from Australia, including reports of a further four species: Tulasnella allantospora, T. calospora, T. deliquescens, and T. violea. According to Roberts (1994), T. asymmetrica was morphologically indistinguishable from T. pinicola, and was listed by Roberts (1999) as a synonym of the latter species. Furthermore, Roberts (1999) noted that the Australian report of T. allantospora by Warcup & Talbot (1971) was possibly misidentified, and might represent T. rubropallens; and T. calospora in the sense of Warcup & Talbot (1967) was deemed to be T. deliquescens. In making redispositions of Australian Tulasnella names, Roberts (1999) noted that he had not examined type material or voucher collections for reports by Warcup & Talbot (1967, 1971) of T. allantospora and T. calospora. Indeed, type material of T. cruciata or T. irregularis could not be located in ADW (Roberts 1999), and although the type of T. asymmetrica is listed by Roberts (1999) as housed at ADW, it was not examined. Warcup s collections were originally in ADW and subsequently transferred to AD (macrofungi) and DAR (microfungi). There are ex-type cultures of T. asymmetrica (Warcup 085, MAFF ) and T. irregularis (Warcup 0632, CBS = JCM 9996), but apparently none of T. cruciata. Tulasnella isolates have been obtained from 21 terrestrial orchid genera and one lithophytic/epiphytic orchid genus (Dendrobium) in Australia (Table 3) (Warcup & Talbot 1967, 1971, 1980, Warcup 1971, 1973, 1981, 1990). For the genera Arthrochilus, Caleana (or from Paracaleana, under which name Caleana was formerly placed), Chiloglottis, and Drakaea that were the source of the apparently novel phylogenetic species delimited by Linde et al. (2014), the only previous reports are of unidentified Tulasnella isolates. An exception is a report of Tulasnella violea from Drakaea, identified only from cultural characteristics (Warcup 1981). However, for Chiloglottis there are reports of T. allantospora, T. asymmetrica, T. cruciata and also an unidentified species (Warcup 1973, 1981). For the two Tulasnella species described from Australia (T. asymmetrica and T. cruciata), the types are from other orchid genera, and the isolates of these two species from Chiloglottis were collected after the species were described. Phylogenetic analysis of isolates from Arthrochilus, Caleana, Chiloglottis, and Drakaea GenBank BLAST searches using query ITS sequences of Tulasnella isolates in this study revealed these sequences were related to T. eichleriana, T. tomaculum, and two Tulasnella lineages (ECU 5 and ECU 6) isolated from decaying branches in Ecuador. Representative sequences of these four lineages were added to the 72 fungal sequences from the Australian orchid genera Arthrochilus, Chiloglottis, Drakaea, and Caleana. The resulting phylogram show high bootstrap (100 %) and posterior probability (1) support for the three phylogenetic species of Tulasnella from (a) Arthrochilus oreophilus, (b) Chiloglottis, and (c) Drakaea and Caleana, that had previously been delimited on multi-gene data by Linde et al. (2014). Further sequences from Chiloglottis (exclusively associated with Chiloglottis growing in Sphagnum) formed a well-separated clade, sister to the other sequences from Chiloglottis (Fig. 1). ITS sequences from the ex-type culture of T. asymmetrica fall outside of the clades depicted in Fig. 1, as do all other sequences from Australian orchids (data not shown) with the exception of HQ386778, HQ and JX from Drakaea, that fell within the clade of isolates from Drakaea and Caleana and JN from an Australian terrestrial orchid that is sister to the two clades consisting of isolates from Chiloglottis. A number of additional sequences from Drakaea (Phillips et al. 2011) all cluster within the clade from Drakaea with 100% bootstrap support. Those matching sequences were subsequently excluded from the final analysis. The LSU sequence from the ex-type culture of T. irregularis is only distantly related to LSU sequences 36 IMA FUNGUS

104 New orchid associated Tulasnella species T. eichleriana AY T eichleriana KC /1 72/ /1 ECU5 KC ECU5 KC /1 EC6 KC ECU6 KC JN Terrestrial orchid C. turfosa 13102_2 C. turfosa 13102_1 C. turfosa 13065_2 Chiloglottis sp _1 Chiloglottis sp. Tulasnella sphagneti 12033_1 C. aff. valida 100/ Chiloglottis sp _1 Chiloglottis sp _1 Chiloglottis sp. KF C. formicifera 100/1 HM C. aff. jeanesii HM C. aff. jeanesii HM C. aff. jeanesii HM C. aff. jeanesii HM C. aff. jeanesii HM C. aff. jeanesii HM C. trapeziformis HM C. aff. jeanesii HM C. aff. jeanesii HM C. aff. jeanesii HM C. valida CLM306 C. formicifera 100/1 CLM308 C. formicifera HM C. aff. jeanesii HM C. aff. jeanesii Tulasnella prima HM C. valida HM C. seminuda HM C. trapeziformis HM C. trapeziformis KF C. trilabra KF C. diphylla HM C. trapeziformis HM C. trapeziformis HM C. trapeziformis HM C. trapeziformis HM C. reflexa HM C. diphylla HM C. trapeziformis 99/1 HM C. seminuda HM C. trapeziformis HM C. seminuda HM C. trapeziformis KF P. lyonsii KF P. minor KF P. terminalis KF D. elastica 100/1 KF D. gracilis KF D. glyptodon JX D. glyptodon KF D. confluens KF D. gracilis KF D. concolor KF D. livida Tulasnella secunda KF D. glyptodon KF P. hortiorum KF D. isolata KF D. confluens KF D. elastica 0.76 HQ D. livida HQ D. glyptodon KF D. glyptodon KF D. livida KF P. triens KF CLM031 A. oreophilus KF CLM027 A. oreophilus 0.74 KF CLM028 A. oreophilus 1/100 KF CLM092 A. oreophilus KF CLM091 A. oreophilus Tulasnella warcupii 1/100 KF CLM007 A. oreophilus 0.75 KF CLM022 A. oreophilus 1/100 KF CLM084 A. oreophilus KF CLM085 A. oreophilus 1/100 T. tomaculum AY T. tomaculum KC Fig. 1. Rooted MrBayes tree for Tulasnella obtained for ITS. The tree with the highest log likelihood is shown. The numbers above the branches are maximum likelihood bootstrap values/bayesian posterior probabilities. Bootstrap values of 70 % and Bayesian posterior probabilities of 0.70 are shown. The branch length is proportional to the inferred divergence level. Host from which the Tulasnella isolate was collected from is indicated after the isolate number or GenBank number. Sequences from the holotype of each species is indicated in bold. from Australian isolates of Tulasnella from Arthrochilus, Chiloglottis, Drakaea, and Caleana (data not shown). The percentage sequence divergence between the two lineages from Chiloglottis was 6.3 %. Sequence divergence between all other Australian Tulasnella lineages and close relatives ranged from % (Table 4). The natural ITS barcode gap between all Tulasnella lineages studied here is between 4 6 % sequence divergence (Fig. 2). Recognition of novel taxa Support for three of the novel taxa was high across the eight loci analysed by Linde et al. (2014) (Table 5) and all VOLUME 8 NO. 1 37

105 Linde et al. Table 4. Within host group and between host group Kimura -2P distances for Tulasnella as calculated from ITS. All positions containing gaps and missing data were eliminated. There were a total of 601 positions in the final dataset. Within taxa T. prima T. sphagneti T. warcupii T. secunda T. tomaculum T. eichleriana T. ECU5 Tulasnella prima 1.2 ± 0.3 Tulasnella sphagneti 0.1 ± ± 1.0 Tulasnella warcupii 3.8 ± ± ± 1.6 Tulasnella secunda 0.2 ± ± ± ± 1.5 Tulasnella tomaculum ± ± ± ± 1.3 Tulasnella eichleriana 2.2 ± ± ± ± ± ± 1.4 Tulasnella ECU5 0.2 ± ± ± ± ± ± ± 1.4 Tulasnella ECU6 1.5 ± ± ± ± ± ± ± ± 1.6 clades had long subtending basal stems in the phylogenies generated. Base-pair differences and their positions for each lineage are given in Table 6. Therefore we conclude that each can be regarded as a well-supported phylogenetic species. The additional clade consisting of isolates from Chiloglottis associated with Sphagnum was also well-supported in the ITS tree (Fig. 1) and well-separated from the sister clade, above the divergence established between the three phylogenetic species delimited on multi-locus concordance, and is therefore recognised as a fourth phylogenetic species. None of the clades for these four phylogenetic species contain sequences from material of Tulasnella previously described from Australia, or indeed any other sequences of described species in GenBank. In addition, the ITS sequences from ex-type cultures of T. asymmetrica and T. irregularis do not cluster with or are close to any of the four phylogenetic species described here. Therefore, we conclude that these four phylogenetic species are previously unrecognized, and consequently they are formally described below. Two further putative new phylogenetic species (Tulasnella sp. Arthrochilus II and Tulasnella sp. Arthrochilus III) (Linde et al. 2014) that were represented by only two and one isolates respectively, are not formally described here pending discovery of further isolates. Previous morphology-based identifications of various Tulasnella species from hosts in Drakaeinae (Table 3) will all need to be re-visited and confirmed with sequence data, if voucher specimens or cultures still exist. Here we diagnose the new species on the basis of both sequence-based synapomorphies and clade-based definitions from molecular phylogenies (Hibbett et al. 2011, Renner 2016). This is because it is not possible to be certain as to which morphological characters are actually diagnostic. In time, certain morphological features may turn out to be unique for particular taxa, but this can only be known if morphological data are comprehensive across known species of the genus. In addition, it is sequence data that are routinely used to identify isolates of Tulasnella, and hence we are providing both rigorous species delimitation and the means to identify further isolates with certainty. Therefore, our descriptions of the morphology of cultures and of hyphal characters are provided for completeness, rather than as species characteristics. g. 2. Barcode gap; percentage sequence divergence among isolates. The vertical Fig. 2. Barcode gap; percentage sequence divergence among Tulasnella isolates. The vertical arrow indicates the ~3.3 to 5.7 % ITS sequence divergence threshold for this dataset. 38 IMA FUNGUS

106 New orchid associated Tulasnella species Table 5. Presence of and support for clades of six phylogenetic species of Tulasnella from Australian orchids in the genera Arthrochilus, Chiloglottis, Drakaea and Caleana in phylogenetic trees constructed separately for each of eight loci, as indicated on trees presented as Fig. 2 and Supporting information figures S2-S8 of Linde et al. (2014). Values are bootstrap/bayesian posterior probability. +: clade present (support less than BS 70% and BPP 0.80); *: one isolate (CLM417) fell outside of the clade, basal to all other sequences; **: support values are from Fig. 1 of the present work (all other clades in this tree representing the phylogenetic species are also 100/1.00); n=1: one isolate only, falls outside of other clades, and separate to any other singletons; na: not present in analysis. no. isolates ITS mtlsu C4102 C12424 C14436 C3304 C4722 C10499 T. prima / / / / /0.99* 100/ / /1.00 T. sphagneti 9 100/1.00** na na na na na na na T. secunda / / /0/96 96/ / / /1.00 T. warcupii 6 100/ / / /1.00 -/ /1.00 n=1 100/1.00 T. sp. Arthrochilus II 2 100/ / / / / n=1 na T. sp. Arthrochilus III 1 n=1 n=1 n=1 n=1 n=1 n=1 na n=1 TAXONOMY Tulasnella prima Linde & T.W. May, sp. nov. MycoBank MB (Fig. 3A) Etymology: Referring to the first Tulasnella found in the host, Chiloglottis. Type: Australia: New South Wales: Blue Mountains, Mt Werong, Ranger Fire Trail, isolated from Chiloglottis trilabra, 22 Mar. 2007, C.C. Linde & R. Peakall CLM159 (MEL holotype; ex-type culture VPRI 42810). Diagnosis: Tulasnella prima can be diagnosed by the following nucleotide characters, which are fixed between T. tomaculum and T. prima respectively: Locus ITS: ITS1 upstream from the 18S at position 18 (G:T), (TGCT:CTGA), (CG:--), 38 (G:T/A), 41 (A:T/C), 44 (G:T), 58 (T:C), 61 (-:T), 68 (-:T), 80 (G:T), 101 (T:A), 117 (T:C), 123 (G:A), 127 (C:T), 130 (G:T), (CT:TC), 140 (A:T), (AG:TT), 152 (C:T), 156 (T:G), 158 (-:A/G), 163 (A:G), 165 (C:-), 168 (T:C), 180 (G:A/T), 190 (A:T/C), 192 (C:T), (CT:TC), (AC:GT), (TG:CT), (TA:--), 237 )C:A/G). 5.8S starting from ITS1 end: Position 4 (-:T/-), 22 (T:C), 139 (T:C), 141 (T:C), 154 (C:T). ITS2 starting from 5.8S end: Position 14 (T:-), 16 (C:A), 25 (T:C), 27 (A:T/C), (CT:TC), 37 (C:T), 49 (T:C), (CT:TC), 59 (C:T), 69 (A:G), (CA:TG), 75 (T:C), (TCTGA:CTAT/CG), 84 (T:C), 88 (A:G), 91 (G:C/T), 93 (C:T), (GTT:AAA), 105 (A:-), 107 (A:T), (--:CT), (TA:--), 120 (T:C), 126 (T:C), 136 (G:T), 139 (C:T), (AT:GA), (CC:TA/G), 150 (-:C/T), 154 (G:A), 157 (T:G), 163 (C:T), 180 (T:G), 187 (-:T/C), 191 (G:T/C), 215 (T:G), 222 (T:-), 237 (T:-), 241 (-:G/ A), 251 (C:G), 253 (G:A), 256 (G:T, 259 (T:A), 262 (C:T), 285 (G-:AC), (TCCG:CTGC), (CG:TT), (TG:AT), (CG:GT/C), 305 (A:G), (AC:TT), 328 (T:C), 331 (G:A), 338 (G:T). Clade-based diagnosis: The least inclusive clade in the ITS phylogeny in Fig. 1 containing HM and HM Substrate or host: Roots and underground stem-collars of Chiloglottis orchid species. Distribution: High rainfall parts of south-eastern Australia and Tasmania in Eucalyptus woodlands and forests. Current known distribution coincides with that of the Chiloglottis hosts. Notes: Cultures on quarter strength PDA show fine concentric rings. Culture edges lack concentric rings, are broad and diffused. Culture appearance is quite variable, with some cultures showing aerial mycelium. Not all cultures grow on 3MN +A-Z. Hyphae from cultures are cylindrical, 2 5 μm diam, branched, often at right angles, septate, lacking clamp connections; wall slightly thickened (to 0.25 μm); rarely with refractive internal bodies, and then small; sometimes uneven (with undulate outline); sometimes with swollen elements to 9.5 μm diam that are thick-walled (to <0.5 μm thick), clavate, terminal or intercalary, sometimes in short chains. Additional material examined: See Table 1. Tulasnella secunda Linde & T.W. May, sp. nov. MycoBank MB (Fig. 3C) Etymology: Referring to the second Tulasnella described that associates with Drakaeinae orchids. Type: Australia: Western Australia: Paganoni Swamp Reserve, Karnup, isolated from Drakaea elastica, 2008, R.D. Phillips [C.C. Linde CLM009], (MEL holotype; extype culture VPRI 42808). Diagnosis: Tulasnella secunda can be diagnosed by the following nucleotide characters, which are fixed between T. tomaculum and T. secunda respectively. Locus ITS: ITS1 upstream from the 18S at position (TT:-A), 41 (A:C/T), 54 (G:A), 77 (C:T), 128 (G:A), 130 (C:A), 132 (G:T), (-C:TT), (TC:CT), 158 (T:C), 163 (C:-), 166 (T:-), 170 (-:C), 179 (G:A), 181 (C:T), 189 (A:C), 202 (C:A), (AC:--), (CA:AC), 223 (T:C), 238 (T:C). 5.8S no differences. ITS2 starting from 5.8S end: Position 23 (C:T), 27 (A:T), 32 (C:T), (GGC:AAT), 48 (G:A), (CT:T/ VOLUME 8 NO. 1 39

107 Linde et al. Table 6. Basepair differences and their positions, among Tulasnella tomaculum, T. sphagneti, T. prima, T. secunda and T. warcupii. Polymorphic basepair differences in two or more isolates of a species are given by the most common basepair/alternative basepair T. tomaculum G G T G C T C G T T T A G C C G T - - T C G - T T T. sphagneti T. C T G A T C. C C C T... C T C C. T - A. C. prima T. C T G A T T. C. T T... C T T.. T - A. T. secunda.. C A/..... A. C C... A T. -.. T. warcupii. A C.. C. C....../T T C. C T. tomaculum C A C T C G C G G C - T G G C A A G C C G - T C T T. sphagneti T C... A T. - T C C. A. T T. T.. G.. C T. prima T. C C. A T T. T - C../A. T T T T.. G C A. T. secunda A. A -. T. T/..... T T G C T C T. warcupii.... T... T T T -.. C T. tomaculum G A T C T G C T A A C C T T T - A - T C A T A T T T. sphagneti A G C. C A.. T. T T C C. A T T C..... C T. prima. G C../C A/T.. C. T T C../C A T T C A... C C T. secunda.. C T C A T. C.. A... A - - C.. C... T. warcupii.. C. C T. C. G. T. A C A. T.. G. T T. tomaculum T C T C C C C T G A T C T G G C G C G T G C C T C T. sphagneti C T.. A.. C T C. T C A/ T C T T. prima C T.. A.. C A/T T/C. T C A/.. T... C A/. T T C T T. secunda..... T... T. T. A A T.. A... T C. T. warcupii.. C G.. T. A. A..... A T. C.. T C. 40 IMA FUNGUS

108 New orchid associated Tulasnella species T. tomaculum G C G T T A C C A T G T G C A G C C G T T G T. sphagneti.. A. C G.. G. C A - C C.. G. T T. G.. T. prima. T... G. T G C A T -. C. T G T T. A A A./A T. secunda A. A C. G.. G/. C C T C/T..../T G. T T... A T. warcupii. T. C C G T. G - C T -. C A. G A T. tomaculum C G G A G A T G T G T A - G T T T C G C C A T C C T. sphagneti.. T G C T C C A T A G T. C. C. T T. G G T. T. prima. A./T G T T C T G/A C. G T. C. C. T T. G A T/.. T. secunda.... A. C C. T. G - A C.. T.. A G A -. T. warcupii T.. G. G C T. T. G -. C G C.. T. G A. A T. tomaculum C C - - G T G C C C - T C T G A G A T T T C T C T T. sphagneti T A C C A A.... G C T G T G C G.. G. C.. T. prima T A - C A G. T./T. G C T G T. T... G. C T G T. secunda T T C C - C A.. T G /C G. C T G T. warcupii T. - C A C.... G. - C.... C C. T C T. tomaculum T C G G T C - G G T G C C G C T C T C G - G A C T T. sphagneti. G. A.. - T. A C. T A A. A - T. prima. G. A.. - T. A.. T A C A - T. secunda C T. A.. T T. C.... A C T... - A. T G T. warcupii G A T. C G - A A C. T... C T C T A C A T. tomaculum - G C T G T G C - - A - A - C - T C A T G T G T. sphagneti T T T C C. C G - T G G G - - T... C A. T T. prima T T T C A. T G - C G T T... C A C T T. secunda A T T - T G T - - T G -. G. T C T. warcupii - T T C - G T G C G/- G C. T.... VOLUME 8 NO. 1 41

109 Linde et al. A B C D Fig. 3. Tulasnella cultures on quarter strength PDA (left), half strength FIM (middle) and 3MN +A-Z (right) media. A. Tulasnella prima (CLM159); B. T. sphagneti (CLM541); C. T. secunda (CLM009) and D. T. warcupii (CLM027). 42 IMA FUNGUS

110 New orchid associated Tulasnella species AC), 58 (G:A), (GT:AC), 69 (A:G), 73 (T:C), 76 (T:C), 80 (-:C/T), 89 (A:G), 94 (C:T), 96 (C:T), 101 (G:A), 107 (G:A), (TG:CC), 112 (G:-), 115 (A:T/C), (-T:AC), 133 (C:T), 141 (C:-), (TCCC:--GA), 149 (-:T), 151 (T:C), (TG:CA), (-C:TG), 214 (T:G), (TCT:CTG), (TC:CT), 251 (G:A), (G-:TT), 258 (T:C), (CT:AC), 276 (C:T), 284 (G:A), 287 (CT:TG), 291 (G:A), (GC:TT), 300 (C:T), 305 (A:G), 308 (-:C). Clade-based diagnosis: The least inclusive clade in the ITS phylogeny in Fig. 1 containing KF and JX Substrate or host: Underground stem-collars of Caleana and Drakaea orchid species. Distribution: South-western and south-eastern Australia, extending from high rainfall areas to the margin of the arid zone, occurring in open areas within eucalypt forests and woodlands, Banksia woodlands and sandplain heath. Most records are from well-drained grey sandy soils, but also known from yellow sands, laterite, sandy clay soil, etc. Current known distribution coincides with that of Caleana (inclusive of Paracaleana) and Drakaea. Diagnosis: Tulasnella sphagneti can be diagnosed by the following nucleotide characters, which are fixed between between T. tomaculum and T. sphagneti respectively: Locus ITS: ITS1 upstream from the 18S at position 18 (G:T), 23 (-:C), 26 (C:-), 27 (T:A), 33 (C:-), 34 (G:), 39 (G:C), 42 (A:C), 45 (G:T), 59 (T:C), 62 (-:T), 69 (:C), 79 (T:C), 81 (G:T), 102 (T:A), 108 (C:T), 110 (A:C), 124 (G:A), 128 (C:T), 132 (G:T), 134 (T:C), 136 (G:A), 141 (A:T), 145 (A:T), 154 (C:T), 156 (-:G), 161 (:-C), 162 (-:A), 166 (A:-), 167 (T:-), 168 (C:-), 171 (T:C), 174 (T:C), 183 (G:A), 193 (A:T), 195 (C:T), 206 (C:T), 207 (T:C), 216 (C:T), 217 (A:G), 218 (C:T), 220 (T:C), 221 (G:T), 227 (T:), 228 (A:). 5.8S starting from ITS1 end: Position 138 (T:C), 140 (T:C), 153 (C:T). ITS2 starting from 5.8S end: Position 14 (T:), 16 (C:A), 25 (T:C), 26 (G:T), 27 (A:C), 32 (C:T), 33 (T:C), 55 (C:T), 56 (T:C), 57 (G:T), 60 (G:A), 64 (T:C), 69 (A:G), 72 (A:G), 77 (-:C), 80 (T:A), 81 (G:C), 85 (T:C), 89 (A:G), 94 (C:T), 96 (C:T), 98 (T:G), 104 (G:T), (AGATGTGTTA:GCTCCATAGT), 118 (T:C), 124 (T:C), 134 (G:T), 137 (C:T), (-A:GG), (-CC:TAT), 149 (T:C), 153 (G:A), 156 (T:A), 179 (T:G), (CA:TC), (A-:CG), 214 (T:G), 221 (T:), 236 (T:C), 249 (C:G), 251 (G:A), 254 (G:T), (TG:AC), 260 (C:T), 264 (G:A), 283 (G:A), (CT:AC), (G-- :TTC), (GT:CG), 301 (C:T), (AA:GG), 307 (C:T), 327 (T:C), 330 (G:A), 337 (G:T). Notes: This taxon was referred to as Tulasnellaceae sp. RP by Phillips et al. (2011). Cultures often have a rosepink colour due to bacterial associates that are not affected by streptomycin in the isolation medium. Application of tetracyclin eliminates bacteria and cultures then assume an off-white colour. On quarter strength PDA, cultures show some aerial mycelium giving it a velvety look. Cultures also have concentric rings with culture edges diffused. Cultures often show scalloped edges. Hyphae from cultures are cylindrical, 2-5 μm diam, frequently branched, often at right angles, septate, lacking clamp connections; wall slightly thickened to thickened (to 0.25 μm); often with refractive internal bodies; sometimes uneven (with undulate outline); often with swollen elements to 10.5 μm diam that are thick-walled (to 0.5 μm thick) and globose to clavate when terminal, and globose to ellipsoid when intercalary; when terminal, subtended by one or two swollen, clavate elements, but not in chains. Refractive bodies within the hyphae are more common and obvious in this and T. warcupii than in the other two species. Additional material examined: See Table 1. Tulasnella sphagneti Linde & T.W. May, sp. nov. MycoBank MB (Fig. 3B) Etymology: Referring to the Sphagnum habitat of the orchid host. Type: Australia: New South Wales: Kosciuszko NP, alongside Tantangara Road, isolated from Chiloglottis aff. valida growing in a Sphagnum hummock, 19 Jan. 2012, C.C. Linde CLM541 & E. Triponez (MEL holotype; extype culture VPRI 42811). Clade-based diagnosis: The least inclusive clade in the ITS phylogeny in Fig. 1 containing and Substrate or host: Roots and collars of Chiloglottis valida, C. aff. valida, and C. turfosa growing in Sphagnum hammocks in alpine areas in eastern Australia. Distribution: South-eastern Australia, occurring in alpine habitats associated with Sphagnum hummocks. Current known distribution coincides with that of Chiloglottis hosts within this particular habitat. Notes: Cultures on quarter strength PDA show fine concentric rings. Culture edges lack concentric rings, are broad and diffused. Hyphae from cultures cylindrical, μm diam, branched, often at right angles, septate, lacking clamp connections; wall slightly thickened (to 0.25 μm); rarely with refractive internal bodies, and then in narrower hyphae; sometimes uneven (with undulate outline); rarely with swollen elements to 7 μm diam that are slightly thickwalled, subglobose and terminal. Swollen elements are less common in this species than in the other three. Additional material examined: See Table 1. Tulasnella warcupii Linde & T.W. May, sp. nov. MycoBank MB (Fig. 3D) Etymology: After J. H. Warcup who was instrumental in studying mycorrhizal fungi associated with orchids in Australia. Type: Australia: Queensland: Atherton Tablelands, Herberton Range State Forest, Atherton, isolated from Arthrochilus VOLUME 8 NO. 1 43

111 Linde et al. oreophilus, 1 Apr. 2010, C.C. Linde & D. Gomez CLM027 (MEL holotype; MEL isotype; ex-holotype culture VPRI 42809). Diagnosis: Tulasnella warcupii can be diagnosed by the following nucleotide characters, which are fixed between T. tomaculum and T. warcupii respectively: Locus ITS: ITS1 upstream from the 18S at position (GT:AC), 26 (T:C), 33 (G:C), 37 (C:-), 47 (C:T), 101 (T:C), 120 (C:T), 130 (G:T/A), 131 (C:T), 154 (G:T), 158 (T:C), 163 (C:-), 166 (T:C), 169 (T:C), 178 (G:T), 186 (T:C), 189 (A:G), 201 (C:T), 203 (T:A), (--:G/AT), (GTCA:----), 238 (A:G), 240 (A:T). 5.8S starting from ITS1 end: Position 1 (T:-). ITS2 starting from 5.8S end: Position 13 (C:G), 24 (C:T), 26 (G:A), 28 (T:A), 39 (G:A), 46 (C:T), 49 (T:C), (CT:TC), 59 (C:T), 61 (T:C), 64 (T:C), (AC:GT), 72 (A:G), 76 (T:C), 84 (T:C), 86 (G:A), 88 (A:G), 92 (-:-/G), 93 (-:-/C), (GC:AT), 107 (A:G), (ATG:GCT), 113 (G:T), 116 (A:G), (TT:CG), 125 (T:C), 138 (C:T), (AT:GA), (CC:AT), 149 (-:C), 153 (G:A), 156 (T:C), 179 (T:G), 181 (C:T), 184 (T:C), (TT:CC), (CT:TC), (TCG:GAT), (TCG:CGA), (GT:AC), 258 (C:T), 269 (T:C), 274 (-:T), 277 (C:A), 278 (G:C/T), 283 (G:A/-), 287 (T:-), (GCT:TTC), 302 (-:- /G), 303 (A:-/G), 307 (T:C), 309 (A:T). Clade-based diagnosis: the least inclusive clade in the ITS phylogeny in Fig. 1 containing KF and KF Substrate or host: Roots and collars of Arthrochilus oreophilus. Distribution: Atherton Tablelands in Queensland, Australia, in association with Arthrochilus oreophilus in Eucalyptus woodland. Notes: On PDA cultures show fine concentric rings with a velvety edge. Usually no aerial mycelium is visible. Cultures are off-white to yellowish. Of the four Tulasnella species described here, it is the slowest growing. Hyphae from cultures are cylindrical, 1 2.5( 4) μm diam, branched, often at right angles, septate, lacking clamp connections; wall slightly thickened to thickened (to 0.5 μm); often with refractive internal bodies; sometimes uneven (with undulate outline) to distinctly monilioid, with short, repeated, globose to subglobose elements to 7 μm diam. The minimum diameter of hyphae is noticeably thinner than in the other three species, and this is the only one of the four species to show chains of globose elements. Additional material examined: See Table 1. DISCUSSION Here we describe four new species of Tulasnella that are found in association with Australian terrestrial orchids belonging to the Diurideae, using diagnostic DNA characters as advocated by Renner (Renner 2016). Three of these species (T. prima, T. secunda, and T. warcupii) were initially revealed by an in depth study using eight sequence loci and three different methods of species delimitation (Linde et al. 2014). These three species were shown to successfully germinate seed of members of the orchid genus they associate with (Linde et al. 2014). Our addition of Tulasnella isolates from Chiloglottis growing in Sphagnum hummocks in alpine areas in eastern Australia revealed the fourth species, T. sphagneti. This represents the second Tulasnella species to be found associated with Chiloglottis orchids. Based on the widely accepted 3 % sequence divergence cut-off value for species delimitation (Nilsson et al. 2008), or the 3 5 % divergence proposed for delimiting Tulasnella species (Girlanda et al. 2011, Jacquemyn et al. 2011), the sequence divergence between the four new species described in this study, exceeds these cut-off thresholds (6.3 %). Tulasnella is representative of the complexity of contemporary taxonomic mycology. Some species are rigorously defined on multi-gene data, or on the single region (ITS) that has been confirmed as having utility as a barcode in this genus, while other species have been and are being described with excellent details of morphological characters. Unfortunately, few species are well known from both morphology and molecular sequence data. Ideally, all type material should be sequenced, which would allow integration of the two approaches. However, Cruz et al. (2016) point out that sequencing of old fungarium specimens of Tulasnella spp. has been unsuccessful probably due to inappropriate conservation of DNA and they consider this could well remain the case even with improvements in techniques. Therefore, sequenced epitypes will need to be designated where the strict application of names without sequences is ambiguous, but the challenge will be to match modern cultures or collections to old names. Apart from their association with orchids, the ecology and distribution of the new Tulasnella species described here remains poorly known. Interestingly, all orchid species investigated, within the orchid genera Chiloglottis, Drakaea, and Caleana, associate with a single Tulasnella species with one exception. The one exception is in Chiloglottis where both T. prima and T. sphagneti associate with Chiloglottis orchids, but T. sphagneti is so far only found in Chiloglottis species growing in Sphagnum. Where orchids in the Drakaeinae are host to multiple Tulasnella species, the fungi are closely related. Tulasnella prima and T. sphagneti from Chiloglottis are sister taxa and the three Tulasnella species from Arthrochilus form a clade. However, the overall phylogeny of the Tulasnella species from Drakaeinae does not appear to match that of the hosts (Miller & Clements 2014), where Chiloglottis is sister to Drakaea, and these form a clade sister to the remaining genera, including Arthrochilus and Caleana. Remarkably, in Caleana, the only orchid genus in this group to be found in both eastern and western Australia, this association extends across the continent. In contrast to the orchid genus-wide association of most Tulasnellas in this study, mycorrhizal associations of the tropical Arthrochilus oreophilus appear far more diverse. Previously, three Tulasnella OTUs were shown to occur in a narrow sample of this subtropical species (Linde et al. 2014). Because two of the OTUs are represented by only one or two sequences, and lack living cultures, only one (T. warcupii) is described here. Our findings raise the question of why only a small diversity 44 IMA FUNGUS

112 New orchid associated Tulasnella species of Tulasnella fungi associates with a large number of orchid species across a vast geographic range. The pattern of one fungal species to many orchid species appears to be in stark contrast to studies of orchid-mycorrhizal interactions outside Australia, which have consistently found a number of mycorrhizal OTUs associating with sympatric as well as allopatric orchid congeners (Jacquemyn et al 2015). For example, 15 OTUs from Tulasnellaceae were associated with four species of Anacamptis orchids. Of those 15 OTUs, 13 associated with seven species of Ophrys and two Orchis species, whereas nine OTUs associated with three Serapias species (Pellegrino et al. 2014). The high diversity of Tulasnella was such that within sites up to 15 OTUs were co-occurring and 85 % of plants associated with more than three different OTUs (Pellegrino et al. 2014). A corresponding result was found along a single 1000 m transect with the same orchid genera where 16 Tulasnellaceae OTUs were recovered for 20 species of orchids (Jacquemyn et al. 2015). The same pattern is found in Andean tropical rainforests where up to six Tulasnella OTUs may associate with Stelis orchid species and Pleurothallis lilijae (Suarez et al. 2006, Kotte et al. 2008). Consistent among these studies and ours, is the finding that multiple species of an orchid genus can share the same fungal OTU. However, the ability to germinate orchid seed was not shown in other studies, making it difficult to ascertain the real mycorrhizal diversity associating with the orchids. Our description of four new species of Tulasnella, all associated with Australian orchids, extends the number of formally described species known as mycorrhizal agents of orchids. However, it is evident that more Tulasnella species await DNA analysis and formal description. For example, previous studies on Tulasnella ITS diversity associated with Diuris orchids have uncovered a large number of OTUs (Smith et al. 2010), likely to represent many undescribed Tulasnella species. It is further evident that earlier morphologically based studies by Warcup and co-workers prior to the advent of DNA sequencing grossly underestimated the Tulasnella species diversity associated with orchids in Australia. Although Tulasnella is most commonly detected in association with orchids, orchids are not essential for Tulasnella existence. To understand issues such as the ecology, habitat, and geographic range of these fungi, it is essential to develop detection methods that are independent of the orchid, such as a metagenomic approach. This may not only uncover further Tulasnella diversity, but will also shed light on the lives of these fungi independent of orchids. ACKNOWLEDGMENTS The research was supported by the Australian Research Council (LP and LP ) to CCL and RP. 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114 New orchid associated Tulasnella species Wright MM, Cross R, Cousens RD, May TW, McLean CB (2010) Taxonomic and functional characterisation of fungi from the Sebacina vermifera complex from common and rare orchids in the genus Caladenia. Mycorrhiza 20: Yang ZH, Rannala B (2010) Bayesian species delimitation using multilocus sequence data. Proceedings of the National Academy of Sciences, USA 107: VOLUME 8 NO. 1 47

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116 doi: /imafungus IMA FUNGUS 8(1): (2017) Elaphomyces species (Elaphomycetaceae, Eurotiales) from Bartlett Experimental Forest, New Hampshire, USA Michael A. Castellano 1 and Ryan B. Stephens 2 1 US Department of Agriculture, Forest Service, Northern Research Station, 3200 Jefferson Way, Corvallis, OR 97331, USA 2 Natural Resources and the Environment, University of New Hampshire, 114 James Hall, 56 College Road, Durham, NH 03824, USA; corresponding author ryan.stephens@unh.edu Abstract: We describe five new species of Elaphomyces from Bartlett Experimental Forest, New Hampshire, USA (E. americanus, E. bartlettii, E. macrosporus, E. oreoides, and E. remickii) and revise the description of a sixth previously published species (E. verruculosus). Of the five new species, E. bartlettii and E. remickii are only known from New Hampshire whereas E. americanus, E. macrosporus, and E. oreoides are widely distributed in eastern North America. Elaphomyces verruculosus is the most widespread and abundant Elaphomyces species in eastern North America with a distribution extending from eastern Canada south to northeastern Mexico. All six Elaphomyces species are putatively associated with Tsuga canadensis, a tree species in regional decline. For five of the six Elaphomyces species, we report partially consumed ascomata or rodent fecal samples containing spores, indicating that small mammals play a key role in dispersing these Elaphomyces species and that the Elaphomyces are an important part of the small mammals diet. Key words: Ascomycota hypogeous fungi mycophagy sequestrate fungi truffles Tsuga canadensis White Mountain National Forest Article info: Submitted: 11 June 2016; Accepted: 23 February 2017; Published: 10 March INTRODUCTION The truffle genus Elaphomyces (Elaphomycetaceae, Eurotiales, Ascomycota) is characterized by a gleba composed of powdery ascospores encased in a thick peridium. The genus is ectomycorrhizal with many vascular plant taxa and occurs in both the Northern and Southern Hemispheres (Castellano et al. 2011, 2012a, b, c, 2016, Reynolds 2011, Trappe et al. 2009). In northeastern North America, the genus is ecologically important because it forms mycorrhizal associations with a diversity of tree species and comprises a substantial portion of the diets of mycophagous mammals (Vernes et al. 2004, Vernes & Poirier 2007). Recent truffle ascomata sampling conducted at Bartlett Experimental Forest, New Hampshire, USA, in stands dominated by eastern hemlock (Tsuga canadensis) and American beech (Fagus grandifolia) indicates a high diversity of Elaphomyces in the Northeast. Using a systematic sampling approach across forest types we sampled m 2 plots, detecting six Elaphomyces species. Here we describe five of the Elaphomyces species as new, and provide a revised description of a sixth previously published species. Of the five new species, E. bartlettii and E. remickii are only known from New Hampshire whereas E. americanus, E. macrosporus, and E. oreoides are widely distributed in eastern North America. Elaphomyces verruculosus was previously described and is the most abundant Elaphomyces species in eastern North America with a distribution extending from eastern Canada south through the eastern USA to northeastern Mexico. All six Elaphomyces species are putatively associated with eastern hemlock, with E. bartlettii and E. macrosporus exclusively associated with that tree. Declines of eastern hemlock throughout the eastern USA, due to the introduced hemlock woolly adelgid, Adelges tsugae (Orwig et al. 2002), may reduce local diversity or abundance of Elaphomyces species. Additionally, we report partially consumed ascomata for four Elaphomyces species (E. americanus, E. bartlettii, E. macrosporus, and E. verruculosus). Moreover, these observations, coupled with rodent fecal samples containing spores of E. bartlettii and E. oreoides, suggest that small mammals and their predators play a key role in dispersing spores and increasing local diversity of Elaphomyces and that small mammals depend in part on the Elaphomyces as a food source. MATERIALS AND METHODS Ascomata were collected from 6 June to 5 October 2014 from Bartlett Experimental Forest, Northern Research Station, USDA Forest Service, New Hampshire, USA ( N, W; m elevation). Additional ascomata were collected during directed searches in 2015 and ascomata that had been partially consumed by small mammals were collected opportunistically during 2014 and We sampled for ascomata on 12 established small mammaltrapping grids (105 x 105 m; m 2 ) stratified across hardwood (n = 4), softwood (n = 4), and mixed (n = 4) forest stands (R. Stephens, unpubl.). Within each m 2 grid, we 2017 International Mycological Association You are free to share - to copy, distribute and transmit the work, under the following conditions: Attribution: You must attribute the work in the manner specified by the author or licensor (but not in any way that suggests that they endorse you or your use of the work). Non-commercial: You may not use this work for commercial purposes. No derivative works: You may not alter, transform, or build upon this work. For any reuse or distribution, you must make clear to others the license terms of this work, which can be found at Any of the above conditions can be waived if you get permission from the copyright holder. Nothing in this license impairs or restricts the author s moral rights. VOLUME 8 NO. 1 49

117 Castellano and Stephens sampled at 64 plots (4-m 2 ; 16 plots each month) raked to a depth of 10 cm or until mineral soil was reached (Luoma & Frenkel 1991). In total, we sampled m 2 plots across all grids (256 in each forest type). Hardwood forest stands were dominated by American beech, red maple (Acer rubrum), sugar maple (A. saccharum), yellow birch (Betula alleghaniensis), white birch (B. papyrifera), and white ash (Fraxinus americana), whereas softwood stands were dominated by eastern hemlock, red spruce (Picea rubens), and balsam fir (Abies balsamea). The coarse-loamy, well-drained soils are spodosols developed from glacial till and are underlain by granite (Schaller et al. 2010). Colours are given in general terms, and not by reference to a colour chart. Ascomata were initially air dried in the field and then thoroughly dried for h at 60 C in the laboratory. Dried specimens were rehydrated and examined in 3 % KOH, Melzer s reagent, and Cotton blue. Microscopic descriptions and micrographs are based on 3 % KOH mounts unless otherwise specified. Twenty ascospores were measured from the holotype collection of each species; dimensions include ornamentation. Dried ascospores from holotypes were mounted on aluminum pegs with double-sided tape and sputter coated with gold for scanning electron microscopy (SEM) with an AmRay 3300 FE field emission scanning electron microscope. Specimens were deposited in OSC (Oregon State University), FLAS (Florida State University), and BPI (US National Fungus Collection, Beltsville, MD). TAXONOMY Key to the Elaphomyces species of Bartlett Experimental Forest 1 Peridial surface yellow to yellow-brown... 2 Peridial surface dark brown to nearly black (1) Inner layer of peridium marbled... E. americanus Inner layer of peridium homogenous in colour (2) Gleba medium brown at maturity... E. remickii Gleba dark brown to nearly black at maturity... E. verruculosus 4 (1) Peridial surface with dark yellow-brown spots; spores µm diam... E. macrosporus Peridial surface an even dark brown to black colour; spores < 31 µm diam (4) Spore ornamentation a complete reticulum; odour sweet... E. oreoides Spore ornamentation incomplete and slight with scattered smooth areas; odour garlicy, acrid... E. bartlettii Elaphomyces americanus Castellano, sp. nov. MycoBank (Fig. 1) Etymology: americanus, describing the North American, particularly the eastern portion, distribution of this species. Diagnosis: Differs from all North American species in the marbled inner peridium and from E. muricatus of Europe by being slightly smaller in overall spore size, having a coarser ornamentation with more lines then rods, and having a taller ornamentation. Type: USA: West Virginia: Pocahontas Co., Middle Mt., Iron Bridge, 25 July 2001, S. Stephenson (OSC holotype). Description: Ascomata to 16 mm broad, more or less globose, completely embedded in a pale mycelial mat which forms a poorly to well-developed husk around individual ascomata incorporating soil, ectomycorrhizal roots, and debris; mycelium not staining when handled. Peridium overall mm thick when mature, epicutis µm thick, of distinct, yellow-brown warts, most acutely pointed, some blunt, 4 5 sided, bases more or less contiguous, to 500 µm tall, µm broad at the base, paler within and at the base, darker along the outline in section; inner layer ±1000 µm thick, marbled with veins which are white to off-white when young to pale brown when mature, matrix pale tan near surface grading to tan to brown to finally dark brown or black near the gleba, overall dark brown when mature, hyphal strands invaginating into the gleba from the inner peridium, pale tan to rose-tan to white to grey-brown, disintegrating at maturity. Gleba spore mass at maturity powdery, dark brown, with tan, spider web-like hyphae, when immature stuffed with bright white mycelium, then as spores form and mature the mycelium becomes off-white to pale grey. Odour indistinct, mild. Taste not recorded. Peridium two-layered, outer (warty) layer of yellow-brown, compact, curly, contorted, agglutinated hyphae, walls ±2 µm broad, with a low wall of septate, hyaline, parallel hyphae between warts ±7 µm broad, walls ±3 µm thick, beneath the warts is a yellow-brown layer that rapidly grades into hyaline hyphae of a similar texture and size as the warts, then grading into a red-brown layer of similar texture except with amorphous pigmented granules scattered across the 50 IMA FUNGUS

118 Elaphomyces species from New Hampshire Fig. 1. Elaphomyces americanus. A. Ascomata showing peridial surface, gleba, and peridium in section (OSC ). B. Ascoma that has been chewed upon by a small mammal (OSC ). C. Peridial warts. D. Outer peridial warts with stacked compact hyphae between warts. E. Ascospore in cross-section showing height and pattern of the ornamentation). F. Ascospores in surface view. G. Scanning electron micrograph of ascospores showing variable ornamentation pattern. H. Scanning electron micrograph of an ascospore showing the complex structure to the wart structure. C H (OSC 8113 holotype). Bars A = 1 mm, B = 5 mm, C = 500 µm, D G = 10 µm, H = 5 µm. VOLUME 8 NO. 1 51

119 Castellano and Stephens layer, these dark areas in the inner layer matrix attributable to numerous pigment granules, macroscopically the entire area is dark making it difficult to determine the transition from white veins to dark veins. Gleba of spores and hyaline hyphae that are, smooth, septate, extensively branched, occasionally somewhat curly, loosely interwoven, 2 3 µm broad, walls <0.5 µm thick. Asci globose, hyaline, mm broad, walls 3-4 µm thick, 4 8-spored, arising from acrogenous hyphae which are puzzle-like, ±5 mm broad; spores with ascus smooth at first then soon with tall spines. Ascospores globose, mm (mean = 30.9 mm) diam, ornamented, walls ±1 mm thick, brown in KOH when mature singly and in mass, ornamentation of tufts consisting of convalesced rods, not reticulate, µm tall; with small, apparently aborted, globose spores measuring µm broad. Distribution, habit, habitat and season: Known from Canada (New Brunswick, Ontario, and Quebec) and the USA (Connecticut, Maine, Massachusetts, Michigan, New Hampshire, New York, North Carolina, Tennessee, Virginia, and West Virginia); hypogeous in sandy or clay soils; under Abies balsamea, Picea rubens, Pinus banksiana, P. strobus, and Tsuga canadensis; mainly June through October, but also found once in December. Other collections examined: USA: Connecticut: New Haven Co., New Haven, Nov. 1889, R. Thaxter (BPI ); Maine: Piscataquis Co., Baxter State Park, Katahdin Lake trail, 1 Sept. 1962, H.E. Bigelow 3738 (NY); Massachusetts: Hampshire Co., University of Massachusetts campus, near Brown Dorm, 10 Aug. 1986, M. Castellano (OSC , OSC ); Middlesex Co., Middlesex Falls, 19 Nov. 1981, J. LaFrankie (FH ); Norfolk Co., Canton, Sept. 1933, D. Linder (BPI , BPI ); Michigan: Chippewa Co., Tahquamenon Falls, 12 Aug. 1951, A.H. Smith (OSC , MICH); New Hampshire: Carroll Co., White Mountain National Forest, Bartlett Experimental Forest, 2.8 km south and 0.06 km west of the intersection of Bear Notch rd. and Hwy 302, 2 Aug. 2014, R. Stephens & T. Remick (OSC ); same data except 2.8 km south and 0.35 km west of the intersection of Bear Notch rd. and Hwy 302, 3 Aug (OSC , FLAS-F59189); same data except 1.5 km south and 0.25 km east of the intersection of Bear Notch rd. and Hwy 302, 15 July 2014 (OSC ); same data except 2.25 km south and 0.75 km east of the intersection of Bear Notch rd. and Hwy 302, 20 June 2014 (OSC ); same data except 20 Sept (BPI ); same data except 2.5 km south of the intersection of Bear Notch rd. and Hwy 302, 28 July 2014 (OSC ); same data except 25 July 2014 (OSC ); same data except 2.25 km south and 0.8 km west of the intersection of Bear Notch rd. and Hwy 302, 11 Sept (OSC ); same data except 2 km south and 0.2 east of the intersection of Bear Notch rd. and Hwy 302, 18 June 2014 (OSC ); Strafford Co., Durham, College Woods, University of New Hampshire, 26 Sept. 2016, R. Stephens (OSC ); New Jersey: Clinton Township, Lebanon, 9 South Deer Hill Rd., 25 Sept. 2003, R. Balsey (OSC ); same data, 1 April 2005 (OSC ); New York: Warren Co., Lake Sherman, 24 Sept. 1971, C. Rogerson (OSC 33331, NY); North Carolina: Clay Co., Nantahala National Forest, Bearpen Gap, 8 July 1996, S. Loeb & F.H. Tainter (OSC ); Haywood Co., Nantahala National Forest, Haywood Gap, 18 July 1996, S. Loeb and F.H. Tainter (OSC ); same data, 10 July 1996, S. Loeb (OSC ); Pisgah National Forest, Sweetwater Creek, 24 July 1996, S. Loeb & F.H. Tainter (OSC ); Macon Co., Nantahala National Forest, Standing Indian Mountain campground, 14 July 2000, M. Castellano (OSC 80489); same data except 31 July 1992, R. Petersen (E ); Rutherford Co., Painters Gap rd. between Whitehouse and Gilkey, 22 July 2012, T. Elliott (OSC ); off Painters Gap rd., Chalk Mountain, 13 June 2012, T. Elliott (OSC ); Tennessee: Great Smokey Mountain National Park, Kephart Prong trail, a few hundred yards up trail, 15 July 2000, E. Cázares (OSC 80490); Montmorency Co., White Rocks Recreation Area, 10 June 1983, S. Miller 686 (OSC ); Virginia: Fairfax Co., Oakton, corner of Jermantown rd. and Chain Bridge rd., 12 July 2015, M. Castellano (OSC ); Gilles Co., north fork of Stoney Creek, 25 Oct. 1983, S. Miller 714 (OSC ); West Virginia: Pocahontas Co., near Cranberry Glades, along Scenic Hwy, 20 Sept. 1981, S. Miller 485 (OSC ); Cromer Run, 23 July 2001, S. Stephenson (OSC 81119); Three Forks of the Williams River, 3 Oct. 1981, S. Miller 0503 (OSC ); Randolph Co., Stuart Knob, 31 Oct. 1995, D. Mitchell (OSC ); 7 miles from Elkins, Shavers Fork of the Cheat River, 22 Aug. 2007, T. Elliott (OSC ); Tucker Co., Moore Run, 6 Aug. 2001, S. Stephenson (OSC 81116). Canada: Quebec: St Anaclee de Rimouski, 14 July 1981, C. Godbout (OSC 40569); Montreal, 1 Oct. 1992, F. Marzitelli (OSC 51100); Rawdon, 13 Sept. 1992, F. Marzitelli (OSC 51037); Beaupre, 19 Aug. 1981, J. Trappe (OSC 42456); Baie St. Paul, 19 Aug. 1981, J. Trappe (OSC 40572); Duchesnay, Las Jaune, 24 Aug. 1938, R. Cain (BPI ); New Brunswick: Fundy National Park, Upper Salmon River, 3 June 1999, K. Vernes (OSC 62335, , , , , , , ); Point Wolfe, 14 July 1999, K. Vernes (OSC , ); same data except 24 June 1999 (OSC , ); Kinnie Brook, 23 June 1999, K. Vernes (OSC , ); same data except 10 July 1999 (OSC , ); same data except 12 Aug (OSC ); Devils Half Acre, 25 June 1999, K. Vernes (OSC , , , , , ); same data except 10 Aug (OSC ); same data except 12 July 1999 (OSC ); same data except 8 Sept (OSC ); Ontario: Holland River Marsh, 6 May 1936, R. Cain (BPI ); Gull Lake, 9 Sept. 1935, R. Cain (BPI ). Discussion: The marbled inner peridium separates this species easily from all other Elaphomyces species in North America. Elaphomyces americanus resembles E. muricatus from Europe, but the spores of E. americanus have a coarser ornamentation with more lines then rods, are slightly smaller, and have a taller ornamentation. The spores of E. americanus enlarge as they mature due to maturation of the ornamentation. The Canadian ascomata examined were younger and the spores in many specimens only µm broad. Ascomata with a loose powdery gleba (the most mature specimens) have spores µm broad. Two specimens collected at Bartlett Experimental Forest on 11 Sept (OSC ) and 21 July 2014 (OSC ) were partially consumed by small mammals. Both specimens had been excavated and discarded after consumption of most of the peridium, leaving the gleba exposed. Characteristics of the incisor marks on OSC indicate consumption by a red squirrel (Tamiasciurus 52 IMA FUNGUS

120 Elaphomyces species from New Hampshire hudsonicus) or eastern chipmunk (Tamias striatus). In addition, collection OSC was also consumed in part and piles of spores were left on a fallen tree. Beug et al. (2014) list E. americanum nom. prov. which actually refers to this species. Elaphomyces bartlettii Castellano & R.B. Stephens, sp. nov. MycoBank (Fig. 2) with a stout pedicel, ±60 mm broad, walls 1-2 µm thick, hyaline, 8-spored. Ascospores globose, often with some slightly flattened sides, mm (mean = 27.7 mm) diam, ornamented; walls ±1 mm thick, in KOH dark brown to black when mature both singly and in mass, ornamentation 1 2 µm tall, appearing as a dimpled or punctate surface with scattered smooth areas, spores can be somewhat coarse looking in cross-section, particularly the darker spores; in SEM with a fine-structured reticulum overlain with patches of flattened, plate-like material. Etymology: For Bartlett Experimental Forest; named after Governor Josiah Bartlett ( ), a New Hampshire politician and signer of the Declaration of Independence. Distribution, habit, habitat and season. New Hampshire; hypogeous in humic soils, scattered to gregarious; under Tsuga canadensis; June through October. Diagnosis: Distinguished from E. leveillei and E. morettii of Europe by the dark brown hyphae that envelope the ascoma and spores which are coarse in ornamentation and mm in size. Type: USA: New Hampshire: Carroll Co., White Mountain National Forest, Bartlett Experimental Forest, 1.5 km south and 0.25 km east of the intersection of Bear Notch rd. and Hwy 302, 22 June 2015, R. Stephens & T. Remick (OSC holotype; BPI isotype). Description: Ascomata subglobose to slightly lobed, to 22 mm broad x 17 mm tall, individual ascomata enclosed in a well-developed husk incorporating dark brown hyphae, soil, ectomycorrhizal roots, and debris; hyphae not staining when handled. Peridium overall ±2 mm thick when mature, epicutis to 500 µm thick, of distinct, closely spaced, black warts, mostly bluntly rounded, variously shaped, to 200 µm tall, bases contiguous and to 250 µm broad, with abundant, dark brown hyphae packed between warts and often with a thin or thick layer of brown hyphae on wart apices, in section the outer layer appearing duplex because of the black warts and inter-wart dark brown hyphae; inner layer to 1.5 mm thick in places, off-white to pale tan, homogeneous (not marbled), pale tan hyphal strands invaginating into the gleba from the inner peridium. Gleba spore mass powdery, dark brown to black, scattered, pale tan, spider web-like hyphae, when immature gleba stuffed with bright white hyphae. Odour of garlic with an acrid component to skunky, extremely pungent and often detectible when surface soil is disturbed. Taste not recorded. Peridium two-layered, outer (warty) layer of dark to nearly black, compact, septate, agglutinated, thick-walled (±2 µm broad), parallel hyphae, grading into dark brown then pale brown textura epidermoidea, of compactly interwoven hyphae, 4 7 µm broad, walls ±1 µm thick, between warts packed with dark brown hyphae arranged perpendicularly to the hyphae of the warts at times, these hyphae sometimes swollen, inner layer a textura intricata, of hyaline, septate, interwoven hyphae, 4 5 µm broad, to 12 µm broad when swollen, walls <0.5 µm thick, with abundant, large, amorphous crystalline particles scattered across the entire layer. Gleba composed of spores and hyphae, that are hyaline, smooth, septate, extensively branched, loosely interwoven, 2 4 µm broad. Asci globose Other collections examined: USA: New Hampshire: Carroll Co., White Mountain National Forest, Bartlett Experimental Forest, 1.5 km south and 0.25 km east of the intersection of Bear Notch rd. and Hwy 302, 21 Sept. 2014, R. Stephens & T. Remick (OSC , BPI ); same data except 2.8 km south and 0.06 km west of the intersection of Bear Notch rd. and Hwy 302, 7 June 2014 (OSC ); same data except 3 Aug (OSC ); same data except 30 June 2014 (OSC ); same data except 14 Sept (FLAS-F59190); same data except 2.25 km south and 0.75 km east of the intersection of Bear Notch rd. and Hwy 302, 14 Aug (OSC ); same data except 2 km south and 1.4 km west of the intersection of Bear Notch rd. and Hwy 302, 28 Sept (OSC ); same data except 2.25 km south and 0.8 km west of the intersection of Bear Notch rd. and Hwy 302, 8 July 2014 (OSC ); Strafford Co., Durham, East Foss Farm, 1 km south of the intersection of Mill rd. and Foss Farm rd., 2 Oct. 2016, R. Stephens & K. Kwasnik (OSC ). Discussion: Immature ascomata of Elaphomyces bartlettii have a bright white inner peridium which grades to off-white towards the centre at maturity. The interior is initially hollow, filling with white cottony hyphae, becoming black and powdery as the spores develop and mature. Elaphomyces bartlettii bears some resemblance to E. leveillei and E. morettii from Europe. The ascomata of E. leveillei, however, are covered with a greenish mycelium and a green mycelium patch persists at the base after handling. Elaphomyces leveillei has similar sized spores but the ornamentation appears smoother in cross-section and under SEM (not shown). The ascomata of E. morettii have a brownish red mycelium and spores which are much smaller (17 22 µm) and have a coarser ornamentation. One specimen collected at Bartlett Experimental Forest on 30 June 2014 (OSC ) was partially consumed by a small mammal. The mature ascoma was left in situ at a depth of approximately 7 cm. Soil was excavated to reach the ascoma and animal activity had left the gleba partially exposed. Additionally, spores of E. bartlettii were detected in fecal samples from a southern red-backed vole (Myodes gapperi) and T. striatus live-trapped at Bartlett Experimental Forest. The M. gapperi sample was collected on 3 Aug just 17 m from OSC , and the Tamias striatus sample was collected on 1 July 2014 approximately 90 m from OSC VOLUME 8 NO. 1 53

121 Castellano and Stephens Fig. 2. Elaphomyces bartlettii. A. Ascomata showing peridial surface, gleba, and peridium in section. B. Ascoma showing small warts on peridial surface. C. Epicutis with the darker outer area grading into a paler inner area. D. Subcutis with abundant amorphous, crystalline particles scattered across the layer. E. Thick-walled, pedicellate ascus. F. Ascospores in surface view showing the clumpy ornamentation. G. Scanning electron micrograph of ascospores showing the labyrinthine pattern to the clumpy ornamentation. H. Scanning electron micrograph showing the fine detail of the ornamentation. A H (OSC holotype). Bars A, B = 10 mm, C E = 20 µm, F, G = 10 µm, H = 5 µm. 54 IMA FUNGUS

122 Elaphomyces species from New Hampshire Elaphomyces macrosporus Castellano & T.F. Elliott, sp. nov. MycoBank (Fig. 3) Etymology: macrosporus macro large, sporus spore, in reference to the large spores. Diagnosis: This species differs from other Elaphomyces species by its large spores which are ornamented with rods and clumps of rods that form broad based cones or ridges and the black hyphae that cover the brown peridial warts. Type: USA: West Virginia: Tucker Co., Canaan Heights, along Hwy 32, south of Davis, 6 July 2015, M. Castellano (OSC holotype). Description: Ascomata to 2 cm broad, irregular, often completely enveloped in a dark sordid yellow mycelium, organic debris, and ectomycorrhizal roots, occasionally ascomata are dislodged from the substrate without adherent mycelium, debris and roots; mycelium not staining when handled; base with yellow mycelial tuft. Peridium to 1000 µm thick, outer surface of yellow-brown warts ±200 µm tall by µm broad at the base, warts embedded in a black mycelium to give the appearance of a black ascoma with dark yellow-brown dots that are the tips of the warts emerging from the black mycelium, subcutis ±800 µm thick, uniform, grey, leathery, homogeneous (not marbled). Gleba spore mass powdery, dark brown to black at maturity, with dark spider web-like hyphae, when immature gleba of stuffed bright white hyphae and pale brown tissue. Odour initially a faint hint of citrus, increasing as ascomata begin to decay underground. Taste not recorded. Peridium two-layered, epicutis, ±200 µm thick, of compact, septate, yellow-brown, disorganized, compact hyphae, ±5 mm broad, walls ±1 µm thick; subcutis ±800 µm thick, composed of hyaline, septate, compact, interwoven hyphae, to 10 µm broad, walls ±1 µm thick; also an ill-defined inner layer of pale tan, loosely interwoven hyphae, to 4 mm broad near the gleba. Asci subglobose to globose with a stiptitate base, µm broad, walls ±2.5 µm thick, hyaline, 4- or 8-spored, stipitate base to 11 µm long. Ascospores globose, (39.5 ) 41 44( 45.5) mm (mean = 42.2 mm) diam, ornamented, walls ±1 mm thick, in KOH singly and in mass dark brown to red-brown when mature, ornamentation of rods and clumps of rods that form broad based cones or ridges, to 3 µm tall, immature spores abundant, dense, dark red-brown, mm broad. Distribution, habit, habitat and season: Known from Maine, Massachusetts, New Hampshire, North Carolina, and West Virginia; hypogeous, usually clustered; under Tsuga canadensis; June through November. Collections examined: USA: Maine: Cumberland Co., Bradbury Mountain State Park, at campground, 17 Oct. 2015, R. Stephens (OSC ); Massachusetts: Hampshire Co., University of Massachusetts campus, near Brown Dormitory, 10 Aug. 1986, M. Castellano (OSC ); Worcester Co., Wachusett Mountain, 16 Nov. 1984, J. Trappe (OSC ); New Hampshire: Carroll Co., White Mountain National Forest, Bartlett Experimental Forest, 1.5 km south and 0.25 east of the intersection of Bear Notch rd. and Hwy 302, 17 June 2014, R. Stephens & T. Remick (OSC ); same data except 2.25 km south and 0.75 east of the intersection of Bear Notch rd. and Hwy 302, 20 June 2014 (OSC ); same data except 3 Aug (OSC ); same data except 19 June 2014 (FLAS-F59192); same data except 2.8 km south and 0.06 west of the intersection of Bear Notch rd. and Hwy 302, 14 Sept (OSC ); same data except 2 Aug (OSC ); same data except 2 Aug (BPI ); same data except 2.25 km south and 0.8 west of the intersection of Bear Notch rd. and Hwy 302, 4 Aug (OSC ): same data except 2 km south and 1.4 km west of the intersection of Bear Notch rd. and Hwy 302, 28 Sept (OSC ); same data except 3 km south and 1.7 km west of the intersection of Bear Notch rd. and Hwy 302, 31 July 2014 (OSC ); same data except 21 Aug (OSC ); Strafford Co., Durham, College Woods, 13.5 metres north of the intersection of Mill rd. and Hemlock Way, 29 Sept. 2015, R. Stephens & T. Remick (OSC ); North Carolina: Haywood, Co., Nantahala National Forest, Buckeye Creek, 13 Aug. 1996, S. Loeb & F.H. Tainter (OSC ); Mitchell Co., Pisgah National Forest, Carver s Gap, 19 Sept. 1995, S. Loeb & F.H. Tainter (OSC ); Transylvania Co., Pisgah National Forest, near Botd, trail to Pink Beds, 27 Nov. 2011, T. Elliott (OSC , ); Pisgah National Forest, Devil s Courthouse, 31 Aug. 1995, S. Loeb & F.H. Tainter (OSC ); West Virginia: Pocahontas Co., Monongahela National Forest, along FS rd. 44, 30 July 2013, C. Diggins (OSC ); Randolph Co., Kumbrabow State Forest, Oxley Creek, 18 July 2013, C. Diggins (OSC ); same data, 22 July 2013 (OSC ); same data, 16 July 2013 (OSC ); Tucker Co., Blackwater State Park, near lodge, between parking lot and road, 6 July 2015, M. Castellano (OSC , ); Canaan Heights, along Hwy 32, south of Davis, 6 July 2015, M. Castellano (OSC , ). Discussion: Elaphomyces macrosporus nom. prov. was listed by Beug et al. (2014) and had been previously identified as E. leveillei by Loeb et al. (2000) from North Carolina. The peridial surface of Elaphomyces macrosporus resembles that of E. aculeatus from Europe which has much smaller spores (21 24 µm broad). The peridial surface and spore ornamentation of E. spinoreticulatus (Zhang & Minter 1989) is also similar to E. macrosporus but E. spinoreticulatus spores are demonstrably smaller (35 38, mean = 36.1 µm) than E. macrosporus spores (41 44, mean = 42.2 µm). Elaphomyces macrosporus is also similar (macroscopically and in spore characteristics) to E. verruculosus and both occur in eastern North America. Elaphomyces verruculosus lacks any black mycelium surrounding the brown peridial warts and on average the spores are smaller (36 45 µm, mean = 41.1 µm broad) and have a more coarse appearance. Immature ascomata of E. macrosporus have a bright white inner peridium which grades to pale tan towards the gleba at maturity, often creating a two-tone appearance. The gleba of immature ascomata is solid and white, becoming black and powdery as the spores mature. Two specimens collected at Bartlett Experimental Forest on 2 Aug (OSC ) and 3 Aug were partially consumed by small mammals. The peridium of OSC (maturing specimen) was partially consumed. The soil was VOLUME 8 NO. 1 55

123 Castellano and Stephens Fig. 3. Elaphomyces macrosporus. A. Ascomata showing peridial surface, gleba, and peridium in section. B. Ascoma showing peridial warts embedded in dark hyphae. C. Microscopic view of sectioned outer peridium showing the pale, stacked hyphae between the darker wart tissue. D. Subcutis showing the thick-walled, interwoven hyphae. E. Ascospores in surface view showing the ornamentation pattern. F. Ascospores in cross-sectional view showing the height and pattern of the ornamentation. G. Scanning electron micrograph of ascospores. H. Scanning electron micrograph of ascospores showing the fine detail of the ornamentation. A, B (OSC ); C F (OSC holotype); G, H (OSC ). Bars A = 1 cm, B = 5 mm, C G = 20 µm, H = 10 µm. 56 IMA FUNGUS

124 Elaphomyces species from New Hampshire excavated to reach the ascoma at a depth of approximately 3 cm; the ascoma was left in situ. The peridium and gleba of OSC (immature specimen) were consumed and left in situ at approximately 3 cm. Elaphomyces oreoides Castellano, sp. nov. MycoBank (Fig. 4) Etymology: oreoides resembling Oreo, in reference to the resemblance of the profile of an Oreo cookie with three inner peridial layers, dark/light/dark, in addition to the slightly sweet odor. Diagnosis: Differs from other eastern North American species of the genus by having yellow-brown spores with a complete reticulum ornamentation and the inclusion of ectomycorrhizas within the lower portion of the inner peridium. It differs from the E. persoonii of Europe by the brown mycelium surrounding ascomata. first inner layer, µm thick, composed of brown, compact hyphae, 3 7 µm broad, arranged in a textura intricata, walls ±1 µm thick; middle inner layer, µm thick, composed of hyaline, compact hyphae, 2 3 µm broad, arranged in a textura porrecta, walls <0.5 µm thick, occasional streaks of pale brown hyphae present in this middle inner layer; third inner layer (closest to the gleba) µm thick, composed of brown, compact hyphae, to 10 µm broad, arranged in a textura epidermoidea, walls ±1 µm broad, somewhat restricted in diameter at the septa. Gleba composed of spores and thin-walled, hyaline, septate, sinuous to curly, slightly encrusted hyphae, ±4 µm broad. Asci globose to irregularly globose (from pressure of enlarging spores), µm broad, walls 0.5 µm thick, 8-spored. Ascospores globose, (27 )28 30( 31) mm (mean = 29.3 mm) diam, ornamented, walls ±1 mm thick, in KOH singly and in mass yellow-brown when mature, ornamentation a complete reticulum with coarse projections at the top of the alveoli, to 2 µm tall, alveoli to 5 µm broad. Type: USA: North Carolina: Buncombe Co., milepost along Blue Ridge Parkway, 4 Sept. 1994, M. Castellano, Z. Miller & W. Miller (OSC holotype). Description: Ascomata to 22 x 27 mm diam, subglobose, often with broad irregular pits, flattened to somewhat turbinate, with a persistent, large basal tuft, to 19 mm broad where attached, 6 7 mm long, composed of a tangle of dark yellowish grey to brown mycelium, ectomycorrhizal roots, soil particles, and debris; dense pale brown to dark brown to greyish black mycelium adherent to the entire peridial surface; mycelium not staining when handled, unchanged when dried. Peridium subcartilaginous, mm thick, four-layered, somewhat zonate with a very dark outer layer underlain by a medium-dark layer grading into a more or less hyaline layer with occasional pale tan streaks, underlain by another dark hyphal layer adjacent to the gleba; outer surface µm thick, very dark brown, carbonaceous, of mostly flattened, irregularly shaped or plate-like warts, to 0.5 mm broad at the base, warts simplex in structure, warts larger at the ascoma apex, smaller along the sides and near the base, often nearly smooth near the base; three inner layers mm thick combined, inner layers uneven in thickness, zonate to somewhat mottled, the middle layer off-white to pale tan, exterior of the inner layers brown; numerous ectomycorrhizas embedded within the inner two layers closest to the gleba, concentrated in the lower half of the specimen, more numerous at the base, absent at the ascoma apex. Gleba spore mass powdery, very dark blue-grey, off-white, spider web-like hyphae invaginating from the inner peridial layer into the gleba towards the centre, thicker patches of off-white hyphae scattered around the gleba-peridium interface from which the spider web-like hyphae emerge. Odour slightly sweet. Taste not recorded. Peridium four-layered, outer layer µm thick, of dark brown, short-segmented, compact hyphae, 3 7 µm broad, to 20 µm long, walls 1 2 µm broad, grading into the Distribution, habit, habitat and season: Known from New Hampshire, North Carolina, and West Virginia; hypogeous, single to clustered; under Betula alleghaniensis, Carya sp., Fagus grandifolia, Gymnocladus diocus, Pinus resinosa, P. strobus, Quercus prinus, Q. ruber, Q. virginiana, and Tsuga canadensis; June, September, and October. Collections examined: USA: New Hampshire: Carroll Co., White Mountain National Forest, Bartlett Experimental Forest, 2.25 km south and 0.8 west of the intersection of Bear Notch rd. and Hwy 302, 8 June 2014, R. Stephens & T. Remick (OSC , FLAS-F59191); North Carolina: Buncombe Co., Mountain to Sea trail, 1 Sept. 1991, W. Sturgeon (OSC ); milepost along Blue Ridge Parkway, 4 Sept. 1994, M. Castellano, J. Roberts, G. Semlak, J. Shubzda & P. Perret (T13589); Haywood Co., Great Smokey Mountain National Park, near summit of Purchase Knob, 22 Sept. 2012, T. Elliott (OSC ); Rutherford Co., off Painters Gap rd., 23 Sept. 2007, T. Elliott (OSC ); West Virginia: Barbour Co., Laurel Mountain, 16 Oct. 1997, D. Mitchell 597 (OSC ). Discussion: Elaphomyces oreoides resembles E. persoonii from Europe in the possession of a basal tuft of mycelium, a dark brown to black, warty peridium and reticulate spores. Elaphomyces persoonii has a green mycelium covering the ascomata whereas E. oreoides has a brown mycelium. Spores of E. oreoides were detected in the fecal samples of two woodland jumping mice (Napaeozapus insignis) livetrapped at Bartlett Experimental Forest. These N. insignis were trapped on 4 June 2014 and 30 July 2014 on the same grid where the ascomata of E. oreoides (OSC , FLAS-F59191) was collected. One additional collection, OSC , is parasitized by an unidentified Tolypocladium species. Beug et al. (2014) list E. fallax nom. prov. which is actually this species. VOLUME 8 NO. 1 57

125 Castellano and Stephens Fig. 4. Elaphomyces oreoides. A. Ascomata showing peridial surface, gleba, and peridium in section (OSC , photo T.F. Elliott). B. Surface of the peridium showing the warts. C. Ectomycorrhizas embedded within the inner peridium. D. Outer peridium showing the gradation of the darkcolored outer hyphae into the paler colored inner hyphae. E. Ascospores with ornamentation in surface view showing the reticulate pattern of the ornamentation. F. Ascospores in cross-sectional view showing the height and pattern of the ornamentation. G. Scanning electron micrograph of ascospores. H. Scanning electron micrograph of ascospores showing the fine detail of the ornamentation. B H (OSC ). Bars A = 20 mm, B, C = 500 µm, D = 20 µm, E G= 10 µm, H = 5 µm. 58 IMA FUNGUS

126 Elaphomyces species from New Hampshire Elaphomyces remickii Castellano & R.B. Stephens, sp. nov. MycoBank (Fig. 5) Etymology: Named for Tyler J. Remick, undergraduate student who aided R. Stephens immensely with collection and preparation of mycological samples at Bartlett Experimental Forest. Diagnosis: Distinguished by the medium brown gleba, unusual in the genus, at maturity, tall pyramidal warts on peridial surface, and the hyaline to pale tan spore colour at maturity. Type: USA: New Hampshire: Carroll Co., White Mountain National Forest, Bartlett Experimental Forest, 1.5 km south and 0.25 km east of the intersection of Bear Notch rd. and Hwy 302, 15 July 2014, R. Stephens & T. Remick (OSC holotype). Description: Ascomata subglobose, to 20 mm broad x 12 mm tall. Peridium overall µm thick when mature, in surface view wart tips variably-shaped, emerging from dense, brown hyphal layer between warts, warts acute, brown, pyramidalshaped overall, bases contiguous, base to µm broad, many smaller, in section warts red-brown in outline, yellow at the base, with abundant, brown hyphae packed between the warts; inner layer homogeneous (not marbled), mm thick in places, mostly off-white but occasionally somewhat tan near gleba, pale tan hyphal strands invaginating into the gleba from the inner peridium. Gleba spore mass powdery, medium brown, scattered, white, hyphal strands invaginating from the inner peridium into the gleba and reaching the centre. Odour not recorded. Taste not recorded. Peridium two-layered, outer (warty) layer of yellow-redbrown, compact, septate, agglutinated, stacked hyphae forming a textura prismatica, µm broad, walls 2 3 µm broad, hyphae between the warts arranged in a tightly stacked manner, sometimes perpendicular to the hyphae within warts, of hyaline, septate, compact hyphae, 3 4 µm broad, walls ± 1µm thick, inner layer a textura epidermoidea, of hyaline, compact, tangled hyphae, 4 5 µm broad, walls ±1 µm broad. Gleba of spores of hyaline, smooth, septate, extensively branched, loosely interwoven hyphae, 2 4 µm broad, walls 0.5 µm thick. Asci not seen. Ascospores globose, mm (mean = 33.2 mm) diam, ornamented, walls ±1 µm thick, in KOH hyaline to pale tan at maturity singly and in mass, ornamentation to 1 µm tall, appearing as a compact undulate surface, in cross-sectional view lumpy; in SEM appearing as a coarse, rugose spore surface. Distribution, habit, habitat and season: Known only from a single locality in New Hampshire; hypogeous in humic soils, clustered; under Abies balsamea, Betula alleghaniensis, or Tsuga canadensis; July. Discussion: Macromorphological characteristics of Elaphomyces remickii are unusual in the genus due to the medium brown, powdery spore mass and the tall, pyramidal warts of the peridial surface. Elaphomyces verruculosus Castellano, Rev. Mex. de Micol. 35: 19 (2012). (Fig. 6) Description: Ascomata irregularly subglobose to reniform, to 22 x 27 mm. Peridial outer surface of semi-rounded to angular or elongate warts, to 500 µm wide and 300 µm tall, rounded to acute or even flattened at the apices, contiguous with each other at the base. Pale yellow-brown when young then mottled brown and yellow-brown when mature with much pale brown, brown or yellow-brown hyphae, soil and debris covering the warts. Warts often obscured by interwart hyphal structures making the surface appear nearly smooth or papillate; in section warts are outlined with a ±140 µm thick layer of red-brown to dark red-brown cells underlain with paler yellow-brown to off-white tissue, µm thick. Below at the base of the warts there is an off-white layer sometimes tinged pale grey-blue; subcutis to 2 mm broad, off-white to pale grey, uniform (not marbled) or sometimes zoned off-white above and pale grey or pale grey-tan below, more readily apparent on mature specimens; often a distinct, brown layer, ±100 µm thick present at the interface with the gleba. This layer is contiguous and concolorous with the dissepiments that invaginate into the gleba. Gleba off-white and cottony when immature, then the spore mass becoming powdery, dark brown to nearly black when mature, numerous pale grey to brown mycelial strands arising from the inner peridial wall and traversing the gleba. Odour indistinct to musky or skunky. Taste of mild cultivated mushrooms. Peridium inter-wart spaces filled with hyaline, pale yellow to pale yellow-brown, septate, parallel hyphae 3 8 µm broad, walls 2 3 µm thick, also at times covering the wart apices; overall the peridium is two-layered, epicutis a warty layer, ±450 µm broad, with a red to red-brown layer near the wart surface then yellow-brown then grading to off-white near the wart base, composed of compact, interwoven hyphae 3-8 µm broad, similar in structure to the inter-wart hyphae and sometimes contiguous in organization but not in colour; subcutis to ±2 mm broad, also similar in structure to the outer inter-wart layer except that the hyphae are hyaline with amorphous, hyaline granules or pigments interspersed across this layer, the scattered granules much less dense near the gleba and dark grey or dark red-brown. Gleba constituted of spores and hyaline, septate, smooth or slightly encrusted, sinuous, loosely interwoven hyphae, to 5 µm broad, walls µm thick. Asci globose, hyaline, µm broad, (2 )4 8-spored, walls ±2 µm thick, arising from acrogenous hyphae of clustered knots of hyaline, short-segmented hyphae, ±3 µm broad, walls ±1 µm thick. Ascospores globose, (35 )36 45( 46) µm broad (mean = 41.1 µm) diam, ornamented; in KOH brown to red-brown to dark red-brown singly and in mass; walls ±1 µm thick; ornamentation of tall, hyaline spines or rods when immature (in asci), of dense rods and tufts of rods when mature, 2 3 x 2 3 µm, giving the spore surface a more or less coarse appearance, appearing fuzzy in section, as spores mature they darken and the tufts become darker and more distinct, mature spores dark redbrown and appearing much more coarse, aborted spores (23 28 µm broad) present and much darker. VOLUME 8 NO. 1 59

127 Castellano and Stephens Fig. 5. Elaphomyces remickii. A. Ascomata showing gleba and peridium in section. B. Outer peridial layer showing the dark coloured tissue of the warts embedded within pale stacked hyphae. C. Pale, thick-walled, stacked hyphae occurring between wart tissue. D. Inner peridial tissue forming a textura epidermoidea. E. Ascospores with ornamentation in surface view showing the undulate pattern of the ornamentation. F. Ascospores in cross-sectional view showing the height and pattern of the ornamentation. G. Scanning electron micrograph of ascospores. H. Scanning electron micrograph of ascospores showing the fine detail of the ornamentation. A H (OSC holotype). Bars A = 10 mm, B = 50 µm, C = 10 µm, D G = 20 µm, H = 10 µm. 60 IMA FUNGUS

128 Elaphomyces species from New Hampshire Fig. 6. Elaphomyces verruculosus. A. Ascomata showing peridial surface, gleba, and peridium in section (OSC , photo T.F. Elliott). B. Ascomata embedded in ectomycorrhizas, soil, and debris; shown in surface and cross-section view (OSC ). C. Ascoma showing the peridial surface in finer detail. D. Outer peridium showing the pale, stacked hyphae between the darker coloured wart tissue. E. Ascospores with ornamentation in surface view showing the rods and spines of the ornamentation. F. Ascospores in cross-sectional view showing the height and pattern of the ornamentation. G. Scanning electron micrograph of ascospores showing the fine detail of the ornamentation. H. Scanning electron micrographof ascospores. D F (OSC 49670); C, G, H (OSC 48377). Bars A = 20 mm, B = 15 mm, C = 5 mm, D = 30 µm, E F= 20 µm, G = 1 µm, H = 10 µm. VOLUME 8 NO. 1 61

129 Castellano and Stephens Distribution, habit, habitat and season: Known from Quebec, Canada south to Florida and northeastern Mexico; hypogeous, single to clustered; under Fagus grandifolia, Picea abies, P. rubens, Pinus elliotii, P. palustris., P. pungens, P. resinosa, P. strobus, P. taeda, P. virginiana, and Tsuga canadensis; June through October but also found once in December. Collections examined: USA: New Hampshire: Carroll Co., White Mountain National Forest, Bartlett Experimental Forest, 2.5 km south and 1.7 km west of the intersection of Bear Notch rd. and Hwy 302, 4 Oct. 2014, R. Stephens & T. Remick (OSC , , ); same data except (OSC ); same data except 2.8 km south and 0.06 km west of the intersection of Bear Notch rd. and Hwy 302, 2 Aug (OSC ); same data except 6 June 2014 (BPI ); same data except 2.5 km south and 0.5 km east of the intersection of Bear Notch rd. and Hwy 302, 15 Aug (OSC ), same data except 2.8 km south and 0.35 km west of the intersection of Bear Notch rd. and Hwy 302, 31 July 2014 (OSC ); same data except 2.25 km south and 0.8 km west of the intersection of Bear Notch rd. and Hwy 302, 8 July 2014 (FLAS-F59193); Rockingham Co., Kwaks Sanctuary, Audubon Society land on Route 152, 1.5 miles west of intersection with Route 108, in Newmarket, 26 Dec. 2015, R. Stephens & K. Kwasnik (OSC ). Discussion: This species was described by Castellano et al. (2012b) from Connecticut, Georgia, Louisiana, Massachusetts, Mississippi, New York, North Carolina, Vermont, Virginia, West Virginia, northeastern Mexico, and Quebec in Canada. This is the most commonly encountered and abundant Elaphomyces species in eastern North America. On a sampling grid dominated by old growth Tsuga canadensis at Bartlett Experimental Forest, 80 % (51 of 64) of the 4 m 2 -plots had 1 ascomata (collections including: OSC , , , ). On another grid, one sampling plot had 173 sporocarps in 4 m 2 (BPI ). The abundance of this species may make it an important food source for small mammals. Vernes & Poirier (2007) found E. verruculosus cashed by a Tamias hudsonicus in a bird s nest in New Brunswick, Canada. We found an air-drying ascoma (OSC ) on a broken-off snag which was putatively placed there by a T. hudsonicus based on the incisor marks. Additionally, we found a single ascoma on 21 June 2014 (OSC ) with incisor marks from a vole or mouse-sized small mammal. The warty, brown, leathery peridial surface and off-white to pale grey, homogeneous inner peridial layer (occasionally tinged bluish) in cross-section clearly distinguish E. verruculosus from other Elaphomyces species in North America. It is similar to E. granulatus from Europe, but E. granulatus has somewhat smaller spores (35 41µm, mean = 37.9 µm) and a coarser pattern to the ornamentation of agglutinated rods and spines that form larger patches. The statements about spore size in E. granulatus in Castellano et al. (2012b) are erroneous. ACKNOWLEDGMENTS We thank Steve Miller (University of Wyoming), Susan Loeb (US Forest Service, South Carolina), Todd F. Elliott (North Carolina), Frances Marzitelli (Quebec), and Donna Mitchell (West Virginia) for sharing their fungal collections. We thank Tyler Remick for his help collecting and curating ascomata. Andrew Uccello and Chris Burke assisted in the fieldwork with additional help from Karrah Kwasnik, among other volunteers, at Bartlett Experimental Forest. Rebecca Rowe provided sampling supplies and constant encouragement and Mariko Yamasaki granted access to sampling at Bartlett Experimental Forest. Scanning electron micrographs of E. bartlettii ascospores were produced at the N.C. Brown Center for Ultrastructure studies using a JEOL JSM- 5800LV scanning electron microscope by Austin Frewert. Todd F. Elliott provided photographs of E. oreoides and E. verruculosus. We thank Terry Henkel, Todd F. Elliott, and an anonymous reviewer for comments which improved the manuscript. Partial funding was provided by the New Hampshire Agricultural Experiment Station. This is Scientific Contribution Number This work was supported by the USDA National Institute of Food and Agriculture McIntire-Stennis Project ( & ). Additional ascomata sampling in 2015 was supported by a grant from the North American Truffling Society. REFERENCES Beug MW, Bessette AE, Bessette AR (2014) Ascomycete Fungi of North America: a mushroom reference guide. Austin: University of Texas Press. Castellano MA, Beever RE, Trappe JM (2012a) Sequestrate fungi of New Zealand: Elaphomyces (Ascomycota, Eurotiales, Elaphomycetaceae). New Zealand Journal of Botany 50: Castellano MA, Dentinger TM, Séne O, Elliott TF, Truong C, et al. (2016) New species of Elaphomyces (Elaphomycetaceae, Eurotiales, Ascomycota) from tropical rainforests of Cameroon and Guyana. IMA Fungus 7: Castellano MA, Guerrero GG, Jiménez, JG, Trappe JM (2012b) Elaphomyces appalachiensis and E. verruculosus sp. nov. (Elaphomycetaceae, Eurotiales, Ascomycota) from eastern North America. Revista Mexicana de Micologia 35: Castellano MA, Henkel TW, Miller SL, Smith ME, Aimee MC (2012c) New Elaphomyces species (Elaphomycetaceae, Eurotiales, Ascomycota) from Guyana. Mycologia 104: Castellano MA, Trappe JM, Luoma DL (2004) Sequestrate fungi. In: Biodiversity of Fungi: inventory and monitoring methods (Mueller GM, Bills GE, Foster MS, eds): New York: Academic Press. Castellano MA, Trappe JM, Vernes K (2011) Australian species of Elaphomyces (Elaphomycetaceae, Eurotiales, Ascomycota). Australian Systematic Botany 24: Loeb SC, Tainter FH, Cazares E (2000) Habitat associations of hypogeous fungi in the southern Appalachians: implications for the endangered northern flying squirrel (Glaucomys sabrinus coloratus). American Midland Naturalist 144: Luoma DL, Frenkel RE (1991) Fruiting of hypogeous fungi in Oregon Douglas-fir forests: seasonal and habitat variation. Mycologia 83: Orwig DA, Foster DR, Mausel DL (2002) Landscape patterns of hemlock decline in New England due to the introduced hemlock woolly adelgid. Journal of Biogeography 29: IMA FUNGUS

130 Elaphomyces species from New Hampshire Reynolds HT (2011) Systematics, phylogeography, and ecology of Elaphomycetaceae. PhD thesis, Duke University. Schaller M, Blum JD, Hamburg SP, Vadeboncoeur MA (2010) Spatial variability of long-term chemical weathering rates in the White Mountains, New Hampshire, USA. Geoderma 154: Trappe JM, Molina R, Luoma DL, Cázares E, Pilz D, et al. (2009) Diversity, Ecology and Conservation of Truffle Fungi in Forests of the Pacific Northwest. [USDA Forest Service General Technical Report PNW GTR 772.] Portland: Pacific Northwest Research Station. Vernes K, Blois S, Bärlocher F (2004) Seasonal and yearly changes in consumption of hypogeous fungi by northern flying squirrels and red squirrels in old-growth forest, New Brunswick. Canadian Journal of Zoology 82: Vernes K, Poirier N (2007) Use of a robin s nest as a cache site for truffles by a red squirrel. Northeastern Naturalist 14: Zhang B-C, Minter DW (1989) Elaphomyces spinoreticulatus sp. nov., with notes on Canadian species of Elaphomyces. Canadian Journal of Botany 67: VOLUME 8 NO. 1 63

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132 doi: /imafungus IMA FUNGUS 8(1): (2017) Pleiocarpon gen. nov. and a new species of Ilyonectria causing basal rot of Strelitzia reginae in Italy Dalia Aiello 1, Giancarlo Polizzi 1, Pedro W. Crous 2,3,4, and Lorenzo Lombard 2 1 Dipartimento di Agricoltura, Alimentazione e Ambiente, sezione Patologia Vegetale, University of Catania, Via S. Sofia 100, Catania, Italy; corresponding author dalia.aiello@live.it 2 Westerdijk Fungal Biodiversity Institute, Uppsalalaan 8, 3584CT Utrecht, The Netherlands 3 Microbiology, Department of Biology, Utrecht University, Padualaan 8, 3584 CT Utrecht, The Netherlands 4 Department of Microbiology and Plant Pathology, Tree Protection Co-operative Programme, Forestry and Agricultural Biotechnology Institute, University of Pretoria, Pretoria 0002, South Africa Abstract: During 2015, a new basal rot disease was observed on potted plants of Strelitzia reginae in an ornamental nursery located in eastern Sicily. Isolations from symptomatic parts of these diseased plants consistently yielded cylindrocarpon-like isolates. Multigene analyses of the partial gene regions of 28S large subunit nrdna, β-tubulin, histone H3, translation elongation factor 1-alpha, internal transcribed spacer region and intervening 5.8S nrrna gene, and RNA polymerase II second largest subunit genes, supported by morphological characters supported the recognition of a new genus, Pleiocarpon based on P. strelitziae sp. nov., and a new Ilyonectria species, described here as I. strelitziae sp. nov. The pathogenicity of both I. strelitziae and P. strelitziae were confirmed on young plants cultivated under controlled conditions in a growth chamber. Both cylindrocarpon-like fungi were pathogenic to S. reginae and reproduced symptoms similar to those observed in the nursery. Of the two species, P. strelitziae was more aggressive than I. strelitziae, resulting in the death of all inoculated plants. Key words: Nectriaceae multigene phylogeny pathogenicity taxonomy Article info: Submitted: 16 December 2017; Accepted: 15 March 2017; Published: 5 April INTRODUCTION Strelitzia reginae, also well-known as bird of paradise, is one of the most commercially cultivated species of Strelitzia in the world (Karsten 2009). This species was first introduced into Europe from South Africa in 1770 for its unique flowers, which are characterized by vivid orange and bright purple or blue inflorescens, making it a highly sought-after cut flower crop (Van Jaarsveld 2008, Xaba 2011). This plant is also widely used in landscaping as a focal point for many European gardens (Xaba 2011). However, commercial cultivation of S. reginae, as cut flowers or as pot plants, is limited by fungal diseases, which are poorly reported in the literature. In Hawaii, a Pythium sp. has been reported as causal agent of root rot on S. reginae, and a similar disease was reported in Egypt and Florida associated with Rhizoctonia solani and Fusarium spp. (Raabe et al. 1981, Alfenas et al. 1984, Hilal & Helmy 1998). In Italy, the most commonly reported fungal diseases of S. reginae are root and foot rot caused by Phytophthora nicotianae (Frisullo et al. 1987, Luongo et al. 2010), wilting associated with root rot caused by Cylindrocarpon destructans (Grasso & Cutuli 1972) and Armillaria mellea (Davino 1984), and southern blight caused by Sclerotium rolfsii (Polizzi et al. 2007). During 2015, a new basal stem rot was detected on approximately 20 % of potted S. reginae plants in a commercial nursery in Carrubba, Riposto (Catania province, eastern Sicily, Italy). The diseased plants displayed a dry rot of the basal stem which resulted in the detachment of the roots from the stem. In addition, symptoms of general wilting and rot of internal foliage were observed on the affected plants. The aims of this study were, therefore, to identify the pathogen(s) associated with the disease via morphological and molecular characterization, and to verify the pathogenicity of the organism(s) associated with these disease symptoms. MATERIALS AND METHODS Field survey and isolation During 2015, a survey undertaken in an ornamental nursery in eastern Sicily led to the discovery of Strelizia reginae plants displaying symptoms of dry basal stem rot (Fig. 1). Some affected plants were randomly selected and brought to the laboratory for further analyses. The symptomatic stem tissues were surface-sterilized with 1.2 % (v/v) sodium hypochlorite for 2 min, rinsed three times in sterile distilled water and dried on sterile absorbent paper. Isolations were done by transferring fragments of symptomatic and the bordering healthy plant tissue onto potato dextrose agar (PDA, Oxoid TM Lda, UK) plates (amended with streptomycin sulphate at 100 ppm) and Phytophthora selective medium PARPH (Jeffers 2017 International Mycological Association You are free to share - to copy, distribute and transmit the work, under the following conditions: Attribution: You must attribute the work in the manner specified by the author or licensor (but not in any way that suggests that they endorse you or your use of the work). Non-commercial: You may not use this work for commercial purposes. No derivative works: You may not alter, transform, or build upon this work. For any reuse or distribution, you must make clear to others the license terms of this work, which can be found at Any of the above conditions can be waived if you get permission from the copyright holder. Nothing in this license impairs or restricts the author s moral rights. 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133 Aiello et al. Fig. 1. Dry basal rot symptoms of Strelitzia reginae observed in the nursery. A C. Wilting and dying S. reginae plants. D. Dry basal stem rot. E. Rot of internal leaf. & Martin 1986). The plates were incubated at 25 C and examined daily for 1 wk. From these primary isolations, single-conidial isolates were derived and maintained on PDA slants for further study. DNA isolation, sequencing and phylogenetic analyses Total genomic DNA was extracted from 7-d-old fungal strains grown on PDA at room temperature (20 24 ºC) using the Wizard Genomic DNA Purification Kit (Promega Corporation, Madison, WI) according to the manufacturer s protocol. Six genomic gene regions were targeted for amplification and sequencing using the primer pairs and protocols described by Lombard et al. (2015): 28S large subunit (LSU) nrdna, internal transcribed spacer regions and intervening 5.8S nrrna (ITS), translation elongation factor 1-alpha (tef1), histone H3 (his3), β-tubulin (tub2), and the RNA polymerase II second largest subunit (rpb2) gene regions. Amplicons were sequenced in both directions using the same primers used for amplification and the BigDye Terminator Cycle Sequencing Kit v. 3.1 (Applied Biosystems Life Technologies, Carlsbad, CA) following the protocol provided by the manufacturer. Sequences were determined on an Applied Biosystems 3730 DNA Analyzer (Life Technologies, Carlsbad, CA). The generated sequences were analysed and consensus sequences were determined using Seqman (DNAStar, Madison, WI). All sequences were manually corrected and the arrangement of nucleotides in ambiguous positions corrected by comparisons of the sequences generated from both the forward and reverse primers. In addition to the sequences generated in this study, other sequences of closely related Nectriaceae were obtained from NCBI s GenBank nucleotide database and added to the sequence datasets generated in this study. The sequences for each locus were aligned using MAFFT v. 7 (Katoh & Standley 2013). The alignments were manually checked using MEGA v. 7 (Kumar et al. 2015) and improved where necessary. Novel sequences were lodged in GenBank (Table 1), and the alignments and phylogenetic trees in TreeBASE (S20598). Congruency of the six loci was tested using the 70 % reciprocal bootstrap criterion (Mason-Gamer & Kellogg 1996) following the protocol of Lombard et al. (2015) for each locus. Phylogenetic analyses were based on Bayesian inference (BI), Maximum Likelihood (ML), and Maximum Parsimony (MP). For both BI and ML, the evolutionary model for each partition was determined using MrModeltest (Nylander 2004) and incorporated into the analyses. For the BI analysis, MrBayes v (Ronquist & Huelsenbeck 2003) was used to generate phylogenetic trees under the optimal model per partition. A Markov Chain Monte Carlo (MCMC) algorithm of four chains was started in parallel from a random tree topology with the heating parameter set to 0.3. The MCMC analysis lasted until the average standard deviation of split frequencies decreased below 0.01 with trees saved each generations. The first 25 % of saved trees were discarded as burn-in and posterior probabilities determined from the remaining trees. The ML analysis was made with RAxML (randomized axelerated [sic] maximum likelihood for high performance computing; Stamatakis 2014) through the CIPRES website ( to obtain a second measure of branch support. The robustness of the analysis was evaluated by bootstrap support (BS) analysis with the bootstrap replicates automatically determined by the software. The MP analysis was carried out with PAUP (Phylogenetic Analysis Using Parsimony, v. 4.0b10; Swofford 2003) with phylogenetic relationships estimated by heuristic searches with random sequence-additions. Tree bisectionreconnection was implemented, with the branch swapping option set on best tree only. All characters were weighted equally and alignment gaps were treated as fifth state. Measures calculated for parsimony included tree length (TL), consistency index (CI), retention index (RI) and rescaled consistency index (RC). The bootstrap support analysis was based on replications. Taxonomy Axenic cultures were grown on synthetic nutrient-poor agar (SNA; Nirenburg 1981) amended with 1-cm 2 sterile filter paper and carnation leaf pieces, and on PDA as described by Cabral et al. (2012a). Gross morphological characteristics were studied by mounting the fungal structures in 85 % lactic acid and 30 measurements were made for all taxonomically informative characters at 1000 magnification using a 66 IMA FUNGUS

134 Pleiocarpon gen. nov. and Ilyonectria on Strelitizia Table 1. Strains included in the phylogenetic analyses. GenBank Accession no. 2 Species Isolate nr. 1 Substrate Locality ITS LSU his3 rpb2 tef1 tub2 Campylocarpon fasciculare CBS ; CPC 3970 Vitis vinifera South Africa AY HM JF AY C. pseudofasciculare CBS ; CPC 5472 V. vinifera South Africa AY HM JF AY Cinnamomeonectria cinnamomea CBS ; PC 1222 Bark Brazil KJ KJ KJ KJ IMI ; G.J.S Bark of living liana French Guiana KJ KJ KJ KJ Cylindrocarpostylus gregarius CBS Hylurgops palliatus Germany KM JQ KM KM CBS Pinus sylvestris Germany KM JQ KM KM Cylindrodendrum album CBS ; ATCC 46842; IMI Fucus distichus Canada KM KM KM KM CBS Soil The Netherlands KM KM KM KM Dactylonectria alcacerensis CBS V. vinifera Portugal JF KM JF AM D. estremocensis CBS V. vinifera Portugal JF KM JF JF D. macrodidyma CBS ; CPC 3976 V. vinifera South Africa AY KM JF AY D. torresensis CBS V. vinifera Portugal JF KM JF JF Ilyonectria capensis CBS ; CPC Protea sp. South Africa JX KM JX JX I. destructans CBS Cyclamen persicum Sweden AY KM JF AY I. leucospermi CBS ; CPC Leucospermum sp. South Africa JX KM JX JX I. liriodendri CBS V. vinifera Portugal DQ KM JF DQ I. mors-panacis CBS Panax quinquefolium Canada JF JF JF I. palmarum CBS Howea forsteriana Italy HF HF HF I. strelitziae CBS ; ST6 Strelitzia reginae Italy KY KY KY KY KY KY CBS ; ST8 S. reginae Italy KY KY KY KY KY KY ST7 S. reginae Italy KY KY KY KY KY KY ST9 S. reginae Italy KY KY KY KY KY KY ST14 S. reginae Italy KY KY KY KY KY ST15 S. reginae Italy KY KY KY KY KY KY ST25 S. reginae Italy KY KY KY KY KY KY ST26 S. reginae Italy KY KY KY KY KY KY ST27 S. reginae Italy KY KY KY KY KY KY ST28 S. reginae Italy KY KY KY KY KY KY Macronectria jungneri CBS Bark Brazil KJ KJ KJ KJ M. magna CBS Theobroma cacao Guatemala KJ KJ KJ KJ M. venezuelana CBS ; G.J.S Wood Venezuela KJ KJ KJ KJ Neonectria candida CBS ; IMI Malus sylvestris England JF HM JF JF N. ditissima CBS M. domestica Ireland KM KM KM DQ N. lugdunensis CBS ; DAOM Populus fremontii USA KM KM KM KM VOLUME 8 NO. 1 67

135 Aiello et al. Table 1. (Continued). GenBank Accession no. 2 Species Isolate nr. 1 Substrate Locality ITS LSU his3 rpb2 tef1 tub2 N. neomacrospora CBS Abies concolor The Netherlands JF HM HM DQ Pleiocarpon strelitziae CBS ; ST1; CPC S. reginae Italy KY KY KY KY KY KY CBS ; ST20 S. reginae Italy KY KY KY KY KY KY CPC S. reginae Italy KY KY KY KY KY ST3 S. reginae Italy KY KY KY KY ST4 S. reginae Italy KY KY KY KY KY ST5 S. reginae Italy KY KY KY KY KY KY ST10 S. reginae Italy KY KY KY KY KY KY ST11 S. reginae Italy KY KY KY KY KY KY ST12 S. reginae Italy KY KY KY KY KY KY ST13 S. reginae Italy KY KY KY KY KY ST17 S. reginae Italy KY KY KY KY KY KY ST18 S. reginae Italy KY KY KY KY KY KY ST19 S. reginae Italy KY KY KY KY KY KY ST21 S. reginae Italy KY KY KY KY KY KY ST22 S. reginae Italy KY KY KY KY KY KY ST23 S. reginae Italy KY KY KY KY KY KY ST24 S. reginae Italy KY KY KY KY KY KY Rugonectria neobalansae CBS Dead tree Indonesia KM HM KM HM R. rugulosa CBS Dead tree Venezuela KM KM KM KM Thelonectria discophora CBS ; AR 4742 Tepualia stipularis Chile KC KC KC KC T. olida CBS ; ATCC 16548; IMI Asparagus officinalis Germany KJ KJ KM CBS S. reginae Italy KY KY KY KY KY KY T. rubi CBS ; IMI Rubus idaeus KC KC KC KC T. trachosa CBS ; IMI Bark of conifer Scotland AY HM KM AY T. veuillotiana CBS ; AR 1751 Eucalyptus sp. Azores JQ JQ JQ JQ Tumenectria laetidisca CBS Bamboo Japan KJ KJ KJ KJ Xenogliocladiopsis cypellocarpa CBS ; CPC CBS Bamboo Jamaica KJ KJ KJ KJ Eucalyptus cypellocarpa Australia KM KM KM KM AR: Amy Y. Rossman working collection; ATCC: American Type Culture Collection, Virginia, USA; CBS: Centraalbureau voor Schimmelcultures, Utrecht, The Netherlands; CPC: Pedro Crous working collection housed at CBS;DAOM: Agriculture and Agri-Food Canada National Mycological Herbarium, Canada; G.J.S.: G.J. Samuels working collection; IMI: International Mycological Institute, CABI- Bioscience, Egham, Bakeham Lane, UK; PC: P. Chaverri working collection; ST: D. Aiello personal culture numbers. 2 ITS = internal transcribed spacers and intervening 5.8S rdna, LSU = 28S large subunit ribosomal rdna, his3 = histone H3, rpb2 = RNA polymerase II largest subunit, tef1 = translation elongation factor 1-alpha, tub2 = β-tubulin. Epi- and ex-type isolates indicated in bold. Sequences generated in this study indicated in italics. 68 IMA FUNGUS

136 Pleiocarpon gen. nov. and Ilyonectria on Strelitizia Plan-Apochromat 100/1.4 oil immersion lens (Carl Zeiss, Germany) mounted on a Zeiss Axioscope 2 microscope, with differential interference contrast (DIC) illumination. The 95 % confidence levels were determined for the conidial measurements with extremes given in parentheses. For all other fungal structures measured, only the extremes are provided. Colony colour was assessed using 7-d-old cultures on PDA incubated at room temperature and the colour charts of Rayner (1970). All descriptions, illustrations and nomenclatural data were deposited in MycoBank (Crous et al. 2004). Optimal and cardinal growth temperatures were determined by inoculating 90 mm diam PDA plates with a 4 mm diam plug cut from the edge of an actively growing colony. Each isolate was incubated at 4, 10, 15, 20, 25, 30, and 35 C with three replicate plates per strain at each temperature. Colony diameter of each isolate was determined after 1 wk by measuring the orthogonal directions. Pathogenicity Two representative isolates for each phylogenetically resolved species (CBS ) were selected for pathogenicity tests on 6-mo-old Strelizia reginae plants. Twenty plants were used for each isolate and the same number of plants was used as control. All plants were removed from their original planting substrate, rinsed with water, dipped for 1 min in a 0.3 % (v/v) mixed solution of 5 % (v/v) peracetic acid and 20 % (v/v) hydrogen peroxide (JetFive, Certis Europe), and then rinsed twice with sterile distilled water. Each plant was inoculated with a 4 mm mycelium plug obtained from the margin of an actively growing 14-d-old culture grown on PDA. Each plug was applied to a wound made at the base of the stem using a 4 mm diam cork borer as previously reported (Aiello et al. 2014, 2015). Control plants were treated similarly but inoculated with sterile PDA plugs. After inoculation, each plant was wrapped with Parafilm around the inoculation point to prevent desiccation and transplanted into pots containing sterilized growth substrate. All plants were covered with a plastic bag, and maintained in a growth chamber at 25 ºC under a 12 h fluorescent light/dark regimen. All plants were irrigated 2 4 times every week and fertilised every 30 d with 2 g/pot of complex NPK fertilizer Nitrophoska special (BASF). Plants were evaluated for disease symptoms after 2 and 4 mo. RESULTS Field survey and isolation During the survey, dry basal stem rot symptoms were observed on Strelizia reginae in a nursery where these plants are commercially cultivated (Fig. 1). These symptoms were observed on approximately of potted 2- to 8-yr-old potted plants. Isolations from the symptomatic and bordering healthy tissues consistently yielded cylindrocarponlike asexual fungi and no Phytophthora or any other fungi previously reported from S. reginae were isolated. Phylogenetic analyses Approximately 325 bases for tub2, bases for his3, ITS and tef1, and bases for LSU and rpb2 were determined in this study. Due to the limited sequence data available in GenBank for cylindrocarpon-like fungi for the his3 and rpb2 gene regions, both loci could not be included in the phylogenetic inference. The 70 % reciprocal bootstrap tree topologies for the remaining four loci revealed no conflicts and were therefore combined. The combined alignment of the ITS, LSU, tef1 and tub2 contained characters from 63 taxa, including Xenogliocladiopsis cypellocarpa (CBS ; Lombard et al. 2015) as outgroup. The number of unique site patterns per data partition, including alignment gaps, was 292 from 570 characters for ITS, 139 from 828 characters for LSU, 377 from 577 characters for tef1, and 124 from 305 characters for tub2. MrModeltest determined that all four partitions had dirichlet base frequencies. A GTR+I+G model with inverse gamma-distributed rates was used for ITS, LSU and tef1, while HKY+I+G with inverse gamma-distributed rates was implemented for tub2. The Bayesian analysis lasted generations and the consensus tree, with posterior probabilities, was calculated from 662 trees left after 220 trees were discarded as burnin. For the MP analysis characters were constant, 115 parsimony-uninformative, and 848 parsimony-informative, yielding 216 equally most parsimonious trees (TL = 3566; CI = 0.508; RI = 0.848; RC = 0.431). ML analysis resulted in a single best ML tree with InL = The best ML tree confirmed the consensus tree topologies obtained from the BI and MP analyses, and therefore only the best ML tree is presented (Fig. 2). In the phylogenetic tree (Fig. 2), the majority of the cylindrocarpon-like isolates obtained from Strelizia reginae clustered into two highly-supported clades (both with MP and ML bootstrap support (BS) of 100 % and posterior probabilities (PP) of 1.0) with a single isolate (CBS ) clustering (MP-BS & ML-BS = 100 %; PP = 1.0) with the extype of Thelonectria olida (CBS ). The first group of isolates (including CBS and CBS ) formed a highly-supported clade within the Ilyonectria clade, closely related but distinct from the ex-type sequence of I. palmarum (CBS ), thus representing a novel phylogenetic species within the cylindrocarpon-like genus Ilyonectria. The second group of isolates (including CBS and CBS ) formed a highly-supported clade closely related but distinct from the Thelonectria clade, representing a previously unrecognized phylogenetically-supported genus. Pathogenicity After 10 d, both isolates of the species named below as Pleiocarpon strelitziae (CBS and CBS ) induced dry basal stem rot symptoms on Strelizia reginae similar to those observed in the nursery (Fig. 3). As a consequence, after 2 mo, all S. reginae plants inoculated with P. strelitziae died. Both isolates of the species named here as Ilyonectria strelitziae (CBS and CBS ; Fig. 3) also induced symptoms of dry basal stem rot as observed in the nursery after 2 mo, but the plants remained alive after 4 mo. All control plants remained healthy throughout the pathogenicity test and none of the test fungi were isolated from them. Re-isolations from the symptomatic plants only yielded the respective inoculated fungi. VOLUME 8 NO. 1 69

137 Aiello et al. x5 x5 x10 Xenogliocladiopsis cypellocarpa CBS Neonectria lugdunensis CBS Neonectria candida CBS Neonectria neomacrospora CBS Neonectria ditissima CBS Dactylonectria estremocensis CBS Dactylonectria alcacerensis CBS Dactylonectria torresensis CBS Dactylonectria macrodidyma CBS Cylindrodendrum album CBS Cylindrodendrum album CBS Ilyonectria capensis CBS Ilyonectria leucospermi CBS Ilyonectria mors-panacis CBS Ilyonectria liriodendri CBS Ilyonectria destructans CBS Ilyonectria palmarum CBS ST26 ST14 ST7 ST28 ST9 Ilyonectria strelitziae CBS ST25 ST27 CBS ST15 Cinnamonectria cinnamomea CBS Cinnamonectria cinnamomea IMI Macronectria magna CBS Macronectria jungneri CBS x2 Macronectria venezolana CBS Tumenectria laetidisca CBS x2 Tumenectria laetidisca CBS Cylindrocarpostylus gregarius CBS Cylindrocarpostylus gregarius CBS Campylocarpon pseudofasciculare CBS x2 Campylocarpon fasciculare CBS Rugonectria rugulosa CBS Rugonectria neobalansae CBS Thelonectria trachosa CBS Thelonectria rubi CBS Thelonectria discophora CBS Thelonectria veuillotiana CBS Thelonectria olida CBS Thelonectria olida CBS ST13 ST17 ST19 ST18 ST5 CBS ST22 CBS CPC Pleiocarpon strelitziae ST4 ST3 ST10 ST12 ST11 ST23 ST ST21 Fig. 2. The ML consensus tree inferred from the combined ITS, LSU, tef1 and tub2 sequence alignments. Thickened branches indicate branches present in the ML, MP and Bayesian consensus trees. Branches with ML-BS & MP-BS = 100 % and PP = 1.0 are in blue. Branches with ML- BS & MP-BS 75 % and PP 0.95 are in red. The scale bar indicates 0.1 expected changes per site. The tree is rooted to Xenogliocladiopsis cypellocarpa (CBS ). Epi- and ex-type strains are indicated in bold. 70 IMA FUNGUS

138 Pleiocarpon gen. nov. and Ilyonectria on Strelitizia Fig. 3. Symptoms induced during the pathogenicity test on Strelitzia reginae. A D. Basal rot and wilting of plant caused by Pleiocarpon strelitziae. E F. Basal stem rot and rot of external leaf caused by Ilyonectria strelitziae. TAXONOMY Morphological observations supported by phylogenetic inference showed that the isolates obtained from the diseased Strelizia reginae included a new taxon in the genus Ilyonectria, and a novel cylindrocarpon-like genus, both of which are described below. Description: Perithecia formed homothallically in vitro, solitary or in groups of 2 3, developing directly on the SNA agar surface, ovoid to obpyriform, orange-red, becoming purple-red in 3 % KOH, finely warted, μm diam, to 375 μm high; with simple conidiophores arising directly from the surface of the ascomatal wall, and without a recognisable stroma; perithecial wall consisting of two poorly distinguishable regions; outer region 8 12 μm thick, composed of 2 4 layers of textura angularis to textura globosa; inner region 3 5 μm thick, composed of 3 4 layers of textura angularis. Asci subcylindrical to clavate, μm, 8-spored; apex truncate to bluntly rounded, with a visible ring. Ascospores ellipsoidal, hyaline, tapering towards both ends, divided into two equal sized cells, smooth, (8 ) μm. Conidiophores simple, solitary or aggregated into sporodochial-like structures, arising laterally or terminally from aerial mycelium or erect, arising from the agar surface, unbranched or sparsely branched, 1 4-septate, μm long, bearing one or rarely two conidiogenous cells. Conidiogenous cells monophialidic, cylindrical, tapering slightly towards the apex, long, and 2 4 μm wide at the base; micro- and macroconidia produced by simple conidiophores. Microconidia abundant, aseptate, ellipsoidal to ovoidal or subcylindrical, straight to slightly curved, with a clearly laterally displaced hilum, 5 7( 9) 2 3 μm (av. 6 2 μm), formed in heads on conidiophores. Macroconidia 1 3-septate, straight to slightly curved, base sometimes with a visible, centrally located to laterally displaced hilum; 1-septate macroconidia (9 )11 17( 20) 2 4 μm (av μm); 2-septate macroconidia (14 )18 22( 24) 3 4 μm (av μm); 3-septate macroconidia (19 )21 29( 37) 3 4 μm (av μm). Chlamydospores not observed on SNA or PDA. Culture characteristics: Colonies after 10 d at 24 C on PDA with cottony, white aerial mycelium in the centre, lacking zonation; centre dark brick becoming cinnamon to honey towards the margins; reverse dark brick in centre, becoming cinnamon towards the margins. Cardinal growth temperatures: No growth was observed at 4 and 35 C. Optimal growth was observed at 20 C, with colonies reaching mm diam. Ilyonectria strelitziae L. Lombard & D. Aiello, sp. nov. MycoBank MB (Fig. 4) Additional culture examined: Italy: Sicily: Catania Province, Carrubba, Riposto, isolated from Strelitzia reginae, 2015, D. Aiello (CBS ). Etymology: Name derived from the host, Strelitzia reginae, from which this fungus was isolated. Diagnosis: Perithecia orange-red with simple conidiophores arising directly from the surface of the ascomatal wall. Asexual morph producing abundant micro- and macroconidia, but no chlamydospores in culture. Type: Italy: Sicily: Catania Province, Carrubba, Riposto, isolated from Strelitzia reginae, 2015, D. Aiello (CBS H [dried culture] holotype; CBS ex-type culture). Notes: Based on the phylogenetic inference obtained in this study, Ilyonectria palmarum (Aiello et al. 2014) is the closest phylogenetic neighbour to I. strelitziae (Fig. 2). Ilyonectria strelitziae can be distinguished from I. palmarum by the simple conidiophores arising from the ascomatal wall, a character not previously reported in Ilyonectria (Chaverri et al. 2011, Cabral et al. 2012a, b, Lombard et al. 2013, 2014, 2015). Additionally, I. strelitziae readily produced 1-septate macroconidia, not reported for I. palmarum (Aiello et al. 2014). VOLUME 8 NO. 1 71

139 Aiello et al. Fig. 4. Ilyonectria strelitziae (ex-type culture CBS ). A. Perithecia on PDA surface. B. Perithecium with simple conidiophores arising from the ascomatal wall. C. Perithecium exuding asci and ascospores. D E. Simple conidiophores arising from the ascomatal wall. F. Ascomatal wall colour reaction in KOH. G H. Asci. I. Ascospores. J K. Simple conidiophores on aerial mycelium. L. Aggregation of simple conidiophores on carnation leaf. M. Aggregation of simple conidiophores. N. Microconidia. O. 1- and 2-septate macroconidia. P. 3-septate macroconidia. Bars A B = 500 μm; C = 100 μm; D E = 50 μm; F K and M P = 10 μm. 72 IMA FUNGUS

140 Pleiocarpon gen. nov. and Ilyonectria on Strelitizia Fig. 5. Pleiocarpon strelitziae (ex-type CBS ). A B. Simple conidiophores. C-D. Sporodochia. E. Microconidia. F. Macroconidia. Bars = 10 μm Pleiocarpon L. Lombard & D. Aiello, gen. nov. MycoBank MB (Fig. 5) Etymology: Named after the highly variable conidial shapes this fungus produces in culture. Diagnosis: Sexual morph unknown. Asexual morph cylindrocarpon-like; microconidia abundant, aseptate, ellipsoid to ovoid or subcylindrical, straight to slightly curved, with clearly laterally displaced hilum; macroconidia cylindrical to subcylindrical, straight to curved, 1 5-septate. Type species: Pleiocarpon strelitziae L. Lombard & D. Aiello Description: Ascomata not observed. Conidiophores simple or aggregated to form sporodochia; simple conidiophores arising laterally or terminally from aerial mycelium, solitary to loosely aggregated, unbranched or sparsely branched, septate, bearing up to two conidiogenous cells. Conidiogenous cells monophialidic, cylindrical, tapering slightly towards the apex. Microconidia abundant, aseptate, hyaline, ellipsoid to ovoid or subcylindrical, straight to slightly curved, with clearly laterally displaced hilum. Macroconidia cylindrical to subcylindrical, hyaline, straight to curved, 1 5-septate, apex or apical cell typically slightly bent to one side and minutely beaked, base with sometimes visible, centrally located or laterally displaced hilum. Chlamydospores not observed. Notes: Pleiocarpon is a new cylindrocarpon-like monotypic genus, phylogenetically closely related to the genus Thelonectria (Chaverriet al. 2011). The asexual morph of Thelonectria rarely produces microconidia (Chaverri et al. 2012, Salgado-Salazar et al. 2016) in contrast to Pleiocarpon. Additionally, the macroconidia of the asexual morph of Thelonectria are large and up to 9-septate (Chaverri et al. 2012, Salgado-Salazar et al. 2016), whereas those of Pleiocarpon are more intermediate in size and up to 5-septate. Pleiocarpon strelitziae L. Lombard & D. Aiello, sp. nov. MycoBank MB (Fig. 5) Etymology: Name derived from the host, Strelitzia reginae, from which this fungus was isolated. Type: Italy: Sicily: Catania Province, Carrubba, Riposto, isolated from Strelitzia reginae, 2015, D. Aiello (CBS H [dried culture] holotype; CBS ex-type culture). Description: Ascomata not observed. Conidiophores simple or aggregating to form sporodochia. Simple conidiophores solitary, arising laterally or terminally from aerial mycelium, to loosely aggregated, unbranched or sparsely branched, 1 3-septate, μm long, bearing one, rarely two conidiogenous cells. Conidiogenous cells monophialidic, cylindrical, tapering slightly towards the apex, μm long, 2 3 μm wide at the base. Sporodochia consisting of a pulvinate mass of short conidiophores, the conidiogenous cells monophialidic, cylindrical, tapering towards the apex, μm long, and 2 4 μm wide at the base. Microconidia aseptate, with a minute or clearly laterally displaced hilum, ellipsoid to ovoid or subcylindrical, straight to slightly curved, (6 ) μm (av. 8 3 μm), formed in heads on simple conidiophores or in hyaline, slimy masses on sporodochia. Macroconidia formed by both types of conidiophores, cylindrical to subcylindrical, hyaline, straight to curved, 1 5-septate, apex or apical cell typically slightly bent to one side and minutely beaked, base with sometimes visible, centrally located or laterally displaced hilum; 1-septate macroconidia (19 )27 40 (3 )5 7 μm (av μm); 2-septate macroconidia 23 29( 31) 5 μm (av μm); VOLUME 8 NO. 1 73

141 Aiello et al. 3-septate macroconidia (28 )30 40( 46) 5 6 μm (av μm); 4-septate macroconidia (36 ) μm (av μm); 5-septate macroconidia (41 )42 47( 50) 5 7 μm (av μm). Chlamydospores not observed. Culture characteristics: Colonies after 10 d at 24 C on PDA with sparse cottony, white aerial mycelium, lacking zonation; surface and reverse cinnamon to honey. Cardinal growth temperatures: No growth observed at 4 C, while colonies grew at C, with optimal growth at C with colonies reaching mm diam in one week. Additional culture examined: Italy: Sicily: Catania Province, Carrubba, Riposto, isolated from Strelitzia reginae, 2015, D. Aiello (CBS ). DISCUSSION Basal stem and root rot of Strelitzia reginae can be caused by different fungal species, including Armillaria mellea, Fusarium spp., Phytophthora spp., Pythium spp., Rhizoctonia solani, and Sclerotium rolfsii. In our study, a new dry basal stem rot was detected on potted S. reginae plants cultivated in a nursery in eastern Sicily. Three different cylindrocarpon-like species were consistently isolated from diseased plants, while the above-mentioned pathogens were not found associated with the symptomatic tissues. According to phylogenetic inference supported by morphological characters, some of the isolates associated with this new dry basal rot of S. reginae were identified as a new species of Ilyonectria, described here as I. strelitziae. Species of this genus are commonly found in soil and cause collar and root diseases on a wide range of plant hosts worldwide (Sánchez et al. 2002, Seifert et al. 2003, Halleen et al. 2004, 2006, Alaniz et al. 2007, 2009, Agustí-Brisach et al. 2011, Dart & Weeda 2011, Petit et al. 2011, Cabral et al. 2012a, b, Özben et al. 2012, Úrbez-Torres et al. 2012, Erper et al. 2013, Lombard et al. 2013, 2014). In Italy, several species have been reported from ornamental nurseries in eastern Sicily. Dactylonectria pauciseptata, I. novozelandica, and I. torresensis, were reported on potted Laurus tinus plants associated with root and crown rot symptoms (Aiello et al. 2015); I. palmarum associated with dry basal rot of Arecaceae (Aiello et al. 2014); and I. macrodidyma associated with root rot of avocado (Persea americana; Vitale et al. 2012). The pathogenicity test in this study showed that I. strelitziae induced similar dry basal rot symptoms on S. reginae plants to those observed in the nursery. However, nothing is yet known on the origin of this fungal pathogen or its distribution in southern Sicily, which requires further investigation Some of the remaining isolates obtained from S. reginae also had a cylindrocarpon-like morphology, which phylogenetic inference in this study showed to represent a separate genus, here named Pleiocarpon, based on the new species P. strelitziae. Phylogenetic inference showed that these fungi formed a highly-supported, but distinct, clade closely related to the genus Thelonectria. The genus Thelonectria was introduced by Chaverri et al. (2011) to accommodate fungi with cylindrocarpon-like asexual morphs belonging to Booth s Group 2 (Booth 1966), characterized by lacking microconidia and chlamydospores. Recently, Salgado-Salazar et al. (2016) segregated Thelonectria by introducing three new genera (Cinnamomeonectria, Macronectria, and Tumenectria) based on phylogenetic inference. Pleicarpon can be distinguished from these four genera by the abundant microconidia it produces in culture. Moreover, these genera morphologically resemble Thelonectria, and are mostly found on bark or exposed wood of dead, dying or diseased trees, and are rarely associated with small cankers and root rots (Chaverri et al. 2011, Salgado-Salazar et al. 2016). Pathogenicity tests undertaken in this study demonstrated the aggressive pathogenic nature of P. strelitziae, resulting in mortality of all inoculated test plants within 2 mo, which should be of great concern to the cut-flower industry. However, the origin and distribution of this aggressive pathogen still requires further investigation. One isolate obtained from diseased S. reginae plants was identified as T. olida based on phylogenetic inference. However, the morphology could not be confirmed, as the isolate did not sporulate on any of the media used. Past phylogenetic studies (Chaverri et al. 2011, Salgado-Salazar et al. 2016) have shown that this atypical species belongs in the genus Thelonectria, although this species produces chlamydospores and has shorter macroconidia. Thelonectria olida has been isolated from rotting roots of several plant hosts, but its pathogenicity has never been confirmed (Salgado-Salazar et al. 2016). The T. olida isolate obtained was excluded from the pathogenicity test undertaken in this study, as its identity was only determined after these had been carried out. On the basis of the disease incidence and severity observed in the ornamental nursery, we believe that this disease represents a serious threat to potted field-grown plants of S. reginae. The cultivation method of pot production could play an important role in promoting infections, since the plants are stressed by frequently being replanting into pots, and wounds can be incurred during transplanting. Moreover, the use of infected soil could represent a further inoculum source for these fungi (Aiello et al. 2014). A wilt disease associated with root rot of S. reginae caused by Cylindrocarpon destructans was reported in Sicily, but that fungus was identified based only on morphological features (Grasso & Cutuli 1972) and requires critical confirmation. 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144 doi: /imafungus IMA FUNGUS 8(1): (2017) New endophytic Toxicocladosporium species from cacti in Brazil, and description of Neocladosporium gen. nov. Jadson D.P. Bezerra 1,2 *, Marcelo Sandoval-Denis 3,4 *, Laura M. Paiva 1, Gladstone A. Silva 1,2, Johannes Z. Groenewald 3, Cristina M. Souza-Motta 1,2, and Pedro W. Crous 3,4,5 1 Departamento de Micologia Prof. Chaves Batista, Universidade Federal de Pernambuco, Av. Prof. Moraes Rego, s/n, Centro de Biociências, Cidade Universitária, CEP: , Recife, PE, Brazil; corresponding author jadsondpb@gmail.com 2 Programa de Pós-Graduação em Biologia de Fungos (PPG-BF), Departamento de Micologia Prof. Chaves Batista, Universidade Federal de Pernambuco, Av. Prof. Moraes Rego, s/n, Centro de Biociências, Cidade Universitária, CEP: , Recife, PE, Brazil 3 Westerdijk Fungal Biodiversity Institute, Uppsalalaan 8, 3584 CT, Utrecht, The Netherlands; corresponding author p.crous@ westerdijkinstitute.nl 4 Faculty of Natural and Agricultural Sciences, Department of Plant Sciences, University of the Free State, P.O. Box 339, Bloemfontein 9300, South Africa 5 Microbiology, Department of Biology, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands *These authors contributed equally to this work Abstract: Brazil harbours a unique ecosystem, the Caatinga, which belongs to the tropical dry forest biome. This region has an important diversity of organisms, and recently several new fungal species have been described from different hosts and substrates within it. During a survey of fungal endophyte diversity from cacti in this forest, we isolated cladosporium-like fungi that were subjected to morphological and multigene phylogenetic analyses including acta, ITS, LSU, rpb2 and tub2 gene sequences. Based on these analyses we identified two new species belonging to the genus Toxicocladosporium, described here as T. cacti and T. immaculatum spp. nov., isolated from Pilosocereus gounellei subsp. gounellei and Melocactus zehntneri, respectively. To improve the species recognition and assess species diversity in Toxicocladosporium we studied all ex-type strains of the genus, for which acta, rpb2 and tub2 barcodes were also generated. After phylogenetic reconstruction using five loci, we differentiated 13 species in the genus. Toxicocladosporium velox and T. chlamydosporum are synonymized based on their phylogenetic position and limited number of unique nucleotide differences. Six strains previously assigned to T. leucadendri, including the ex-type strain (CBS ) of that species, were found to belong to an undescribed genus here named as Neocladosporium gen. nov., with N. leucadendri comb. nov. as type species. Furthermore, this study proposes the acta, ITS, rpb2 and tub2 as main phylogenetic loci to recognise Toxicocladosporium species. Key words: Cladosporiaceae Endophytic fungi Multigene phylogeny Taxonomy Article info: Submitted: 16 March 2017; Accepted: 24 April 2017; Published: 1 May INTRODUCTION The genus Toxicocladosporium (Cladosporiaceae, Capnodiales) was described by Crous et al. (2007) to accommodate cladosporium-like fungi having distinct dark, thick-walled conidial and conidiophore septa, and lacking the typical coronate Cladosporium scar type. The type species of this genus, T. irritans, was isolated from mouldy paint in Suriname and named irritans because of the production of several volatile metabolites in culture, causing skin irritation when there is exposure to the fungus (Crous et al. 2007). After its introduction, several new species were described in the genus, which currently comprises 13 species reported from different host plants in studies from America (Suriname and USA), Africa (Madagascar and South Africa), Asia (China), and Oceania (Australia). Similar to Cladosporium, Toxicocladosporium exhibits a widespread distribution and the capacity to colonise distinct substrates and plant families. Almost all species in this genus were described from plant species belonging to the families Asteraceae, Cyperaceae, Myrtaceae, Pinaceae, Proteaceae, Rubiaceae, and Strelitziaceae (Crous et al. 2009a, 2010a, b, 2011a, 2012a, b, 2013, 2014, Crous & Groenewald 2011), the exception being T. irritans and T. hominis described from mouldy paint (Crous et al. 2007) and a human bronchoalveolar lavage fluid specimen (Crous et al. 2016), respectively. In addition to species descriptions in this genus, few reports of isolation of Toxicocladosporium species, mainly T. irritans, have been published in different countries. For example, studying ancient laid-paper documents of the 17th century in Portugal, Mesquita et al. (2009) reported the isolation of T. irritans. Similar results were obtained in Italy by Piñar et 2017 International Mycological Association You are free to share - to copy, distribute and transmit the work, under the following conditions: Attribution: You must attribute the work in the manner specified by the author or licensor (but not in any way that suggests that they endorse you or your use of the work). Non-commercial: You may not use this work for commercial purposes. No derivative works: You may not alter, transform, or build upon this work. For any reuse or distribution, you must make clear to others the license terms of this work, which can be found at Any of the above conditions can be waived if you get permission from the copyright holder. Nothing in this license impairs or restricts the author s moral rights. VOLUME 8 NO. 1 77

145 Bezerra et al. al. (2015) who used culture-independent molecular methods and scanning electron microscopy (SEM) to verify the fungi colonizing parchment manuscripts, and by Bonadonna et al. (2014), who reported T. irritans colonising tattoo inks. These reports may show similarities because the first isolation of T. irritans was associated with mouldy paint in Suriname (Crous et al. 2007). Toxicocladosporium irritans was also reported associated with patients having atopic dermatitis in Japan (Zhang et al. 2011), and it was isolated from human blood and a fingernail by Sandoval-Denis et al. (2015) in the USA. This species was also reported by Cruywagen et al. (2015) on baobab trees in southern Africa, and on equipment used in the International Space Station or Space Shuttle in Japan (Satoh et al. 2016). There are also reports from other unusual substrates or hosts, including coffee scale insects in Vietnam (Nhạ et al. 2011), the vector of visceral leishmaniasis (Lutzomyia longipalpis) in Brazil (McCarthy et al. 2011), an unidentified sponge from Korea (Cho et al. 2016), patients with seborrheic dermatitis in Japan (Tanaka et al. 2014), outdoor dust samples in the USA (Barberán et al. 2015), and as a plant pathogen on African olive (Olea europaea subsp. cuspidata, Oleaceae) in Australia (Australian Government Department of Agriculture 2015). Toxicocladosporium and Cladosporium were also suggested as candidates for fungal structures found in the fossilized extinct aquatic angiosperm Eorhiza arnoldii in Canada (Klymiuk et al. 2013). These reports show that Toxicocladosporium host associations are not specific and may differ from Cladosporium, in which species tend to have confined host ranges, but with some exceptions (Bensch et al. 2012). Toxicocladosporium chlamydosporum and T. rubrigenum were, however, described from a single leaf spot of Eucalyptus camaldulensis (Myrtaceae) growing in Madagascar (Crous et al. 2009a). This example demonstrates that specimens from a single host and location can be colonized by genotypes representing different species (Bensch et al. 2012). Toxicocladosporium species may be recovered from inconspicuous substrates and extreme habitats, showing a lack of environmental preference and an ability to be associated with unusual materials and ecological conditions (McCarthy et al. 2011, Nhạ et al. 2011, Cho et al. 2016, Satoh et al. 2016). Dematiaceous fungi isolated from different plant species in extreme environments generally live as endophytes (Redman et al. 2002, Suryanarayanan et al. 2011, Loro et al. 2012, Sun et al. 2012, Knapp et al. 2015). Although Cladosporium species are widely reported as endophytes (Bensch et al. 2012), the closely related genus Toxicocladosporium has not previously been reported as endophytic. All presently known associations of Toxicocladosporium species with plant material were as an epiphyte, saprobe, or phytopathogen, or with unusual substrates or hosts. Plants living in dry environments are an important host for fungi with widespread distributions, and have always shown a great diversity of species (Fisher et al. 1994, Suryanarayanan et al. 2005, Khidir et al. 2010, Silva-Hughes et al. 2015, Fonseca-García et al. 2016). The Caatinga, one of the most important tropical dry forests in Brazil, harbours several cacti that prove to have a great diversity of endophytic fungi (Bezerra et al. 2012, 2013, Freire et al. 2015). Recently, Bezerra et al. (2017) described a new order in the class Dothideomycetes for endophytes isolated from the cactus Tacinga inamoena collected in the Caatinga. We studied all ex-type strains of Toxicocladosporium species isolated from different substrates and hosts in order to report on the isolation and to describe those we recovered as endophytes from the cacti Melocactus zehntneri and Pilosocereus gounellei subsp. gounellei growing in the Caatinga. Using morphological characters and multigene phylogenetic analyses (acta, ITS, LSU, rpb2 and tub2), the genus Toxicocladosporium and its respective species were reevaluated. We aimed to determine the phylogenetic relationship of endophytes from cacti with species of Toxicocladosporium, provide an overview of hosts and substrates amongst Toxicocladosporium species, and propose new loci to assist with species differentiation in the genus. MATERIALS AND METHODS Endophytic fungi from cacti Endophytic fungi were isolated as described by Bezerra et al. (2013) from the cacti Melocactus zehntneri and Pilosocereus gounellei subsp. gounellei growing in the Brazilian tropical dry forest (Caatinga), Catimbau National Park, Buíque municipality, Pernambuco state, Brazil (8º36 35 S, 37º14 40 W), and sustainable family farming plots, Itaíba municipality, Pernambuco state, Brazil (9 o S, 37 o W). The collections were authorized by the Ministério do Meio Ambiente (MMA)/Instituto Chico Mendes de Conservação da Biodiversidade (ICMBio); permission number: / authentication code issued on 4 November, In addition, 32 isolates selected on the basis of genetic and morphological relatedness with cacti endophytes, were obtained from the collection of the Westerdijk Fungal Biodiversity Institute (formerly CBS-KNAW Fungal Biodiversity Centre, Utrecht, The Netherlands) and the CPC collection (collection of P.W. Crous, held at CBS) and included in the analyses (Table 1). Morphology Endophytes previously identified as belonging to Toxicocladosporium were cultured on malt extract agar (MEA), oatmeal agar (OA), potato dextrose agar (PDA), and synthetic nutrient deficient agar (SNA) (Crous et al. 2009c), and incubated at 22 C under a natural day-night cycle. Macro- and micro-morphological features, and reproductive structures were visualized after 3 wk on MEA, OA, PDA, and/ or SNA culture media. Culture colours were evaluated using the charts of Rayner (1970). Slide preparations were mounted as described by Bensch et al. (2012) in clear lactic acid and/ or in Shear s solution. Endophytic strains are deposited in the culture collections of Micoteca URM Prof. Maria Auxiliadora Cavalcanti (Federal University of Pernambuco, Recife, Brazil WCDM 604) and the CBS collection at Westerdijk Fungal Biodiversity Institute (under Material Transfer Agreement MTA N o 05/2015/Micoteca URM, issued on 14 April, 2015). Nomenclatural and taxonomic information were deposited in MycoBank ( org) (Crous et al. 2004). 78 IMA FUNGUS

146 New Toxicocladosporium species from cacti in Brazil DNA extraction, amplification (PCR) and sequencing Genomic DNA extraction was performed using the Wizard Genomic DNA Purification Kit (Promega, Madison, WI) according to the manufacturer s instructions. The primers LR0R and LR5 (Vilgalys & Hester 1990), ITS5 and ITS4 (White et al. 1990), ACT-512F and ACT-783R (Carbone & Kohn 1999), Bt2a and Bt2b or Bt10 (Glass & Donaldson 1995) and 5f2 and 7cr (O Donnell et al. 2010) were used to amplify part of the nuclear ribosomal large subunit (LSU) of the rdna, the ITS region (first and second ITS regions and intervening 5.8S nrdna), the partial actin gene (acta), partial β-tubulin gene (tub2), and a fragment of the RNA polymerase second largest subunit gene (rpb2) respectively. Amplification and sequencing reactions, sequences analyses, and consensus sequences were performed as described by O Donnell et al. (2010) and Bezerra et al. (2017). In addition, 136 DNA sequences representing 57 taxa were retrieved from GenBank and included in the phylogenetic analyses (Table 1). Phylogenetic analyses Following blast searches of the NCBI s GenBank nucleotide database for preliminary identifications, an initial backbone tree was constructed using ITS, LSU and rpb2 sequences from Cladosporiaceae (Schubert et al. 2007a, b, Zalar et al. 2007, Crous et al. 2007, 2009b, 2011a, Bensch et al. 2010, 2012, 2015) and from the other six families in Capnodiales following Quaedvlieg et al. (2014) and Videira et al. (2016). Parastagonospora nodorum (CBS ) was used as outgroup. Firstly, the alignments for each locus were performed using the online MAFFT interface (Katoh & Standley 2013) followed by manual adjustments using MEGA v. 7 (Kumar et al. 2015). These alignments were used to infer preliminary phylogenetic relationships for Toxicocladosporium species in Cladosporiaceae. A second, more inclusive analysis included acta, LSU, ITS, rpb2 and tub2 sequences derived from ex-type cultures of Toxicocladosporium species and endophytes isolated from cacti (Crous et al. 2007, 2009a, 2010a, b, 2011a, 2012a, b, 2013, 2014, 2016, Crous & Groenewald 2011). Neocladosporium leucadendri (CPC = CBS ), previously published as Toxicocladosporium leucadendri, was used as outgroup for that analysis. Maximum Parsimony analyses (MP) were performed with PAUP v. 4.0b10 (Swofford 2003) and involved 1000 replicates of heuristic search with random addition of sequences. The tree bisection-reconnection option was used, with the branch swapping option set to best-trees only. Gaps were treated as missing data and all characters were unordered and given equal weight. The tree length (TL), consistency index (CI), Retention index (RI), and rescaled consistence index (RC) were calculated. Maximum parsimony bootstrap analyses (MP-BS) were performed using 1000 replicates. Maximum likelihood analyses (ML) were performed using RAxML-HPC2 v (Stamatakis 2014) on XSEDE in the CIPRES science gateway ( The robustness of the trees obtained was evaluated according to the level of bootstrap support (ML- BS), with the number of replicates determined automatically by the software. Bayesian analyses (BI) were performed using MrBayes v (Huelsenbeck & Ronquist 2001). The program was executed with four Markov chains in two simultaneous runs for 5 M generations with the stopval option on and saving trees every 1000 generations. The analyses were stopped when the two runs converged and the average standard deviation of split frequencies came below The 50 % majority-rule consensus tree and the Bayesian posterior probabilities (BPP) were calculated after discarding the first 25 % of saved trees as burn-in. The best fit evolutionary models were calculated independently for each gene data partition using MrModelTest v. 2.3 (Nylander 2004) following the Akaike information criterion and included in the analyses, in all cases selecting the GTR+I+G model. All resulting trees were plotted using FigTree v ( All the analyses were first made independently for each locus and visually inspected for topological incongruences between nodes with significant statistical support before being combined into multigene datasets (Mason-Gamer & Kellogg 1996, Wiens 1998). The new sequences generated in this study were deposited in the NCBI s GenBank nucleotide database and the European Nucleotide Archive (Table 1) and the alignments and phylogenetic trees in TreeBASE (Study ID S20701). RESULTS In order to verify the relationship of Toxicocladosporium with other genera in the family Cladosporiaceae, we used ITS, LSU and rpb2 sequences from representatives of 19 genera from seven families in Capnodiales. Parastagonospora nodorum (CBS ) was used as outgroup. The final combined alignment contained 68 isolates and 1750 characters (ITS: 427, LSU: 621 and rpb2: 702) of which 770 were parsimony-informative (ITS: 178, LSU: 161 and rpb2: 431), 117 were variable and parsimony-uninformative (ITS: 37, LSU: 59 and rpb2: 21), and 848 were constant (ITS: 427, LSU: 621 and rpb2: 702). Because of the high degree of sequence conservation, the LSU analysis alone was not able to resolve the generic limits in Cladosporiaceae, i.e. the Toxicocladosporium species did not form a monophyletic clade but were intermixed with species of Cladosporium (data not shown); thus, the combined ITS, LSU, and rpb2 sequences were more informative when used in a combined alignment. Fig. 1 shows a RaxML tree and node support values obtained using MP, ML and BI analyses. Parsimony analysis resulted in 68 trees (TL = 4735; CI = 0.347; RI = 0.705; RC = 0.245). These analyses show that all Toxicocladosporium species cluster together in a clade (MP-BS 100 %, ML-BS 91 %, BPP 0.98) closely related to Cladosporium, with the exception of T. leucadendri. The ex-type strain of the latter species (CBS ) and five other isolates formed a distinct linage phylogenetically, close but unrelated to, the genera Graphiopsis and Verrucocladosporium, representing a different genus we describe here as Neocladosporium, with N. leucadendri as the type species. The second alignment included ITS and LSU sequences from all the available ex-type strains of Toxicocladosporium species with N. leucadendri as outgroup. To further improve VOLUME 8 NO. 1 79

147 Bezerra et al. Table 1. GenBank accession numbers and details of strains used in this study. Species Strain/isolate number 1 Substrate/host (country) GenBank accession numbers 2 Acrodontium crateriforme CBS T (ex-type of Chloridium crateriforme) ITS LSU acta rpb2 tub2 Tuberculina maxima (The Netherlands) FN KX KX A. luzulae CBS T On leaf of Carex sp. (The Netherlands) KX KX KX Cercospora beticola CBS On Beta vulgaris (Italy) DQ DQ KT C. capsici CBS Unknown host, on calyx attached to GU KF KT fruit (Fiji) Cladosporium allicinum CBS ET = CPC On Hordeum vulgare (Belgium) EF KJ C. chalastosporoides CBS ET = CPC On Protea arborea (South Africa) HM KJ LT C. fusiforme CBS T Hypersaline water of Secovlje salterns DQ KJ (Slovenia) C. herbarum CBS ET = CPC On Hordeum vulgare (The Netherlands) EF KJ LT C. hillianum CBS T = CPC On Typha orientalis (New Zealand) HM KJ C. iridis CBS ET (ex-epitype of On Iris sp. (The Netherlands) EF EU KT Scolicotrichum iridis) Dissoconium aciculare CBS On Astragalus sp. (Germany) AY GU KX D. aciculare CBS T On Medicago lupulina (Germany) NR_ EU D. eucalypti CBS = CPC On Malus domestica fruit (USA) JQ JQ D. proteae CBS T = CPC On leaves of Protea sp. (Spain) EU EU Extremus adstrictus CBS T = TRN96 (ex-type of Rock sample (Spain) NR_ KF Devriesia adstricta) E. antarcticus CBS T = CCFEE 451 (ex-type of Devriesia antarctica) Rock sample (Antarctica) NR_ GU CBS = CCFEE 5207 Rock sample (Antarctica) KF KF Graphiopsis chlorocephala CBS = CPC On leaves of Paeonia delavayi EU EU LT (Germany) CBS On leaf and stem lesions on Paeonia EU EU KT sp. (New Zealand) Mycodiella eucalypti CBS = CPC On leaves of Eucalyptus diversicolor KY KY KY (Australia) M. laricis-leptolepidis MAFF On Larix leptolepis (Japan) JX JX M. sumatrensis CBS T = CPC (ex-type of On Eucalyptus sp. (Indonesia) KF KF LT Mycosphaerella sumatrensis) 80 IMA FUNGUS

148 New Toxicocladosporium species from cacti in Brazil Table 1. (Continued). Species Strain/isolate number 1 Substrate/host (country) GenBank accession numbers 2 ITS LSU acta rpb2 tub2 Neocladosporium leucadendri gen. sp. nov. CBS T = CPC (ex-type of Toxicocladosporium leucadendri) On leaves of Leucadendron sp. (South Africa) JQ JQ LT LT KY CPC CPC On leaves of Kunzea pauciflora (Australia) On leaves of Hakea marginata (Australia) LT LT LT LT LT LT CPC On leaves of Hakea sp. (Australia) LT LT LT CPC On leaves of Banksia media (Australia) LT LT LT CPC On leaves of Petrophile sp. (Australia) LT LT LT Neodevriesia hilliana CBS T = CPC (ex-type of Devriesia hilliana) On leaves of Macrozamia communis (New Zealand) NR_ GU LT Neodevriesia sp. CBS = TRN142 Rock sample (Spain) AY GU N. xanthorrhoeae CBS T = CPC (ex-type of Devriesia zantheria) On leaves of Xanthorrhoea australis (Australia) NR_ HQ Parastagonospora nodorum CBS On Lolium perenne (Denmark) KF EU Pseudocercospora eucalyptorum CBS = CPC On Eucalyptus sp. (Portugal) KF KF LT P. robusta CBS T = CPC 1269 On Eucalyptus robur (Malaysia) KF KF KX P. schizolobii CBS T = CPC (ex-type of Passalora schizolobii) On Schizolobium parahybum (Ecuador) KF KF Rachicladosporium cboliae CBS T = CPC On twig debris (USA) GU GU LT R. luculiae CBS T = CPC On leaf of Luculia sp. (New Zealand) EU EU R. pini CBS T = CPC On needles of Pinus monophylla (The Netherlands) JF JF LT Ramichloridium apiculatum CBS Soil (Pakistan) EU EU KX R. cucurbitae CBS T = CPC On fruit of Cucurbita maxima (USA) NR_ NG R. luteum CBS T = CPC On fruit of Malus domestica (China) NR_ JQ Ramularia endophylla CBS ET (ex-epitype of Sphaeria punctiformis) On dead leaves of Quercus robur KF KF KP From human bronchoalveolar lavage R. glennii CBS T fluid (The Netherlands) Readeriella menaiensis CBS T = CPC R. tasmanica CBS T = CPC On leaves of Eucalyptus oblonga (Australia) On leaves of Eucalytus delegatensis (Australia) KJ KJ KJ KX KF KX KF KF KX VOLUME 8 NO. 1 81

149 Bezerra et al. Table 1. (Continued). Species Strain/isolate number 1 Substrate/host (country) GenBank accession numbers 2 ITS LSU acta rpb2 tub2 Schizothyrium pomi CBS On Polygonum sachalinense (The Netherlands) EF KF CBS Unknown host (Italy) EF KF Teratosphaeria fibrillosa CBS ET = CPC On leaves of Protea sp. (South Africa) KF KF LT T. fimbriata CBS T = CPC (ex-type of Mycosphaerella fimbriata) On leaves of Corymbia sp. (Australia) KF KF LT T. molleriana CBS = CMW On Eucalyptus globulus (Australia) KF KF KX Toxicocladosporium banksiae CBS T = CPC T. cacti sp. nov. URM 7489 T = CBS URM 7490 = CBS JB 191 JB 192 JB JB 225 JB 226 JB 231 JB 235 JB 236 JB JB T. chlamydosporum CBS T = CPC (ex-type of T. chlamydosporum) On leaves of Banksia emulata (Australia) Endophyte from Pilosocereus gounellei subsp. gounellei (Brazil) Endophyte from Pilosocereus gounellei subsp. gounellei (Brazil) Endophyte from Pilosocereus gounellei subsp. gounellei (Brazil) Endophyte from Pilosocereus gounellei subsp. gounellei (Brazil) Endophyte from Pilosocereus gounellei subsp. gounellei (Brazil) Endophyte from Pilosocereus gounellei subsp. gounellei (Brazil) Endophyte from Pilosocereus gounellei subsp. gounellei (Brazil) Endophyte from Pilosocereus gounellei subsp. gounellei (Brazil) Endophyte from Pilosocereus gounellei subsp. gounellei (Brazil) Endophyte from Pilosocereus gounellei subsp. gounellei (Brazil) Endophyte from Pilosocereus gounellei subsp. gounellei (Brazil) Endophyte from Pilosocereus gounellei subsp. gounellei (Brazil) On leaf of Eucalyptus camaldulensis (Madagascar) HQ HQ LT LT KY KY KY LT LT KY KY KY LT LT KY KY KY LT LT KY KY KY LT LT KY KY KY LT LT KY KY KY LT KY KY KY LT LT KY KY KY LT LT KY KY KY LT LT KY KY KY LT LT KY KY KY LT LT KY KY KY LT KY FJ FJ LT LT KY CBS T = CPC (ex-type of T. velox) On leaf of Eucalyptus camaldulensis (Madagascar) FJ FJ LT LT KY IMA FUNGUS

150 New Toxicocladosporium species from cacti in Brazil Table 1. (Continued). Species Strain/isolate number 1 Substrate/host (country) GenBank accession numbers 2 ITS LSU acta rpb2 tub2 T. ficiniae CBS T = CPC T. hominis CBS T = FMR T. immaculatum sp. nov. URM 7491 T = CBS On leaves of Ficinia indica (South Africa) From human bronchoalveolar lavage fluid (USA) Endophyte from Melocactus zehntneri (Brazil) KF KF LT LT KY LN KY LT LT KY KY KY LT LT KY T. irritans CBS T From mouldy paint (Suriname) EU EU LT LT KY T. pini CBS T = CPC On needles of Pinus sp. (China) KJ KJ LT LT KY T. posoqueriae CBS T = CPC T. protearum CBS T = CPC T. pseudoveloxum CBS T = CPC T. rubrigenum CBS T = CPC T. strelitziae CBS T = CPC On leaves of Posoqueria latifolia (Australia) On leaves of Protea burchellii (South Africa) On leaf bracts of Phaenocoma prolifera (South Africa) On leaf of Eucalyptus camaldulensis (Madagascar) On leaves of Strelitzia reginae (South Africa) NR KC LT LT KY HQ HQ LT LT KY JF JF LT KY FJ FJ LT LT KY NR JX LT LT KY Undescribed species CPC On Cyperaceae (Australia) LT LT LT Undescribed species CPC On Cyperaceae (Australia) LT LT LT Uwebraunia australiensis CBS T = CPC On Eucalyptus platyphylla (Australia) KF GQ LT U. commune CBS T = CPC 830 On Eucalyptus nitens (South Africa) AY KJ U. dekkeri CPC On Eucalyptus molucana (Australia) GQ GQ LT On lichen Dirina massiliensis (United Verrucocladosporium dirinae CBS T Kingdom) EU EU CBS: Westerdijk Fungal Biodiversity Institute, Uppsalalaan 8, 3584 CT, Utrecht, The Netherlands; CCFEE: Culture collection from extreme environments of the Dipartamento di Scienze Ambientali, University of Tuscia, Viterbo, Italy; CMW: Culture collection of the Forestry and Agricultural Biotechnology Institute (FABI) of the University of Pretoria, Pretoria, South Africa; CPC: Culture collection of P.W. Crous, held at the Westerdijk Fungal Biodiversity Institute; FMR: Facultat de Medicina i Ciències de la Salut, Universitat Rovira i Virgili, Reus, Spain; JB: Collection of J.D.P. Bezerra, housed at URM; TRN: C. Ruibal private collection, currently in MAF; URM: Micoteca URM Profa. Maria Auxiliadora Cavalcanti, Departamento de Micologia Prof. Chaves Batista, Universidade Federal de Pernambuco, Brazil. T ex-type strain, ET ex-epitype strain. 2 ITS: first and second internal transcribed spacer regions and intervening 5.8S nrdna; LSU: nuclear ribosomal large subunit of the rdna; acta: actin gene; rpb2: RNA polymerase second largest subunit gene; tub2: β-tubulin gene. Names of the new taxa and sequences newly obtained in this study are shown in bold. VOLUME 8 NO. 1 83

151 Bezerra et al. 8x 99/96/ /100/1 71/87/- 100/100/1 P. robusta CBS T P. eucalyptorum CBS Pseudocercospora P. schizolobii CBS T C. beticola CBS C. capsici CBS /98/1 Cercospora 85/99/0.99 M. sumatrensis CBS T 100/100/1 M. eucalypti CBS Mycodiella 80/72/- M. laricis-leptolepidis MAFF /-/- 100/100/1 R. glennii CBS T Ramularia R. endophylla CBS ET S. pomi CBS /100/1 S. pomi CBS Schizothyrium D. eucalypti CBS D. proteae CBS T 100/100/1 Dissoconium D. aciculare CBS D. aciculare CBS T U. commune CBS T 100/100/1 99/99/1 U. dekkeri CPC Uwebraunia U. australiensis CBS T 91/99/1 R. apiculatum CBS /99/1 R. cucurbitae CBS T Ramichloridium R. luteum CBS T 100/100/0.99 E. antarcticus CBS T 100/100/1 E. antarcticus CBS Extremus 97/100/1 E. adstrictus CBS T N. hilliana CBS T 100/100/1 N. xanthorrhoeae CBS T Neodevriesia Neodevriesia sp. CBS /98/1 100/100/1 T. fimbriata CBS T T. molleriana CBS Teratosphaeria 93/100/1 T. fibrillosa CBS ET 98/100/1 100/100/1 R. tasmanica CBS T R. menaiensis CBS T 100/100/1 A. luzulae CBS T A. crateriforme CBS T Readeriella Acrodontium 93/99/1 71/78/0.98 T. chlamydosporum CBS T T. velox CBS T 100/99/1 T. protearum CBS T T. pini CBS T 100/78/1 70/90/1 95/82/- T. pseudovelox CBS T T. posoqueriae CBS T Toxicocladosporium T. rubrigenum CBS T 88/100/1 T. strelitziae CBS T 100/91/0.98 T. hominis CBS T T. irritans CBS T T. ficiniae CBS T 95/89/0.96 T. banksiae CBS T C. fusiforme CBS T C. chalastosporoides CBS ET 100/98/1 C. hillianum CBS T -/84/0.96 C. allicinum CBS ET Cladosporium 80/87/1 C. iridis CBS ET C. herbarum CBS ET N. leucadendri CPC N. leucadendri CPC N. leucadendri CPC /100/1 N. leucadendri CPC Neocladosporium gen. nov. N. leucadendri CPC /95/1 N. leucadendri CPC T 99/75/- 94/77/- 100/100/1 CPC CPC Incertae sedis -/90/1 T V. dirinae CBS Verrucocladosporium G. chlorocephala CBS /100/1 G. chlorocephala CBS Graphiopsis 91/95/0.97 R. pini CBS T 100/95/.097 R. luculiae CBS T Rachicladosporium R. cboliae CBS T Parastagonospora nodorum CBS /-/0.99 Mycosphaerellaceae Schizothyriaceae Dissoconiaceae Extremaceae Neodevriesiaceae Teratosphaeriaceae Cladosporiaceae Fig. 1. Maximum likelihood (RaxML) tree obtained by phylogenetic analysis of the combined ITS and LSU rdna and rpb2 sequences of 67 taxa belonging to Capnodiales. The new genus, Neocladosporium, is shown in bold. Bootstrap support values from Maximum Parsimony (MP-BS) and Maximum Likelihood (ML-BS), and Bayesian posterior probabilities (BPP) above 70 % and 0.95, respectively, are indicated at the nodes (MP-BS/ML-BS/BPP). Parastagonospora nodorum (CBS ) was used as outgroup. T = ex-(holo-)type strain, ET = ex-epitype strain. 84 IMA FUNGUS

152 New Toxicocladosporium species from cacti in Brazil 70/73/ JB (Pilosocereus gounellei subsp. gounellei, Brazil) JB (Pilosocereus gounellei subsp. gounellei, Brazil) 225 JB (Pilosocereus gounellei subsp. gounellei, Brazil) URM 7489 T (Pilosocereus gounellei subsp. gounellei, Brazil) 188 JB (Pilosocereus gounellei subsp. gounellei, Brazil) 70/88/ /100/ JB (Pilosocereus gounellei subsp. gounellei, Brazil) URM 7490 (Pilosocereus gounellei subsp. gounellei, Brazil) 231 JB (Pilosocereus gounellei subsp. gounellei, Brazil) 236JB (Pilosocereus gounellei subsp. gounellei, Brazil) T. cacti sp. nov. 99/100/1 235 JB (Pilosocereus gounellei subsp. gounellei, Brazil) 192 JB (Pilosocereus gounellei subsp. gounellei, Brazil) x 100/100/1 -/94/1 100/100/1 93/91/1 93/99/ /-/- -/81/ /100/1 226JB (Pilosocereus gounellei subsp. gounellei, Brazil) 94/98/1 CBS T (Banksia emulata, Australia) CBS T (Phaenocoma prolifera, South Africa) 100/100/1 CBS (Eucalyptus camaldulensis, Madagascar) CBS T (Eucalyptus camaldulensis, Madagascar) CBS T (Protea bruchelii, South Africa) CBS T (Pinus sp., China) CBS T (Strelitzia reginae, South Africa) CBS T (Bronchoalveolar lavage fluid, USA) CBS T (Mouldy paint, Suriname) CBS T (Eucalyptus camaldulensis, Madagascar) T. banksiae T. pseudovelox T. chlamydosporum T. protearum T. pini T. strelitziae T. hominis T. irritans T. rubrigenum URM 7491 T (Melocactus zehnteri, Brazil) T. immaculatum sp. nov. CBS T (Posoqueria latifolia, Australia) T. posoqueriae CBS T (Ficinia indica, South Africa) T. ficiniae CBS T (Leucadendron sp., South Africa) Neocladosporium leucadendri Fig. 2. Maximum likelihood (RaxML) tree obtained by phylogenetic analysis of the combined ITS rdna, LSU rdna, acta, rpb2 and tub2 datasets of the genus Toxicocladosporium. Newly introduced species are shown in bold. Bootstrap support values from Maximum Parsimony (MP-BS), Maximum Likelihood (ML-BS), and Bayesian posterior probabilities (BPP) above 70 % and 0.95, respectively, are indicated at the nodes (MP-BS/ ML-BS/BPP). Neocladosporium leucadandri (CBS ) was used as outgroup. T ex-(holo-)type strain. the species resolution, acta, rpb2 and tub2 sequences were also included in this analysis. This second phylogeny included sequences from 26 isolates (including the outgroup) and characters (acta: 247, ITS: 396, LSU: 780, rpb2: 724 and tub2: 415) of which 482 were parsimony-informative (acta: 25, ITS: 22, LSU: 28, rpb2: 230 and tub2: 125), 215 were variable and parsimony-uninformative (acta: 35, ITS: 48, LSU: 28, rpb2: 68 and tub2: 36), and were constant (acta: 123, ITS: 311, LSU: 726, rpb2: 420 and tub2: 240). The results of this analysis are shown in Fig. 2. Parsimony analysis resulted in a single tree showing the best score (TL = 1870; CI = 0.570; RI = 0.651; RC = 0.371). The endophytic isolates grouped in two linages; 12 isolates formed a fullysupported clade close to T. banksiae (CBS ) (MP-BS 100 %, ML-BS 100 %, BPP 1) while one isolate (URM 7491 = CBS ) formed a moderately supported monotypic linage closely related to but distinct from T. ficiniae (CBS ) and T. posoqueriae (CBS ) (MP-BS < 70 %, ML-BS 81 %, BPP 0.95). These two groups are described here as the new species Toxicocladosporium cacti (ex-type culture URM 7489 = CBS ) and Toxicocladosporium immaculatum (ex-type culture URM 7491 = CBS ), respectively. In addition, the ex-type strains of T. chlamydosporum (CBS ) and T. velox (CBS ) always clustered together with high support values (MP-BS 100 %, ML-BS 100 %, BPP 1.00). From these phylogenetic results and based on the few nucleotide differences between the two species (acta: 1 nt and 1 gap, ITS: 5 nt, LSU: 1 nt, rpb2:0 nt and TUB: 0 nt) and given that both species show similar morphological and ecological features, we treat the name T. velox as a synonym of T. chlamydosporum. LSU and ITS were informative loci to verify the relationship between genera and species groups. However, the acta, rpb2 and tub2 sequences were more informative to distinguish related species, especially in the case of T. cacti, which is closely related to T. banksiae. VOLUME 8 NO. 1 85

153 Bezerra et al. Table 2. Morphological features of Neocladosporium and Toxicocladosporium species included in this paper. Newly described species names are shown in bold. Conidia in μm [number of septa] References Species Conidiophores in μm [number of septa] Conidiogenous cells in μm Ramoconidia in μm [number of septa] Macroconidiophores Microconidiophores intercalary terminal N. leucadendri [6 15] [1 2]; secondary [0 1] T. banksiae [3 7] (14 )17 25 (2.5 )3 4 [0 1] 10 12( 20) (2.5 )3 3.5 [0 1] T. cacti to [2 6] [0 1] ( 20.5) 2 3 [0 1]; secondary 7 10( 14) 2 3 [0 1] T. chlamydosporum [1 4] to 15 5 [0 1] (15 )16 17( 18) (2.5 )3 4 [0 1]; secondary (9 )10 14( 16) (2.5 )3 4 [0 1-] T. ficiniae [1 15] [0]; secondary [0 1] T. hominis [0 2]; secondary [0 1] T. immaculatum to [2 5] [0 1] [0 1]; secondary (7 )8 14( 18.5) 2 3 [0 1] T. irritans [2 7] [0(1 2)] 9 11( 15) (2.5 )3( 4) (6 )7 8( 9) (2.5 )3( 4) Crous et al. (2011) (7 )8 10( 11) (2 )2.5 3 [0] Crous et al. (2010) {0 1] [0] This paper (8 )9 11( 12) 2.5 3( 3.5) [0 1] ( 3) [0] This paper and Crous et al. (2009) (9 )10 11 (2.5 )3 (7 )8 9 (2.5 )3 Crous et al. (2013) [0 1] Crous et al. (2016) ( 11) 2 3 This paper [(0 )1( 3)] - (5 )6 8( 10) (3 )4( 5) [0 1] Crous et al. (2007) T. pini [2 8] ( 3.5) [0 1] [0 1] 8 10( 11) 2.5( 3) [0 1] Crous et al. (2014) T. posoqueriae [1 3] [0] - (4 )6 7 (3 )4 Crous et al. (2012) T. protearum [1 8] [0 1] - (9 )11 13( 16) (2 )2.5( Crous et al. (2010) 3) [0 1] T. pseudovelox [2 5] [0 1] - (6 )7 10( 11) (2 )2.5( 3) [0] Crous & Groenewald (2011) T. rubrigenum to [1 8] to [0 1] (13 )14 15( 16) 7 8( 9) 2( 2.5) (4 )6 7 2( 2.5) Crous et al. (2009) 2.5 3( 3.5); secondary (9 )10 12( 14) 2.5 3( 3.5) T. strelitziae [2 5] [0]; secondary [0] (5 )7 8( 9) 2( 2.5) Crous et al. (2012) 86 IMA FUNGUS

154 New Toxicocladosporium species from cacti in Brazil TAXONOMY Our phylogenetic analyses revealed that the endophytic fungi from cactus species previously identified as Toxicocladosporium represent two new species in this genus. These newly proposed species are established based on phylogenetic analyses and morphological features. In addition, we introduce a new generic name, Neocladosporium to accommodate Toxicocladosporium leucadendri, which is not congeneric with Toxicocladosporium. In this section a bibliographic synopsis of the genus is compiled including key morphological features for identification, known host affiliations, substrates, and geographic distribution for all the currently accepted species of Toxicocladosporium. Table 2 summarises key morphological features of the Neocladosporium and Toxicocladosporium species included here. Neocladosporium J.D.P. Bezerra, Sandoval-Denis, C.M. Souza-Motta & Crous, gen. nov. MycoBank MB Etymology: Named because of its similarity to the genus Cladosporium. Diagnosis: Differs from Toxicocladosporium by its verruculose to warty ramoconidia, and from Cladosporium s. str. by its dark, thick-walled conidial and conidiophore septa, also lacking the typical coronate Cladosporium scar. Type species: Neocladosporium leucadendri (Crous) J.D.P. Bezerra et al (syn. Toxicocladosporium leucadendri Crous 2011). Description: Mycelium consisting of pale brown, smooth, branched, septate hyphae. Conidiophores solitary, erect, unbranched or branched above, subcylindrical, straight to flexuous, apical septum becoming dark brown and thickened. Conidiogenous cells integrated, polyblastic, terminal and lateral, subcylindrical, smooth, brown; scars truncate, thickened and darkened. Ramoconidia medium brown, verruculose to warty, giving rise to branched chains of conidia, subcylindrical, polyblastic, brown, verruculose to warty, 0 1-septate, frequently forking close to apex; scars darkened, thickened. Intercalary conidia subcylindrical to fusoid-ellipsoidal, brown, smooth to somewhat warty. Small terminal conidia fusoid-ellipsoidal, brown, smooth; hila thickened and darkened. Neocladosporium leucadendri (Crous) J.D.P. Bezerra, Sandoval-Denis, C.M. Souza-Motta & Crous, comb. nov. MycoBank MB (Fig. 3) Basionym: Toxicocladosporium leucadendri Crous, Persoonia 27: 157 (2011). Type: South Africa: Western Cape Province: Hermanus, Fernkloof Nature Reserve, on leaves of Leucadendron sp. (Proteaceae), 4 May 2010, P.W. Crous (CBS H holotype; CPC = CBS culture ex-type). Description: Crous et al. (2011a). Substrate and distribution: On leaves of Leucadendron sp. (Proteaceae) in the Western Cape province of South Africa (Crous et al. 2011a). On leaves of Kunzea pauciflora (Myrtaceae), and Banksia media, Hakea sp., and Petrophile sp. (Proteaceae) in Western Australia. Other material examined: Australia: Western Australia: Albany, Fitzgerald River National Park, Point Ann, on leaves of Banksia media (Proteaceae), 21 Sep. 2015, P.W. Crous (CPC 29237); Denmark, Lights Beach, on leaves of Hakea sp. (Proteaceae), 19 Sep. 2015, P.W. Crous (CPC 29166); Wellstead, Cape Riche, on leaves of Hakea marginata (Proteaceae), 21 Sep. 2015, P.W. Crous (CPC 29092); ibid., on leaves of Kunzea pauciflora (Myrtaceae), 21 Sep. 2015, P.W. Crous (CPC 29090); Williams, Williams Nature Reserve, on leaves of Petrophile sp. (Proteaceae), 18 Sep. 2015, P.W. Crous (CPC 29545). Notes: Crous et al. (2011a) published the strain CPC = CBS as T. leucadendri based on phylogenetic analyses using LSU and ITS sequences, and morphological characters. According to these authors, based on a combination of culture characteristics, conidiophore and conidial dimensions, it differs from known taxa, many of which also occur in the fynbos vegetation (Crous et al. 2011b). A megablast search of the NCBIs GenBank nucleotide sequence database using the ITS and LSU sequences of N. leucadendri retrieved as closest hits Graphiopsis chlorocephala and Verrucocladosporium dirinae, amongst others. In our phylogenetic analyses this strain appeared in a single lineage closely related to Graphiopsis chlorocephala and Verrucocladosporium dirinae as shown before by Crous et al. (2011a), but clearly separated from members of Toxicocladosporium (Fig. 1). Morphologically, Neocladosporium leucadendri is very similar to Toxicocladosporium species, but can be distinguished from it by size and ornamentation of ramoconidia (verruculose to warty) and ramoconidia frequently forking close to the apex. Sequences of ITS and LSU rdna or rpb2 are the best approach to separate N. leucadendri from Toxicocladosporium species and related genera. Also very similar to Cladosporium s. str., but differing in the size and ornamentation of the ramoconidia (verruculose to warty), which are frequently forking close to apex, dark, thick-walled conidial and conidiophore septa, and lacking the typical coronate Cladosporium scar type similar to Toxicocladosporium (David 1997, Crous et al. 2007); it differs from Graphiopsis which has morphological peculiarities on its conidiophores (cladosporioid and periconioid morphs), conidiogenous loci and hila (Schubert et al. 2007a, Braun et al. 2008); from Rachicladosporium which has an apical conidiophore rachis with inconspicuous to subconspicuous scars and unthickened, not darkened-refractive conidial hila (Crous et al. 2007); and from Verrucocladosporium which has mainly an unusual conidial and hyphal ornamentation (Crous et al. 2007). Because of our phylogenetic results and morphological observations, the new generic name Neocladosporium, is proposed to accommodate N. leucadendri. VOLUME 8 NO. 1 87

155 Bezerra et al. Fig. 3. Neocladosporium leucadendri (CBS ex-type culture). A. Colony sporulating on MEA. B F. Conidiophores giving rise to chains of conidia. Bars = 10 μm. Toxicocladosporium Crous & U. Braun, Stud. Mycol. 58: 39 (2007). Type species: Toxicocladosporium irritans Crous & U. Braun Notes: Toxicocladosporium was introduced by Crous et al. (2007) to accommodate cladosporium-like fungi having distinct dark, thick-walled conidial and conidiophore septa, and lacking the typical coronate Cladosporium scar type. After this original publication, several new species isolated from different substrates and hosts were introduced in this genus using morphological characters and phylogenetic analyses of ITS and LSU sequences (Crous et al. 2007, 2009a, 2010a, b, 2011a, 2012a, b, 2013, 2014, 2016, Crous & Groenewald 2011). Toxicocladosporium banksiae Crous et al., Persoonia 25: 147 (2010). Type: Australia: Queensland: Noosa National Park, 26 o S 153 o E, on leaves of Banksia sp., 13 July 2009, P.W. Crous et al. (CBS H holotype; CPC 17281, CPC = CBS culture ex-type). Description and illustration: Crous et al. (2010). Substrate and distribution: On leaves of Banksia sp. (Proteaceae), Australia (Crous et al. 2010b). Notes: According to Crous et al. (2010b), the ITS and LSU sequences of T. banksiae are close to those of T. chlamydosporum and T. irritans. The ITS sequences of T. banksiae also differ from those of T. protearum. Morphologically, T. banksiae differs from these three species in the size and shape of the intercalary and terminal conidia, ramoconidia, and presence or absence of chlamydospores. In our phylogeny using five different loci, this species is closely related to the new species T. cacti which differs in microconidiophore size [ µm (aseptate) in T. banksiae vs μm (0 1-septate) in T. cacti], ramoconidia ( µm vs μm), intercalary conidia ( µm vs µm), terminal conidia ( µm vs μm), and culture characteristics (colonies olivaceous grey reaching up to 7 mm diam in 2 wk in T. banksiae vs. colonies pale grey to grey, growing up to 30 mm diam in 3 wk and presence of a pale brown to brown exudate in T. cacti). Toxicocladosporium cacti J.D.P. Bezerra, C.M. Souza-Motta & Crous, sp. nov. MycoBank MB (Fig. 4) Etymology: Named after the nature of the host, a cactus, from which it was isolated. Diagnosis: Differs from T. banksiae in its slightly smaller and less septate microconidiophores and conidia, and by its pale grey to grey colonies. Type: Brazil: Pernambuco: Catimbau National Park, 8º36 35 S 37º14 40 W, as endophytic fungus from cactus Pilosocereus gounellei subsp. gounellei, Sep. 2013, J.D.P. Bezerra (URM holotype; URM 7489 = CBS culture ex-type). Other material examined: Brazil: Pernambuco: Catimbau National Park, 8º36 35 S 37º14 40 W, as endophytic fungus from cactus Pilosocereus gounellei subsp. gounellei, Sep. 2013, J.D.P. Bezerra 88 IMA FUNGUS

156 New Toxicocladosporium species from cacti in Brazil Fig. 4. Toxicocladosporium cacti (URM 7489 = CBS ex-type culture). A. Colony sporulating on PDA. B. Colony sporulating on OA. C. Colony sporulating on MEA. D H. Conidiophores and conidia. I. Ramoconidia and conidia. Bars = 10 μm. (URM 7490 = CBS , 188 JB, 191 JB, 192 JB, JB, 225 JB, 231 JB, 235 JB, 236 JB, 226 JB, JB). Description: Mycelium consisting of branched, septate, smooth, brown, μm wide hyphae; wall and septa becoming dark brown and thickened with age. Conidiophores dimorphic. Macroconidiophores solitary, arising from superficial mycelium, erect, brown, unbranched or branched above, finely verruculose, subcylindrical, straight to flexuous, up to μm, 2 6-septate. Microconidiophores reduced to conidiogenous cells, rarely with one supporting cell, pale brown, smooth, erect, subcylindrical, μm, 0 1-septate. Conidiogenous cells integrated, terminal or lateral, smooth, brown, μm, proliferating sympodially with 1 2 apical loci; scars truncate, thickened and darkened, μm wide. Conidia catenate in branched or unbranched chains, pale brown, thick-walled, septa dark and thick or inconspicuous, finely verruculose. Primary ramoconidia brown, finely verruculose, 0 1-septate, ellipsoidal to subcylindrical, 10 14( 20.5) 2 3 μm; secondary ramoconidia brown, finely verruculose, 0 1-septate, ellipsoidal to subcylindrical, 7 10( 14) 2 3 μm; scars darkened, thickened, μm wide. Intercalary conidia subcylindrical to fusoid-ellipsoidal, 0 1-septate, brown, finely verruculose, µm. Small terminal conidia fusoid-ellipsoidal, aseptate, brown, finely verruculose, μm; hila thickened and darkened, μm wide. Culture characteristics (in a day-night cycle, 22 C after 3 wk): Colonies on MEA are slightly folded and sulcate, velvety, pale grey to grey with a pale grey rim, reverse dark grey, reaching 30 mm diam; on OA flat to semi erumpent, spreading, with sparse to moderate aerial mycelium, smooth, surface and reverse pale grey to grey, to 29 mm; and on PDA surface and reverse olivaceous grey, to 25 mm. Exudate pale brown to brown observed on cultures growing on MEA and PDA. Substrate and distribution: An endophytic fungus isolated from the cactus Pilosocereus gounellei subsp. gounellei (Cactaceae), Brazil. Notes: Toxicocladosporium cacti is phylogenetically related to T. banksiae but differs morphologically from it in microconidiophore size and septation [ μm (0 1-septate) vs µm], smaller ramoconidia ( μm vs µm), intercalary conidia ( µm vs µm), and small terminal conidia ( μm vs µm). VOLUME 8 NO. 1 89

157 Bezerra et al. Furthermore, the culture characteristics are different from those of T. banksiae, colonies pale grey to grey, growing to 30 mm diam in 3 wk with exudate pale brown to brown in T. cacti vs. colonies olivaceous grey reaching up to 7 mm diam after 2 wk in T. banksiae. Toxicocladosporium chlamydosporum Crous & M.J. Wingf., Persoonia 22: 90 (2009). Synonym: Toxicocladosporium velox Crous & M.J. Wingf., Persoonia 22: 92 (2009); as veloxum. Types: Madagascar: Morondavo, on leaf of Eucalyptus camaldulensis, Aug. 2007, M.J. Wingfield (CBS H holotype of T. chlamydosporum; CPC = CBS culture ex-type); ibid. (CBS H holotype of T. velox; CPC = CBS culture ex-type). Description: Mycelium consisting of branched, septate, smooth, brown, 2 3 μm wide hyphae, containing swollen, globose, dark brown chlamydospore-like cells to 12 μm diam. Conidiophores dimorphic. Macroconidiophores solitary, erect, arising from superficial mycelium, penicillate, subcylindrical, straight to once geniculate-sinuous, medium to dark brown, smooth to finely verruculose, μm long, 3 5 μm wide at base, 1 4-septate, not swollen, and lacking rhizoids. Microconidiophores erect, subcylindrical, to 15 μm tall and 5 μm wide, 0 1-septate, medium brown. Conidiogenous cells terminal, integrated, subcylindrical, straight, medium brown, μm, smooth to finely verruculose; loci terminal and lateral, flat tipped, thickened, darkened, at times subdenticulate, (0.5 )1 2 μm wide. Conidia in branched chains, brown, smooth to finely verruculose, ellipsoid to cylindrical-oblong. Primary ramoconidia rarely observed, 0 1-septate, fusoid-ellipsoidal to subcylindrical, (15 )16 17( 18) (2.5 )3 4 μm. Secondary ramoconidia 0 1-septate, fusoid-ellipsoidal, (9 )10 14( 16) (2.5 )3 4 μm. Intercalary conidia 0 1-septate, fusoid-ellipsoidal, (8 )9 11( 12) 2.5 3( 3.5) μm. Small terminal conidia aseptate, fusoid-ellipsoidal, ( 3) μm (conidia dark brown and verruculose on MEA) (based on Crous et al. 2009a). Culture characteristics (in the dark, at 25 C after 1 mo): Colonies on MEA erumpent, spreading, with sparse aerial mycelium; surface folded, irregular and sectored, with feathery margin, centre pale olivaceous grey to fuscousblack, outer region olivaceous grey to greyish sepia; reverse iron-grey to dark grey; reaching up to 25 mm diam. Black sclerotial bodies on MEA, consisting of an agglomeration of chlamydospore-like cells; they remain sterile, and eventually resemble hollow fruiting bodies, although they lack an ostiole or defined wall. On OA spreading, flat, with sparse aerial mycelium, and even catenulate margin; surface iron-grey with patches of pale olivaceous grey to smoke-grey; colonies reaching up to 30 mm diam (Crous et al. 2009a). Substrate and distribution: On leaves of Eucalyptus camaldulensis (Myrtaceae), Madagascar (Crous et al. 2009a). Notes: Crous et al. (2009a) described this species using ITS and LSU sequences, and morphological characters to differentiate it from T. irritans. Toxicocladosporium chlamydosporum differs from other species in the genus in the presence of larger ramoconidia, and longer, narrower intercalary conidia, and in that it forms chlamydospores and sclerotial bodies in culture. Toxicocladosporium velox was isolated from the same leaf spot (Crous et al. 2009a). Based on the limited nucleotide differences and their morphological similarity, we consider T. velox a synonym of T. chlamydosporum. A revised description is provided to enable T. chlamydosporum in its expanded circumscription to be distinguished from other species in the genus. This species is closely related to T. protearum which differs from it mainly in the size and degree of septation of its conidiophores [20 60 μm 3 5 μm (1 4-septate) in T. chlamydosporum vs µm 3 4 µm (1 8-septate) in T. protearum], ramoconidia ( μm vs µm), and intercalary and terminal conidia ( μm vs µm). Toxicocladosporium ficiniae Crous & A.R. Wood, Persoonia 31: 191 (2013). Type: South Africa: Western Cape Province: Brackenfell, Cape Town, Bracken Nature Reserve, on leaves of Ficinia indica (Cyperaceae), 18 Aug. 2012, A.R. Wood (CBS H holotype; CPC 21283, CPC = CBS culture ex-type). Description and illustration: Crous et al. (2013). Substrate and distribution: On leaves of Ficinia indica (Cyperaceae), South Africa (Crous et al. 2013). Notes: Toxicocladosporium ficiniae is phylogenetically related to T. posoqueriae which differs in conidiophore size and septation [ μm (1 15-septate) vs μm (1 3-septate) in T. posoqueriae], and sizes of the conidiogenous cells ( μm vs μm), primary ramoconidia ( μm vs μm), and terminal conidia ( μm vs μm). Toxicocladosporium hominis Sandoval-Denis et al., Persoonia 36: 421 (2016). Type: USA: Florida: Daytona Beach, from human bronchoalveolar lavage fluid, D.A. Sutton (FMR H holotype; CBS H isotype; FMR = UTHSCSA DI = CBS cultures ex-type). Description and illustration: Crous et al. (2016). Substrate and distribution: From human bronchoalveolar lavage fluid, USA (Crous et al. 2016). Notes: Toxicocladosporium hominis is phylogenetically related and morphologically similar to T. strelitziae (Crous et al. 2012b), but differs from T. strelitziae in the production of larger conidiogenous cells ( µm vs µm) and intercalary conidia ( µm vs µm). In addition, the latter species has smooth to verruculose ramoconidia, secondary ramoconidia and intercalary conidia, 90 IMA FUNGUS

158 New Toxicocladosporium species from cacti in Brazil Fig. 5. Toxicocladosporium immaculatum (URM 7491 = CBS ex-type culture). A. Colony sporulating on PDA. B. Colony sporulating on OA. C. Colony sporulating on MEA. D. Conidiophores. E G. Conidiophores and conidia. H. Conidiophore and conidia after 1 mo on SNA at 22 C. I. Ramoconidia and conidia. Bars = 10 μm. without constrictions in the medial portion or at the septum (Crous et al. 2016). Phylogenetically, this species is a distinct taxon closely related to T. strelitziae (Fig. 2). Toxicocladosporium immaculatum J.D.P. Bezerra, C.M. Souza-Motta & Crous, sp. nov. MycoBank MB (Fig. 5) Etymology: Named after its pristine, well-developed, penicillate conidiophores. Diagnosis: Differs from most Toxicocladosporium species by its red to dark red pigmented colonies when grown on OA. Different from T. ficiniae mainly by the larger and less septate conidiophores with shorter primary and secondary ramoconidia. Distinguished from T. posoqueriae by the slightly reduced conidiophores and conidia, and from T. rubrigenum by its less septate macroconidiophores, shorter microconidiophores and somewhat larger conidia. Type: Brazil: Pernambuco: Itaíba, Curral Velho Farm, 9 o S 37 o W, as endophyte from cactus Tacinga inamoena, Sep. 2013, J.D.P. Bezerra (URM holotype; URM 7491 = CBS culture ex-type). Description: Mycelium on SNA consisting of branched, septate, smooth to verruculose, pale brown, 2 3 μm wide hyphae. Conidiophores dimorphic, arising from superficial mycelium, erect to sinuous, brown, unbranched, finally verruculose, subcylindrical, straight to flexuous. Macroconidiophores up to μm, 2 5-septate. Microconidiophores sometimes reduced to conidiogenous cells on hyphae, pale brown, smooth to finally verruculose, flexuous, subcylindrical, μm, 0 1-septate. Conidiogenous cells integrated, polyblastic, terminal and lateral, smooth, becoming verruculose, brown, μm; scars truncate, thickened and darkened, μm wide. Primary ramoconidia medium brown, finely verruculose, 0 1-septate, subcylindrical, μm. Secondary ramoconidia giving rise to branched chains of conidia, subcylindrical, polyblastic, brown, finely verruculose, 0 1-septate, (7 )8 14( 18.5) 2 3 μm; scars darkened, thickened, μm wide. Intercalary conidia subcylindrical to fusoid-ellipsoidal, brown, finely verruculose to verruculose, μm. Small terminal conidia fusoid-ellipsoidal, VOLUME 8 NO. 1 91

159 Bezerra et al. brown, finely verruculose, 8 10( 11) 2 3 μm; hila thickened and darkened, μm wide. Culture characteristics (in a day-night cycle, at 22 C after 3 wk): Colonies on MEA are folded and sulcate, velvety, pale grey to olive-yellowish with a very light grey rim, reverse dark brown, reaching 33 mm diam; on OA flat, spreading, with sparse to moderate aerial mycelium, smooth, surface olive, with a light grey rim, reverse dark brown, red to dark red pigmentation produced, growing up to 33 mm diam; and on PDA surface olivaceous to olivaceous yellowish, reverse dark green, with sparse to moderate aerial mycelium, reaching up to 33 mm diam. Exudate pale brown to brown on MEA and PDA. Substrate and distribution: As an endophyte isolated from the cactus Tacinga inamoena (Cactaceae), Brazil. Notes: Toxicocladosporium immaculatum is phylogenetically closely related to T. ficiniae, T. posoqueriae and T. rubrigenum (Fig. 2). It differs morphologically from T. ficiniae in conidiophore size and septation [up to μm (2 5-septate) vs μm (1 15-septate) in T. ficiniae], conidiogenous cells ( μm vs μm in T. ficiniae), ramoconidia size (primary μm and secondary μm vs. primary μm and secondary μm in T. ficiniae) and intercalary conidia ( μm vs μm in T. ficiniae). It differs from T. posoqueriae in the size of the conidiophores [to μm (2 5-septate) vs μm (1 3-septate) in T. posoqueriae], conidiogenous cells ( μm vs μm in T. posoqueriae), ramoconidia (primary μm and secondary μm vs μm in T. posoqueriae) and terminal conidia ( μm vs μm in T. posoqueriae). Toxicocladosporium rubrigenum differs in the size of the conidiophores [macroconidiophores to μm (2 5-septate) vs. to 100 μm 2 4 μm (1 8-septate) in T. rubrigenum and microconidiophores μm vs. to μm in T. rubrigenum], conidiogenous cells ( μm vs μm in T. rubrigenum), ramoconidia (primary μm and secondary μm vs. primary μm and secondary μm in T. rubrigenum), intercalary conidia ( μm vs μm in T. rubrigenum), and terminal conidia ( μm vs μm in T. rubrigenum). Furthermore, T. immaculatum also differs from these species in colony colour, the presence of a red to dark red pigmentation in OA medium, and slower growth rates. Toxicocladosporium irritans Crous & U. Braun, Stud. Mycol. 58: 39 (2007). Type: Suriname: Paramaribo: isolated from mouldy paint, Feb. 1958, M.B. Schol-Schwarz (CBS H holotype; CBS culture ex-type). Description and illustration: Crous et al. (2007). Portugal (Mesquita et al. 2009); associated with patients with atopic dermatitis, Japan (Zhang et al. 2011); colonizing tattoo inks, Italy (Bonadonna et al. 2014); on parchment manuscripts, Italy (Piñar et al. 2015); from human blood and finger nail, USA (Sandoval-Denis et al. 2015); on Adansonia digitata, South Africa (Cruywagen et al. 2015); and on equipment used in the International Space Station or Space Shuttle, Japan (Satoh et al. 2016). Notes: Crous et al. (2007) described Toxicocladosporium irritans as producing volatile metabolites, which cause a skin rash within minutes of opening an inoculated dish for microscopic examination. Morphologically and phylogenetically it is very similar to Cladosporium s. str., and produces dimorphic conidiophores, which is also a feature commonly observed in that genus. It is distinct in having dark, thick-walled conidial and conidiophore septa, and lacking the typical coronate Cladosporium scar type (David 1997, Crous et al. 2007). In our phylogenetic analyses (Fig. 2), T. irritans forms a lineage related to T. rubrigenum and T. hominis. It differs from T. rubrigenum in the size and septation of the conidiophores [ μm (2 7-septate) vs. to 100 μm 2 4 μm (1 8-septate)], conidiogenous cells ( μm vs μm), ramoconidia ( μm vs. primary μm and secondary μm) and terminal conidia ( μm vs μm). It differs from T. hominis in conidiophore size ( μm), conidiogenous cells ( μm), ramoconidia (primary μm and secondary μm), and intercalary conidia ( μm). Toxicocladosporium pini Crous & Y. Zhang ter, Persoonia 32: 269 (2014). Type: China: Beijing, Badaling, 40 o N 116 o E, on needles of Pinus sp. (Pinaceae), 1 Sept. 2013, P.W. Crous & Y. Zhang (CBS H holotype; CPC = CBS culture ex-type). Description and illustration: Crous et al. (2014). Substrate and distribution: On needles of Pinus sp. (Pinaceae), China (Crous et al. 2014). Notes: According to Crous et al. (2014), Toxicocladosporium pini is morphologically similar to T. pseudovelox (ramoconidia 0 1-septate, broadly ellipsoid to subcylindrical, μm; intercalary and terminal conidia ellipsoid, μm) and T. protearum (ramoconidia 0 1-septate, subcylindrical, μm; intercalary and terminal conidia subcylindrical to narrowly fusoid-ellipsoidal, μm). Based on conidial dimensions, T. pini can be distinguished from T. protearum, but because of its morphological similarity to T. pseudovelox it can only be distinguished from that species by DNA data. Phylogenetically, this species is positioned as a distinct lineage between T. protearum and T. strelitziae (Fig. 2). Substrate and distribution: Isolated from mouldy paint, Suriname (Crous et al. 2007); ancient laid-paper documents, Toxicocladosporium posoqueriae Crous & R.G. Shivas, Persoonia 29: 181 (2012). 92 IMA FUNGUS

160 New Toxicocladosporium species from cacti in Brazil Type: Australia: Northern Territory: Darwin, on leaves of Posoqueria latifolia (Rubiaceae), 12 Apr. 2011, R.G. Shivas (CBS H holotype; CPC = CBS culture ex-type). Description and illustration: Crous et al. (2012b). Substrate and distribution: On leaves of Posoqueria latifolia (Rubiaceae), Australia (Crous et al. 2012b). Notes: According to Crous et al. (2012b), Toxicocladosporium posoqueriae differs from other members of the genus in that it has whorls of conidiogenous cells, resembling those of Parapericoniella asterinae (Heuchert et al. 2005, Bensch et al. 2012). This species is closely related to T. ficiniae which differs in conidiophore size and septation [ μm (1 3-septate) vs μm (1 15-septate)], conidiogenous cells ( μm vs μm), ramoconidia ( μm vs. primary μm and secondary μm), intercalary conidia ( μm) and terminal conidia ( μm vs μm). It is also similar to the newly described T. immaculatum which differs from in the conidiophores [macroconidiophores to μm (2 5-septate) and microconidiophores μm], conidiogenous cells ( μm), ramoconidia (primary μm and secondary μm), intercalary conidia ( μm), and terminal conidia ( μm). P.W. Crous (CBS H holotype; CPC = CBS culture ex-type). Description and illustration: Crous & Groenewald (2011). Substrate and distribution: On leaf bracts of Phaenocoma prolifera (Asteraceae), South Africa (Crous & Groenewald 2011). Notes: Crous & Groenewald (2011) showed that Toxicocladosporium pseudovelox was similar to T. chlamydosporum and other Toxicocladosporium species, but has shorter ramoconidia ( μm) than T. chlamydosporum ( μm). Toxicocladosporium pseudovelox is closely related to T. pini, which has larger conidiophores [macroconidiophores μm (2 8-septate) and microconidiophores μm], conidiogenous cells ( μm), and intercalary conidia ( μm, 0 1-septate). Toxicocladosporium pseudovelox was placed in a basal position at a highly supported node, which clustered it with T. pini, T. protearum, and T. chlamydosporum (Fig. 2). Toxicocladosporium rubrigenum Crous & M.J. Wingf., Persoonia 22: 91 (2009). Type: Madagascar: Morondavo, on leaf of Eucalyptus camaldulensis, Aug. 2007, M.J. Wingfield (CBS H holotype; CPC = CBS culture ex-type). Toxicocladosporium protearum Crous & Roets, Persoonia 25: 135 (2010). Type: South Africa: Western Cape Province: Stellenbosch, J.S. Marais Garden, on leaves of Protea sp., 22 Apr. 2008, F. Roets (CBS H holotype; CPC = CBS culture ex-type). Description and illustration: Crous et al. (2010a). Substrate and distribution: On leaves of Protea sp. (Proteaceae), South Africa (Crous et al. 2010a). Notes: Blast analyses of the LSU and ITS sequences of Toxicocladosporium protearum showed that it is closely related to T. chlamydosporum and T. irritans (Crous et al. 2010a). Morphologically it differs from T. chlamydosporum which has smaller intercalary ( μm) and terminal ( ) conidia. Our phylogenetic analyses place T. protearum as a distinct lineage between T. chlamydosporum and T. pini which has larger macroconidiophores ( μm), intercalary conidia ( μm, 0 1-septate), and smaller terminal conidia ( μm, 0 1-septate) (Crous et al. 2014). Toxicocladosporium pseudovelox Crous, Persoonia 26: 81 (2011); as pseudoveloxum. Type: South Africa: Western Cape Province: Hermanus, Fernkloof Nature Reserve, 34 o S 19 o E, on leaf bracts of Phaenocoma prolifera, 2 May 2010, K.L. Crous & Description and illustration: Crous et al. (2009a). Substrate and distribution: On leaf of Eucalyptus camaldulensis (Myrtaceae), Madagascar (Crous et al. 2009a). Notes: This species differs from other Toxicocladosporium species in the production of densely branched penicillate conidiophores, and colonies that form a prominent red pigment on OA (Crous et al. 2009a). Toxicocladosporium rubrigenum is phylogenetically related to T. irritans and the new species T. immaculatum (Fig. 2). It differs from T. irritans in having longer and narrower conidiophores and conidiogenous cells (to 100 µm 2 4 µm and µm), as well as narrower ramoconidia ( µm); and from T. immaculatum in the size of the conidiophores [macroconidiophores to μm (2 5-septate) and microconidiophores μm], conidiogenous cells ( μm), ramoconidia (primary μm and secondary μm), intercalary conidia ( μm), terminal conidia ( μm), and culture characteristics (culture colour, pigmentation in the culture medium, production of exudates, and growth rates). Toxicocladosporium strelitziae Crous, Persoonia 28: 179 (2012). Type: South Africa: Mpumalanga Province: Kruger Game Reserve, Satara Rest Camp, on leaves of Strelitzia reginae (Strelitziaceae), 11 July 2011, P.W. Crous (CBS H holotype; CPC 19763, CPC = CBS culture ex-type). VOLUME 8 NO. 1 93

161 Bezerra et al. Description and illustration: Crous et al. (2012b). Substrates and distribution: On leaves of Strelitzia reginae (Strelitziaceae), South Africa (Crous et al. 2012b). Notes: In a previous phylogenetic analysis, Toxicocladosporium strelitziae was placed in close proximity to T. pseudovelox (Crous et al. 2012b), but in the present analysis is placed in a lineage distant from that species with T. hominis as the closest relative (Fig. 2). Toxicocladosporium strelitziae is distinct from T. pseudovelox in having longer, narrower conidiophores ( μm vs μm in T. pseudovelox), and larger, aseptate ramoconidia ( μm vs μm, 0 1-septate in T. pseudovelox), and from T. hominis which has larger conidiophores ( μm in T. strelitziae vs μm in T. hominis), conidiogenous cells ( μm in T. strelitziae vs μm), ramoconidia [primary μm (aseptate) and secondary μm (aseptate) in T. strelitziae vs. primary μm (0 2-septate) and secondary μm (0 1-septate) in T. hominis], and intercalary conidia [ μm in T. strelitziae vs μm (0 1-septate) in T. hominis]. DISCUSSION The generic name Toxicocladosporium was introduced by Crous et al. (2007) to accommodate fungi similar to Cladosporium species but with different conidiophore and conidium morphology and phylogeny. Following this description, several new species were reported mainly as epiphytic, saprobic or phytopathogenic fungi from all continents (Crous et al. 2009a, 2010a, b, 2011a, 2012a, b, 2013, 2014, 2016, Crous & Groenewald 2011). However, in contrast to Cladosporium, Toxicocladosporium species had not previously been reported as endophytic fungi (Bensch et al. 2012, Bezerra et al. 2012, 2013). The isolation of novel Toxicocladosporium species as endophytic fungi from cacti in a tropical dry forest (Caatinga) in Brazil is reported here for the first time, and illustrates the diversity of fungi present as endophytes in different hosts and ecosystems. In this study we revisited all currently published species of Toxicocladosporium using morphology and phylogenetic analyses (including three new loci). Based on these data we proposed two new species and one new closely related genus. Using a multigene phylogeny to recognise taxa in Dothideomycetes, Schoch et al. (2006) showed that Cladosporium belongs to the family Cladosporiaceae (an older name for the previously published Davidiellaceae). Later, during the investigation of cladosporium-like taxa, Crous et al. (2007) studied several isolates and proposed different genera based on their morphology and phylogeny, using sequences of part of the LSU nrdna. In their phylogenetic reconstruction, six new genera were proposed, including Rachicladosporium, Toxicocladosporium, and Verrucocladosporium as incertae sedis. Bensch et al. (2012) monographed the genus Cladosporium and showed that it belongs to the family Cladosporiaceae (Capnodiales, Dothideomycetes) along with other four genera, Graphiopsis, Rachicladosporium, Toxicocladosporium, and Verrucocladosporium. Using ITS, LSU and rpb2 sequences from these genera, from all Toxicocladosporium species and from the other six families in Capnodiales we reconstructed the phylogenetic relationships of Cladosporiaceae and determined the phylogenetic position of each genus, including the newly described genus Neocladosporium (Fig. 1). Our results are similar to those of Bensch et al. (2012), who used LSU sequences to verify the relationship among these genera of the Cladosporiaceae. Sandoval-Denis et al. (2015) studied clinical samples from the USA and reported the isolation of Cladosporium and Toxicocladosporium mainly obtained from respiratory specimens. These authors used phylogenetic analyses from all the available ITS and LSU sequences of Toxicocladosporium species except T. leucadendri, as well as morphological characters to identify two isolates as T. irritans, while a third isolate was unidentified, but phylogenetically positioned in a lineage between T. rubrigenum and T. strelitziae. In a subsequent paper the unidentified isolate was published as a new species, T. hominis (Crous et al. 2016). Sandoval-Denis et al. (2015) may not have included sequences from T. leucadendri in their analyses because this species appeared as a different genus, not belonging to Toxicocladosporium s. str. Toxicocladosporium leucadendri (CPC = CBS ) was published by Crous et al. (2011a) based on megablast searches in combination with culture characteristics, and conidiophore and conidial dimensions. Also, the phylogenetic analyses of the ITS and LSU sequences showed this strain in a single clade between Graphiopsis chlorocephala and Verrucocladosporium dirinae. The same result was observed in our phylogenetic analyses using the same loci, and also using acta, rpb2 and tub2 sequences. Based on these results, we introduced the new genus, Neocladosporium, with N. leucadendri as type species. Furthermore, based on the phylogenetic position and the small nucleotide differences between T. chlamydosporum and T. velox, we treat them as conspecific. These similarities can be also observed in the phylogenetic reconstruction published by Crous & Groenewald (2011), where T. velox and T. chlamydosporum are placed in the same clade with a high bootstrap support value. In addition, these authors used few morphological characters, such as the colour and size of conidia (darker brown and somewhat larger), absence and/or presence of chlamydospores and growth in culture to separate these species. These features are now combined in the revised circumscription of T. chlamydosporum presented here. To improve the discrimination of species in the genus Toxicocladosporium, we generated acta, rpb2 and tub2 sequences from all the available ex-type strains as well as endophytic isolates generated in this study (Fig. 2). In our analyses using a combined matrix of ITS, LSU, acta, rpb2 and tub2 sequences, we recognise 13 species in this genus, including the two new species, T. cacti and T. immaculatum. As previously demonstrated in Cladosporium by different authors (Braun et al. 2003, Schubert et al. 2007b, Zalar et al. 2007, Bensch et al. 2010, 2012, 2015, Sandoval-Denis et al. 2016), ITS, and LSU sequences are less informative than acta, rpb2 and tub2 sequences to separate species in 94 IMA FUNGUS

162 New Toxicocladosporium species from cacti in Brazil Toxicocladosporium. In our analyses, rdna sequences were very similar among some species, but are useful to separate genera (LSU) and species groups (ITS). After inclusion of acta sequences, the third most informative region after rpb2 and tub2, respectively, the separation of species was improved. Sequences of rpb2, followed by tub2 were the best loci to recognise species in our analyses. We therefore recommend these markers as barcoding targets for species recognition as well as for the description of new taxa in addition to ITS and acta sequences in this genus. The inclusion of acta, rpb2 and tub2 sequences in our analyses was crucial to facilitate the separation of T. cacti from T. banksiae, since rdna sequences from endophytic isolates were closely related to T. banksiae, but could not unambiguously resolve both species. In contrast, the acta, rpb2 or tub2 loci consistently separate these two taxa with high statistical confidence (data not shown). A similar situation was observed in Cladosporium for which a combined phylogenetic analysis including ITS, translation elongation factor 1-alpha (tef1) and acta loci has been adopted in order to separate species within that genus, with ITS being the least informative locus (Bensch et al. 2012, 2015, Sandoval-Denis et al. 2016). Our study shows that Toxicocladosporium species, as those of Cladosporium (Bensch et al. 2012, Bezerra et al. 2012, 2013), may be isolated as endophytic fungi from plants growing in tropical dry regions. This report also expands our knowledge about endophytes associated with cacti and highlights the mostly underestimated fungal diversity associated with this little-studied group of host plants, and as well as the importance of protecting them in their natural habitats. ACKNOWLEDGEMENTS We thank the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) (Process /2014-9), the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) and the Fundação de Amparo à Ciência e Tecnologia de Pernambuco (FACEPE) of Brazil for financial support and scholarships. We also thank Konstanze Bensch and David L. Hawksworth for the valuable comments and suggestions to improve the manuscript. We extend our thanks to the Universidade Federal de Pernambuco and to the technical staff, Eliane Silva-Nogueira and Luan Amim from the URM Culture Collection, and to Marjan Vermaas, Arien van Iperen and Mieke Starink-Willemse from the Westerdijk Fungal Biodiversity Institute. We also thank Tamara Caldas, Greicilene Albuquerque, Gianne Rizzuto, Karla Freire and other students of the Laboratório de Micologia Ambiental/UFPE for their technical help and processing of samples. 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166 doi: /imafungus IMA FUNGUS 8(1): (2017) Asexual-sexual morph connection in the type species of Berkleasmium Joey Tanney 1, and Andrew N. Miller 2 1 Biodiversity (Mycology & Botany), Eastern Cereal and Oilseed Research Centre, Agriculture and Agri-Food Canada, Ottawa, ON K1A 0C6, Canada; corresponding author jtanney@lakeheadu.ca 2 University of Illinois at Urbana-Champaign, Illinois Natural History Survey, 1816 South Oak Street, Champaign, IL 61820, USA Abstract: Berkleasmium is a polyphyletic genus comprising 37 dematiaceous hyphomycetous species. In this study, independent collections of the type species, B. concinnum, were made from Eastern North America. Nuclear internal transcribed spacer rdna (ITS) and partial nuc 28S large subunit rdna (LSU) sequences obtained from collections and subsequent cultures showed that Berkleasmium concinnum is the asexual morph of Neoacanthostigma septoconstrictum (Tubeufiaceae, Tubeufiales). Phylogenies inferred from Bayesian inference and maximum likelihood analyses of ITS-LSU sequence data confirmed this asexual-sexual morph connection and a re-examination of fungarium reference specimens also revealed the co-occurrence of N. septoconstrictum ascomata and B. concinnum sporodochia. Neoacanthostigma septoconstrictum is therefore synonymized under B. concinnum on the basis of priority. A specimen identified as N. septoconstrictum from Thailand is described as N. thailandicum sp. nov., based on morphological and genetic distinctiveness. Key words: Ascomycota molecular systematics morphology new species taxonomy Tubeufia Tubeufiales Article info: Submitted: 16 February 2017; Accepted: 26 April 2017; Published: 3 May INTRODUCTION Naturalist Thomas Gibson Lea botanized extensively in Ohio until his death in 1844, after which his fungal specimens were sent to his correspondent Miles J. Berkeley (Lea 1849). Berkeley (1845) described Sporidesmium concinnum ( A very pretty object under the microscope ) from Lea s specimen growing on dead wood from a rotten tree trunk (host unknown) in Ohio. Zobel (in Corda 1854) later described the new genus Berkleasmium to accommodate S. concinnum, illegitimately devising the name Berkleasmium cordeanum. More than one century later, Moore (1958) re-established the generic name Berkleasmium to accommodate sporodochial species previously placed in Sporidesmium, later accepting ten species (Moore 1959). Berkleasmium currently comprises 37 species characterized by sporodochial conidiomata bearing macronematous conidiophores and monoblastic conidiogenous cells that give rise to brown or black, dry, rhexolytically-seceding, dictyoconidia (Ellis 1971, Bussaban et al. 2001, Seifert et al. 2011, org/). Berkleasmium species are associated with a variety of decaying above-ground tissues of monocot and dicot plants from terrestrial and aquatic habitats. No known sexual morph has yet been connected to Berkleasmium. Available sequences in GenBank are restricted to species described from Thailand or Micronesia, including Berkleasmium crunisia, B. micronesicum, B. nigroapicale, and B. typhae. Previous phylogenetic analyses based on SSU and LSU sequences of these species indicate the polyphyly of Berkleasmium, with B. micronesicum and B. nigroapicale placed incertae sedis sister to Sporormiaceae, and B. crunisia and B. typhae placed incertae sedis sister to Mycopepon smithii (Pinnoi et al. 2007, Wang et al. 2007, Hyde et al. 2016). This polyphyly is not surprising given the treatment of Berkleasmium as a genus characterised by the sporodochia bearing dark brown dictyoconidia. For example, Monodictys monilicellularis was transferred to Berkleasmium because of sporodochia formation, despite the presence of distinctive sterile moniliform appendages, while B. papillatum was placed within the genus despite proliferating conidiophores; the taxonomic informativeness of these characters remains to be tested by molecular phylogenetic analyses (Raghuveer Rao & Rao 1964, Whitton et al. 2012). Determining the phylogenetic placement of the type species, B. concinnum, is crucial to delineate taxonomic boundaries within this polyphyletic and morphologically heterogeneous genus. In this study, ITS and LSU sequences were analyzed with other taxa in Tubeufiales to estimate phylogenetic relationships. MATERIALS AND METHODS Sampling and isolation of fungi Field collections of Berkleasmium concinnum were made independently in Gatineau, Quebec, Canada and the Great Smoky Mountains National Park, North Carolina and Tennessee, USA. Single conidium cultures were generated by transferring individual conidia to 6 cm diam Petri dishes containing MEA using an electrolytically sharpened tungsten needle (Brady 1965). Conidial germination was visually verified and cultures were incubated at 16 o C under 12: International Mycological Association You are free to share - to copy, distribute and transmit the work, under the following conditions: Attribution: You must attribute the work in the manner specified by the author or licensor (but not in any way that suggests that they endorse you or your use of the work). Non-commercial: You may not use this work for commercial purposes. No derivative works: You may not alter, transform, or build upon this work. For any reuse or distribution, you must make clear to others the license terms of this work, which can be found at Any of the above conditions can be waived if you get permission from the copyright holder. Nothing in this license impairs or restricts the author s moral rights. VOLUME 8 NO. 1 99

167 Tanney & Miller h dark:light conditions. Specimens were accessioned in the Canadian National Mycological Herbarium (Ottawa, Canada; DAOM) or the Illinois Natural History Survey Fungarium (Champaign, USA; ILLS) and living cultures were deposited in the Canadian Collection of Fungal Cultures (Ottawa, Canada; DAOMC). Morphological observations Conidiomata and ascomata from fresh and dried specimens were mounted in deionized water, 5 % KOH, or lactic acid. Observations were made using an Olympus BX50 light microscope (Olympus, Tokyo) and an Olympus SZX12 stereomicroscope and micrographs were captured using an InfinityX-32 camera (Lumenera, Ottawa) and Infinity Analyze (Lumenera) software. Photographic plates were assembled using Adobe Photoshop 5.5 (Adobe Systems, San Jose, CA). DNA extraction, PCR amplification, and sequencing Total genomic DNA was extracted from 12-wk-old cultures or directly from conidiomata of B. concinnum using the Ultraclean Microbial DNA Isolation Kit (Mo Bio Laboratories, Carlsbad, CA) or NucleoSpin Plant II Kit (Macherey-Nagel, Düren, Germany) following the manufacturers protocol. The entire ITS and the first two domains of LSU were amplified and sequenced following the methods of Promputtha & Miller (2010) and Tanney et al. (2015). Phylogenetic analyses Sequence contigs were assembled and trimmed using Geneious R8 v (Biomatters, Auckland, New Zealand). ITS and LSU sequences were concatenated to create a dataset containing 37 sequences that was aligned using MAFFT v. 7 (Katoh & Standley 2013) and visually inspected in Geneious. The most suitable sequence evolution model (GTR+I+G) was determined based on the optimal Akaike information criterion scores in MrModeltest v (Nylander 2004). The ex-type culture of Botryosphaeria corticis (CBS119047; NR ) was selected as outgroup based on previous analyses (Sanchez et al. 2012). Bayesian inference (BI) phylogenetic reconstruction was performed with MrBayes v. 3.2 (Ronquist et al. 2012). Three independent Markov Chain Monte Carlo (MCMC) samplings were performed with 12 chains (11 heated and one cold) with sampling every 500 generations until the standard deviation of split frequencies reached a value < The first 25 % of trees were discarded as burn-in and the remaining trees kept and combined into one 50 % majority rule consensus tree. Convergence was assessed from the three independent runs using Tracer v. 1.6 (Rambaut et al. 2014). Maximum likelihood (ML) analysis was performed using RAxML v (Stamatakis 2014) in PAUP v. 4.0b10 (Swofford 2003) starting from a random starting tree with 1000 bootstrap replicates. Consensus trees were visualized in FigTree (available at uk/software/figtree/) and exported as SVG vector graphics for assembly in Adobe Illustrator v10 (Adobe Systems, San Jose, CA). All novel sequences used in this study were accessioned in GenBank (Table 1) and taxonomic novelties and associated metadata were deposited in MycoBank (www. mycobank.org). RESULTS The concatenated ITS-LSU alignment consisted of 37 sequences and 1277 positions. All Berkleasmium concinnum specimens shared identical ITS sequences except for a single A-to-G transition (position 441 in alignment) in specimen ILLS Berkleasmium concinnum ITS sequences were identical to the ITS sequence of the holotype of Neoacanthostigma septoconstrictum (ILLS 59356), except for the single bp difference in ILLS LSU sequences for all Berkleasmium concinnum isolates were identical and 99 % similar to the N. septoconstrictum type (ILLS 59356; NR_119758), with one A-to-T transversion (position 758) and two C-to-T transitions (positions 806 and 1213). Phylogenies inferred from BI and ML analyses were generally concordant. However, the backbone topology was polytomous in the BI analysis versus dichotomous in the ML analysis, albeit with low-supported branch nodes in both analyses. The placement of Acanthostigma filiforme was also discordant among both analyses, probably a result of the poorly-supported backbone, and PP values were generally higher than BS values. Berkleasmium concinnum and N. septoconstrictum, including the type species N. fusiforme (MFLUCC ; KF301529) and a specimen identified as N. septoconstrictum (MFLUCC ; KX454176) but with a distinct ITS sequence, formed a wellsupported clade in both analyses (BS = 93; PP = 1) (Fig. 1). The Berkleasmium-Neoacanthostigma clade was weaklysupported (BS = 37; PP = 0.57) sister to a clade comprising Helicoma conicodentatum, Helicosporium linderi, Tubeufia khunkornensis, and a strain identified as Chlamydotubeufia cf. huaikangplaensis. Sequence similarity and ITS-LSU phylogenetic analyses reveal that B. concinnum is the asexual morph of N. septoconstrictum (Tubeufiaceae, Tubeufiales) and that the type species of both genera occur in a well-supported clade (BS = 100; PP = 1). Although not originally reported with the description of N. septoconstrictum (Promputtha & Miller 2010), re-examination of the type specimen revealed numerous conidiomata of B. concinnum covering the woody substrate along with ascomata of N. septoconstrictum. Examining additional reference specimens of B. concinnum confirmed the co-occurrence of the ascomatal sexual morph (i.e. N. septoconstrictum) among the asexual B. concinnum conidiomata (DAOM 24916, DAOM 29377, DAOM 41029, DAOM 75764, DAOM , DAOM , DAOM , ILLS 80802; Fig. 2 A C). Since Berkleasmium Zobel 1854 is an earlier name than Neoacanthostigma Boonmee et al. 2014, N. septoconstrictum must be synonymized under B. concinnum, on the assumption that all priority currently accorded to generic names typified by a sexual morph is ended in July In addition, the specimen previously identified by Hyde et al. (2016) as N. septoconstrictum (MFLUCC ; KX454176) is not conspecific with the type specimen of this species (ILLS59356; NR119758) based on asexual morph dissimilarity (hyaline helicoid conidia) and genetic (13 % ITS sequence divergence) differences and is therefore described as a new species, N. thailandicum. 100 IMA FUNGUS

168 Asexual-sexual morphs in Berkleasmium 93 Berkleasmium concinnum ILLS A Berkleasmium concinnum ILLS A Berkleasmium concinnum DAOM A Berkleasmium concinnum NB-789 A 100 Berkleasmium concinnum ILLS A 99 Berkleasmium concinnum ILLS A Berkleasmium concinnum ILLS S Botryosphaeria corticis CBS Neoacanthostigma thailandicum MFLUCC Neoacanthostigma fusiforme MFLUCC Helicosporium linderi NBRC Tubeufia khunkornensis MFLUCC Helicoma conicodentatum UBC F14998 Chlamydotubeufia cf. huaikangplaensis MFLUCC Acanthostigma filiforme ILLS Helicomyces lilliputeus NBRC Tubeufia javanica MFLUCC Tubeufia roseohelicospora MFLUCC Helicosporium talbotii MUCL Helicomyces roseus CBS Tubeufia amazonensis ATCC Helicosporium pallidum CBS Tubeufia aurantiella ANM 718 Helicosporium panachaeum CBS Tamhinispora indica AMH Helicosporium aureum NBRC 7098 Acanthostigma scopulum ANM Helicosporium guianense UAMH Chlamydotubeufia helicospora MFLUCC Chlamydotubeufia khunkornensis MFLUCC Helicoön gigantisporum BCC 3550 Acanthostigma multiseptatum ILLS Acanthostigma perpusillum UAMH 7237 Acanthostigma chiangmaiensis MFLUCC Helicosporium guianense CBS Helicosporium gracile CBS * Thaxteriellopsis lignicola MFLUCC * * * * * * 0.07 Fig. 1. Most likely tree from a RAxML analysis of ITS-LSU dataset containing representative Tubeufiales species. Culture collection accession numbers or specimen identifiers follow the species name, with type or ex-type strains in bold and asterixes (*) denoting generic types. A denotes sequences derived from Berkleasmium concinnum asexual morph and S denotes sequences derived from B. concinnum sexual morphs. RAxML bootstrap support percentages 50 from a summary of 1000 replicates are presented at the branch nodes. Thickened branches indicate Bayesian posterior probability values The tree was rooted with Botryosphaeria corticis and the scale bar represents the number of substitutions per site. TAXONOMY Berkleasmium concinnum (Berk.) S. Hughes, Canad. J. Bot. 36: 740 (1958). (Fig. 2; Promputtha & Miller 2010: figs 15 22) Basionym: Sporidesmium concinnum Berk., London J. Bot. 4: 309 (1845). Synonyms: Berkleasmium cordeanum Zobel, in Corda, Icon. Fung. 6: 4 (1854); nom. illegit. (Art. 52.1). Neoacanthostigma septoconstrictum (Promp. & A.N. Mill.) S. Boonmee & K.D. Hyde, Fungal Diversity 68: 279 (2014). Acanthostigma septoconstrictum Promp. & A.N. Mill., Mycologia 102: 579 (2010). VOLUME 8 NO

169 Tanney & Miller Type: USA: Ohio: on dead wood, T.G. Lea 168 (K(M) - holotype, n.v.; K(M)-IMI slide ex-holotype). Other specimens examined: Canada: Quebec: Gatineau, Aylmer, Boucher Forest, on large, rotten, decorticated hardwood log, 17 Jul. 2015, J.B. Tanney (DAOM ); ibid., 29 Aug. 2015, J.B. Tanney (DAOM ); ibid., 10 Jul. 2015, J.B. Tanney (DAOM , DAOMC , DAOMC ); ibid., fallen hardwood log, 5 Sep. 2016, J.B. Tanney (NB-789). Russia: Primorsky Krai, Khasansky District, Kedrovaya Pad Nature Preserve, log of an unknown deciduous species, 3 Oct. 1987, I.M. Bacuebeba (DAOM ). USA: Louisiana: St. Tammany Parish, Honey Island Swamp near Pearl River, rotten wood, 6 Jun. 1976, S.J. Hughes (DAOM ); ibid., 6 Jun. 1976, W.B. Cooke (DAOM ). Massachusetts: Salem, on Salix wood, 1834 (DAOM 43686). Missouri: St Louis County, Benbush, on dead wood, 15 Nov. 1941, G.D. Darker (DAOM 75764). New York: Flatbush, 1890, I.L. Zabriskie (DAOM 34307); Lloyd-Cornell Preserve, Ringwood, on rotten wood, 6 Sep. 1952, S.J. Hughes (DAOM 29052, DAOM 29377); ibid., on decorticated wood (old), 6 Sep. 1952, W.I. Illman (DAOM ); Table 1. Sequences used in phylogenetic analyses in this study. Species Specimen/strain GenBank No. ITS LSU Acanthostigma chiangmaiense MFLUCC T JN JN Acanthostigma filiforme ILLS T GQ GQ Acanthostigma multiseptatum ILLS T NR_ GQ Acanthostigma perpusillum UAMH 7237 AY AY Acanthostigma scopulum ANM 386 GQ GQ Berkleasmium concinnum ILLS (T) NR_ NG_ Berkleasmium concinnum DAOM KY KY Berkleasmium concinnum NB-789 KY KY Berkleasmium concinnum ILLS KY Berkleasmium concinnum ILLS KY Berkleasmium concinnum ILLS KY Berkleasmium concinnum ILLS KY Botryosphaeria corticis CBS T NR_ NG_ Chlamydotubeufia cf. huaikangplaensis MFLUCC KF KF Chlamydotubeufia helicospora MFLUCC T KX KX Chlamydotubeufia khunkornensis MFLUCC T NR_ JN Helicoma conicodentatum UBC F14998 AY AY Helicomyces lilliputeus NBRC AY AY Helicomyces roseus CBS AY AY Helicoön gigantisporum BCC 3550 AY AY Helicosporium aureum NBRC 7098 AY AY Helicosporium gracile CBS AY AY Helicosporium guianense UAMH 1699 AY AY Helicosporium guianense CBS AY AY Helicosporium linderi NBRC 9207 AY AY Helicosporium pallidum CBS AY AY Helicosporium panachaeum CBS AY AY Helicosporium talbotii MUCL AY AY Neoacanthostigma fusiforme MFLUCC T KF KF Neoacanthostigma thailandicum MFLUCC T KX Tamhinispora indica AMH 9555 T NR_ KC Thaxteriellopsis lignicola MFLUCC JN JN Tubeufia amazonensis ATCC T AY AY Tubeufia aurantiella ANM 718 GQ GQ Tubeufia javanica MFLUCC T KJ KJ Tubeufia khunkornensis MFLUCC T JN JN Tubeufia roseohelicospora MFLUCC KX KX New sequences generated in this study are in bold, T denotes sequence from type or ex-type material, (T) refers to the holotype of Neoacanthostigma septoconstrictum, not B. concinnum. 102 IMA FUNGUS

170 Asexual-sexual morphs in Berkleasmium Fig. 2. Berkleasmium concinnum conidiomata and ascomata. A C. Co-occurring ascomata and sporodochia, arrows point to ascomata and asterisks (*) denote sporodochia. D E. Sporodochia in situ. F. Sporodochium. G I. Conidiogenous cells. J. Conidia mounted in H 2 O. K. Developing and mature conidia mounted in lactic acid. A C DAOM , D-H NB-789, I J DAOM , K DAOM Bars: A E = 500 µm, F = 100 µm, G K = 10 µm. Buffalo, G.W. Clinton (DAOM 34362). North Carolina: Haywood Co., Great Smoky Mountains National Park, Big Creek, Baxter Creek Trail, o N, o W, 610 m elev., decorticated 3 cm diam branch on ground, 16 Jun. 2007, A.N. Miller et al. (ANM 1141, ILLS 80802); Cullowhee, on rotten wood, Jun. 1887, R. Thaxter (DAOM 29026). South Carolina: on wood, (DAOM 34361). Tennessee: Blount County, Bote Mountain, Great Smoky Mountains National Park, on rotten wood and bark, 23 Aug. 1977, S.J. Hughes (DAOM , DAOM ); Cocke County, Great Smoky Mountains National Park, Cosby, Low Gap Trail, o N, o W, 716 m elev., decorticated wood on ground, 15 Jul. 2005, A.N. Miller & A.M. Stchigel (ANM536.1, ILLS holotype of Acanthostigma septoconstrictum); ibid., decorticated 5 cm diam. branch on ground, 3 Nov. 2007, A.N. Miller et al. (ANM 1407, ILLS 80805); ibid., Cosby Nature Trail, o N, o W, 716 m elev., 50 cm diam log on ground, 20 May 2008, A.N. Miller et al. (ANM 1701, ILLS 80806); Sevier Co., Great Smoky Mountains National Park, 5 miles east of Gatlinburg, Greenbrier, Old Settlers Trail, o N, o W, 457 m elev., decorticated 6 cm diam branch on ground, 19 Jun. 2007, A.N. Miller et al. (ANM 1171, ILLS 80803); Twin Creeks, Twin Creeks Nature Trail, near ATBI plot, o N, o W, 549 m elev., decorticated 5 cm diam branch on ground, 18 Jun. 2007, A.N. Miller et al. (ANM 1227, ILLS 80804). West Virginia: Giles County, Mountain Lake, on well decayed log of deciduous tree, 2 Sep. 1936, D.H. Linder (DAOM 24916); Nuttallburg, Nov. 1893, on rotten wood, L.W. Nuttall (DAOM 34290). Wisconsin: Devil s Lake near Madison, 4 Sep. 1953, R.F. Cain (DAOM 41029). Neoacanthostigma thailandicum Tanney & A.N. Mill. sp. nov. MycoBank MB Etymology: Named for the country where the type specimen was collected. Diagnosis: Neoacanthostigma thailandicum is distinguished from all other species in the genus by macronematous conidiophores and larger, pale brown to brown, multi-septate conidia (to 920 µm long). Description: For a complete description of this taxon see Hyde et al. (2016: 125; N. septoconstrictum MFLU ). Type: Thailand: Prachuap Khiri Khan, Bang Sapan, Ron Thai, on decaying wood in flowing freshwater stream, 30 Jul. 2015, K.D. Hyde KH02 (MFLU holotype, n.v.; BBH isotype, n.v.; MFLUCC , TBRC cultures ex-type). Illustration: Hyde et al. (2016: fig. 76). Notes: Neoacanthostigma thailandicum (MFLU ) was identified as N. septoconstrictum in Hyde et al. (2016). VOLUME 8 NO

171 Tanney & Miller DISCUSSION Both ITS and LSU sequences confirm that the hyphomycete morph Berkleasmium concinnum and the pyrenomycete morph Neoacanthostigma septoconstrictum are the same species and therefore must be given a single name. Thus, N. septoconstrictum is synonymized under B. concinnum because of the priority of Berkleasmium over Neoacanthostigma. Re-examination of the N. septoconstrictum type specimen and additional B. concinnum reference specimens confirmed the frequent cooccurrence of both the asexual and sexual morphs on the same substrate, providing additional circumstantial support for this connection (Fig. 2 A C). Neoacanthostigma was proposed to accommodate a new species, N. fusiforme, which was chosen as the type species, along with N. filiforme and N. septoconstrictum, two species previously included in Acanthostigma but demonstrated to be phylogenetically distinct from the type species, A. perpusillum (Boonmee et al. 2014). Based on the ITS-LSU phylogeny presented here, the transfer of Acanthostigma filiforme to Neoacanthostigma appears unwarranted given its phylogenetic distance from N. fusiforme (Fig. 1). However, these two species were weakly clustered together based on the LSU phylogeny from Promputtha & Miller (2010) and the combined ITS and LSU phylogeny from Boonmee et al. (2014). The placement of N. filiforme was not resolved in this study because of low support. Hyde et al. (2016) reported a helicomyces-like asexual morph for N. septoconstrictum based on a specimen from decaying wood in a flowing freshwater stream in Thailand; however, the ITS sequence (KX454176) of this specimen (MFLUCC ) clearly distinguishes it from the N. septoconstrictum type specimen [identities = 465/537 (87 %), gaps = 42/537 (7 %)]. The ITS dissimilarity and distinctive asexual morph morphology from B. concinnum resulted in our description of this specimen as a novel species, N. thailandicum. Tubeufiaceae (Tubeufiales) asexual morphs are morphologically diverse, containing helicosporous genera such as Helicoma, Helicomyces, Helicoön, and Helicosporium, and staurosporous genera including Araneomyces and Tetracrium (Réblová & Barr 2000, Kodsueb et al. 2004, Tsui & Berbee 2006). Reports of dictyosporous conidial morphs in Tubeufiaceae more reminiscent of B. concinnum include Chlamydotubeufia spp., Manoharachariella tectonae, Tubeufia amazonensis, and T. khunkornensis (Rossman & Müller 1979, Boonmee et al. 2011, 2014, Rajeshkumar & Sharma 2013, Doilom et al. 2016). The close phylogenetic relationship between B. concinnum and the helicosporous N. fusiforme and N. thailandicum is unexpected but supported by molecular evidence generated from independently collected and sequenced specimens and isolates. The phylogenetic placement and identification of a sexual morph for B. concinnum presented in this study demonstrates the significance of culturing and sequencing named-butunsequenced dematiaceous hyphomycetes. ACKNOWLEDGEMENTS We are grateful to Scott A. Redhead for nomenclatural advice and to Olga Koppel for Russian translation of the DAOM specimen label. This study was partially supported by a National Science Foundation grant (DEB ) to ANM and a Natural Sciences and Engineering Research Council of Canada scholarship (PGSD ) to JBT. JBT is grateful to the Microbiology Molecular Technologies Laboratory (MMTL) group of ORDC-AAFC, Ottawa, for processing DNA sequencing, and Keith A. Seifert for his comments on this manuscript. REFERENCES Berkeley MJ (1845) Decades of fungi. Dec. VIII. X. London Journal of Botany 4: Boonmee S, Zhang Y, Chomnunti P, Chukeatirote E, Tsui CK, et al. (2011) Revision of lignicolous Tubeufiaceae based on morphological reexamination and phylogenetic analysis. Fungal Diversity 51: Boonmee S, Rossman AY, Liu J-K, Li W-J, Dai D-Q, et al. (2014) Tubeufiales, ord. nov., integrating sexual and asexual generic names. Fungal Diversity 68: Brady J (1965) A simple technique for making very fine, durable dissecting needles by sharpening tungsten wire electrolytically. Bulletin of the World Health Organization 32: 143. Bussaban B, Lumyong S, Lumyong P, McKenzie EHC, Hyde KD (2001) A synopsis of the genus Berkleasmium with two new species and new records of Canalisporium caribense from Zingiberaceae in Thailand. Fungal Diversity 8: Corda ACJ (1854) Icones Fungorum hucusque cognitorum. Vol. 6. Prague: J.G. Calve. Doilom M, Dissanayake AJ, Wanasinghe DN, Boonmee S, Liu JK, et al. (2016) Microfungi on Tectona grandis (teak) in Northern Thailand. Fungal Diversity 82: Ellis MB (1971) Dematiaceous Hyphomycetes. Kew: Commonwealth Mycological Institute. Hyde KD, Hongsanan S, Jeewon R, Bhat DJ, McKenzie EHC, et al. (2016) Fungal diversity notes : taxonomic and phylogenetic contributions to fungal taxa. Fungal Diversity 80: Katoh K, Standley DM (2013) MAFFT multiple sequence alignment software version 7: improvements in performance and usability. Molecular Biology and Evolution 30: Kodsueb R, Lumyong S, Lumyong P, McKenzie EH, Ho WH, et al. (2004) Acanthostigma and Tubeufia species, including T. claspisphaeria sp. nov., from submerged wood in Hong Kong. Mycologia 96: Lea TG (1849) Catalogue of Plants, native and naturalized, collected in the vicinity of Cincinnati, Ohio, during the years Philadelphia: Collins. Moore RT (1958) Deuteromycetes I: the Sporidesmium complex. Mycologia 50: Moore RT (1959) The genus Berkleasmium. Mycologia 51: Nylander J (2004) MrModeltest 2.2. Program distributed by the author, Evolutionary Biology Centre, Uppsala University, Sweden. Pinnoi A, Jeewon R, Sakayaroj J, Hyde KD, Jones EG (2007) Berkleasmium crunisia sp. nov. and its phylogenetic affinities to the Pleosporales based on 18S and 28S rdna sequence analyses. Mycologia 99: IMA FUNGUS

172 Asexual-sexual morphs in Berkleasmium Promputtha I, Miller AN (2010) Three new species of Acanthostigma (Tubeufiaceae, Dothideomycetes) from Great Smoky Mountains National Park. Mycologia 102: Raghuveer Rao P, Rao D (1964) Berkleasmium Zobel from india. Mycopathologia 22: Rajeshkumar K, Sharma R (2013) Tamhinispora a new genus belongs to family Tubeufiaceae from the Western Ghats, India based on morphology and phylogenetic analysis. Mycosphere 4: Rambaut A (2014) FigTree v University of Edinburgh, Scotland. Reblova M, Barr ME (2000) The genus Acanthostigma (Tubeufiaceae, Pleosporales). Sydowia 52: Ronquist F, Teslenko M, van der Mark P, Ayres DL, Darling A, et al. (2012) MrBayes 3.2: efficient Bayesian phylogenetic inference and model choice across a large model space. Systematic Biology 61: Rossman A, Müller E (1979) Life history studies of Brazilian ascomycetes 6. Three species of Tubeufia with, respectively, dictyosporous-pycnidial and helicosporous anamorphs. Sydowia 31: Sanchez RM, Miller AN, Bianchinotti MV (2012) A new species of Acanthostigma (Tubeufiaceae, Dothideomycetes) from the Southern Hemisphere. Mycologia 104: Seifert KA, Morgan-Jones G, Gams W, Kendrick B (2011) The Genera of Hyphomycetes. [Biodiversity Series no. 9] Utrecht. CBS-KNAW Fungal Biodiversity Centre. Stamatakis A (2014) RAxML version 8: a tool for phylogenetic analysis and post-analysis of large phylogenies. Bioinformatics 30: Swofford DL (2003) PAUP*: phylogenetic analysis using parsimony (*and other methods). Version 4. Sunderland, MA: Sinaeuer Associates. Tanney JB, Nguyen HD, Pinzari F, Seifert KA (2015) A century later: rediscovery, culturing and phylogenetic analysis of Diploöspora rosea, a rare onygenalean hyphomycete. Antonie van Leeuwenhoek 108: Tsui C, Berbee M (2006) Phylogenetic relationships and convergence of helicosporous fungi inferred from ribosomal DNA sequences. Molecular Phylogenetics and Evolution 39: Wang H-K, Aptroot A, Crous PW, Hyde KD, Jeewon R (2007) The polyphyletic nature of Pleosporales: an example from Massariosphaeria based on rdna and RBP2 gene phylogenies. Mycological Research 111: Whitton SR, McKenzie EH, Hyde KD (2012) List of fungi associated with Pandanaceae. In: Fungi Associated with Pandanaceae (SR Whitton, EH McKenzie & KD Hyde KD, eds): Dordrecht: Springer. VOLUME 8 NO

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174 doi: /imafungus IMA FUNGUS 8(1): (2017) Anther smuts of Silene acaulis and S. uniflora in the Outer Hebrides, including an assessment of ITS genotypes of Microbotryum silenes-acaulis Paul A. Smith 1, Matthias Lutz 2, Rebekka Ziegler 2, and Marcin Piątek 3 1 Statistical Sciences Research Institute, University of Southampton, Highfield, Southampton, SO17 1BJ, UK; corresponding author p.a.smith@soton.ac.uk 2 Plant Evolutionary Ecology, Institute of Evolution and Ecology, University of Tübingen, Auf der Morgenstelle 5, D Tübingen, Germany 3 Department of Mycology, W. Szafer Institute of Botany, Polish Academy of Sciences, Lubicz 46, PL Kraków, Poland; corresponding author m.piatek@botany.pl Abstract: Anther smuts on Silene acaulis and S. uniflora from the Outer Hebrides, Scotland, UK), are analysed using morphological and molecular techniques, and found to represent Microbotryum silenes-acaulis and M. silenes-inflatae, respectively. This is the first identification of caryophyllaceous anther smuts in the Outer Hebrides according to modern species concepts and the first report of Microbotryum silenes-acaulis confirmed by molecular analysis from the British Isles. Additionally, the genetic structure of Microbotryum silenesacaulis, based on all currently available ITS sequences, is analysed and discussed. Seven ITS genotypes are determined for Microbotryum silenes-acaulis, including three genotypes in North America and four genotypes in Europe. Compared to European accessions, all North American accessions share specific nucleotides and are genetically divergent. Key words: arctic and alpine fungi Basidiomycota genotypes Microbotryum molecular phylogenetics plant pathogens Ustilaginales Article info: Submitted: 19 December 2016; Accepted: 30 April 2017; Published: 11 May INTRODUCTION The anther smuts of Caryophyllaceae, in the past ascribed to eight species (Vánky 1994, 1998), but most commonly to Microbotryum violaceum s.lat., have recently been reassessed taxonomically. As a result, host specificity and sometimes subtle morphological differences has led to the division of this complex into a group of species, whose differences have been supported by DNA analyses (Freeman et al. 2002, Lutz et al. 2005, 2008, Le Gac et al. 2007, Refrégier et al. 2008, Piątek et al. 2012, 2013). Following a consolidated species concept approach (Quaedvlieg et al. 2014), anther smuts on Caryophyllaceae are currently divided into 23 species that are usually highly host-specific (Deml & Oberwinkler 1982, 1983, Scholz & Scholz 1988, Vánky 1988, Chlebicki & Suková 2005, Lutz et al. 2005, 2008, Denchev 2007a, 2007b, Denchev et al. 2009, Denchev & Denchev 2011, Piątek et al. 2012, 2013). Yet, the process of disentangling this species complex is still not finished and several groups, such as species on Dianthus (Kemler et al. 2013) or species on diverse, mostly non-european Silene species (Vánky 2012) still need to be resolved. Likewise, the geographical distribution of known species is still poorly known, not only in the non-european regions, but also in European countries. In Great Britain and Ireland, 29 Microbotryum s. lat. (including Bauhinus and Haradaea) species (Fungal Records Database for Britian and Ireland, FRDBI; fieldmycology.net/frdbi/frdbi.asp) have been reported so far, of which 11 are anther smuts on Caryophyllaceae. The FRDBI has no records for Microbotryum species from the Outer Hebrides in Scotland, but there are published records of anther smuts under the aggregate name Ustilago violacea (s.lat.) in Campbell (1936) on Silene uniflora and Dennis (1986) on S. flos-cuculi. Recent field surveys in the Outer Hebrides have already resulted in discoveries of several smut fungi (Piątek et al. 2011, Smith & Lutz 2013, 2014), and enlarged our knowledge of this group of plant pathogens compared to that, for example, in Dennis (1986). During fieldwork in the Outer Hebrides between 2008 and 2016, anther smuts on Silene acaulis and S. uniflora were collected. This work aims to identify the causative agents using morphological and molecular analyses and to compare them with the current state of knowledge of Microbotryum species on these hosts in Great Britain and Ireland. Additionally, the genetic structure of M. silenes-acaulis, based on all currently available ITS sequences, is analysed and discussed. MATERIALS AND METHODS Specimen sampling, documentation and analysis of infection rate Specimens used for morphological and molecular analyses were of Microbotryum species infecting the anthers of Silene acaulis and S. uniflora, collected in , and preserved by pressing. Specimens used for morphological 2017 International Mycological Association You are free to share - to copy, distribute and transmit the work, under the following conditions: Attribution: You must attribute the work in the manner specified by the author or licensor (but not in any way that suggests that they endorse you or your use of the work). Non-commercial: You may not use this work for commercial purposes. No derivative works: You may not alter, transform, or build upon this work. For any reuse or distribution, you must make clear to others the license terms of this work, which can be found at Any of the above conditions can be waived if you get permission from the copyright holder. Nothing in this license impairs or restricts the author s moral rights. VOLUME 8 NO

175 Smith et al. analyses only and used to summarise distributions were collected in 2008, and between , and also preserved by pressing. The collection details are given in Table 1. The voucher specimens are deposited in the Herbarium of the Eberhard-Karls-Universität Tübingen, Germany (TUB), and in the W. Szafer Institute of Botany, Polish Academy of Sciences, Kraków, Poland (KRAM F). In selected populations of S. acaulis, the numbers of infected and uninfected clumps were counted in defined areas to assess the rate of infection; only plants flowering at the time of field work could be assessed. Morphological analyses Morphological characters were studied using dried material. All material was analysed by light microscopy (LM). The spores were mounted in 80 % lactic acid, heated to boiling point, and then examined under a Nikon Eclipse 80i light microscope (Nikon) at x1000. Spore sizes were counted using the NIS-Elements BR 3.0 imaging software (Nikon). The extreme measurements were adjusted to the nearest 0.5 µm. The spore size range, mean and standard deviation of 30 spore counts from each specimen are given in Table 1. The morphological description of species includes combined values from all measured specimens. LM micrographs were taken with a Nikon DS-Fi1 camera (Nikon). The spore ornamentation of the anther smut on Silene uniflora (KRAM F-59020) was additionally analysed by scanning electron microscopy (SEM). For this purpose, dry spores were mounted on carbon tabs and fixed to an aluminium stub with double-sided transparent tape. The tabs were sputtercoated with carbon using a Cressington carbon-coater and viewed with a Hitachi S-4700 scanning electron microscope (Hitachi High-Technologies Corporation), at 10 kv and 9,500 na, with a working distance of ca. 12 mm, in the secondary electron imaging mode. SEM micrographs were taken in the Laboratory of Field Emission Scanning Electron Microscopy and Microanalysis at the Institute of Geological Sciences, Jagiellonian University, Kraków (Poland). Phylogenetic analyses Molecular phylogenetic analyses followed the techniques used in previous work (Piątek et al. 2013). ITS genotypes of Microbotryum silenes-acaulis were assessed using the MAFFT-alignment (available in TreeBASE: S20963) that included all ITS sequences available in GenBank (Freeman et al. 2002, Lutz et al. 2008, Hood et al. 2010) and obtained in this study. To elucidate the phylogenetic position of the Microbotryum specimens from the Outer Hebrides, their concatenated ITS + LSU sequences were analysed within the dataset of Piątek et al. (2013) that covers all caryophyllaceous anther smut species for which ITS and LSU sequences are available and comprises 21 of the 23 currently recognized species. For the final analyses, the dataset (alignment available in TreeBASE: S20963) was reduced to a maximum of two sequences per species except for the clades in which the specimens from the Outer Hebrides clustered. GenBank accession numbers of sequences generated in this study are given in Table 1. RESULTS AND DISCUSSION This study provides the first identification of caryophyllaceous anther smuts in the Outer Hebrides according to modern species concepts, and the first report of anther smuts on Silene acaulis in the Outer Hebrides. The anther smuts on Silene acaulis and S. uniflora could be assigned to two species as follows. Anther smut on Silene acaulis Silene acaulis is a member of the European arctic-montane element of the British flora (Preston & Hill 1997), with only a few sites in England, Wales and Ireland, all in the higher mountains. In Scotland it is more widespread on the mountains, and also descends to sea level in a number of places. The Outer Hebrides have examples of both altitude types, with scattered plants on exposed rocky summits of a few of the higher hills in North Harris from m, and more extensive populations in very exposed situations on the west coast of Lewis from 0 50 m; there are also outlying populations on St Kilda and old records from Mingulay and Berneray, where it has not been found since the 1940s (Fig. 1). The pattern of S. acaulis genotypes in Europe and across the whole range has been investigated by Mikhaylova et al. (2010) and Gussarova et al. (2015), respectively. Anther smut on S. acaulis has been found in both highaltitude and sea level populations in the Outer Hebrides, from six populations in total (Table 1, Figs 2 3). Since many of the plant records have been made by botanists who have not been looking for smuts, it is likely that many other populations have infected individuals which have not yet been recognized. The same phenomenon has been found by Hood et al. (2010), even in herbarium specimens. The specimens in anthers of S. acaulis produced a violaceous, powdery spore mass replacing the pollen in pollen sacs. Spores were pale violet to violet in transmitted light, globose, subglobose, slightly ellipsoid to ovoid, ( 12.0) (5.5 ) ( 10.0) µm [av. ± SD, 7.9 ± ± 0.7 µm, n = 150/5]; the wall was reticulate, ca µm high, the meshes were more or less polyhedral, usually irregular, rarely regular, and the number of meshes per spore diameter was 4 8 (usually 6 8), interspaces were smooth as observed by LM. The colour of the spore mass in the anthers, and the morphology and spore sizes of the analysed specimens agree well with those reported in the original description of Microbotryum silenes-acaulis (Lutz et al. 2008). The ITS and LSU sequences of the analysed smut specimens on S. acaulis from the Outer Hebrides were identical and clustered together with different accessions of anther smut on S. acaulis ascribed to Microbotryum silenes-acaulis (Fig. 4). Therefore, the anther smut on Silene acaulis in the Outer Hebrides is identified as M. silenes-acaulis. Microbotryum silenes-acaulis is a rarely reported species in Great Britain and Ireland, with the FRDBI having only two records from Scotland (Mid-Perthshire: Ben Lawers, 1942; and North Ebudes: Isle of Skye, 1986), presumably re-annotated automatically in line with the changed nomenclature in Lutz et al. (2008). The records from Lewis are therefore a useful confirmation of the occurrence of this species, and constitute the first identification of M. silenes-acaulis from Great Britain and Ireland based on morphological and molecular evidence. 108 IMA FUNGUS

176 Microbotryum smuts on Silene Table 1. Microbotryum specimens, with host plants, spore size range, mean spore size with standard deviation, GenBank accesion numbers, and reference specimens used in this study. All sites are in vice-county 110, Outer Hebrides (Scotland). Species Host species Spore size range (µm) Mean spore size with standard deviation (µm) Microbotryum silenes-acaulis Silene acaulis (6.0 ) ( 10.0) ( 8.5) Silene acaulis (7.0 ) (6.0 ) ( 8.5) Silene acaulis (6.0 ) ( 11.0) (5.5 ) ( 8.0) GenBank acc. no. (ITS/LSU) 7.8 ± ± 0.6 JN223408/ JN ± ± 0.5 JN223406/ JN ± ± 0.6 JN223407/ JN Reference specimens 1 Loch a Bheannaich, Lewis, NB , 7 May 2010, P.A. Smith, TUB Mangersta sands, Lewis, NB , 1 May 2010, P.A. Smith, TUB Mangersta Radio Station, Lewis, NB , 4 May 2010, P.A. Smith, TUB Silene acaulis ± ± 0.4 N/A Druim a Bheannaich, Lewis, NB0338, 7 May 2010, leg. P.A. Smith, KRAM F Silene acaulis ( 12.0) ( 10.0) 8.3 ± ± 0.9 N/A Ceartabhal, summit, N. Harris, 550 m, NB043127, 17 June 2013, P.A. Smith, KRAM F Silene acaulis No specimen available No specimen available No specimen available Camas Geodhachan an Duilisg, Aird Uig, Lewis, 20 m, NB , 29 April 2010, P.A. Smith, not preserved Microbotryum silenes-inflatae Silene uniflora (5.5 ) (5.5 ) Silene uniflora ( 11.0) (6.0 ) ( 10.0) Silene uniflora (6.0 ) (5.5 ) ± ± 0.6 JN223404/ JN ± ± 0.8 JN223405/ JN ± ± 0.5 KY321304/ KY Sgiogarstaigh, Lewis, NB56L, 31 July 2009, P.A. Smith, TUB Leum Langa, Lewis, NB560572, 6 Aug. 2009, P.A. Smith, TUB Gasgeir, NA8711, 13 July 2012, M. Inglis, KRAM F Silene uniflora (6.0 ) ± ± 0.7 N/A Pabaigh Mor, Lewis, NB13E, 30 June 2008, P.A. Smith, KRAM F Silene uniflora ( 9.0) 7.7 ± ± 0.7 N/A Lingail Mor, Uig, Lewis, NB027345, 1 July 2008, P.A. Smith, KRAM F Silene uniflora (5.5 ) ± ± 0.4 N/A Bearasaigh, Loch Roag, Lewis, NB14G, 3 July 2008, P.A. Smith, KRAM F Silene uniflora ± ± 0.7 N/A Flodaigh, Loch Roag, Lewis, NB14F, 3 July 2008,. P.A. Smith, KRAM F Silene uniflora (6.0 ) ± ± 0.4 N/A Seana Cnoc, Loch Roag, Lewis, NB14B, 3 July 2008,. P.A. Smith, KRAM F Silene uniflora (5.5 ) ( 8.0) 7.1 ± ± 0.6 N/A Knoch Ibidale, sea cliff, Lewis, NB , 24 April 2014, P.A. Smith, KRAM F Silene uniflora ( 8.5) 7.6 ± ± 0.5 N/A Loch Eirearaigh, shingle bank, Lewis, NB , 30 July 2016, P.A. Smith, KRAM F Silene uniflora (6.5 ) ( 10.0) ( 9.5) Silene uniflora (5.5 ) ( 8.5) (5.5 ) ( 8.0) 7.9 ± ± 0.7 N/A Aird Bheag Bhragair, cliff, Lewis, NB , 3 August 2016, P.A. Smith, KRAM F ± ± 0.5 N/A Liongam, by loch, NA , 11 August 2016, P.A. Smith, KRAM F KRAM F, Mycological Collections, W. Szafer Institute of Botany, Polish Academy of Sciences, Kraków, Poland; TUB, Eberhard-Karls-Universität Tübingen, Germany. Infection rates Bueker et al. (2016) discussed the correlation between isolation in populations and the incidence of infection by Microbotryum silenes-acaulis. They found that the incidence of infection was greater in more northern latitudes and in areas where the densities of Silene acaulis locations were lower. The Outer Hebrides populations of Silene acaulis are isolated, both on a medium scale within the islands (Fig. 1), and on a larger scale in Great Britain and Ireland (Preston et al. 2002: 177). We therefore expect rates of infection to VOLUME 8 NO

177 Smith et al. be relatively high, and infection rates within populations in 2010 were estimated at 9 % at Loch a Bheannaich, 4 % at Mangersta Sands, and 25 % at Mangersta Radio Station (see Table 1 for locality details). These are all higher than the average 2.9 % found by Bueker et al. (2016) across the known distribution range based on herbarium specimens, although the populations represented by these specimens will have varying infection rates, some higher and some lower than the average. The population infection rate is likely to be much higher, as to date all populations which have been searched specifically for infections have been found to contain infected individuals. Abbate & Antonovics (2014) investigated a related hostpathogen system with Microbotryum species and Silene vulgaris. They found that disease was more likely to be found where overall temperatures were lower, fluctuations in diurnal and seasonal temperatures were smaller, and where precipitation totals were greater and more stable. This succinctly describes conditions in the Outer Hebrides, which have mild winters and cool summers because of their extreme oceanicity, and one of the smallest annual temperature ranges in Britain (8.8 C in Stornoway). More information on climate in the Outer Hebrides is provided by Angus (1997: ). The high infection rates observed are therefore consistent with both isolation and climate. Fig. 1. Distributions of Silene acaulis and S. uniflora in the Outer Hebrides based on all records from any date. Localities mentioned in the text are named. Genetic diversity The ITS sequences of Microbotryum silenes-acaulis from different geographic areas differed. Based on all ITS sequences available in GenBank (Freeman et al. 2002, Lutz et al. 2008, Hood et al. 2010) and those obtained in this study, and assuming that there were no sequencing errors, seven ITS genotypes were determined for M. silenes-acaulis (Figs 4 5). First, these genotypes reveal a disjunction between M. silenes-acaulis specimens from North America and from Europe. Compared to European accessions, all North American accessions share specific nucleotides and differ genetically. This is congruent with the genetic structure of the host plant Silene acaulis. North American populations are genetically distinct from other world populations (from Europe and Siberia) having nine unique plastid DNA haplotypes of the 17 haplotypes detected across Fig. 2. Microbotryum silenes-acaulis on Silene acaulis. A. The habitat at Loch a Bheannaich, Lewis. B. Infected and uninfected flowers at the same locality. Note that this is a female tussock, but the fungus has induced infected flowers to produce stamens. (Corresponds to TUB ; scale bar approx 1 cm). 110 IMA FUNGUS

178 Microbotryum smuts on Silene microsatellite markers derived from herbarium specimens. There are clear patterns with reduced diversity in northern latitudes. Bueker et al. (2016) did not include any specimens from Great Britain, Ireland or the Tatra Mts, and their genetic analyses were based on microsatellites rather than ITS, so it is not clear how the Outer Hebrides or the Tatra specimens would fit into this structure. Nevertheless, the residual structure in the phylogenetic hypothesis for M. silenesacaulis (Fig. 4) agrees with Bueker et al. (2016) s pattern, with distinctions between North America and Europe, and within Europe between northern and southern latitudes. The grouping of the Outer Hebrides specimens with those from Sweden and Switzerland is also consistent with Bueker et al. (2016) s groups, which show some genotypes in common between Scandinavia and the Alps. The homology of these groupings, however, remains to be demonstrated. Fig. 3. Distribution of Microbotryum silenes-acaulis on Silene acaulis in the Outer Hebrides. the whole geographical range of the species (Gussarova et al. 2015). Future studies including a larger sampling and infection trials should reveal whether specific nucleotides and the genetic divergence between North American and European accessions of M. silenes-acaulis indicate that the anther smuts on S. acaulis in these areas belong to two distinct species. Recent studies of different fungi revealed that morphologically similar species in North America and Europe, earlier unified under the same specific epithet, are divergent genetically and represent distinct species (e.g. Hughes et al. 2014, Spirin et al. 2015, 2016). The European accessions of M. silenes-acaulis show a phylogenetic substructure that separates specimens from Austria (Eastern Alps) and Italy (Central Apennines), ones from Poland (Tatra Mts), and those from Switzerland (western part of the Eastern Alps), Sweden (Lapland), and the Outer Hebrides (Fig. 4). The sequences from the Outer Hebrides specimens differ in one nucleotide position from sequences obtained from Swedish and Swiss specimens (Fig. 5). The existence of a distinct genotype of M. silenes-acaulis in the Tatra Mts is noteworthy and may indicate a long period of isolation of the anther smut population in this mountain range. Previous studies detected the occurrence of distinct genotypes of the alpine vascular plants Carex atrofusca (Schönswetter et al. 2006) and Ranunculus glacialis (Ronikier et al. 2012) in the Tatra Mts, but our new work is the first indication of the occurrence of distinct genotypes amongst representatives of Fungi. More intensive sampling of ITS sequences (and maybe other markers) from other mountain and arctic populations of M. silenes-acaulis is needed; this may confirm or modify the currently inferred phylogenetic structure within this species. Bueker et al. (2016) have recently analysed the genetic pattern of M. silenes-acaulis across its range using Anther smut on Silene uniflora Silene uniflora is a common, gynodioecious, coastal plant in Great Britain and Ireland, but also has some inland populations, usually at higher altitudes in mountains or in heavy metal contaminated sites (Marsden-Jones & Turrill 1957), giving a distribution which is qualitatively similar to that of S. acaulis, though it is a much more frequent species (Preston et al. 2002: 176). Silene uniflora is closely related to S. vulgaris, which is also widely distributed in Great Britain and Ireland, but has a generally more southern distribution and is often found inland. Two species of anther smut, Microbotryum silenes-inflatae and M. lagerheimii, have been reported as infecting Silene uniflora from south-west England by Chung et al. (2012), who report sympatric populations of these two Microbotryum species in Somerset and Suffolk (SW and SE England, respectively). In the Outer Hebrides, Silene uniflora is the only one of the S. uniflora/s. vulgaris species pair so far known, and it is exclusively coastal, usually growing on shingle banks, rocks and cliffs where it is protected from grazing (Fig. 1). Smutted flowers are occasional, and were first reported in the Outer Hebrides from Barra by Campbell (1936). Fieldwork since 2008 has identified smutted flowers from 12 locations (Table 1, Fig. 6), and these specimens have been analysed in more detail below. In the same way as for S. acaulis, smutted flowers are almost certainly much more frequent than has been recorded, as plant records are generally made by people who are not looking out for smuts. The specimens in anthers of S. uniflora produced a relatively dark violaceous, occasionally light violaceous (one specimen: from Gasgeir, KRAM F-59020), powdery spore mass replacing the pollen in pollen sacs. Spores were pale violet in transmitted light, globose, subglobose, slightly ellipsoid to ovoid, (5.5 ) ( 11.0) (5.5 ) ( 10.0) µm [av. ± SD, 7.5 ± ± 0.7 µm, n = 360/12]; the wall was reticulate, ca. 1.0 µm high, the meshes were more or less polyhedral, usually irregular, rarely regular, and the number of meshes per spore diameter was 5 9 (usually 6), interspaces were smooth as observed by LM, smooth or rugulose as observed by SEM (Fig. 7). The colour of the spore mass in the anthers, and the morphology and spore sizes of the analysed specimens agree well with those reported in the description of M. silenes-inflatae (Denchev VOLUME 8 NO

179 Smith et al. 1 substitution/site 74/93 M. silenes-inflatae on S. vulgaris AY588105/DQ M. silenes-inflatae on S. vulgaris AY588106/DQ M. silenes-inflatae on S. uniflora JN223405/JN M. silenes-inflatae on S. uniflora JN223404/JN M. silenes-inflatae on S. uniflora KY321304/KY M. silenes-inflatae on S. uniflora AY877408/- M. silenes-saxifragae on S. saxifraga AY588102/JN /- M. silenes-saxifragae on S. saxifraga JN000071/JN M. sp. on S. campanula JN942212/JN M. coronariae on Lychnis flos-cuculi KC684887/KC M. coronariae on Lychnis flos-cuculi AY877417/KC M. sp. on S. ciliata AF038833/- M. violaceum s.str. on S. nutans DQ640065/DQ M. violaceum s.str. on S. nutans DQ640071/DQ M. lychnidis-dioicae on S. latifolia ssp. alba AY588096/DQ M. lychnidis-dioicae on S. latifolia ssp. alba AY588097/DQ M. silenes-dioicae on S. dioica AY877416/DQ M. silenes-dioicae on S. dioica AY588094/DQ M. minuartiae on Minuartia recurva DQ366853/DQ M. minuartiae on Minuartia recurva DQ366852/DQ M. bardanense on S. moorcroftiana DQ366856/DQ M. violaceo-irregulare on S. vulgaris AY588104/DQ M. silenes-acaulis on S. acaulis JN223408/JN Outer Hebrides M. silenes-acaulis on S. acaulis JN223407/JN Outer Hebrides M. silenes-acaulis on S. acaulis JN223406/JN Outer Hebrides M. silenes-acaulis on S. acaulis DQ366850/- Switzerland (W part of Eastern Alps) M. silenes-acaulis on S. acaulis DQ366849/- Sweden (Lapland) M. silenes-acaulis on S. acaulis DQ366846/DQ Poland (Tatra Mountains) M. silenes-acaulis on S. acaulis DQ366851/- Poland (Tatra Mountains) M. silenes-acaulis on S. acaulis DQ366855/- Poland (Tatra Mountains) M. silenes-acaulis on S. acaulis DQ366854/DQ Austria (Eastern Alps) M. silenes-acaulis on S. acaulis GQ150515/- Italy (Central Apennines) M. silenes-acaulis on S. acaulis GQ150516/- USA (Colorado) M. silenes-acaulis on S. acaulis GQ150524/- USA (Alaska) M. silenes-acaulis on S. acaulis AY014226/- Canada M. majus on S. otites AY877419/DQ M. majus on S. otites AY877418/EF M. adenopetalae on S. adenopetala DQ366848/DQ M. chloranthae-verrucosum on S. chlorantha AY877421/DQ M. chloranthae-verrucosum on S. chlorantha AY877404/DQ /91 88/59 64/- 94/61 90/ 65 53/- 79/- 80/81 72/74 M. shykoffianum on D. sylvestris AY588082/DQ M. shykoffianum on D. carthusianorum AY588079/DQ M. dianthorum s.l. on D. monspessulanus AY588080/DQ M. superbum on D. superbus AY588081/DQ M. dianthorum s.l. on Petrorhagia saxifraga DQ366845/DQ M. dianthorum s.l. on D. jacquemontii DQ366844/DQ M. saponariae on Saponaria officinalis AY588089/DQ M. saponariae on Saponaria pumila AY588091/DQ M. stellariae on Stellaria graminea AY588108/DQ M. stellariae on Stellaria graminea AY588109/DQ M. heliospermae on Heliosperma pusillum HQ832084/HQ M. heliospermae on Heliosperma pusillum HQ832082/HQ M. lagerheimii s.str. on Viscaria vulgaris AY877413/AY M. lagerheimii s.l. on Atocion rupestre HQ832090/HQ M. violaceo-verrucosum on S. viscosa AY588103/DQ M. violaceo-verrucosum on S. italica AF045874/- M. sp. on S. virginica AY014235/- M. sp. on S. caroliniana ssp. caroliniana AY014239/- M. scabiosae AY588083/DQ Fig. 4. Bayesian inference of phylogenetic relationships between the sampled Microbotryum species: Markov chain Monte Carlo analysis of an alignment of concatenated ITS + LSU base sequences using the GTR+I+G model of DNA substitution with gamma distributed substitution rates and estimation of invariant sites, random starting trees, and default starting parameters of the DNA substitution model. A 50 % majority-rule consensus tree is shown computed from trees that were sampled after the process had reached stationarity. The topology was rooted with Microbotryum scabiosae. Numbers on branches before slashes are estimates for a posteriori probabilities; numbers on branches after slashes are ML bootstrap support values. Branch lengths were averaged over the sampled trees. They are scaled in terms of expected numbers of nucleotide substitutions per site. The colour dots refer to ITS genotypes of Microbotryum silenes-acaulis (see also Fig. 5). D. = Dianthus, M. = Microbotryum, S. = Silene. 112 IMA FUNGUS

180 Microbotryum smuts on Silene Fig. 5. Nucleotide differences between aligned ITS sequences, and the geographical distribution of corresponding specimens of Microbotryum silenesacaulis. Locations depicted with a question mark are not precise as they were included in GenBank as Alaska and Canada, respectively. Colours refer to ITS genotypes (see also Fig. 4). Fig. 6. Records of Microbotryum silenesinflatae on Silene uniflora from the Outer Hebrides since b). Based on the relatively dark violaceous spore mass from specimens in locations where ITS and LSU sequences were not investigated, all of the remaining collected smuts on S. uniflora were assigned to M. silenes-inflatae. The colour of the spore mass, pale violaceous (low colour intensity) vs. dark violaceous (high colour intensity), differentiates M. lagerheimii and M. silenes-inflatae (Denchev 2007b). The LM and SEM-imaging ornamentation of anther smut spores VOLUME 8 NO

181 Smith et al. Fig. 7. Microbotryum silenes-inflatae on Silene uniflora from the Outer Hebrides. A. The habitat at Loch Eirearaigh, Lewis. B. Infected flower at the same locality (corresponds to KRAM F-59027; scale bar approx 1 cm). C D. Spores seen by LM, median and superficial views (TUB ). E F. Spores seen by SEM (KRAM F-59020). Bars: C D = 10 µm, E = 5 µm, F G = 4 µm. from S. uniflora is shown here for the first time, and is roughly congruent with those published for specimens of M. silenesinflatae on S. vulgaris (Deml & Oberwinkler 1983, Vánky 1985, 1994). The ITS and LSU sequences of the three smut specimens on S. uniflora analysed from the Outer Hebrides, including the specimen from Gasgeir (KRAM F-59020) with light violaceous sori, were identical and clustered together with the anther smut specimens on S. vulgaris from Austria and Switzerland (ITS/LSU: AY588105/DQ366884, AY588106/ DQ366879; Lutz et al. 2008). The ITS sequence of the smut specimen on S. uniflora from Norway (AY877408; Lutz et al. 114 IMA FUNGUS

182 Microbotryum smuts on Silene 2005) differed in 3 bp (0.48 %). All these specimens were assigned to M. silenes-inflatae (Fig. 4). Therefore, the anther smut on S. uniflora in the Outer Hebrides is identified as that species. Microbotryum silenes-inflatae is already known from Great Britain and Ireland, with more than 40 records in FRDBI on S. uniflora and S. vulgaris assigned to this species, mostly on the basis of host plant information. It is likely that other collections of anther smuts on this group of hosts, tentatively assigned in FRDBI to Microbotryum sp. or M. violaceum, will prove to be the same species. Caution is necessary, however, as some of these records may represent M. lagerheimii, which could also infect S. uniflora and S. vulgaris (Denchev 2007b, Chung et al. 2012). Most records are from England and Wales, with a few records from Scotland, the Channel Islands, and Ireland. In addition to the reports of M. silenes-inflatae on Silene uniflora from south-west England (Chung et al. 2012) based on a molecular identification, our study provides the first reports of this species from the Outer Hebrides (Scotland) supported by both morphological and molecular identifications. Both studies provide reliable, DNAbased, evidence for the occurrence of this fungus in Great Britain and Ireland. ACKNOWLEDGEMENTS We thank Anna Łatkiewicz (Kraków) for technical help with scanning electron microscopy. Marcin Piątek was supported by the statutory funds of the W. Szafer Institute of Botany, Polish Academy of Sciences, Kraków, Poland. REFERENCES Abbate JL, Antonovics J (2014) Elevational disease distribution in a natural plant pathogen system: insights from changes across host populations and climate. Oikos 123: Angus S (1997) The Outer Hebrides: the shaping of the islands. Cambridge: White Horse Press. Bueker B, Eberlein C, Gladieux P, Schaefer A, Snirc A, et al. 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184 doi: /imafungus IMA FUNGUS 8(1): (2017) Asexual and sexual morphs of Moesziomyces revisited Julia Kruse 1, 2, Gunther Doehlemann 3, Eric Kemen 4 1, 2, 5, and Marco Thines 1 Goethe University, Department of Biological Sciences, Institute of Ecology, Evolution and Diversity, Max-von-Laue-Str. 13, D Frankfurt am Main, Germany; corresponding author thines@smut-fungi.net 2 Biodiversität und Klima Forschungszentrum, Senckenberg Gesellschaft für Naturforschung, Senckenberganlage 25, D Frankfurt am Main, Germany 3 Botanical Institute and Center of Excellence on Plant Sciences (CEPLAS), University of Cologne, BioCenter, Zülpicher Str. 47a, D-50674, Köln, Germany 4 Max Planck Institute for Plant Breeding Research, Carl-von-Linne-Weg 10, Köln, Germany 5 Integrative Fungal Research Cluster (IPF), Georg-Voigt-Str , D Frankfurt am Main, Germany Abstract: Yeasts of the now unused asexually typified genus Pseudozyma belong to the smut fungi (Ustilaginales) and are mostly believed to be apathogenic asexual yeasts derived from smut fungi that have lost pathogenicity on plants. However, phylogenetic studies have shown that most Pseudozyma species are phylogenetically close to smut fungi parasitic to plants, suggesting that some of the species might represent adventitious isolations of the yeast morph of otherwise plant pathogenic smut fungi. However, there are some species, such as Moesziomyces aphidis (syn. Pseudozyma aphidis) that are isolated throughout the world and sometimes are also found in clinical samples and do not have a known plant pathogenic sexual morph. In this study, it is revealed by phylogenetic investigations that isolates of the biocontrol agent Moesziomyces aphidis are interspersed with M. bullatus sexual lineages, suggesting conspecificity. This raises doubts regarding the apathogenic nature of asexual morphs previously placed in Pseudozyma, but suggests that there might also be pathogenic sexual morph counterparts for those species known only from asexual morphs. The finding that several additional species currently only known from their yeast morphs are embedded within the genus Moesziomyces, suggests that the yeast morph might play a more dominant role in this genus as compared to other genera of Ustilaginaceae. In addition, phylogenetic reconstructions demonstrated that Moesziomyces bullatus has a narrow host range and that some previously described but not widely used species names should be applied for Moesziomyces on other host genera than Echinochloa. Key words: ecology evolution phylogeny plant pathogens pleomorphic fungi Ustilaginomycotina yeast Article info: Submitted: 7 December 2017; Accepted: 4 May 2017; Published: 15 May INTRODUCTION Ustilaginales is the largest order within the smut fungi (Ustilaginomycetes), including species forming a blackish to brownish powdery spore mass in different organs of monocotyledonous, and exceptionally dicotyledonous plants (Vánky 2012, Begerow et al. 2006). The order includes nine families, encompassing 54 genera. The Anthracoideaceae with the occurrence on Cyperaceae and Juncaceae, and Ustilaginaceae, with few exceptions parasitic to the Poaceae, are the largest families within the order. The latter contains the three largest smut genera, Anthracocystis, Sporisorium, and Ustilago. The difference between these three closely related genera is the almost complete lack of a plant-derived columella within sori formed by Ustilago species (Vánky 2012, McTaggart et al. 2012). Within Ustilago some species are economically important pathogens, like corn smut (Ustilago maydis) or wheat smut (Ustilago nuda). Ustilago maydis is a species for which one of the first fungal genomes was sequenced (Kämper et al. 2006). Smut fungi of the Ustilaginales usually feature both an asexual yeast morph and a sexual morph infecting host plants in a biotrophic manner. In rare cases yeasts of the Ustilaginales could also be found to be affecting humans (McNeil & Palazzi 2012, Teo & Tay 2006). The earliest case of an invasive infection with an Ustilago species, possibly U. maydis, was reported in 1946 (Moore et al. 1946). But spores of the Ustilaginales potentially also cause pneumonias, allergic reactions, or asthma (Valverde et al. 1995). There are several studies dealing with the phylogeny of Ustilaginomycotina, mostly based on the LSU or ITS locus and some of them include asexual morphs as well (e.g. Begerow et al. 2000, 2006, Stoll et al. 2005, Wang et al. 2006, 2015, Boekhout 2011). Wang et al. (2015) link many asexual yeasts to their corresponding sexual morphs, an important step within the naming of pleomorphic fungi, as dual naming of sexual and asexual morphs is now discontinued (Hawksworth et al. 2011). Pseudozyma has been used for species of ustilaginomycetous yeasts belonging to Ustilaginales which are mostly 2017 International Mycological Association You are free to share - to copy, distribute and transmit the work, under the following conditions: Attribution: You must attribute the work in the manner specified by the author or licensor (but not in any way that suggests that they endorse you or your use of the work). Non-commercial: You may not use this work for commercial purposes. No derivative works: You may not alter, transform, or build upon this work. For any reuse or distribution, you must make clear to others the license terms of this work, which can be found at Any of the above conditions can be waived if you get permission from the copyright holder. Nothing in this license impairs or restricts the author s moral rights. VOLUME 8 NO

185 Kruse et al. believed to be apathogenic (Begerow et al. 2000, 2006). The genus was described in 1985, by Bandoni (1985), and later refined by Boekhout (1995). After Sampaio (2004) and finally Wang et al. (2015) established that the type species Pseudozyma prolifica was a synonym of Ustilago maydis, the name Pseudozyma should no longer be used. However, the phylogenetic position of some species referred to Pseudozyma is still unclear. Wang et al. (2015) suggested using the name Pseudozyma now with the addition pro tempore for five Pseudozyma species with an unclear phylogenetic position. In the current study we give Pseudozyma names with reference to the new combinations recently made, where possible. To date, about 20 Pseudozyma species names were validly published ( Fifteen are now linked to a corresponding sexually typified genus (Wang et al. 2015). Of these, Pseudozyma antarctica, P. aphidis, P. parantarctica, and P. rugulosa were transferred to Moesziomyces (Wang et al. 2015). Ustilaginalean yeasts are isolated from diverse habitats, for Ustilaginales, but mostly from grasses (Boekhout 1995, Avis et al. 2001). Some were isolated from flowers, leaves or fruits of other plants, but it is also possible to isolate them from soil or even human blood or secretory fluid (Sugita et al. 2003, Arendrup et al. 2014). Apart from the few clinical cases of Ustilago maydis infestation, it was not known until 2003 that species referred to Pseudozyma could also infect humans (Sugita et al. 2003). However, such species can cause invasive infections, especially in immunosuppressed individuals (Arendrup et al. 2014). The infection risk, according to Prakash et al. (2014), is the same as being colonised by nonalbicans Candida infections. Furthermore, Avis et al. (2001) and Gafni et al. (2015) reported antifungal properties of some pseudozyma-like yeast species, including Moesziomyces aphidis, and some strains have been reported to be natural antagonists of powdery mildews (Erysiphales). Colonization of leaf surfaces by these yeasts provides a natural source of protection against some plant pathogenic fungi (Gafni et al. 2015). Of the ustilaginalean yeasts, especially Moesziomyces antarcticus and M. aphidis, formerly treated as Pseudozyma species, have been frequently isolated from various substrates. Moesziomyces antarcticus was isolated from plants and soil, but also from blood of humans (Boekhout 2011). Moesziomyces aphidis was described in 1995 and first isolated from the secretion of an aphid, but it has later been isolated from water (Boekhout 2011) and various other sources, including soil and human blood. Wang et al. (2015) showed that these pseudozyma-like species, together with M. parantarcticus and M. rugulosus, belong to the genus Moesziomyces, which before had been generally regarded as monotypic (Vánky 2005). Moesziomyces mainly differs from other smut fungi in having remnants of ruptured sterile cells (Vánky 1977). Vánky (1977) initially included four species in the genus Moesziomyces. Three of them, occurring on different genera of grasses (Leersia, Paspalum, Pennisetum) were later considered to be conspecific and united under the oldest name, M. bullatus (Vánky 1986, 2005). The other species, M. eriocauli (Vánky 1986), was transferred to a new genus Eriomoeszia, because of a thin cortex of sterile cells, which surrounds the spore balls (Vánky 2005). After the transfer of four pseudozyma-like species (M. antarcticus, M. aphidis, M. parantarcticus and M. rugulosus) to this genus, it now contains five species (Wang et al. 2015). Given the high host specificity observed for most Ustilaginales species (McTaggart et al. 2012, Escudero 2015, Li et al. 2017), it seemed doubtful that Moesziomyces bullatus was parasitic to seven not closely related genera, suggesting that more species might be present in the genus, some of which might be conspecific with smuts in the past named in Pseudozyma. It was the aim of the current study to clarify the relationships of asexual and sexual morphs in the genus Moesziomyces. MATERIAL AND METHODS Fungal material The fungal material used in this study is listed in Tables 1 and 2. The nomenclature of the hosts is derived from the latest version of The International Plant Names Index ( org), the nomenclature of the fungi follows Vánky (2012) and MycoBank ( Yeast cultivation Fresh material of Moesziomyces bullatus collected in 2015 (GLM-F105817) was used for yeast cultivation. A suspension of spores in 2 ml water was prepared. Three tubes with 200 µl spore suspension were exposed to three different conditions: (1) heating of the suspension on a thermomixer at 45 C for 5 min (Shetty & Safeeulla 1979); (2) chilling the suspension overnight on ice; and (3) incubation for 5 min at room temperature (ca. 20 C). From each tube 20 µl suspension was each plated on two plates of SAM (Thines lab Standard Agar Medium, consisting of 20 g agar, 20 g PDB, 10 g yeast extract, 10 g malt extract, 40 ml clarified vegetable juice, 960 ml water) with the addition of 75 mg Rifampicin/L. One set of plates was incubated at 30 C, the other set at room temperature. After 3 d on every plate abundant yeast growth was recognized. Pure cultures were produced by picking and transferring individual single colonies to the SAM medium (isolate A1 A10). To isolate pseudozyma-morphs associated with Albugo laibachii on Arabidopsis thaliana, A. laibachii spores were suspended in water and treated with antibiotics to remove bacteria. Subsequently, the suspension was plated on PDA at 20 C and colonies were singled out after 7 d. DNA extraction, PCR and sequencing In total 5 20 mg of infected plant tissue from herbarium specimens and yeast colonies were disrupted in a mixer mill (MM2, Retsch), using two iron beads of 3 mm and 5 8 iron beads of 1 mm diam per sample and shaking at 25 Hz for 5 10 min. Genomic DNA was extracted using the BioSprint 96 DNA Plant Kit (Qiagen, Hilden) on a KingFisher Flex robot (Thermo Scientific, Dreieich). PCR amplification of the complete ITS nrdna was performed using the M-ITS1 forward primer (Stoll et al. 2003) and the ITS4 (White et al. 1990) or smits-r1 reverse primer (Kruse et al. 2017). 118 IMA FUNGUS

186 Moesziomyces revisited Table 1. List of Moesziomyces specimens used in the present study. Species Host Host family Location Year Collector Fungarium no. ITS GenBank acc. no. Moesziomyces aphidis Arabidopsis thaliana infected with Albugo laibachii Brassicaceae UK, Norwich 2007 E. Kemen GLM-F KY Moesziomyces bullatus Echinochloa crus-galli Poaceae Germany, Bavaria 2013 J. Kruse GLM-F KY Echinochloa crus-galli Poaceae Germany, Bavaria 2013 J. Kruse GLM-F KY Echinochloa crus-galli Poaceae Germany, Bavaria 2013 J. Kruse GLM-F KY Echinochloa muricata Poaceae Germany, Saxony 2000 H. Jage GLM-F KY Echinochloa muricata Poaceae Germany, Saxony 2000 D. Schulz, B. Huber & F. Klenke GLM-F KY Echinochloa muricata Poaceae Germany, Saxony-Anhalt 2003 H. Jage GLM-F KY Echinochloa muricata Poaceae Germany, Saxony-Anhalt 2005 H. Jage GLM-F KY Echinochloa crus-galli Poaceae Germany, North Rhine-Westphalia 2010 J. Kruse GLM-F KY Echinochloa crus-galli Poaceae Germany, Schleswig-Holstein 2011 J. Kruse GLM-F KY Echinochloa crus-galli Poaceae Poland 1979 K. Vánky HUV No. 283, ex TUB KY Echinochloa crus-galli Poaceae Germany, Hesse 2015 J. Kruse ex-glm-f KY culture No. A1 GLM-F Echinochloa crus-galli Poaceae Germany, Hesse 2015 J. Kruse ex-glm-f KY culture No. A3 GLM-F Echinochloa crus-galli Poaceae Germany, Hesse 2015 J. Kruse ex-glm-f KY culture No. A10 GLM-F Moesziomyces penicillariae Pennisetum glaucum Poaceae Westafrica, Gambia 1973 K. Vánky HUV No. 154, ex TUB KY VOLUME 8 NO

187 Kruse et al. Table 2. List of additional sequences used in the phylogenetic tree, downloaded from GenBank. Species Source ITS GenBank acc. no. Citation Eriomoeszia eriocauli Eriocaulon cinereum AY Stoll et al. (2005) Macalpinomyces eriachnes Eriachne helmsii AY Stoll et al. (2005) Moesziomyces bullatus Paspalum distichum AY74015)3 Stoll et al. (2005) human preterm low birth weight infant KF Okolo et al. (2015) - DQ Matheny et al. (2006) human preterm low birth weight infant KF Okolo et al. (2015) - DQ Matheny et al. (2006) Pseudozyma antarctica- JX Gujjari et al. (unpublished) - JN An (unpublished) unpolished Japanese rice AB Sugita et al. (2003) Antarctica sediment AF Avis et al. (2001) Albizia julibrissin flower AY6415)57 Wei et al. (2005) lake sediment AB Sugita et al. (2003) Pseudozyma aphidis Japanese pear fruit AB Yasuda et al. (2007) human pulmonary infection Q Parahym et al. (2013) Saccharum officinarum AB Morita et al. (2012) Leucaena glauca HQ Wei et al. (2011) human EU105)207 Lin et al. (2008) human blood AB Sugita et al. (2003) human HQ Xie et al. (unpublished) Fallopia japonica KC Wang & Liu (unpublished) blood culture from hospitalized patient KM Bosco-Borgeat & Taverna (unpublished) Leucaena glauca HQ Wei et al. (2011) Saccharum officinarum AB Morita et al. (2012) poplar leaf KM Sun & Yan (unpublished) Forcipomia taiwana KM Chen (unpublished) seaweeds KP Wang et al. (unpublished) aphid secretion AF Avis et al. (2001) Neoreglia cruenta FN Garcia et al. (unpublished) Saccharum officinarum AB Morita et al. (2012) giant panda secrete KF Li et al. (unpublished) Camellia sinensis foliar lesions HQ Li et al. (unpublished) Echinochloa crus-galli GU Hamayun & Ahmad (unpublished) aphid secretion on Solanum pseudocapsicum JN An (unpublished) mulberry leaf KF Qiu et al. (unpublished) Citrus leaf JQ Soliman (unpublished) - JN An (unpublished) Pseudozyma hubeiensis Magnolia denudata wilting leaf DQ Wang et al. (2006) Pseudozyma parantarctica- JN Chen (unpublished) yam tuber steep water KF Babajide et al. (2015) - KP Irinyi et al. (2015) human blood AB Sugita et al. (2003) - NR An (unpublished) Pseudozyma rugulosa mouldy Zea mays leaf AB Sugita et al. (2003) ex-leaf of corn AF Avis et al. (2001) plant leaf HE Han et al. (2002) Pseudozyma sp. Hyoscyamus muticus AB Abdel-Motaal & Itu (unpublished) 120 IMA FUNGUS

188 Moesziomyces revisited Table 2. (Continued). Species Source ITS GenBank acc. no. Citation Coffea arabica EU Vega et al. (unpublished) Hyoscyamus muticus AB Abdel-Motaal & Itu (unpublished) Coffea arabica DQ Vega et al. (2008) Saccharum officinarum leaves LC05)3989 Surussawadee & Limtong (unpublished) shoot of tip pepper GU Sim et al. (unpublished) marine environment DQ Chang et al. (2008) Helicoverpa armigera caterpilla gut AM Molnar & Prillinger (unpublished) marine sediment KC Qu et al. (unpublished) - KR Wang et al. (unpublished) pharmaceutical effluent KF Selvi & Das (unpublished) barley kernels and leaf HG Korhola et al. (2014 Uncultured fungus Ericaceae roots HQ Walker et al. (2011) cleaned rice AB Ikeda et al. (2007) Uncultured fungus clone Axonopus compressus soil HQ Kee & Chia (unpublished) Uncultured Ustilago tomato rhizosphere KF Johnston-Monje et al. (unpublished) * type collections are highlighted in bold The reaction was performed in a thermocycler (Eppendorf Mastercycler 96 vapo protect, Eppendorf, Hamburg) with an initial denaturation at 95 C for 4 min, 36 PCR cycles of denaturation at 95 C for 40 s, annealing at 56 C for 40 s and elongation at 72 C for 60 s, followed by a final elongation at 72 C for 4 min. The resulting amplicons were sequenced at the Biodiversity and Climate Research Centre (BiK-F) laboratory using the PCR primers. Sequences were deposited in GenBank (NCBI, Table 1). Phylogenetic reconstructions The dataset included sequences of Moesziomyces species sexual and asexual morphs, both newly sequenced (Table 1) and downloaded from GenBank (Table 2). First all available sequences were extracted from NCBI on the basis of sequence similarity. Subsequently sequences were removed that were: (1) highly redundant with already-included ITS genotypes; or (2) of doubtful sequence quality, i.e. with mutations at positions highly conserved or with nucleotide changes only towards one end of the sequences. Macalpinomyces eriachnes was selected as an outgroup, based on the phylogenetic tree in Shivas et al. (2013). Alignments were made using mafft v. 7 (Katoh & Standley 2013) employing the Q-INS-I algorithm and removing leading and trailing gaps. The resulting total alignment length was 576 bp. For phylogenetic analyses, Minimum Evolution (ME) analysis was done with Mega v (Tamura et al. 2013), using the Tamura-Nei substitution model, assuming partial deletion at a cut-off of 80 % and 1000 bootstrap replicates. Maximum Likelihood (ML) analysis was done using RAxML on the webserver TrEase ( trease) with all parameters were set to default values. For Bayesian analysis also the webserver TrEase was used for calculating 10 M tree generations on four incrementally heated MC chains. The first 30 % of the trees obtained this way were discarded to ensure sampling of the stationary phase. All other parameters were set to default. Morphological examination For light microscopy, the herbarium specimens GLM-F and GLM-F were transferred to distilled water on a slide. Morphological examination was carried out using a Zeiss Imager M2 AX10 microscope (Carl Zeiss, Göttingen). Measurements of the spore balls and spores were performed at 400. Measurements are reported as maxima and minima in parentheses, and the mean plus and minus the standard deviation of a number of measurements given in parenthesis; the means are given in italics (Table 3). RESULTS The isolated yeasts from fresh Moesziomyces bullatus samples from Echinochloa crus-galli and E. muricatus were fast-growing on SAM medium. The colour of the yeasts was cream to light reddish, and the shape of the colonies was regular and roundish. A phylogenetic hypothesis for the sampled Moesziomyces species and the cultivated yeast asexual morphs is given in Fig. 1. The results of the Minimum Evolution, Maximum Likelihood and Bayesian Analyses were congruent. The clade comprising the type of M. aphidis also includes M. bullatus s. str. from Echinochloa crus-galli and E. muricata, as well as the sequence of the type of M. rugulosa, an isolate of Moesziomyces from Albugo laibachii on Arabidopsis thaliana and many other isolates not determined to the species level from various sources. While visual inspection of the alignments revealed that there was some sequence variation within the Moesziomyces bullatus clade, the relationships of the four subgroups was not resolved apart from the clustering VOLUME 8 NO

189 Kruse et al. Table 3. Measurements from 25 spore balls and 100 teliospores for collections of Moesziomyces bullatus on Echinochloa crus-galli from two different clades. Moesziomyces bullatus ex Echinochloa crus-galli, GLM-F sporeballs spores sporeballs Moesziomyces bullatus ex Echinochloa crus-galli, GLM-F spores No. length breadth l/b length breadth l/b length breadth l/b length breadth l/b 1 148, ,49 7,5 7, ,5 76,5 1,58 8,5 7 1, ,22 8 7,5 1, ,5 1,18 8 7,5 1, , ,5 55,5 1,16 7,5 7 1, ,5 97,5 1,22 8,5 7 1,21 58,5 53 1, , ,5 58, , ,5 1,81 7,5 7, ,5 83,5 1,2 8 7,5 1, ,5 1, , ,11 7,5 7 1, ,08 7,5 6,5 1, ,5 1,04 7,5 7 1, ,39 7,5 6,5 1, ,5 59 1, ,33 101,5 69 1,47 7,5 6,5 1, , ,36 8,5 6 1, ,5 1, ,29 144,5 73,5 1,97 8 6,5 1, ,5 1, , , , ,22 8,5 6,5 1, ,5 1,29 8 6,5 1, ,5 1,28 7,5 6,5 1,15 83,5 61 1,37 8,5 6,5 1, ,5 96,5 1,44 8,5 6,5 1,31 96,5 51,5 1,87 8,5 7,5 1, ,5 1, ,5 78,5 1,42 7,5 6,5 1, ,26 7 6,5 1,08 91,5 68,5 1,34 7 6,5 1, ,5 1,43 7,5 6,5 1,15 68,5 49,5 1,38 7 5,5 1, , ,07 7,5 7 1, ,5 1,16 8,5 7 1, ,5 95,5 1,23 7,5 7,5 1 84,5 78,5 1,08 7,5 7 1, ,5 75,5 1,23 7,5 7 1,07 86,5 62 1,4 7,5 6,5 1, ,5 68 1, ,14 122,5 86 1,42 8,5 7,5 1, ,5 97,5 1, ,14 113,5 78,5 1,45 7,5 5,5 1, ,5 53,5 1,09 8,5 7 1, , , ,5 52 1,01 8,5 6 1,42 105,5 84,5 1, , ,5 1, ,5 7,5 1 6,5 6, ,5 8 1,06 6,5 5,5 1, ,5 6 1,08 8,5 7,5 1, ,17 8 6,5 1, ,5 6 1, , ,5 1,07 7,5 6,5 1, ,5 1,08 7 6,5 1, ,5 1,08 8,5 8 1, ,5 7,5 1,13 8 7,5 1, ,17 8 7,5 1, ,5 6 1,25 8,5 7 1, , , ,5 6,5 1,15 8 6,5 1, ,5 1,23 6,5 6 1, ,5 6 1, ,5 6 1,08 8,5 6 1, ,5 6,5 1,31 7 6,5 1, ,5 7,5 1, ,5 6,5 1,15 8 6,5 1, ,5 7 1,21 8,5 7 1, IMA FUNGUS

190 Moesziomyces revisited Table 3. (Continued). Moesziomyces bullatus ex Echinochloa crus-galli, Moesziomyces bullatus ex Echinochloa crus-galli, GLM-F GLM-F sporeballs spores sporeballs spores No. length breadth l/b length breadth l/b length breadth l/b length breadth l/b ,5 6,5 1, ,5 1,08 8 7,5 1, ,5 6, , ,29 7,5 7 1, ,33 6 5,5 1, ,33 7,5 6 1, ,5 7 1, ,5 1,08 8,5 6,5 1, ,5 1, , ,5 6,5 1,15 7 6,5 1, ,5 6,5 1, ,5 1,23 7,5 6,5 1, ,5 1,23 7,5 7 1, ,5 6,5 1,15 7,5 6,5 1, ,5 7 1,21 8,5 7 1, ,5 7,5 1, ,5 1,07 8 6,5 1, ,5 7, , ,5 7, ,5 1,2 67 7,5 7 1,07 7,5 7 1, ,5 1,27 7,5 5,5 1, ,5 7 1,21 7,5 6 1, ,5 7 1,07 7,5 6,5 1, ,5 1,07 8,5 6,5 1, ,14 8,5 6,5 1, ,5 1,23 9,5 7 1, ,5 7 1,07 8,5 6,5 1, ,5 7 1,07 8,5 7 1, ,14 9,5 6,5 1, ,5 7 1, , ,5 1,23 7,5 6,5 1, ,5 6,5 1, , ,33 7,5 7 1, ,5 1, , ,5 1,45 8 6,5 1, ,5 1,08 8,5 6,5 1, ,5 1,08 7,5 6,5 1, ,5 7 1,21 7,5 7 1, ,5 5,5 1, , ,5 1, ,5 6,5 1,15 8,5 7 1, ,17 8,5 7 1, ,5 7, ,5 1,38 7,5 6 1, ,5 1,07 8,5 6,5 1,31 VOLUME 8 NO

191 Kruse et al. Table 3. (Continued). Moesziomyces bullatus ex Echinochloa crus-galli, Moesziomyces bullatus ex Echinochloa crus-galli, GLM-F GLM-F sporeballs spores sporeballs spores No. length breadth l/b length breadth l/b length breadth l/b length breadth l/b ,5 1,07 9 6,5 1, ,5 7,5 1 7,5 6 1, ,5 6,5 1,15 6,5 6 1, ,5 1,07 7,5 7 1, ,5 7 1,07 8,5 7,5 1, ,5 7 1,21 7,5 6 1, ,43 7,5 6,5 1, ,5 7, ,14 of Pseudozyma aphidis and the majority of M. bullatus isolates with the clade containing the type of P. rugulosa. Collections from E. crus-galli with smut symptoms were present in two different clades. The morphological investigation of a sexual morph from each clade (GLM-F and GLM-F105814) revealed no morphological differences. Moesziomyces bullatus clustering within the majority of M. aphidis had the following spore characteristics: sporeballs variable in shape and size, x µm, spores ovoid, globose, often irregular, pale yellow-brown, (6.5 ) ( 10) (5.5 ) ( 8) µm, a length/breadth ratio of (n = 100) (Fig. 2). In comparison, the collection of M. bullatus clustering together with the sequence of the type species of M. rugulosus showed the following spore characteristics: sporeballs variable in shape and size, x µm, spores ovoid, globose, often irregular, pale yellow-brown, (6 ) ( 9.5) (5.5 ) ( 8) µm, a length/breadth ratio of (n = 100) (Fig. 2). The sister group to M. bullatus was formed by M. antarcticus. The four lineages of M. bullatus formed an isolated clade with high to maximum support in all analyses together with samples classified as M. antarcticus. Apart from the groups mentioned above, four additional distinct groups were revealed. Two of these corresponded to lineages formed by sexual smuts of the genus Moesziomyces isolated from plants with smut disease symptoms. One of these corresponded to M. bullatus s. lat. on Paspalum distichum, and the other to Eriomoeszia eriocauli. The remaining two lineages formed a monophyletic clade with high support in Minimum Evolution Analysis. One lineage included sequences of yeasts classified as M. parantarcticus, the other a sexual morph of a plant pathogenic fungus of the genus Moesziomyces from Pennisetum glaucum, as well asexual morphs isolated from symptom-free barley and a preterm infant. DISCUSSION Moesziomyces is a morphologically well-defined genus in the smut fungi, mainly characterised by ruptured sterile cells in the sori around the spores. The genus was believed to be monotypic by Vánky (2012), but phylogenetic investigations of the past decade have shown that several species previously assigned to the asexually typified yeast genus Pseudozyma, were closely related to Moesziomyces bullatus (Begerow et al. 2000, 2006, Wang et al. 2006, 2015). In the latest edition of the International Code of Nomenclature for algae, fungi and plants (ICN) it is ruled that the dual naming for asexual and sexual morphs of fungi has been discontinued (McNeill et al. 2012). Consequently, Wang et al. (2015) attempted to resolve the names of species placed in the genus Pseudozyma as far as possible, and combined those related to Moesziomyces bullatus s.lat. into Moesziomyces. The yeast asexual morphs were, for example, found to live epiphytically on different hosts (Boekhout 1995), but also to occur on a variety of other substrates. Due to their asexual reproduction with pullulating and division, it is possible for them to colonize suitable habitats in a short period of time. Of these yeasts, Pseudozyma aphidis is often considered as a biocontrol agent for plant pathogenic fungi (Avis et al. 2001, Buxdorf et al. 2013). Thus it is noteworthy that one isolate of this species co-occurred in Albugo laibachii lesions on Arabidopsis thaliana, indicating only no or only limited antagonism against this specialised white blister rust species (Thines et al. 2009). It is commonly believed that most Pseudozyma species have lost pathogenicity, which is seemingly supported by recent genomic analyses (Lefebvre et al. 2013). However, it should be noted that if a different start codon is taken for translation than the one predicted, all Pseudozyma yeasts included by Lefebvre et al. (2013) have a functional copy of PEP1, a conserved effector among smut fungi of the Ustilaginales (Sharma et al. 2014, Hemetsberger et al. 2015), suggesting the possibility of a misannotation of the start codon. In-depth bioinformatic analyses and functional testing will be needed to clarify this situation. Deducing the conspecificity of Moesziomyces bullatus with Pseudozyma aphidis and P. rugulosa was not possible for Wang et al. (2015), as they did not include sequences from the type host of M. bullatus, Echinochloa crus-galli, but only from M. verrucosus on Paspalum distichum, which they erroneously assumed to be conspecific with M. bullatus. However, the smut sexual morphs from the type host, E. crusgalli from Germany, are placed in two of the four subclusters 124 IMA FUNGUS

192 Moesziomyces revisited -/64/0.76 AB P. aphidis ex Japanese pear fruit JQ P. aphidis ex human pulmonary infection AB P. aphidis ex Saccharum officinarum HQ P. aphidis ex Leucaena glauca EU P. aphidis ex human AB P. aphidis ex human blood HQ P. aphidis ex human KC P. aphidis ex Fallopia japonica KM P. aphidis ex blood culture from hospitalized patient AB P. sp. ex Hyoscyamus muticus EU P. sp. ex Coffea arabica HQ P. aphidis ex Leucaena glauca AB P. aphidis ex Saccharum officinarum KM P. aphidis ex poplar leaf KM P. aphidis ex Forcipomia taiwana KP P. aphidis ex seaweeds HQ Uncultured fungus ex Ericaceae roots AF P. aphidis strain ex aphid secretion, ex-type sequence FN P. aphidis ex Neoreglia cruenta AB P. aphidis ex Saccharum officinarum KF P. aphidis ex giant panda secrete HQ P. aphidis ex Camellia sinensis foliar lesions AB P. sp. ex Hyoscyamus muticus KF P. aphidis ex mulberry leaf KY P. sp. ex Arabidopsis thaliana infected with Albugo laibachii JQ P. aphidis ex Citrus leaf DQ P. sp. ex Coffea arabica KY M. bullatus ex Echinochloa crus-galli isolate A3 KY M. bullatus ex Echinochloa crus-galli KY M. bullatus ex Echinochloa crus-galli KY M. bullatus ex Echinochloa muricatus KY M. bullatus ex Echinochloa muricatus KY M. bullatus ex Echinochloa muricatus KY M. bullatus ex Echinochloa muricatus KY M. bullatus ex Echinochloa crus-galli KY M. bullatus ex Echinochloa crus-galli KY M. bullatus ex Echinochloa crus-galli KY M. bullatus ex Echinochloa crus-galli isolate A1 KY M. bullatus ex Echinochloa crus-galli isolate A10 AB P. rugulosa ex mouldy Zea mays leaf GU P. sp. ex shoot of tip pepper KY M. bullatus ex Echinochloa crus-galli No 1642 HE P. rugulosa ex plant leaf DQ P. sp. ex marine environment AM P. sp. ex Helicoverpa armigera caterpilla gut AB Uncultured fungus ex cleaned rice KF Uncultured Ustilago ex tomato rhizosphere KC P. sp. ex marine sediment 64/72/0.98 JX P. antarctica JN P. antarctica AB P. antarctica ex unpolished Japanese rice AY P. antarctica ex Albizia julibrissin flower 99/98/1 AB P. antarctica ex lake sediment KF P. sp. ex pharmaceutical effluent AY M. bullatus ex Paspalum distichum 99/97/1 -/91/0.99 AY Eriomoeszia eriocauli ex Eriocaulon cinereum 98/65/ /95/0.96 JN P. parantarctica 70/67/0.91 KF P. parantarctica ex yam tuber steep water 97/98/1 99/92/ /-/ /89/0.96 AF P. antarctica ex Antarctica sediment, ex-type sequence 71/-/- KR P. sp. GU P. aphidis ex Echinochloa crus-galli 63/-/- JN P. aphidis ex aphid secretion on Solanum pseudocapsicum 86/-/- JN P. aphidis 94/92/0.99 LC P. sp. ex Saccharum officinarum leaves 76/-/0.64 AF P. rugulosa ex leaf of corn, ex-type sequence 99/97/0.82 KP P. parantarctica AB P. parantarctica ex human blood NR P. parantarctica, ex-type sequence HQ Uncultured fungus clone ex Axonopus compressus soil HG P. sp. ex barley kernels and leaf KY M. bullatus ex Pennisetum glaucum No 833 KF Moesziomyces bullatus ex human preterm low birth weight infant DQ Moesziomyces bullatus AY Macalpinomyces eriachnes ex Eriachne helmsii DQ P. hubeiensis ex Magnolia denudata wilting leaf Moesziomyces bullatus Moesziomyces antarcticus Moesziomyces verrucosus Moesziomyces eriocauli Moesziomyces parantarcticus Moesziomyces penicillariae 0.02 Fig. 1. Phylogenetic tree based on Minimum Evolution analyses of nrits sequences of Moesziomyces spp., rooted with Macalpinomyces eriachnes. Numbers on branches denote bootstrap support in Minimum Evolution, Maximum Likelihood and a posteriori probabilities from Bayesian Analyses. Values below 55 % are not shown. The bar indicates expected substitutions per site. GenBank numbers precede taxon names, and are followed by the name of the host or isolation source of the fungus. VOLUME 8 NO

193 ART I CLE Kruse et al. C A B D E B 10 µm D E 10 µm F 20 µm 20 µm C F 10 µm 10 µm 10 µm Fig. 2. Sori and spores of Moesziomyces bullatus on Echinochloa crus-galli. A, D. Sori. B, E. Teliospore balls. C, F. Teliospores. A C (GLM-F105812), D F (GLM-F105814). of a larger cluster, which is interpreted here as representing M. bullatus. It is noteworthy that three of the four subclusters of M. bullatus contain environmental samples from various sources. In conjunction with the ease of cultivation observed for M. bullatus from E. crus-galli, it is concluded that unlike the vast majority of genera of Ustilaginales, the asexual yeast morph plays a major role as a proliferating life-cycle stage in Moesziomyces and that the plant-parasitic dikaryophase is probably mainly important for maintaining the possibility of sexual recombination. As the two subclades previously referred to as M. aphidis and M. rugulosus are interspersed with the morphologically identically lineages of M. bullatus from E. crus-galli, they are probably better included in M. bullatus until more sequence data become available. It seems probable that, with the inclusion of additional smut samples from Echinochloa, additional smut-causing members of the subclades will be discovered. Sampling in Africa seems to be promising in this respect, as the species diversity of Echinochloa is highest on this continent. Also, the notion that yeasts of the subclade containing the ex-type culture of Pseudozyma aphidis can withstand high temperatures, such as the human body temperature, is suggestive of a subtropical to tropical origin of this lineage. Further, our investigations show that the older name M. eriocauli for Eriomoeszia eriocauli, should be taken up again, as this species was found embedded within Moesziomyces. With the synonymy of the generic name Eriomoeszia and 126 the reappraisal of the hardly used Moesziomyces names of the smut fungi of Paspalum and Pennisetum, Moesziomyces now includes six species. It is, however, likely that additional species will have to be added because smut samples from some Poaceae genera listed as host plants for the M. bullatus complex in Vánky (2012) could not be included in the current study, such as smuts from Leersia, Panicum, Polytrias, and Uranthoecium. Given the apparently high host specificity of Moesziomyces species, it seems likely that these pathogens represent species independent from M. bullatus. TAXONOMY Based on the phylogenetic data presented here, the following nomenclature and taxonomic changes are made. Moesziomyces antarcticus (Goto et al.) Q.M. Wang et al., Stud. Mycol. 81: 81 (2015). Basionym: Sporobolomyces antarcticus Goto et al., Mycologia 61: 759 (1969). Synonyms: Candida antarctica (Goto et al.) Kurtzman et al., Yeasts: 86 (1983). Vanrija antarctica (Goto et al.) R.T. Moore, Bibltheca Mycol. 108: 167 (1987). Pseudozyma antarctica (Goto et al.) Boekhout, J. Gen. Appl. Microbiol. 41: 364 (1995). IMA FUNGUS

194 Moesziomyces revisited Moesziomyces bullatus (J. Schröt.) Vánky, Bot. Notiser 130: 133 (1977). Basionym: Sorosporium bullatum J. Schröt., Abh. Schles. Ges. Vaterl. Cult., Abth. Naturwiss. 72: 6 (1869). Synonyms: Tolyposporium bullatum J. Schröt, in Cohn., Krypt. Fl. Schles. 3(1): 276 (1887). Sterigmatomyces aphidis Henninger & Windisch, Arch. Microbiol. 105: 50 (1975). Tolypoderma bullata (J. Schröt.) Thirum. & M.J. O`Brien, Friesia 11: 190 (1978) ["1977 ]. Sporothrix rugulosa Traquair et al., Canad. J. Bot. 66: 929 (1988). Stephanoascus rugulosus Traquair et al., Canad. J. Bot. 66: 929 (1988). Pseudozyma aphidis (Henninger & Windisch) Boekhout, J. Gen. Appl. Microbiol. 41: 364 (1995). Pseudozyma rugulosa (Traquair, et al.) Boekhout & Traquair, J. Gen. Appl. Microbiol. 41: 364 (1995). Moesziomyces aphidis (Henninger & Windisch) Q.M. Wang et al., Stud. Mycol. 81: 81 (2015). Moesziomyces rugulosus (Traquair, et al.) Q.M. Wang et al., Stud. Mycol. 81: 81 (2015). Moesziomyces eriocauli (G.P. Clinton) Vánky, Nordic J. Bot. 6: 71 (1986). Basionym: Tolyposporium eriocauli G.P. Clinton, Rhodora 2: 82 (1901). Synonyms: Dermatosorus eriocauli (G.P. Clinton) M.D. Whitehead & Thirum., Mycologia 64: 128 (1972). Tolypoderma eriocauli (G.P. Clinton) Thirum., Friesia 11: 191 (1978). Eriomoeszia eriocauli (G.P. Clinton) Vánky, Mycol. Balcanica 2: 106 (2005). Moesziomyces parantarcticus (Sugita et al.) Q.M. Wang et al., Stud. Mycol. 81: 81 (2015). Basionym: Pseudozyma parantarctica Sugita et al., Microbiol. Immun. 47: 186 (2003). Moesziomyces verrucosus (J. Schröt.) J. Kruse & Thines, comb. nov. MycoBank MB Basionym: Ustilago verrucosa J. Schröt., Hedwigia 35: 214 (1896). Synonyms: Tolyposporium evernium Syd., Ann. Mycol. 37: 443 (1939). Tolyposporium paspali Langdon, Univ. Queensland Dept. Biol. Pap 2(9): 4 (1948). Moesziomyces evernius (Syd.) Vánky, Bot. Notiser 130: 135 (1977). Tolyposporidium evernium (Syd.) 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Critical Reviews in Food Science and Nutrition 35: Vánky K (1977) Moesziomyces, a new genus of Ustilaginales. Botaniska Notiser 130: Vánky K (1986) The genus Moesziomyces (Ustilaginales). Nordic Journal of Botany 6: Vánky K (2005) The smut fungi (Ustilaginomycetes) of Eriocaulaceae. I. Eriomoeszia gen. nov. Mycologia Balcanica 2: Vánky K (2012) Smut Fungi of the World. St Paul, MN: American Phytopathological Society Press. Wang Q-M, Jia J-H, Bai F-Y (2006) Pseudozyma hubeiensis sp. nov. and Pseudozyma shanxiensis sp. nov., novel ustilaginomycetous anamorphic yeast species from plant leaves. International 128 IMA FUNGUS

196 Moesziomyces revisited Journal of Systematic and Evolutionary Microbiology 56: Vega FE, Posada F, Aime MC, Peterson SW, Rehner SA (2008) Fungal endophytes in green coffee seeds. Mycosystema 27: Walker JF, Aldrich Wolfe L, Riffel A, Barbare H, Simpson NB, et al. (2011) Diverse Helotiales associated with the roots of three species of Arctic Ericaceae provide no evidence for host specificity. New Phytologist 191: Wang Q.-M, Begerow D, Groenewald M, Liu X-Z, Theelen B, et al. (2015) Multigene phylogeny and taxonomic revision of yeasts and related fungi in the Ustilaginomycotina. Studies in Mycology 81: Wei YH, Fwu-Ling L, Wen-Haw H, Shyue-Ru CHEN, Chien-Cho CHEN, et al. (2005) Pseudozyma antarctica in Taiwan: a description based on morphological, physiological and molecular characteristics. Botanical Bulletin of Academia Sinica 46: Wei YH, Liou GY, Lee FL (2011) Pseudozyma aphidis, a new record of ustilaginomycetous anamorphic fungi in Taiwan. Fungal Science 26: 1 5. White TJ, Bruns T, Lee S, Taylor JW (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: PCR Protocols: a guide to methods and applications (Innis MA, Gelfand DH, Sninsky JJ, White TJ, eds): San Diego: Academic Press. Yasuda F, Yamagishi D, Izawa H, Kodama M, Otani H (2007) Fruit stain of Japanese pear caused by basidiomycetous, yeastlike fungi Meira geulakonigii and Pseudozyma aphidis. Japanese Journal of Phytopathology 73: VOLUME 8 NO

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198 Isolates Freshly collected twigs were placed in damp chambers, and incubated at room temperature (ca. 20 o C) for 1 2 d. Single conidial or ascospore colonies were established from sporulating conidiomata or ascomata in Petri dishes containing 2 % malt extract agar (MEA) as described earlier (Crous et al. 1991). After 1 2 d, single spores were picked up and transferred to fresh MEA plates. Colonies were subdoi: /imafungus IMA FUNGUS 8(1): (2017) The Genera of Fungi G 4: Camarosporium and Dothiora Pedro W. Crous 1,2,3, and Johannes Z. Groenewald 1 1 Westerdijk Fungal Biodiversity Institute, Uppsalalaan 8, 3584 CT Utrecht, The Netherlands; corresponding author p.crous@ westerdijkinstitute.nl 2 Microbiology, Department of Biology, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands 3 Department of Microbiology and Plant Pathology, Forestry and Agricultural Biotechnology Institute (FABI), University of Pretoria, P. Bag X20, Pretoria 0028, South Africa Abstract: The current paper represents the fourth contribution in the Genera of Fungi series, linking type species of fungal genera to their morphology and DNA sequence data. The present paper focuses on two genera of microfungi, Camarosporium and Dothiora, which are respectively epi- and neotypified. The genus Camarosporium is typified by C. quaternatum, which has a karstenula-like sexual morph, and phoma-like synasexual morph. Furthermore, Camarosporomyces, Foliophoma and Hazslinszkyomyces are introduced as new camarosporiumlike genera, while Querciphoma is introduced as a new phoma-like genus. Libertasomycetaceae is introduced as a new family to accommodate Libertasomyces and Neoplatysporoides. Dothiora, which is typified by D. pyrenophora, is shown to produce dothichiza- and hormonema-like synasexual morphs in culture, and D. cactacearum is introduced as a new species. In addition to their typification, ex-type cultures have been deposited in the Westerdijk Fungal Biodiversity Institute (CBS Culture Collection), and species-specific DNA barcodes in GenBank. Authors interested in contributing accounts of individual genera to larger multi-authored papers in this series should contact the associate editors listed on the List of Protected Generic Names for Fungi. Key words: DNA Barcodes fungal systematics ITS LSU typification Article info: Submitted: 15 April 2017; Accepted: 19 May 2017; Published: 23 May INTRODUCTION The present paper clarifies two generic names in the Genera of Fungi project ( Crous et al. 2014a), which has the aim to revise the generic names of fungi accepted in Kirk et al. (2013). The two genera treated are supplemented with recently collected epi- or neotypes, which have been registered in MycoBank (Robert et al. 2013). Furthermore, in keeping with the one fungus = one name initiative for fungi (Hawksworth et al. 2011, Crous et al. 2015a), a single name is indicated for each genus. The aim of the present contribution was to treat two problematic genera, namely Camarosporium, and Dothiora. In recent years, it has become clear that the Camarosporium morphology has evolved several times within Dothideomycetes, and that camarosporium-like morphs have phoma-like synasexual morphs, and pleospora-like sexual morphs. An additional complication lies in that some taxa only produce a single morph (e.g. see below in the present study), and thus it is almost impossible to clarify the generic identification without molecular data. Furthermore, the treatment of Phoma by Boerema et al. (2004) placed genetically distinct genera into one genus, while the same was true for the treatment of Pleospora by De Gruyter et al. (2013), and the assumption that the reference strains of Camarosporium included in Crous et al. (2006) were authentic for the name. The genus Dothiora (based on D. pyrenophora) produces a Dothichiza asexual morph in culture, but the Dothideaceae has several genera that are dothiora-like in morphology, and that produce dothichiza-like asexual morphs in culture. Although we address the status of Dothiora in the present study, many additional collections are still required to resolve the generic boundaries of other old generic names in this family, particularly as we have observed hyaline, 1-septate ascospores to become muriformly septate with age, and eventually to even become pigmented. Given that these characters have traditionally been used to separate genera in the family (Sivanesan 1984, Thambugala et al. 2014), it now appears likely that there are far fewer genera in Dothideaceae than previously assumed (Crous & Groenewald 2016). MATERIALS AND METHODS 2017 International Mycological Association You are free to share - to copy, distribute and transmit the work, under the following conditions: Attribution: You must attribute the work in the manner specified by the author or licensor (but not in any way that suggests that they endorse you or your use of the work). Non-commercial: You may not use this work for commercial purposes. No derivative works: You may not alter, transform, or build upon this work. For any reuse or distribution, you must make clear to others the license terms of this work, which can be found at Any of the above conditions can be waived if you get permission from the copyright holder. Nothing in this license impairs or restricts the author s moral rights. VOLUME 8 NO

199 Crous and Groenewald cultured onto 2 % potato dextrose agar (PDA), oatmeal agar (OA), MEA (Crous et al. 2009), autoclaved pine needles on 2 % tap water agar (PNA) (Smith et al. 1996), and incubated at 25 o C under continuous near-ultraviolet light to promote sporulation. Reference strains and specimens are maintained at the Westerdijk Fungal Biodiversity Institute (CBS Culture Collection) in Utrecht, The Netherlands. at 25 ºC. Colony colours (surface and reverse) were rated according to the colour charts of Rayner (1970). Taxonomic novelties and new typifications were deposited in MycoBank ( Crous et al. 2004). RESULTS DNA isolation, amplification and analyses Genomic DNA was extracted from fungal colonies growing on MEA using the Wizard Genomic DNA purification kit (Promega, Madison, WI), according to the manufacturer s protocol. The primers V9G (de Hoog & Gerrits van den Ende 1998) or ITS5 (White et al. 1990) and LR5 (Vilgalys & Hester 1990) were used to amplify for all isolates part of the nuclear rdna operon (ITS) spanning the 3 end of the 18S nrrna gene, the first internal transcribed spacer (ITS1), the 5.8S nrrna gene, the second ITS region (ITS2) and approximately 900 bp of the 5 end of the 28S nrrna gene. The primers ITS4 (White et al. 1990) and LR0R (Vilgalys & Hester 1990) were used as internal sequence primers to ensure good quality sequences over the entire length of the amplicon. Part of the beta-tubulin gene (tub2) was amplified and sequenced for selected isolates using T1 (O Donnell & Cigelnik 1997) or Bt-2a and Bt-2b (Glass & Donaldson 1995). In addition, part of the first or second half of the translation elongation factor 1-alpha gene (tef1) was amplified and sequenced for selected isolates using EF1-728F (Carbone & Kohn 1999) and EF2 (O Donnell et al. 1998), and EF1-983F and EF1-2218R (Rehner & Buckley 2005), respectively. Part of the 18S small subunit nrrna gene (SSU) was amplified and sequenced for selected isolates using NS1 and NS4 (White et al. 1990). Amplification conditions followed Cheewangkoon et al. (2008) and Quaedvlieg et al. (2012). The programme SeqMan Pro v (DNASTAR, Madison, WI) was used to obtain consensus sequences of each isolate. Blast searches using ITS and LSU sequences were performed for each isolate and the closest matches were retrieved from GenBank and included in the phylogenetic analyses. The SSU, tef1 and tub2 sequences were not included in phylogenetic analyses but were used in blast searches to confirm the species identification where possible. Multiple sequence alignments for ITS and LSU were generated using the online version of MAFFT ( and subsequent phylogenetic analyses were conducted using parsimony in PAUP v. 4.0b10 (Swofford 2003) as described by Cheewangkoon et al. (2008). Sequence data were deposited in GenBank (Table 1) and the alignments and trees in TreeBASE ( Morphology Slide preparations were mounted in clear lactic acid or water from colonies sporulating on the media previously mentioned. Sections of sporocarps were made by hand for examination purposes. Observations were made with a Nikon SMZ25 stereo-microscope, and with a Zeiss Axio Imager 2 light microscope using differential interference contrast (DIC) illumination and a Nikon DS-Ri2 camera and software. Colony characters and pigment production were noted after 2 wk of growth on MEA, PDA and OA (Crous et al. 2009) incubated Phylogeny Four analyses were performed in this study; the first was based on a partial alignment of LSU (Fig. 1) to provide an overview phylogeny of the species and genera treated in the present study, whereas the remaining three alignments were ITS alignments representing Camarosporium and allied genera (Fig. 2), Dothidea and Dothiora (Fig. 3), and Paracamarosporium and Pseudocamarosporium (Fig. 4), respectively. The statistical parameters for these four phylogenies are summarised in Table 2. The overview LSU phylogeny (Fig. 1) shows that Dothiora pyrenophora is nestled inside the Dothiora clade. Based on this LSU phylogeny, the Dothidea clade has a bootstrap support value of 99 %, but the Dothiora clade did not receive any bootstrap support. Libertasomyces and Neoplatysporoides formed a lineage distinct from other families included in the phylogeny and therefore a family name is introduced below to accommodate these genera. Camarosporium quaternatum clustered with Ochrocladosporium, but without any bootstrap support. Megablast searches of the NCBI GenBank nucleotide database failed to reveal any closer matches. Strains previously published as Camarosporium quaternatum (CBS and CBS ) proved to be neither conspecific nor congeneric with C. quaternatum and therefore new combinations are proposed to accommodate these two strains in Libertasomyces and the new genus Hazslinszkyomyces. Two species of Pleospora, P. flavigena and P. fallens, were not congeneric with Pleospora (based on P. herbarum) and therefore the new genera Camarosporomyces and Foliophoma are introduced to accommodate these taxa. The genus Hazslinszkyomyces is erected below to accommodate Camarosporium aloes, while Querciphoma is erected to accommodate Coniothyrium carteri. Several species treated until now as belonging to Camarosporium represent yet another new genus and family, which will be treated in elsewhere. The ITS phylogeny of Camarasporium and allied genera (Fig. 2) resolved most of the included species, except for those in the Camarosporium clade where the ITS sequences were highly similar (97 % similar and higher when compared to Camarosporium sp. 1 strain CPC 12441; a maximum of 14 nucleotides differences). Neoplatysporoides aloicola (GenBank KR476719) clustered inside Libertasomyces, compared to the LSU phylogeny (Fig. 1) where it was basal to the clade. The sister placement of N. aloicola (conidia brown, 1-septate) to L. myopori (conidia hyaline, aseptate) did not receive any bootstrap support and this species is therefore not combined into Libertasomyces. In the LSU phylogeny (Fig. 1), Camarosporomyces flavigenus clustered as a basal lineage in the Hazslinszkyomyces clade, and based on this position was first considered to be a species of that genus. However, the ITS sequence was clearly not congeneric with 132 IMA FUNGUS

200 The Genera of Fungi G4 Hazslinszkyomyces (Fig. 2) and a new generic name is therefore introduced to accommodate it. Both the ITS and LSU sequences of this strain were confirmed by two independent DNA isolations, and subsequent PCR and sequencing. The ITS phylogeny of Dothidea and Dothiora (Fig. 3) resolved the included species, although the lineages in Dothiora were poorly supported. Crous & Groenewald (2016) used a combination of ITS, tef1 and tub2 to resolve the species in this genus. Dothiora cactacearum is introduced below as a new species related to D. buxi while the generic type species, D. pyrenophora, is shown to be sister to D. sorbi. In the Dothidea clade, three isolates representing D. ribesia were not conspecific with two isolates published by Thambugala et al. (2014) as Plowrightia ribesia and which are indicated in our phylogeny as Dothidea sp. pending further elucidation. The ITS phylogeny of Paracamarosporium and Pseudocamarosporium (Fig. 4) resolved most of the included Paracamarosporium species, but did not resolve species in the Pseudocamarosporium clade where the ITS sequences were highly similar (99 % similar and higher when compared to Pseudocamarosporium sp. 1 strain CPC 25926; a maximum of five nucleotides differences). Camarosporium mamanes (GenBank DQ885900) clustered as sister to Paracamarosporium psoraleae and therefore a new combination is provided for it in that genus. Current generic circumscription: Ascomata pseudothecial, single or in clusters, subcorticolous, +/- globose, black, ostiolum central, short papillate and terete, without setae; ascomatal wall of textura angularis. Pseudoparaphyses numerous, filiform, cellular, multi-celled, branched, anastomosing, hyaline, smooth. Asci 8-spored, cylindrical, apically rounded, pedicel short and furcate, thick-walled, bitunicate, fissitunicate, inamyloid. Ascospores 6-celled, ellipsoidal, straight, muriform, golden, with 1 3 longitudinal septa, eguttulate, without a gelatinous sheath and appendages. Conidiomata dimorphic, pycnidial, subcorticolous, single to gregarious, partly caespitose, globose, ostiole central, terete, short papillate; conidiomatal wall few-layered, consisting of a textura globulosa-angularis with red brown, thick-walled, and smooth cells. Paraphyses and conidiophores absent. Conidiogenous cells formed from the inner cells of the pycnidial wall, doliiform, hyaline, thin-walled, annellidic. Conidia multicelled, muriformly septate, with one longitudinal or diagonal septum per cell and 1 2 per conidium, ellipsoidal, pyroid, clavate, straight to slightly curved, yellowish not brown, basal cell often paler or hyaline, wall golden. Synasexual morph: conidiomata separate, pycnidial, immersed to superficial on PNA, brown, globose, with 1 2 papillate ostioles, exuding a crystalline conidial mass. Conidiophores reduced to conidiogenous cells. Conidiogenous cells lining the inner cavity, hyaline, smooth, ampulliform. Conidia solitary, hyaline, smooth, subcylindrical, straight, rarely curved, apex obtuse, base truncate. Type species: Camarosporium quaternatum (Hazsl.) Schulzer THE GENERA Camarosporium complex Coniothyriaceae W.B. Cooke, Revista de Biol. 12: 289 (1983) [ ]. Type species: Coniothyrium palmarum Corda Genera included: Camarosporium, Camarosporomyces, Coniothyrium, Dimorphosporicola, Foliophoma, Hazslinszkyomyces, Neocamarosporium, Ochrocladosporium, Pseudoleptosphaeria. Note: Camarosporiaceae Locq is not validly published (Art. 39.1) in lacking a Latin diagnosis. However, the genus Camarosporium s. str. on which this family was based, falls in Coniothyriaceae (see De Gruyter et al. 2013) in our analysis, thus we refrain from validating Camarosporiaceae to accommodate Camarosporium. Camarosporium Schulzer, Verh. K.K. Zool.-Bot. Ges. Wien 17: 717 (1867). Classification: Coniothyriaceae, Pleosporales, Dothideomycetes. Camarosporium quaternatum (Hazsl.) Schulzer, Verh. K.K. Zool.-Bot. Ges. Wien 17: 717 (1867). (Fig. 5) Basionym: Clinterium quaternatum Hazsl., Verh. K.K. Zool.-Bot. Ges. Wien 15: 450 (1865); as Clinterium (quaternatum) ; type: Hazslinszky (1865: figs 9 12 lectotype designated here, MBT376246; Hungary: near Budapest, private garden, on twigs of Lycium barbarum (Solanaceae), attached, corticated, 2 May 2016, L. Bartalos, det. R.K. Schumacher (CBS H epitype designated here, MBT376247; CPC = CBS culture exepitype, CPC 31518). Synonyms: Cucurbitaria varians Hazsl., Verh. K.K. Zool.-Bot. Ges. Wien 15: 451 (1865); type: Hazslinszky (1865: figs lectotype designated here, MBT376243). Karstenula varians (Hazsl.) Sacc., Syll. Fung. 2: 241 (1883). Pleomassaria varians (Hazsl.) G. Winter, Rabenh. Krypt.-Fl., 2 nd edn 1(2): 552 (1886). Camarosporium hendersonia Schulzer, Verh. K.K. Zool.-Bot. Ges. Wien 17: 716 (1867), nom. illegit. (Art. 53.1, based on C. quaternatum (Hazsl.) Schulzer 1867). Camarosporium hazslinszkyi Sacc., Syll. Fung. 3: 468 (1884), nom. illegit. (Art. 53.1, based on C. quaternatum (Hazsl.) Schulzer 1867). Additional material examined: Germany: Berlin, mixed forest, about 50 m asl, sand, acid, fresh, mesotroph, on twig of Daphne mezereum (Thymelaeaceae), 18 May 2013, R. Jarling, det. R.K. Schumacher (CBS H-23063, culture CPC = CBS ). Description: Ascomata pseudothecial, single or in clusters, subcorticolous, +/- globose with a flattened base, black, smooth, soft, thick, ostiole central, short papillate and terete, VOLUME 8 NO

201 Crous and Groenewald Table 1. Details of the strains included in the taxomic treatments or for which novel sequences were deposited in GenBank. Host or substrate Country GenBank accession number 3 Species name Strain accession Collector and number 1,2 collection date Camarosporium arezzoensis CPC R.K. Schumacher, 22 Jul Cytisus borysthenicus, branch ITS LSU tef1 (first part) tef1 (second part) tub2 SSU Ukraine KY KY Camarosporium sp. 1 CPC W. Gams, Aug Sophora chrysophylla USA: Hawaii KY DQ Camarosporium sp. 2 CPC R.K. Schumacher, Caragana sp., twig Finland KY KY KY Dec CPC R.K. Schumacher, 1 Caragana sp., twig Finland KY KY KY Dec Germany KY KY Camarosporium sp. 3 CPC R.K. Schumacher, 10 May 2016 Camarosporium sp. 4 CPC R.K. Schumacher, 12 Oct Camarosporium sp. 5 CPC A. Usichenko, 25 May 2015 Camarosporium quaternatum Camarosporomyces flavigenus CPC R.K. Schumacher, 2 Apr CPC R.K. Schumacher, 18 May 2013 CPC quaternatum ET of Clinterium R.K. Schumacher, 2 May 2016 CPC R.K. Schumacher, 2 May 2016 Elaeagnus rhamnoides, twig Ulmus laevis, twig Ukraine KY KY Robinia pseudoacacia, dead branch Philadelphus coronarius, twig Daphne mezereum, twig Ukraine KY KY Germany KY KY Germany KY KY KY KY Lycium barbarum, twig Hungary KY KY KY KY Lycium barbarum, twig Hungary KY KY KY KY CBS T K. Fodor, 1980 Water Romania KY GU GU Dothidea puccinioides CBS Viburnum lantana Switzerland KY AY Dothidea ribesia CPC R.K. Schumacher Ribes uva-crispa, stem Germany KY KY KY KY CPC R.K. Schumacher, 4 Ribes rubrum, twig Germany KY KY KY KY May 2016 CPC R.K. Schumacher, 4 Ribes rubrum, twig Germany KY KY KY KY May 2016 Dothiora cactacearum CBS = CPC P.W. Crous, 2013 Cactaceae (ornamental) USA KY KY KY KY CPC P.W. Crous, 2013 Cactaceae (ornamental) USA KY KY KY KY Dothiora pyrenophora CPC NT R.K. Schumacher, Sorbus aucuparia, twig Germany KY KY KY KY KY Apr CPC R.K. Schumacher, Sorbus aucuparia, twig Germany KY KY KY KY KY Apr KY GU GU Foliophoma fallens CBS G.F. Laundon, 1978 Olea europaea, leaf spot New Zealand 134 IMA FUNGUS

202 The Genera of Fungi G4 Table 1. (Continued). GenBank accession number 3 Species name Strain accession Collector and number 1,2 collection date Host or substrate Country ITS LSU tef1 (first part) tef1 (second part) tub2 SSU CBS H.A. van der Aa, Apr Hazslinszkyomyces aloes CBS = CPC M.J. Wingfield, Sept T 2012 Hazslinszkyomyces aptrootii CBS T A. Aptroot, 17 Mar Hazslinszkyomyces lycii CPC T R.K. Schumacher, 2 May 2016 CPC R.K. Schumacher, 1 May 2016 Libertasomyces quercus CBS = INIFAT R.F. Castañeda, 20 C96/108 T Jul Neocamarosporium chersinae CPC T P.W. Crous, 25 May 2015 Paracamarosporium fagi CPC R.K. Schumacher, 10 May 2016 Paracamarosporium sp. 1 CPC R.K. Schumacher, 1 May 2016 Pseudocamarosporium sp. 1 CPC M.J. Wingfield, Nov Nerium oleander, leaf spot Aloe dichotoma, dead bark Italy KY GU GU South Africa KF KF Lycium sp. Netherlands KY DQ GU GU Lycium barbarum, twig Hungary KY KY Lycium barbarum, twig Hungary KY KY Quercus ilex, leaf litter Spain KY DQ KY KY Dead angulate tortoise shell Elaeagnus rhamnoides, twig South Africa KY KY Germany KY KY Tilia platyphyllos, twig Germany KY KY Erica sp. South Africa KY KY Pseudocamarosporium sp. 2 CPC R.K. Schumacher Platanus sp., branch Switzerland KY KY CPC R.K. Schumacher Platanus sp., branch Switzerland KY KY CPC R.K. Schumacher, Betula pendula, twig Germany KY KY Dec CPC R.K. Schumacher, Frangula alnus Germany KY KY May 2015 Germany KY KY Querciphoma carteri CBS = PD 84/74 CPC R.K. Schumacher, 10 May 2016 CPC R.K. Schumacher, 22 Jul Elaeagnus rhamnoides, thorn and twig Malus domestica, twig Ukraine KY KY PD Lisse Quercus sp. Netherlands KF GQ KF KF GQ CBS H. Schill Quercus robur, leaves and twigs Germany KF GQ KF KF GQ CBS: Westerdijk Fungal Biodiversity Institute, Utrecht, The Netherlands; CPC: Culture collection of Pedro Crous housed at CBS; INIFAT: Alexander Humboldt Institute for Basic Research in Tropical Agriculture, Ciudad de La Habana, Cuba; PD: Plant Protection Service, nvwa, Division Plant, Wageningen, The Netherlands. 2 ET: ex-epitype culture; NT: ex-neotype culture; T: ex-type culture. 3 ITS: internal transcribed spacers and intervening 5.8S nrdna; LSU: partial 28S nrdna; SSU: partial 18S nrdna; tef1: partial translation elongation factor 1-alpha gene; tub2: partial beta-tubulin gene. VOLUME 8 NO

203 Crous and Groenewald changes Eurotium chevalieri DAOM JN Pseudosydowia eucalypti CPC GQ Aureobasidium proteae CPC 2824 JN Aureobasidium pullulans CBS DQ Pseudoseptoria collariana CBS KF Pseudoseptoria obscura CBS KF Delphinella strobiligena CBS DQ Sydowia polyspora CBS DQ Neocylindroseptoria pistaciae CBS KF Coniozyma leucospermi CBS EU Endoconidioma populi UAMH EU Dothidea insculpta CBS DQ Dothidea sambuci CBS AY Dothidea ribesia CPC Dothidea ribesia CPC Dothidea ribesia CPC Dothidea ribesia CBS AY Dothidea puccinioides CBS AY Dothidea muelleri CBS EU Dothidea berberidis CBS KC Dothidea sp. MFLU KM Dothiora bupleuricola CBS KU Dothiora agapanthi CPC KU Dothiora viburnicola CBS KU Dothiora maculans CBS KU Dothiora sorbi CBS KU Dothiora ceratoniae CBS KF Dothiora phillyreae CBS EU Dothiora oleae CBS KU Dothiora cannabinae CBS DQ Dothiora laureolae CBS KU Dothiora prunorum CBS KU Dothiora elliptica CBS KU Dothiora phaeosperma CBS KU Dothiora pyrenophora CPC Dothiora pyrenophora CPC Dothiora cactacearum CPC Dothiora cactacearum CPC Dothiora buxi MFLU KX Pleurophoma acaciae CPC KY Keissleriella cladophila CBS GU Keissleriella sparticola MFLUCC KP Keissleriella genistae CBS GU Pleurophoma ossicola CPC KR Pleurophoma pleurospora CBS JF Keissleriella yonaguniensis KT 2604 AB Murilentithecium clematidis IT1078 KM Phragmocamarosporium platani MFLUCC KP Phragmocamarosporium hederae MFLUCC KP Didymosphaeria rubi-ulmifolii MFLUCC KJ Paraconiothyrium brasiliense CBS JX Pseudocamarosporium sp. 2 CPC Pseudocamarosporium sp. 2 CPC Pseudocamarosporium sp. 2 CPC Pseudocamarosporium sp. 2 CPC Pseudocamarosporium sp. 2 CPC Paracamarosporium psoraleae CPC KF Paracamarosporium fagi CPC KR Paracamarosporium fungicola CBS JX Paracamarosporium hawaiiense CBS DQ Paracamarosporium sp. 1 CPC Paracamarosporium fagi CPC Pseudocamarosporium sp. 1 CPC Pseudocamarosporium brabeji CBS EU Pseudocamarosporium sp. 2 CPC Pseudocamarosporium piceae MFLUCC KJ Pseudocamarosporium africanum CBS JX Saccotheciaceae I Aureobasidiaceae Saccotheciaceae II Dothideaceae Lentitheciaceae Didymosphaeriaceae Dothideales Pleosporales Fig. 1. One of equally most parsimonious trees obtained from a maximum parsimony analysis of the LSU sequence alignment. The scale bar shows 10 changes, and parsimony bootstrap support values > 49 % from replicates are shown at the nodes. Thickened lines represent those branches also present in the strict consensus tree and families are indicated with coloured blocks and the taxonomic novelties, or species treated in the present paper, are in bold text. Orders are indicated on the right side of the tree. The tree was rooted to Eurotium chevalieri (GenBank accession number JN938915). 136 IMA FUNGUS

204 The Genera of Fungi G changes Neoplatysporoides aloicola CBS KR Libertasomyces myopori CPC KX Libertasomyces quercus CBS DQ Libertasomyces platani CPC KY Pseudoleptosphaeria etheridgei CBS JF Coniothyrium palmarum CBS EU Ochrocladosporium elatum CBS EU Ochrocladosporium frigidarii CBS EU Camarosporium quaternatum CPC Camarosporium quaternatum CPC Camarosporium quaternatum CPC Coniothyrium telephii CBS GQ Camarosporomyces flavigenus CBS GU Hazslinszkyomyces lycii CPC Hazslinszkyomyces lycii CPC Hazslinszkyomyces aptrootii CBS DQ Hazslinszkyomyces aloes CPC KF Foliophoma fallens CBS GU Foliophoma fallens CBS GU Dimorphosporicola tragani CBS KU Neocamarosporium chichastianum CBS KP Pleospora obiones CBS DQ Neocamarosporium chersinae CPC Neocamarosporium calvescens CBS EU Neocamarosporium goegapense CBS KJ Neocamarosporium betae CBS EU Neocamarosporium betae CBS EU Neoleptosphaeria rubefaciens CBS JF Subplenodomus drobnjacensis CBS JF Querciphoma carteri CBS GQ Querciphoma carteri CBS GQ Pyrenochaeta acaciae CPC KX Pyrenochaeta pinicola CBS KJ Pyrenochaeta protearum CBS JQ Pleospora herbarum CBS GU Stemphylium beticola CBS KX Plenodomus lingam CBS JF Plenodomus biglobosus CBS GU Plenodomus enteroleucus CBS JF Plenodomus libanotidis CBS JF Pyrenochaeta nobilis CBS EU Camarosporium caraganicola MFLUCC KP Camarosporium sp. 3 CPC Camarosporium aureum MFLUCC KP Camarosporium sp. 5 CPC Camarosporium sp. 5 CPC Camarosporium robiniicola MFLUCC KJ Camarosporium clematidis MFLUCC KJ Camarosporium elongata CBS DQ Camarosporium spartii MFLUCC KJ Camarosporium aborescentis MFLUCC KP Camarosporium arezzoensis CPC Camarosporium sp. 4 CPC Camarosporium sp. 1 CPC DQ Camarosporium arezzoensis MFLUCC KP Camarosporium sp. 2 CPC Camarosporium sp. 2 CPC Libertasomycetaceae Coniothyriaceae Leptosphaeriaceae Incertae sedis Pleosporaceae Incertae sedis Incertae sedis Pleosporales (continued) Fig. 1. (Continued). basally with a few red-brown, smooth, thick-walled and eguttulate hyphae, without setae; ascomatal wall consisting of a textura angularis with red-brown, thick-walled, smooth and eguttulate cells. Pseudoparaphyses numerous, cellular, filiform, multi-celled, short celled, branched, anastomosing, hyaline, smooth, thin-walled, eguttulate, µm diam. Asci 8-spored, cylindrical, apically rounded, pedicel short and furcate, thick-walled, bitunicate, fissitunicate, inamyloid (water plus Lugol), spores oblique uniseriate, µm. Ascospores (4 )6-celled, ellipsoidal, straight, muriformly septate, golden, faintly thick-walled and smooth, septa in the middle constricted otherwise +/- faintly constricted, mostly the middle cells and seldom both end cells with 1 3 longitudinal septa, eguttulate, without a gelatinous sheath and appendages, rehydrated and examined in water, mature (20 )22.5( 26.5) (10 )11.5( 13) µm, (1.79 )1.99( 2.2) (l/b). Conidiomata pycnidial, saprobic, subcorticolous, single to gregarious, partly caespitose, globose seldom broad pyroid with a flattened base, ostiole centrally, terete, short papillate, and somewhat erumpent, black, soft, +/-thin, covered with hyaline to ochre and smooth hyphae, to 0.8 mm diam; conidiomatal wall few-layered, consisting of a textura globulosa-angularis with red brown, thick-walled, and smooth cells. Paraphyses VOLUME 8 NO

205 Crous and Groenewald Dothiora oleae CBS KU Leptosphaeria doliolum CBS JF Leptosphaeria conoidea CBS JF Leptosphaeria sclerotioides CBS JF Camarosporomyces flavigenus CBS CPC CPC CPC Camarosporium quaternatum Subplenodomus drobnjacensis CBS JF Subplenodomus apiicola CBS JF Plenodomus enteroleucus CBS NR Paraleptosphaeria dryadis CBS JF Paraleptosphaeria praetermissa CBS JF CBS CBS Foliophoma fallens 99 Libertasomyces platani CPC KY Libertasomyces quercus CBS Neoplatysporoides aloicola CBS KR Libertasomyces myopori CPC KX Neocamarosporium goegapense CBS KJ Dimorphosporicola tragani CBS KU Neocamarosporium chersinae CPC Neocamarosporium chichastianum CBS KP Coniothyrium glycines CBS KF Coniothyrium hakeae CPC KY CBS KF CBS KF Querciphoma carteri Hazslinszkyomyces aptrootii CBS Hazslinszkyomyces aloes CBS KF CPC CPC Hazslinszkyomyces lycii Coniothyrina agaves CBS JX Pyrenochaetopsis microspora NRRL HM Camarosporium sp. 1 CPC MFLUCC KP CPC Camarosporium arezzoensis Camarosporium spartii MFLUCC KJ Camarosporium clematidis MFLUCC KJ CPC CPC Camarosporium sp. 2 Camarosporium sp. 3 CPC Camarosporium sp. 4 CPC Camarosporium caraganicola MFLUCC KP Camarosporium aborescentis MFLUCC KP CPC CPC Camarosporium sp changes Camarosporium aureum MFLUCC NR Camarosporium robiniicola MFLUCC KJ Leptosphaeria Camarosporomyces Camarosporium Subplenodomus Plenodomus Paraleptosphaeria Foliophoma Libertasomyces / Neoplatysporoides Neocamarosporium / Dimorphosporicola Coniothyrium Querciphoma Hazslinszkyomyces Coniothyrina Pyrenochaetopsis Camarosporium Fig. 2. One of 284 equally most parsimonious trees obtained from a maximum parsimony analysis of the ITS sequence alignment of Camarosporium and allied genera. The scale bar shows 20 changes, and parsimony bootstrap support values > 74 % from replicates are shown at the nodes. Thickened lines represent those branches also present in the strict consensus tree and genera are indicated with coloured blocks and the taxonomic novelties, or species treated in the present paper, are in bold text. The tree was rooted to Dothiora oleae (GenBank accession number KU728511). and conidiophores absent. Conidiogenous cells formed from the inner cells of the pycnidial wall, doliiform, hyaline, thin-walled, smooth, eguttulate, annellidic, secession apically and singly. Conidia as brownish, dark olive to black drops in mass, (3 )4( 6)-celled, muriformly septate, with one longitudinal or diagonal septum per cell and 1 2 per conidium, ellipsoidal, pyroid, clavate, straight to slightly curved, apically rounded, base often tapered but never truncate, always yellowish, but never brown, both end cells but mostly the basal cell often paler or hyaline, wall golden, thin and smooth, septa golden, thick-walled and smooth to slightly constricted, eguttulate at maturity, examined in water, living and mature, (18 )26.5( 34) (9 )12( 16) µm, (1.19 )2.26( 3.32) (l/b). Synasexual morph in culture: 138 IMA FUNGUS

206 The Genera of Fungi G changes Pezicula cinnamomea CBS KR CBS AF MFUCC KM Dothidea insculpta AC836 KF CBS AF Dothidea hippophaeos Dothidea puccinioides CBS Dothidea muelleri CBS EU MFLU KM MFLUCC KM Dothidea sp. 100 DAOM DQ CBS AY Dothidea sambuci Dothidea berberidis CBS EU CPC CPC CPC Dothidea ribesia Dothiora buxi MFLU KX CPC Dothiora cactacearum 100 CPC Dothiora cannabinae CBS AJ Dothiora laureolae CBS KU Dothiora phaeosperma CBS KU Dothiora europaea CBS AJ Dothiora elliptica CBS KU Dothiora prunorum CBS AJ Dothiora rhamni-alpinae CBS AJ Dothiora sorbi CBS KU CPC CPC Dothiora pyrenophora CBS KU CBS KU Dothiora maculans Dothiora agapanthi CPC KU Dothiora viburnicola CBS KU Dothiora bupleuricola CBS KU Dothiora phillyreae CBS KU CBS KU CBS KF CBS KU CBS KU Dothiora ceratoniae Dothiora oleae Dothidea Dothiora Fig. 3. One of 140 equally most parsimonious trees obtained from a maximum parsimony analysis of the ITS sequence alignment of Dothidea and Dothiora. The scale bar shows 10 changes, and parsimony bootstrap support values > 74 % from replicates are shown at the nodes. Thickened lines represent those branches also present in the strict consensus tree and genera are indicated with coloured blocks and the taxonomic novelties, or species treated in the present paper, are in bold text. The tree was rooted to Pezicula cinnamomea (GenBank accession number KR859133). Conidiomata separate, pycnidial, immersed to superficial on PNA, brown, globose, µm diam, with 1 2 papillate ostioles, exuding a crystalline conidial mass. Conidiophores reduced to conidiogenous cells. Conidiogenous cells lining the inner cavity, hyaline, smooth, ampulliform, with periclinal thickening at apex, µm. Conidia solitary, hyaline, smooth, aseptate, subcylindrical, straight, rarely curved, apex obtuse, base truncate, (3 )4 5( 6) 1.5 µm. Culture characteristics: On MEA spreading, with fluffy aerial mycelium and feathery margin; surface dirty white with patches of olivaceous grey. On PDA surface and reverse olivaceous grey. On OA surface olivaceous grey with patches of dirty white and umber. Notes: Single conidial colonies of C. quaternatum formed a phoma-like fungus in culture. The single conidial isolation step was redone three times on different media, with the same result. It was only once colonies were subcultured onto SNA plates supplemented with autoclaved pieces of banana leaf, that a few typical Camarosporium conidiomata again developed on leaf pieces. The trigger influencing which morph develops in culture, however, remains unknown, with the phoma-like morph being most commonly encountered, and the Camarosporium morph extremely rare. Ascospores of Camarosporium quaternatum studied here agree with those of the original description of Cucurbitaria varians (figs 13 24), and conidia that of its asexual morph, Clinterium quaternatum (figs 9 12) (Hazslinszky 1865). Because the holotype material could not be traced, the original illustrations are designated as lectotypes, to facilitate epitypification. VOLUME 8 NO

207 Crous and Groenewald changes Keissleriella trichophoricola CBS KJ Paracamarosporium fungicola CBS JX Paracamarosporium sp. 1 CPC Paracamarosporium leucadendri CBS EU Paracamarosporium mamanes CBS DQ Paracamarosporium psoraleae CPC KF Paracamarosporium 100 CBS DQ Paracamarosporium hawaiiense CPC DQ CPC CPC KR Paracamarosporium fagi 99 CPC KR Pseudocamarosporium sp. 1 CPC CBS JX Pseudocamarosporium africanum STE U-6316 EU CPC CPC CPC Pseudocamarosporium sp. 2 Pseudocamarosporium CPC CPC CPC Pseudocamarosporium brabeji CBS EU Pseudocamarosporium corni MFLUCC KJ Pseudocamarosporium cotinae MFLUCC KP Pseudocamarosporium lonicerae MFLUCC KJ Pseudocamarosporium piceae MFLUCC KJ Pseudocamarosporium pinicola MFLUCC KT Pseudocamarosporium propinquum MFLUCC KJ Pseudocamarosporium quercinum MFLUCC KT Pseudocamarosporium tiliicola MFLUCC KJ Pseudocamarosporium brabeji CBS EU Pseudocamarosporium brabeji IMI JX Pseudocamarosporium pini MFLUCC KU Pseudocamarosporium sp. MFLUCC KT Fig. 4. One of 16 equally most parsimonious trees obtained from a maximum parsimony analysis of the ITS sequence alignment of Paracamarosporium and Pseudocamarosporium. The scale bar shows 10 changes, and parsimony bootstrap support values > 69 % from replicates are shown at the nodes. Thickened lines represent those branches also present in the strict consensus tree and genera are indicated with coloured blocks and the taxonomic novelties, or species treated in the present paper, are in bold text. The tree was rooted to Keissleriella trichophoricola (GenBank accession number KJ869113). Table 2. Statistical information on the individual alignments and number of equally most parsimonious trees saved for each dataset analysed 1. LSU overview Camarosporium ITS Dothidea ITS Paracamarosporium ITS Aligned characters (including gaps) Parsimony-informative characters Variable and parsimony-uninformative characters Constant characters Equally most parsimonious trees obtained Tree length Consistency index (Cl) 0,551 0,499 0,691 0,912 Retention index (RI) 0,948 0,764 0,832 0,891 Rescaled Consistency index (RC) 0,522 0,381 0,575 0,812 1 ITS: internal transcribed spacers and intervening 5.8S nrdna; LSU: partial 28S nrdna. 140 IMA FUNGUS

208 The Genera of Fungi G4 Fig. 5. Camarosporium quaternatum (CPC 31081). A. Immersed ascomata on twig (arrows). B E. Asci and pseudoparaphyses. F. Ascospores. G. Conidiomata on twig. H. Camarosporium conidioma on OA. I. Phoma-like conidiomata on SNA. J K. Conidiogenous cells giving rise to Camarosporium conidia. L. Conidia. Bars: A, I = 200 µm, G = 800 µm, H = 300 µm, all others = 10 µm. Saccardo (1883) reported the fungus to occur on Lycium barbarum in Hungary and Germany. In the present study, we also found the sexual morph on Daphne mezereum, a new host for C. quaternatum. The two cultures used in previous studies as representative of this fungus (CBS , CBS ) (e.g. Crous et al. 2006) represent two camarosporium-like fungi that are morphologically distinct from C. quaternatum. Sutton (1980) stated that the genus Camarosporium, which contains several hundred species, is polyphyletic and in need of revision. As we have shown here, Camarosporium s. str. is distinct from other taxa that have been treated as representative of or similar in some respects to Camarosporium, viz. Henfellra, Neocamarosporium, Paracamarosporium, or Xenocamarosporium (Crous et al. 2013, 2014b, 2015b, Wijayawardene et al. 2014, 2016, Hawksworth et al. 2016). Further studies are underway to treat those taxa that are presently known from culture. During the course of evaluating the status of Camarosporium s. str., several other camarosporium-like or pleospora-like isolates also had to be examined and were found to be undescribed. These are treated below: Camarosporomyces Crous, gen. nov. MycoBank MB Etymology: Named after its morphological similarity to Camarosporium and its phoma-like synasexual morph. Diagnosis: Distinct from phoma-like genera in having pycnidial conidiomata with prominent papillate dark brown periphysate ostiole, and thicker wall at apex. Type species: Camarosporomyces flavigenus (Constantinou & Aa) Crous Description: Conidiomata pycnidial, solitary to aggregated, obovoid, medium brown, with prominent papillate ostiole, VOLUME 8 NO

209 Crous and Groenewald Fig. 6. Camarosporomyces flavigenus (CBS ). A. Conidiomata on PDA. B. Conidiomata on SNA. C. Conidiomata showing darker ostiolar area. D. Conidiogenous cells. E. Phoma-like conidia. Bars: A C = 90 µm, D E = 10 µm. dark brown, with extra 1 2 layers of wall at the apex (thicker than pycnidial body), periphysate. Conidiophores reduced to conidiogenous cells lining inner cavity, hyaline, smooth, ampulliform; phialidic with periclinal thickening. Conidia solitary, hyaline, smooth, aseptate, subcylindrical, straight, apex obtuse, base truncate. Camarosporomyces flavigenus (Constant. & Aa) Crous, comb. nov. MycoBank MB (Fig. 6) Basionym: Phoma flavigena Constant. & Aa, Trans. Brit. Mycol. Soc. 79: 343 (1982). Synonym: Pleospora flavigena (Constant. & Aa) Gruyter & Verkley, Stud. Mycol. 75: 25 (2013) [ 2012 ]. Type: Romania: Bucharest, station for water treatment, from water, 1980, K. Fodor (CBS H holotype; CBS culture ex-type). Description: Conidiomata pycnidial, solitary to aggregated, obovoid, medium brown, µm diam, with prominent papillate ostiole, dark brown, mm diam with extra 1 2 layers of wall at the apex (thicker than pycnidial body), periphysate. Conidiophores reduced to conidiogenous cells lining inner cavity, hyaline, smooth, ampulliform, µm; phialidic with periclinal thickening. Conidia solitary, hyaline, smooth, aseptate, subcylindrical, straight, apex obtuse, base truncate, (2.5 )3 1.5 µm. Culture characteristics: Colonies flat, spreading, with sparse aerial mycelium, margins smooth, lobate, reaching 30 mm diam after 2 wk. On OA surface umber with diffuse yellow pigment. On PDA surface umber, outer region pale luteous, reverse umber. Note: Camarosporomyces flavigenus is a phoma-like fungus which was originally described as Phoma flavigena, and later placed in Pleospora by De Gruyter et al. (2013), who used a much wider circumscription of the genus Pleospora than applied here. Foliophoma Crous, gen. nov. MycoBank MB Etymology: Named after its association with leaf spots, and its morphological similarity to Phoma. Diagnosis: Distinct from phoma-like genera in having eustromatic conidiomata, uni- to multi-loculate with 1 3 ostioles. Conidiogenous cells with periclinal thickening or percurrent proliferation at apex. Type species: Foliophoma fallens (Sacc.) Crous Description: Conidiomata globose, eustromatic, uni- to multi-locular with 1 3 ostioles, medium brown, outer surface smooth; wall of 3 6 layers of brown textura angularis. Conidiophores reduced to conidiogenous cells lining inner cavity, hyaline, smooth, doliiform to subcylindrical; phialidic with periclinal thickening or percurrent proliferation at apex. Conidia aseptate, solitary, hyaline, smooth, guttulate to granular, broadly ellipsoidal, thick-walled, apex obtuse, base truncate to bluntly rounded. Foliophoma fallens (Sacc.) Crous, comb. nov. MycoBank MB (Fig. 7) Basionym: Phoma fallens Sacc., Syll. Fung. 10: 146 (1892). Synonyms: Pleospora fallens (Sacc.) Gruyter & Verkley, Stud. Mycol. 75: 25 (2013) ["2012"]. Phyllosticta glaucispora Delacr., Bull. Soc. Mycol. France 9: 266 (1893). Phoma glaucispora (Delacr.) Noordel. & Boerema, Versl. Meded. Plantenziektenk. Dienst Wageningen 166 : 108 (1989) [ 1988"]. Phyllosticta oleandri Gutner, Trudy Bot. Inst. Akad. Nauk S.S.S.R., ser. 2, Sporov. Rast. 1: 306 (1933). Description: Conidiomata globose, eustromatic, uni- to multilocular with 1 3 ostioles, µm diam, medium brown, outer surface smooth; wall of 3 6 layers of brown textura angularis. Conidiophores reduced to conidiogenous cells lining inner cavity, hyaline, smooth, doliiform to subcylindrical, 142 IMA FUNGUS

210 The Genera of Fungi G4 Fig. 7. Foliophoma fallens (CBS ). A. Conidiomata on PDA. B. Conidiomata on SNA. C D. Conidiogenous cells. E. Conidia. Bars: A B = 250 µm, all others = 10 µm µm; phialidic with periclinal thickening or percurrent proliferation at apex. Conidia aseptate, solitary, hyaline, smooth, guttulate to granular, broadly ellipsoidal, thick-walled, apex obtuse, base truncate to bluntly rounded, (5 )5.5 6( 7) (3 )4( 5) µm (based on CBS ). Culture characteristics: Colonies flat, spreading, with moderate aerial mycelium and even, lobate margins, reaching 50 mm diam after 2 wk. On PDA surface sepia to umber, reverse dark mouse grey. On OA surface greyish sepia. Material examined: Italy: Capri, Villa Jovis, leaf spot on Nerium oleander (Apocynaceae), Apr. 1970, H.A. van der Aa (CBS H-16639, 23066, culture CBS ). New Zealand: Levin, from leaf spot of Olea europaea (Oleaceae), 1978, G.F. Laundon (CBS = LEV 1131). Notes: Phoma fallens is associated with leaf spots of Olea europaea in Europe (conidia µm), while Phoma glaucispora (conidia µm) is associated with leaf spots on Nerium oleander in Europe (Boerema et al. 2004). Phylogenetically, however, De Gruyter et al. (2013) found these two species to be closely related, and therefore treated them as a single species, placing them in Pleospora based on the larger circumscription used at the time. However, the present fungus is not congeneric with Stemphylium herbarum (syn. Pleospora herbarum; Rossman et al. 2015), and therefore a new generic name is introduced to accommodate it. Hazslinszkyomyces Crous & R.K. Schumach., gen. nov. MycoBank MB Etymology: Named after Friedrich August Hazslinszky von Hazslin ( ), in recognition for his work on camarosporium-like fungi. Diagnosis: Morphologically similar to Camarosporium, but distinct as conidia are uniformly brown in colour, those of Camarosporium having paler end cells. Type species: Hazslinszkyomyces aloes (Crous & M.J. Wingf.) Crous Description: Ascomata pseudothecial, saprobic, subcorticolous, densely crowded, erumpent, globose to pyroid with a flattened base, ostiole central, terete, and short papillate, black, periphysate; ascomatal wall multi-layered, consisting of a textura angularis. Pseudoparaphyses numerous, basally moniliform, upwards filiform, multi-celled, hyaline, thin-walled, smooth. Asci 8-spored, cylindrical, thick-walled, apically rounded and with an ocular chamber, pedicel short and furcate, bitunicate, fissitunicate. Ascospores transversely septate, constricted at the median septum, becoming muriformly septate, ellipsoidal, golden brown. Phoma-like morph: Conidiomata pycnidial, solitary, brown, globose, with central ostiole; wall of 2 3 layers of brown textura angularis. Conidiophores reduced to conidiogenous cells lining the inner cavity, hyaline, smooth, doliiform to subcylindrical. Conidia solitary, hyaline, smooth, guttulate, subcylindrical, straight, aseptate. Camarosporium-like morph: Occurring in the same or separate conidioma with a phoma-like morph. Conidiogenous cells lining the inner cavity, doliiform to subcylindrical, hyaline, smooth, proliferating percurrently at apex. Conidia solitary, brown, smooth, broadly ellipsoidal to obovoid, transversely septate, becoming muriformly septate. Hazslinszkyomyces aloes (Crous & M.J. Wingf.) Crous, comb. nov. MycoBank MB Basionym: Camarosporium aloes Crous & M.J. Wingf., Persoonia 31: 247 (2013). Type: South Africa: Western Cape Province: Clanwilliam, on dark lesions on dead bark of Aloe dichotoma (Xanthorrhoeaceae), Sept. 2012, M.J. Wingfield (CBS H holotype; CPC = CBS culture ex-type). Hazslinszkyomyces aptrootii Crous, sp. nov. MycoBank MB (Fig. 8) Etymology: Named after André Aptroot, who collected this fungus in The Netherlands. VOLUME 8 NO

211 Crous and Groenewald Fig. 8. Hazslinszkyomyces aptrootii (CBS ). A. Conidiomata on PNA. B. Conidiomata on PDA. C. Hazslinszkyomyces and phoma-like conidia. D E. Conidiogenous cells. F. Phoma-like conidia. Bars: A B = 170 µm, all others = 10 µm. Diagnosis: Similar to Hazslinszkyomyces aloes [conidia (9 ) 11 13( 14) (4 )6 7( 8) μm], but distinct in having larger conidia. Type: The Netherlands: Province of Noord-Holland: Egmond, on Lycium sp., 17 Mar. 1995, A. Aptroot (CBS H holotype; CBS culture ex-type). Description: Conidiomata separate, erumpent, pycnidial, globose, brown, µm diam, with central, non-papillate ostiole, 10 µm diam, exuding a creamy conidial mass; wall of 3 6 layers of brown textura angularis. Conidia dimorphic. Phoma-like morph: Conidiophores reduced to conidiogenous cells lining the inner cavity, ampulliform to subcylindrical, hyaline, smooth, phialidic with periclinal thickening at apex, becoming percurrent with age, µm. Conidia solitary, hyaline, smooth, guttulate, subcylindrical, aseptate, straight to curved, apex obtuse, base truncate, (4 )5 6 2 µm. Camarosporium-like morph: Occurring in the same conidiomata as the phoma-like morph. Conidiophores reduced to conidiogenous cells, hyaline, smooth, doliiform to subcylindrical, proliferating percurrently at apex, µm. Conidia solitary, broadly ellipsoidal, apex subobtusely rounded, base truncate, hilum 2 µm diam, medium brown, smooth, muriformly septate, with 3 transverse septa, and 1 3 oblique septa, (11 )13 14( 16) (7 )8( 9) µm. Culture characteristics: Colonies flat, spreading, with sparse to moderate aerial mycelium, covering dish in 2 wk, with even, lobate margins. On PDA surface umber, reverse brown vinaceous. On OA surface chestnut to brown vinaceous. Notes: This collection was originally identified as Camarosporium quaternatum, based on its general morphology, and the fact that it occurred on Lycium. Morphologically, however, the conidia are much smaller than those of C. quaternatum, and thus it is described as a new species of Hazslinszkyomyces. Phylogenetically the two genera are also quite distinct. Hazslinszkyomyces lycii Crous & R.K. Schumach., sp. nov. MycoBank MB (Fig. 9) Etymology: Named after the host genus from which it was isolated, Lycium. Diagnosis: Similar to Hazslinszkyomyces aptrootii [conidia (11 )13 14( 16) (7 )8( 9) µm], but distinct in having larger conidia. Type: Hungary: near Budapest, on twig of Lycium barbarum (Solanaceae), 2 May 2016, L. Bartalos (CBS H holotype; CPC = CBS culture ex-type). Description: Ascomata pseudothecial, saprobic, subcorticolous, densely crowded, erumpent, globose to pyroid with a flattened base, ostiole central, terete, and short papillate, black, soft, thick, to 130 µm diam, periphysate; ascomatal wall multi-layered, consisting of a textura angularis with red-brown, thick-walled, smooth and small cells. Pseudoparaphyses numerous, basally moniliform, upwards filiform, multi-celled, short-celled, branched, with anastomoses, gnarled, hyaline, thin-walled, smooth. Asci 8-spored, cylindrical, thick-walled, apically rounded and with an ocular chamber, 2 3 µm diam, pedicel short and furcate, bitunicate, fissitunicate, µm, ascospores oblique uniseriate. Ascospores initially with three transverse septa, constricted at the median septum, developing up to five transverse septa, becoming muriformly septate, 1 2 longitudinal or diagonal septa per cell, ellipsoidal, straight, the upper part often wider, end cells mostly bluntly rounded seldom conical, wall faintly thick and smooth, golden brown, septa red-golden and faintly thick, median septum constricted otherwise faintly constricted, eguttulate at maturity, no mucilaginous sheath in all stages of development, in lactic acid (18 )20 21( 23) (8.5 )9 10( 12) µm, in water µm. Phoma-like morph: Conidiomata pycnidial, solitary, brown, globose, µm diam, with central ostiole, µm diam; wall of 2 3 layers of brown textura angularis. Conidiophores reduced to conidiogenous cells lining the inner cavity, hyaline, 144 IMA FUNGUS

212 The Genera of Fungi G4 Fig. 9. Hazslinszkyomyces lycii (CPC 30998). A. Ascoma immersed in twig. B D. Asci and pseudoparaphyses. E. Phoma-like conidiomata on SNA. F G. Conidiogenous cells. H. Conidia. I. Hazslinszkyomyces conidioma on twig. J. Conidiogenous cells. K. Conidia. Bars: A = 130 µm, I = 250 µm, all others = 10 µm. smooth, doliiform to subcylindrical, µm; phialidic, with periclinal thickening at apex. Conidia solitary, hyaline, smooth, guttulate, subcylindrical, straight, aseptate, apex obtuse, base truncate, hilum 1.5 µm diam, (3.5 )4 5( 6) 2 µm. Camarosporium-like morph: Conidiomata pycnidial, solitary, gregarious, brown, globose, with central ostiole, wall few-layered of brown textura globulosa-angularis, completely covered with hyaline to brownish and short hyphae, to 800 µm diam. Conidiogenous cells lining the inner cavity, doliiform to subcylindrical, hyaline, smooth, µm; annellidic, proliferating percurrently at apex. Conidia solitary, pale yellowish with a golden wall, smooth, broadly ellipsoidal to obovoid but also clavate, seldom cylindrical, straight, seldom slightly curved, apex obtuse, base sometimes tapered and never truncate, 4 5 µm diam, transversely (2 )3( 4 5)-septate, becoming muriformly septate, with vertical and oblique septa, septa smooth to slightly constricted and thick, eguttulate, (15 )16 18( 20) (8 )9( 10) µm in vitro, µm in vivo. Culture characteristics: Colonies covering dish in 2 wk, with moderate aerial mycelium, and smooth, lobate margins. On PDA surface fawn to hazel, reverse fawn. On OA surface ochreous to umber. Other material examined: Hungary: near Budapest, on twig of Lycium barbarum (Solanaceae), 1 May 2016, L. Bartalos (HPC 1029 = CPC = CBS ). Notes: Hazslinszkyomyces lycii occurred in association with a camarosporium-like morph which, when cultured, proved to be identical based on DNA sequence data. The sexual morph also produced a phoma-like synasexual morph in culture. Pyrenophora lycii (syn. Pleospora lycii), also described from Lycium barbarum in Hungary, has setose pseudothecia and ascospores that are transversely 5-septate with only one longitudinal septum per cell, µm, and is distinct from the present fungus. A further name to consider is Hendersonia lycii Hazsl. Hazslinszky (1865) referred to a Hendersonia form on Lycium barbarum, but the name Hendersonia lycii was never validly published with any description, and has crept into literature in error, and is thus unavailable for this collection. The taxon is therefore described here as new. VOLUME 8 NO

213 Crous and Groenewald Fig. 10. Neocamarosporium chersinae (CPC 27298). A. Conidiomata on SNA. B. Conidiogenous cells. C D. Conidia. Bars: A = 300 µm, all others = 10 µm. Neocamarosporium chersinae Crous, sp. nov. MycoBank MB (Fig. 10) Etymology: Named after the angulate tortoise or rooipens (Chersina angulata), a tortoise species found in dry areas and scrub forest in South Africa. The angulate tortoise is used as a food source by people in rural areas, and is also often kept as a pet. Diagnosis: Similar to Neocamarosporium chichastianum [conidia (11 )15 19( 22) (6 )8 9( 11) μm], but conidia smaller. Type: South Africa: Western Cape Province: Robben Island, on dead angulate tortoise shell, 25 May 2015, P.W. Crous (CBS H holotype; CPC = CBS culture ex-type). Description: Conidiomata solitary, pycnidial, immersed to erumpent, golden brown, µm diam, with central ostiole, µm diam; wall of 2 3 layers of brown textura angularis. Conidiophores reduced to conidiogenous cells, hyaline, smooth, subcylindrical, µm; proliferating several times percurrently at apex (conidiogenous cells dissolve at maturity). Conidia solitary, golden brown, thickwalled, smooth, broadly ellipsoidal to subcylindrical or irregular, transverse 3-septate, with 0 3 oblique septa, (10 ) 12 13( 15) (5 )6( 7) µm. Culture characteristics: Colonies covering the dish in 2 wk, with fluffy aerial mycelium. On OA, MEA and PDA surface olivaceous grey to pale olivaceous grey, reverse olivaceous grey. Notes: Neocamarosporium chersinae is introduced as a new species of Neocamarosporium (Crous et al. 2014b). Neocamarosporium is camarosporium-like in morphology, and can only be safely distinguished from this genus based on DNA data. Libertasomycetaceae Crous, fam. nov. MycoBank MB Etymology: Named after the genus Libertasomyces. Classification: Libertasomycetaceae, Pleosporales, Dothideomycetes. Diagnosis: Similar to Coniothyriaceae, but distinct in that ascomata are immersed in a brown stroma, and conidiomata are stromatic in culture. Type genus: Libertasomyces Crous & Roets Description: Ascomata immersed in a brown stroma, becoming erumpent, breaking through the host surface, aggregated in clusters, with a central ostiole; wall of 6 10 layers of brown textura angularis. Pseudoparaphyses hyphal-like, intermingled among asci, hyaline, smooth, septate, anastomosing. Asci fasciculate, stipitate, hyaline, smooth, subcylindrical, bitunicate with ocular chamber, containing 8 ascospores. Ascospores fusoid-ellipsoidal, brown, verruculose with obtuse ends, muriformly septate, encased in a mucoid sheath. Conidiomata unilocular, stromatic, separate, globose, immersed, brown, opening via a central ostiole, exuding a brown conidial mass; wall of 3 6 layers of brown textura angularis. Conidiophores reduced to conidiogenous cells. Conidiogenous cells lining the inner cavity, hyaline, smooth, ampulliform to doliiform, with prominent periclinal thickening at the apex, or with tightly aggregated percurrent proliferations at the apex. Conidia solitary, golden brown, subcylindrical to ellipsoidal, straight to curved, 0 1-septate, constricted at median septum, apex obtuse, base truncate, with marginal frill, and longitudinal striations, or hyaline, smooth, granular, thin-walled, ellipsoidal, apex obtuse, base truncate to bluntly rounded, aseptata. Genera included: Libertasomyces and Neoplatysporoides. Libertasomyces quercus Crous, sp. nov. MycoBank MB (Fig. 11) Etymology: Named after the genus from which it was isolated, Quercus. Diagnosis: Similar to Camarosporium quaternatum (conidia µm), but distinct in having smaller, uniformly brown conidia ( µm). 146 IMA FUNGUS

214 The Genera of Fungi G4 Fig. 11. Libertasomyces quercus (CBS ). A. Conidiomata on OA. B, C. Conidiogenous cells. D. Conidia. Bars: A = 200 µm, all others = 10 µm. Type: Spain: location unknown, on leaf litter of Quercus ilex (Fagaceae), 20 Jul. 1996, R.F. Castañeda (CBS H holotype; CBS = INIFAT C96/108 culture ex-type). Description: Conidiomata brown, solitary or aggregated in clusters, stromatic, obovoid, µm diam, with paler brown papillate ostiole, outer region covered with brown hyphae; wall of 6 12 layers of brown textura angularis. Conidiophores reduced to conidiogenous cells lining the inner cavity, hyaline, smooth, doliiform to ampulliform, µm; proliferating percurrently at apex. Conidia solitary, brown, smooth, subcylindrical to broadly ellipsoidal, apex obtuse, base truncate to bluntly rounded, transversely 3 7-septate, becoming muriformly septate with vertical and oblique septa, (15 )17 19( 21) (6 )7 8( 10) µm. Culture characteristics: Colonies flat, spreading, reaching 40 mm diam after 2 wk with sparse to moderate aerial mycelium and even, lobate margins. On PDA surface umber with patches of salmon, reverse hazel. On OA surface dirty white with patches of umber. Notes: Libertasomyces quercus (conidia µm) was originally identified as Camarosporium quaternatum (conidia µm, see above), but differs in conidial dimensions. Phylogenetically, it is better accommodated in Libertasomyces (based on L. myopori; Crous et al. 2016), which is pleomorphic, having a sexual morph, a camarosporium-like and a phoma-like synasexual morph. Leptosphaeriaceae M.E. Barr, Mycotaxon 29: 503 (1987). Type genus: Leptosphaeria Ces. & De Not Querciphoma Crous, gen. nov. MycoBank MB Etymology: Named after the host on which it occurs, Quercus, and its phoma-like morphology. Diagnosis: Morphologically similar to Phoma, but distinct in that conidiomata are eustromatic, uni- to multi-locular, and conidia become brown and verruculose with age. Type species: Querciphoma carteri (Gruyter & Boerema) Crous Description: Conidiomata pycnidial, globose, eustromatic, uni- to multi-locular, with 1 3 papillate ostioles, medium brown, outer surface covered in short, brown, verruculose, septate setae with obtuse ends; wall of 3 6 layers of brown textura angularis. Conidiophores reduced to conidiogenous cells lining inner cavity, hyaline, smooth, doliiform; phialidic with periclinal thickening at the apex. Conidia solitary, hyaline, smooth, guttulate, broadly ellipsoidal, aseptate, becoming brown, verruculose with age. Querciphoma carteri (Gruyter & Boerema) Crous, comb. nov. MycoBank MB (Fig. 12) Basionym: Phoma carteri Gruyter & Boerema, Persoonia 17: 547 (2002) [ 2001 ]. Synonyms: Coniothyrium carteri (Gruyter & Boerema) Verkley & Gruyter, Stud. Mycol. 75: 23 (2013) [ 2012 ]. Pyrenochaeta minuta J.C. Carter, Bull. Illinois Nat. Hist. Surv. 21: 214 (1941). Description: Conidiomata pycnidial, globose, eustromatic, uni- to multi-locular, with 1 3 papillate ostioles, µm diam, medium brown, outer surface covered in short, brown, verruculose, septate setae with obtuse ends, to 80 µm tall; wall of 3 6 layers of brown textura angularis. Conidiophores reduced to conidiogenous cells lining inner cavity, hyaline, smooth, doliiform, µm; phialidic with periclinal thickening at apex. Conidia solitary, hyaline, smooth, guttulate, broadly ellipsoidal, aseptate, becoming brown, verruculose with age, apex obtuse, base truncate to bluntly rounded, (3.5 )4 5( 6) 3( 3.5) µm. Culture characteristics: Colonies flat, spreading, reaching 50 mm diam after 2 wk, with sparse to moderate aerial mycelium, and smooth, lobate margins. On PDA surface umber, reverse chestnut. On OA surface umber. Material examined: Germany: former West-Germany: on leaves and twigs of Quercus robur (Fagaceae), H. Schill (CBS H-23071, culture CBS ). The Netherlands: PD Lisse, on Quercus sp., date and collector unknown (PD 84/74 = CBS ). VOLUME 8 NO

215 Crous and Groenewald Fig. 12. Querciphoma carteri (CBS ). A. Conidiomata on PDA. B. Conidiomata on SNA. C. Setae. D. Conidiogenous cells. E. Conidia. Bars: A, B = 250 µm, all others = 10 µm. Notes: This isolate was deposited as Phoma sp., and reclassified as Coniothyrium carteri (based on Phoma carteri; see Boerema et al. 2004) by De Gruyter et al. (2013). However, as shown here, it clusters separately from Coniothyrium s.str. (having brown, 1-septate conidia), and is best placed in a separate genus. Didymosphaeriaceae Munk, Dansk bot. Ark. 15: 128 (1953). Type genus: Didymosphaeria Fuckel Paracamarosporium mamanes (Crous) Crous, comb. nov. MycoBank MB Basionym: Camarosporium mamanes Crous, Fungal Planet No. 5: 1 (2006). Type: USA: Hawaii: Saddle Road, on stems of Sophora chrysophylla (Leguminosae), Aug. 2005, W. Gams & Y. Degawa (CBS H holotype; CPC = CBS , CPC cultures ex-type). Notes: Camarosporium mamanes is allocated to the genus Paracamarosporium based on its phylogenetic position. However, the distinction between Paracamarosporium and Pseudocamarosporium is highly debatable (Wijayawardene et al. 2014), and although we could separate them here based on ITS sequence data, the genera cannot be separated based on LSU data, and appear to represent a single genus, Paracamarosporium. Furthermore, there have also been an excessive number of species introduced in the genus, which are not supported based on their DNA barcodes. DOTHIORA COMPLEX Dothideaceae Chevall., Fl. gén. env. Paris 1: 446 (1826); as Dothideae. Type genus: Dothidea Fr Dothiora Fr., Summa veg. Scand. 2: 418 (1849). Synonyms: Dothichiza Lib. ex Roum., Fungi Selecti Gallici Exs. cent. 7, no. 627 (1880). Coleonaema Höhn., Mitt. Bot. Lab. TH Wien 1(3): 95 (1924). Cylindroseptoria Quaedvl., et al., Stud. Mycol. 75: 358 (2013). Additional synonyms: See Crous & Groenewald (2016). Classification: Dothideaceae, Dothideales, Dothideomycetes. Current generic circumscription: Ascostromata immersed to erumpent, pulvinate to globose, black, multi-loculate; wall of dark brown textura angularis. Locules globose to subglobose, broadly rounded or papillate with central ostiole. Pseudoparaphyses absent. Asci 8- or more spored, bitunicate, fissitunicate, oblong to clavate, pedicellate, with a small ocular chamber. Ascospores biseriate to multi-seriate, septate, constricted at the primary median septum, at times with a vertical septum, hyaline, rarely pale brown, obovate to ellipsoidal to fusoid, often inequilateral or slightly curved, smooth, at times with a thin mucoid sheath. Conidiomata pycnidial, separate, or aggregated in a stroma. Conidiophores reduced to conidiogenous cells lining the inner cavity, hyaline, smooth, ampulliform to doliiform, phialidic. Conidia aseptate, hyaline, smooth, subcylindrical to ovoid or oblong. Hyphae becoming brown, verruculose and constricted at septa, giving rise to a hormonema-like synasexual morph. Type species: Dothiora pyrenophora (Fr.) Fr Dothiora cactacearum Crous, sp. nov. MycoBank MB (Fig. 13) Etymology: Named after the host family from which the fungus was isolated, Cactaceae. Diagnosis: Morphologically distinct from other species of Dothiora in that conidia become brown and roughened with age. Type: USA: Texas: Austin, on phyllodes of Cactaceae, 2013, P.W. Crous (CBS H holotype; CPC = CBS culture ex-type, CPC 15587). 148 IMA FUNGUS

216 The Genera of Fungi G4 Fig. 13. Dothiora cactacearum (CPC 15585). A. Conidiomata on OA. B. Conidioma. C. Brown hyphae forming on SNA. D, E. Conidiogenous cells. F. Conidia. G. Conidia turning brown and verruculose with age. Bars: A = 150 µm, all others = 10 µm. Description: Conidiomata separate, erumpent, pycnidial, globose, medium brown, µm diam, with a central ostiole, exuding a creamy conidial mass; wall of 3 6 layers of brown textura angularis. Conidiophores reduced to conidiogenous cells lining the inner cavity, hyaline, smooth, ampulliform to doliiform, µm, phialidic, at times with percurrent proliferation and prominent collarette. Conidia hyaline, smooth, guttulate, subcylindrical to broadly ellipsoidal, apex obtuse, tapering to a truncate, protruding hilum, 2 3 µm diam, (12 )14 17( 19) (5 )6 7.5( 8) µm (av µm); conidia becoming brown and roughened with age. Hyphae becoming brown, verruculose and constricted at the septa. Culture characteristics: Colonies flat, spreading, with sparse to moderate aerial mycelium, and even, lobate margin, reaching 40 mm diam after 2 wk at 25 ºC. On MEA surface umber, reverse luteous; on PDA surface fawn, reverse isabelline; on OA surface isabelline. Dothiora pyrenophora (Fr.) Fr., Summa veg. Scand. 2: 418 (1849). (Fig. 14) Basionym: Dothidea pyrenophora Fr., Kongl. Vetensk. Acad. Handl. 40: 88 (1819) : Fr., Syst. mycol. 2(2): 552 (1823). Type: Sweden: without locality, on dead branches of Sorbus sp. (Rosaceae), E.M. Fries (no original material found). Germany: on twig of Sorbus aucuparia, 27 Apr. 2016, R.K. Schumacher (CBS H neotype designated here, MBT376258; CPC 30634, CPC = CBS cultures ex-neotype). Description: Ascostromata solitary to aggregated, black, immersed to erumpent, unilocular, to 400 µm diam, elliptical, pulvinate, opening by an irregular pore, upper layer dissolving with age; wall of 6 10 layers of brown textura angularis. Asci bitunicate, hyaline, oblong to subcylindrical, short stipitate, 8-spored, with apical apiculus, 2 3 µm diam, µm. Ascospores bi- to triseriate in ascus, hyaline, smooth, at times turning yellow-brown with age, fusiform, inequilateral, slightly curved, with prominent mucoid sheath when young (in water), dissolving at maturity, (5 )5( 8) transversely septate, prominently constricted at primary septum, with oblique or vertical septa in central cells, (22 )25 30( 35) (7 )8 µm; ascospores directly giving rise to asexual morph via budding, with ascomata transforming with age into large conidiomata, with apical opening completely dissolving. Conidiomata immersed to erumpent, pycnidial, black, globose, to 300 µm diam, separate or gregarious, unilocular; wall of 3 6 layers of brown textura angularis. Conidiophores lining the inner cavity, reduced to conidiogenous cells. Conidiogenous cells hyaline, smooth, doliiform to ampulliform, µm, with minute periclinal thickening at apex. Conidia solitary, aseptate, hyaline, smooth, ovate to ellipsoidal, with minute guttules, subobtuse apex, and truncate hilum, (5 )7 8( 9) (3 )4 µm. Culture characteristics: Colonies flat, spreading, with sparse aerial mycelium and feathery, undulate margins. On PDA, MEA and OA surface and reverse iron-grey. Other material examined: Germany: on twig of Sorbus aucuparia, 27 Apr. 2016, R.K. Schumacher (CBS H-23072). Notes: The taxonomic position of the several species occurring on Sorbus require further clarification based on fresh material. One culture identified as Dothiora sorbi (CBS ) was included in this study, and clustered slightly apart from D. pyrenophora (nine bp different on ITS), suggesting VOLUME 8 NO

217 Crous and Groenewald Fig. 14. Dothiora pyrenophora (CPC 30632). A. Ascostroma on twig. B. Section through ascoma. C E. Asci. F. Ascospores. G. Ascospores undergoing microcyclic conidiation. H. Conidia undergoing budding. I. Conidia. Bars: A, B = 200 µm, all others = 10 µm. that these may very well be two closely related, but distinct species. Genera such as Sydowia Bres.1895 and Pringsheimia Schulzer 1866 appear to represent younger generic names within Dothideaceae, based on their morphological similarity, and the formation of Dothichiza and hormonemalike morphs in culture, which are commonly observed in the family (Froidevaux 1972, Sivanesan 1984, Crous & Groenewald 2016). Another confusing aspect of many of the taxa in this family is that young ascospores can be hyaline and uniseptate, but become brown and muriformly septate with age (e.g. Dothidea ribesia, Fig. 15), which complicate the generic delimitation within the family (Thambugala et al. 2014), and identification keys based on immature material. Species of Dothiora are commonly isolated from dead branches of woody hosts (Sivanesan 1984), while Crous & Groenewald (2016) also reported some species from dead leaves and fruit of diverse hosts, suggesting that it is a saprobe, possibly acting as a weak pathogen on stressed plant tissues. ACKNOWLEDGEMENTS We acknowledge Laszlo Bartalos and René Jarling for sending us specimens of Camarosporium and similar taxa examined in the present study. We are also thankful to René K. Schumacher for making many valuable collections available to be included in this study, and for bringing the problem related to the nomenclature of Camarosporium to our attention. REFERENCES Boerema GH, De Gruyter J, Noordeloos ME, Hamers MEC (2004) Phoma Identification Manual: Differentiation of specific and infraspecific taxa in culture. Wallingford: CABI Publishing. Carbone I, Kohn LM (1999) A method for designing primer sets for speciation studies in filamentous ascomycetes. Mycologia 91: Cheewangkoon R, Crous PW, Hyde KD, Groenewald JZ, To-anan C (2008) Species of Mycosphaerella and related anamorphs on Eucalyptus leaves from Thailand. Persoonia 21: Crous PW, Gams W, Stalpers JA, Robert V, Stegehuis G (2004) MycoBank: an online initiative to launch mycology into the 21st century. Studies in Mycology 50: Crous PW, Giraldo A, Hawksworth DL, Robert V, Kirk PM, et al. (2014a) The Genera of Fungi: fixing the application of type species of generic names. IMA Fungus 5: Crous PW, Groenewald JZ (2016) They seldom occur alone. Fungal Biology 120: IMA FUNGUS

218 The Genera of Fungi G4 Fig. 15. Dothidea ribesia (CPC 30638). A. Ascostromata on twig. B. Ostioles with exuding ascospores. C E. Asci with ascospores that become muriformly septate. F. One-septate ascospores. G. Muriformly septate ascospore. Bars: A = 3 mm, B = 100 µm, all others = 10 µm. Crous PW, Hawksworth DL, Wingfield MJ (2015a) Identifying and naming plant-pathogenic fungi: past, present, and future. Annual Review of Phytopathology 53: Crous PW, Shivas RG, Quaedvlieg W, van der Bank M, Zhang Y, et al. (2014b) Fungal Planet Description Sheets: Persoonia 32: Crous PW, Slippers B, Wingfield MJ, Rheeder J, Marasas WFO, et al. (2006) Phylogenetic lineages in the Botryosphaeriaceae. Studies in Mycology 55: Crous PW, Verkley GJM, Groenewald JZ, Samson RA (eds) (2009) Fungal Biodiversity. [CBS Laboratory Manual Series no.1.] Utrecht: CBS-KNAW Fungal Biodiversity Centre. Crous PW, Wingfield MJ, Guarro J, Cheewangkoon R, van der Bank M, et al. (2013). Fungal Planet description sheets: Persoonia 31: Crous PW, Wingfield MJ, Guarro J, Hernández-Restrepo M, Sutton DA, et al. (2015b) Fungal Planet description sheets: Persoonia 34: Crous PW, Wingfield MJ, Park RF (1991) Mycosphaerella nubilosa a synonym of M. molleriana. Mycological Research 95: Crous PW, Wingfield MJ, Richardson DM, Leroux JJ, Strasberg D, et al. (2016). Fungal Planet description sheets: Persoonia 36: De Gruyter J, Woudenberg JHC, Aveskamp MM, Verkley GJM, Groenewald JZ, Crous PW (2013) Redisposition of Phoma-like anamorphs in Pleosporales. Studies in Mycology 75: De Hoog GS, Gerrits van den Ende AHG (1998) Molecular diagnostics of clinical strains of filamentous basidiomycetes. Mycoses 41: Froidevaux L (1972) Contribution à l étude des Dothioracées (Ascomycètes). Nova Hedwigia 23: Glass NL, Donaldson G (1995). Development of primer sets designed for use with PCR to amplify conserved genes from filamentous ascomycetes. Applied and Environmental Microbiology 61: Hawksworth DL, Crous PW, Redhead SA, Reynolds DR, Samson RA et al. (2011) The Amsterdam Declaration on Fungal Nomenclature. IMA Fungus 2: Hawksworth DL, Halici MG, Kocakaya Z, Kocakaya M (2016) Henfellra muriformis gen. et sp. nov., a new dictyosporous pycnidial fungus on Candelariella, with a key to the lichenicolous fungi known from that genus. Herzogia 29: Hazslinszky F (1865) Beitrag zur Kenntnis der Sphärien des Lyciums. Verhandlungen der Kaiserlich-Königlichen Zoologisch- Botanischen Gesellschaft in Wien 15: Kirk PM, Stalpers JA, Braun U, Crous PW, Hansen K, et al. (2013) A without-prejudice list of generic names of fungi for protection under the International Code of Nomenclature for algae, fungi and plants. IMA Fungus 4: O Donnell K, Cigelnik E (1997) Two divergent intragenomic rdna ITS2 types within a monophyletic lineage of the fungus Fusarium are nonorthologous. Molecular Phylogenetics and Evolution 7: O Donnell K, Kistler HC, Cigelnik E, Ploetz RC (1998) Multiple evolutionary origins of the fungus causing Panama disease of banana: concordant evidence from nuclear and mitochondrial gene genealogies. Proceedings of the National Academy of Sciences, USA 95: Quaedvlieg W, Groenewald JZ, de Jesús Yáñez-Morales M, Crous PW (2012) DNA barcoding of Mycosphaerella species of quarantine importance to Europe. Persoonia 29: Robert V, Vu D, Amor ABH, van de Wiele N, Brouwer C, et al. (2013) MycoBank gearing up for new horizons. IMA Fungus 4: VOLUME 8 NO

219 Crous and Groenewald Rehner SA, Buckley E (2005) A Beauveria phylogeny inferred from nuclear ITS and EF1-α sequences: evidence for cryptic diversification and links to Cordyceps teleomorphs. Mycologia 97: Rayner RW (1970) A Mycological Colour Chart. Kew: Commonwealth Mycological Institute. Rossman AY, Adams GC, Cannon PF, Castlebury LA, Crous PW, et al. (2015) Recommendations of generic names in Diaporthales competing for protection or use. IMA Fungus 6: Saccardo PA (1883) Sylloge Pyrenomycetum. Sylloge Fungorum 2: Sivanesan A (1984) The Bitunicate Ascomycetes and their Anamorphs. Vaduz: J. Cramer. Smith H, Wingfield MJ, Crous PW, Coutinho TA (1996) Sphaeropsis sapinea and Botryosphaeria dothidea endophytic in Pinus spp. and Eucalyptus spp. in South Africa. South African Journal of Botany 62: Sutton BC (1980) The Coelomycetes: fungi imperfecti with pycnidia, acervuli, and stromata. Kew: Commonwealth Mycological Institute. Swofford DL (2003) PAUP*: phylogenetic analysis using parsimony (*and other methods). Version 4. Sunderland, MA: Sinauer Associates. Thambugala KM, Ariyawansa HA, Li Y-M, Boonmee S, Hongsanan S, et al. (2014) Dothideales. Fungal Diversity 68: Vilgalys R, Hester M (1990) Rapid genetic identification and mapping of enzymatically amplified ribosomal DNA from several Cryptococcus species. Journal of Bacteriology 172: White TJ, Bruns T, Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: PCR Protocols: a guide to molecular methods and applications (Innis MA, Gelfand DH, Sninsky JJ, White JW, eds): San Diego: Academic Press. Wijayawardene NN, Hyde KD, Bhat DJ, Camporesi E, Schumacher RK, et al. (2014) Camarosporium-like species are polyphyletic in Pleosporales; introducing Paracamarosporium and Pseudocamarosporium gen. nov. in Montagnulaceae. Cryptogamie, Mycologie 35: Wijayawardene NN, Hyde KD, Wanasinghe DN, Papizadeh M, Goonasekara ID, et al. (2016) Taxonomy and phylogeny of dematiaceous coelomycetes. Fungal Diversity 77: IMA FUNGUS

220 doi: /imafungus IMA FUNGUS 8(1): (2017) Diaporthe is paraphyletic Yahui Gao 1, 2 *, Fang Liu 1 *, Weijun Duan 3, Pedro W. Crous 4,5, and Lei Cai 1, 2 1 State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, Beijing , P.R. China 2 University of Chinese Academy of Sciences, Beijing , P.R. China; corresponding author cail@im.ac.cn 3 Ningbo Academy of Inspection and Quarantine, Zhejiang , P.R. China 4 Westerdijk Fungal Biodiversity Institute, Uppsalalaan 8, 3584CT Utrecht, The Netherlands 5 Department of Microbiology and Plant Pathology, Tree Protection Co-operative Programme, Forestry and Agricultural Biotechnology Institute, University of Pretoria, Pretoria 0002, South Africa *These authors contributed equally to this work. Abstract: Previous studies have shown that our understanding of species diversity within Diaporthe (syn. Phomopsis) is limited. In this study, 49 strains obtained from different countries were subjected to DNA sequence analysis. Based on these results, eight new species names are introduced for lineages represented by multiple strains and distinct morphology. Twelve Phomopsis species previously described from China were subjected to DNA sequence analysis, and confirmed to belong to Diaporthe. The genus Diaporthe is shown to be paraphyletic based on multi-locus (LSU, ITS and TEF1) phylogenetic analysis. Several morphologically distinct genera, namely Mazzantia, Ophiodiaporthe, Pustulomyces, Phaeocytostroma, and Stenocarpella, are embedded within Diaporthe s. lat., indicating divergent morphological evolution. However, splitting Diaporthe into many smaller genera to achieve monophyly is still premature, and further collections and phylogenetic datasets need to be obtained to address this situation. Key words: Ascomycota Diaporthales Phomopsis phylogeny taxonomy Article info: Submitted: 25 March 2017; Accepted: 22 May 2017; Published: 1 June INTRODUCTION Species of Diaporthe are known as important plant pathogens, endophytes or saprobes (Udayanga et al. 2011, Gomes et al. 2013). They have broad host ranges, and occur on many plant hosts, including cultivated crops, trees, and ornamentals (Diogo et al. 2010, Thompson et al. 2011, Gomes et al. 2013, Huang et al. 2015). Some Diaporthe species are responsible for severe diebacks, cankers, leaf-spots, blights, decay or wilts on different plant hosts, several of which are economically important (Mostert et al. 2001, Van Rensburg et al. 2006, Thompson et al. 2011, Gomes et al. 2013), leading to serious diseases and significant yield losses (Santos et al. 2011). For example, Diaporthe helianthi is the cause of one of the most important diseases of sunflower (Helianthus annuus) worldwide, and has reduced production by up to 40 % in Europe (Masirevic & Gulya 1992, Thompson et al. 2011). Diaporthe neoviticola and D. vitimegaspora, the causal agents of leaf-spot and swelling arm, are known as severe pathogens of grapevines (Vitis vinifera) (Van Niekerk et al. 2005). Úrbez-Torres et al. (2013) indicated that D. neoviticola was one of the most prevalent fungi isolated from grapevine perennial cankers in declining vines. Diaporthe scabra has been reported causing cankers and dieback on London plane (Platanus acerifolia) in Italy (Grasso et al. 2012). Symptoms of umbel browning and necrosis caused by D. angeliace have been regularly observed on carrots in France, resulting in seed production losses since 2007 (Ménard et al. 2014). Avocado (Persea americana), cultivated worldwide in tropical and subtropical regions, is threatened by branch cankers and fruit stem-end rot diseases caused by D. foeniculina and D. sterilis (Guarnaccia et al. 2016). Furthermore, species of Diaporthe are commonly introduced into new areas as endophytes or latent pathogens along with plant produce. For instance, Torres et al. (2016) reported D. rudis causing stemend rot in avocados in Chile, which was imported via avocado fruit from California (USA). Some endophytes have been shown to act as opportunistic plant pathogens. Diaporthe foeniculina (syn. P. theicola), which is a common endophyte, has been shown to cause stem and shoot cankers on sweet chestnut (Castanea sativa) in Italy (Annesi et al. 2015, Huang et al. 2015). Because of this unique ecology and potential role as plant pathogens, it is of paramount importance to accurately identify species of Diaporthe to facilitate disease surveillance, control, and trade. The initial species concept of Diaporthe based on the assumption of host-specificity, resulted in the introduction of more than 1000 names ( Names/Names.asp); (Gomes et al. 2013, Gao et al. 2016). In recent years, however, a polyphasic approach employing multi-locus DNA data together with morphology and ecology has been employed for species delimitation in the genus 2017 International Mycological Association You are free to share - to copy, distribute and transmit the work, under the following conditions: Attribution: You must attribute the work in the manner specified by the author or licensor (but not in any way that suggests that they endorse you or your use of the work). Non-commercial: You may not use this work for commercial purposes. No derivative works: You may not alter, transform, or build upon this work. 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221 Gao et al. (Udayanga et al. 2011, Gomes et al. 2013). The nuclear ribosomal internal transcribed spacer (ITS), the translation elongation factor 1-α (TEF1), β-tubulin (TUB), histone H3 (HIS), and calmodulin (CAL) genes are the most commonly used molecular loci for the identification of Diaporthe spp. (Dissanayake et al. 2015, Udayanga et al. 2015, Huang et al. 2015, Santos et al. 2017). Furthermore, molecular marker aids are being used to rapidly identify Diaporthe species which tend to be morphologically conserved (Udayanga et al. 2012, Tan et al. 2013, Lombard et al. 2014, Thompson et al. 2015, Huang et al. 2015). However, defining species boundaries remains a major challenge in Diaporthe (Huang et al. 2015), which may be a consequence of limited sampling or the use of DNA loci with insufficient phylogenetic resolution (Liu et al. 2016). It has therefore been proposed that new species in the genus should be introduced with caution, and that multiple strains from different origins should be subjected to a multi-gene phylogenetic analysis to determine intraspecific variation (Liu et al. 2016). The generic relationships of Diaporthe with other genera in Diaporthaceae remain unclear. The family name Diaporthaceae was established by Wehmeyer (1926) to accommodate Diaporthe, Mazzantia, Melanconis, and some other genera, mainly based on morphological characters such as the position, structure, and arrangement of ascomata, stroma, and spore shapes. Castlebury et al. (2002) reported that Diaporthaceae comprised Diaporthe and Mazzantia based on LSU DNA sequence data, removing other genera to different families in Diaporthales. Additional genera subsequently placed in the Diaporthaceae include Leucodiaporthe (Vasilyeva et al. 2007), Stenocarpella (Crous et al. 2006), Phaeocytostroma (Lamprecht et al. 2011), Ophiodiaporthe (Fu et al. 2013), and Pustulomyces (Dai et al. 2014). All the above genera were represented by a few species or are monotypic. Although they appeared to be morphologically divergent from Diaporthe, their phylogenetic relationships remain unclear. About 991 names of Diaporthe and 979 of Phomopsis have been established to date ( org/names/names.asp). Among them, many old epithets lack molecular data, and few morphological characters can be used in species delimitation, making it difficult to merge these names to advance to the one name scenario (Rossman et al. 2014, 2015). In China, more than 50 plant pathogenic Phomopsis species have been published to date (Chi et al. 2007). In order to stabilize these species names in the genus Diaporthe, here we introduce 12 new combinations for Phomopsis species that have been subjected to DNA sequencing, and whose phylogenetic position has been resolved in Diaporthe in the present study. The objectives of this study were: (1) to examine the phylogenetic relationships of Diaporthe with other closely related genera in Diaporthaceae; (2) to introduce new species in Diaporthe; and (3) to transfer Phomopsis species described from China to Diaporthe based on morphological and newly generated molecular data. MATERIAL AND METHODS Isolates Strains were isolated from leaves of both symptomatic and healthy plant tissues from Yunnan, Zhejiang, and Jiangxi Provinces in China. A few other strains were obtained via the Ningbo Entry-Exit Inspection and Quarantine Bureau, which were isolated from imported plants from other countries. Single spore isolations were conducted from diseased leaves with visible fungal sporulation following the protocol of Zhang et al. (2013), and isolation from surface sterilized leaf tissues was conducted following the protocol of Gao et al. (2014). Fungal endophytes were isolated according to the method described by Liu et al. (2015). The Diaporthe strains were primarily identified from the other fungal species based on cultural characteristics on PDA, spore morphology, and ITS sequence data. Type specimens of new species were deposited in the Mycological Herbarium, Microbiology Institute, Chinese Academy of Sciences, Beijing, China (HMAS), with ex-type living cultures deposited in the China General Microbiological Culture Collection Center (CGMCC). Morphological analysis Cultures were incubated on PDA at 25 C under ambient daylight and growth rates were measured daily for 7 d. To induce sporulation, isolates were inoculated on PNA (pine needle agar; Smith et al. 1996) containing doubleautoclaved (30 min, 121 C, 1 bar) healthy pine needles and incubated at a room temperature of ca. 25 o C (Su et al. 2012). Cultures were examined periodically for the development of conidiomata and perithecia. Conidia were taken from pycnidia and mounted in sterilized water. The shape and size of microscopic structures were observed and noted using a light microscope (Nikon Eclipse 80i) with differential interference contrast (DIC). At least 10 conidiomata, 30 conidiophores, alpha and beta conidia were measured to calculate the mean size and standard deviation (SD). DNA extraction, PCR amplification and sequencing Isolates were grown on PDA and incubated at 25 C for 7 d. Genomic DNA was extracted following the protocol of Cubero et al. (1999). The quality and quantity of DNA was estimated visually by staining with GelRed after 1 % agarose gel electrophoresis. The primers ITS5 and ITS4 (White et al. 1990) were used to amplify the internal transcribed spacer region (ITS) of the nuclear ribosomal RNA gene operon, including the 3 end of the 18S nrrna, the first internal transcribed spacer region, the 5.8S nrrna gene; the second internal transcribed spacer region and the 5 end of the 28S nrrna gene. The primers EF1-728F and EF1-986R (Carbone & Kohn 1999) were used to amplify part of the translation elongation factor 1-α gene (TEF1), and the primers CYLH3F (Crous et al. 2004) and H3-1b (Glass & Donaldson 1995) were used to amplify part of the histone H3 (HIS) gene. The primers T1 (O Donnell & Cigelnik 1997) and Bt2b (Glass & Donaldson 1995) were used to amplify the beta-tubulin gene (TUB); the additional combination of Bt2a/Bt2b (Glass & Donaldson 1995) was used in case of amplification failure of the T1/Bt2b primer pair. The primer pair CAL228F/CAL737R 154 IMA FUNGUS

222 Diaporthe is paraphyletic Table 1. Sources of isolates and GenBank accession numbers used in the phylogenetic analyses of Diaporthaceae. Species names * Culture collection Isolation sources Country GenBank Accession Numbers References no. ITS LSU TEF1 D. acaciigena CBS (extype) Acacia retinodes Australia KC KC Gomes et al. (2013) D. ampelina FAU 586 Vitis sp. USA: New - AF York D. angelicae CBS Heracleum sphondylium Austria KC KC Gomes et al. (2013) AR 3724 Heracleum sphondylium Austria KC KC Gomes et al. (2013) D. apiculata LC 3418 (ex-type) Camellia sinensis China KP KY KP This study LC 3452 Camellia sinensis China KP KY KP This study D. arecae complex LC 4155 Rhododendron sp. China KY KY KY This study LC 4159 Rhododendron sp. China KY KY KY This study LC 4164 Unknown host China KY KY KY This study D. biguttusis LC 1106 (ex-type) Lithocarpus glaber China KF KY KF This study D. compacta LC 3078 Camellia sinensis China KP KY KP This study LC 3083 (ex-type) Camellia sinensis China KP KY KP This study LC 3084 Camellia sinensis China KP KY KP This study D. decedens CBS Corylus avellana Austria KC KC Gomes et al. (2013) D. detrusa CBS Berberis vulgaris Austria KC KC Gomes et al. (2013) D. discoidispora LC 3503 Camellia sinensis China KY KY KY This study D. elaeagni-glabrae LC 4802 (ex-type) Elaeagnus glabra China KX KY KX This study LC 4806 Elaeagnus glabra China KX KY KX This study D. ellipicola LC 0810 (ex-type) Lithocarpus glaber China KF KY KF This study D. eres LC 3198 Camellia sinensis China KP KY KP This study LC 3205 Camellia sinensis China KP KY KP This study LC 3206 Camellia sinensis China KP KY KP This study CBS Acer campestre Austria KC KC Gomes et al. (2013) D. fusicola LC 1126 Lithocarpus glaber China KF KY KF This study LC 0778 (ex-type) Lithocarpus glaber China KF KY KF This study D. hongkongensis LC 0784 Lithocarpus glaber China KC KY KC This study LC 0812 Smilax china China KC KY KC This study D. incompleta LC 6706 Camellia sinensis China KX KY KX This study LC 1127 (ex-type) Lithocarpus glaber China KF KY KF This study D. mahothocarpi LC 0732 Mahonia bealei China KC KY KC This study LC 0763 (ex-type) Lithocarpus glaber China KC KY KC This study D. masirevicii Diaporthe sp. Camellia sinensis China KY KY KY This study D. neoarctii CBS Ambrosia trifida USA: New Jersey KC KC Gomes et al. (2013) D. oncostoma CBS Robinia pseudoacacia Russia KC KC Gomes et al. (2013) D. oraccinii LC 3166 (ex-type) Camellia sinensis China KP KY KP This study LC 3172 Camellia sinensis China KP KY KP This study LC 3296 Camellia sinensis China KP KY KP This study D. ovoicicola LC 1128 (ex-type) Lithocarpus glaber China KF KY KF This study D. penetriteum LC 3215 Camellia sinensis China KP KY KP This study LC 3353 (ex-type) Camellia sinensis China KP KY KP This study LC 3394 Camellia sinensis China KP KY KP This study D. perjuncta CBS Ulmus glabra Austria KC KC Gomes et al. (2013) VOLUME 8 NO

223 Gao et al. Table 1. (Continued). Species names * Culture collection no. Isolation sources Country GenBank Accession Numbers References ITS LSU TEF1 D. pseudophoenicicola LC 6150 Phoenix canariensis China KY KY KY This study LC 6151 Phoenix canariensis China KY KY KY This study D. pustulata CBS Acer pseudoplatanus Austria KC KC Gomes et al. (2013) CBS Acer pseudoplatanus Austria KC KC Gomes et al. (2013) CBS Prunus padus Austria KC KC Gomes et al. (2013) D. rudis LC 6147 Dendrobenthamia USA KY KY KY This study japonica LC 6145 Ilex aquifolium China KY KY KY This study D. saccarata CBS Protea repens, cankers South Africa KC KC34391 Gomes et al. (2013) D. sclerotioides CBS Cucumis sativus Netherlands KC KC Gomes et al. (2013) D. tectonendophytica LC 6623 Unknown host China KX KY KX This study D. tectonigena LC 6512 Camellia sinensis China KX KY KX This study D. ternstroemiae LC 0777 (ex-type) Ternstroemia China KC KY KC This study gymnanthera D. ueckerae LC 3564 Camellia sinensis China KP KY KP This study D. undulata LC 6624 Unknown host China KX KY KX This study D. velutina LC 4414 Lithocapus sp. China KX KY KX This study LC 4419 Neolitsea sp. China KX KY KX This study LC 4421 (ex-type) Neolitsea sp. China KX KY KX This study D. xishuangbanica LC 6707 Camellia sinensis China KX KY KX This study LC 6744 Camellia sinensis China KX KY KX This study D. yunnanensis LC 6168 Coffea sp. China KX KY KX This study Diaporthe sp. LC 3156 Camellia sinensis China KP KY KP This study LC 6170 Coffea sp. China KY KY KY This study LC 6171 Solanum melongena China KY KY KY This study LC 6232 Theobroma cacao China KX KY KX This study Mazzantia napelli AR 3498 Aconitum vulparia Austria - AF EU Castlebury et al. (2002) Ophiodiaporthe cyatheae Phaeocytostroma ambiguum BCRC Cyathea lepifera Taiwan JX JX KC Fu et al. (2013) CPC Zea mays South Africa FR FR Lamprecht et al. (2011) CPC Zea mays South Africa FR FR FR Lamprecht et al. (2011) Ph. plurivorum CBS Helianthus annuus Portugal FR FR FR Lamprecht et al. (2011) Ph. sacchari CBS Japan FR FR FR Lamprecht et al. (2011) Ph. megalosporum CBS Rice-field soil India FR FR FR Lamprecht et al. (2011) Pustulomyces bambusicola Stenocarpella macrospora MFLUCC CBS on dead culm of bamboo Rain damaged Bt maize hybrid, season S. maydis CBS Traditional/landrace maize from 2003/04 season Thailand - KF KF Dai et al. (2014) South Africa FR DQ Lamprecht et al. (2011) South Africa FR DQ FR Lamprecht et al. (2011) Valsa ambiens CFCC Pyrus bretschneideri China KR KR KU Fan et al. (2014) *New species described in this paper are shown in bold. 156 IMA FUNGUS

224 Diaporthe is paraphyletic (Carbone & Kohn 1999) and LR0R/LR5 primer pair (Rytas & Mark 1990) were used to amplify the calmodulin gene (CAL) and the LSU rdna, respectively. Amplification reactions of 25 μl were composed of 10 EasyTaq buffer (MgCl 2+ included; Transgen, Beijing), 50 μm dntps, 0.2 μm of each forward and reverse primers (Transgen), 0.5 U EasyTaq DNA polymerase (Transgen) and 1 10 ng of genomic DNA. PCR parameters were as follows: 94 C for 5 min, followed by 35 cycles of denaturation at 94 C for 30 s, annealing at a suitable temperature for 30 s (52 C for ITS and LSU, 56 C for CAL, HIS, TEF1 and TUB), extension at 72 C for 30 s and a final elongation step at 72 C for 10 min. DNA sequencing was performed by Omegagenetics Company, Beijing. Phylogenetic analyses The DNA sequences generated with forward and reverse primers were used to obtain consensus sequences using MEGA v. 5.1 (Tamura et al. 2011), and subsequently aligned using MAFFT v. 6 (Katoh & Toh 2010); alignments were manually edited using MEGA v. 5.1 when necessary. Two datasets were employed in the phylogenetic analyses. LSU, ITS and TEF1 loci were selected to infer the generic relationships within Diaporthaceae (Table 1), with Valsa ambiens as outgroup. All available sequences of Diaporthe species were included in the dataset of combined ITS, HIS, TEF1, TUB, and CAL regions to infer the interspecific relationships within Diaporthe (Table 2) with Diaporthella corylina as outgroup. Maximum likelihood (ML) gene trees were estimated using the software RAxML v Black Box (Stamatakis 2006, Stamatakis et al. 2008). The RAxML software selected the GTR model of nucleotide substitution with the additional options of modelling rate heterogeneity (Γ) and proportion invariable sites (I). Bayesian analyses (critical value for the topological convergence diagnostic set to 0.01) were performed on the concatenated loci using MrBayes v (Ronquist et al. 2012) as described by (Crous et al. 2006) using nucleotide substitution models for each data partition selected by jmodeltest (Darriba et al. 2012) and MrModeltest v. 2.3 (Nylander 2004). Bayesian analyses were launched with random starting trees for generations, and Markov chains were sampled every 1000 generations. The first 25 % resulting trees were discarded as burn-in. The remaining trees were summarized to calculate the posterior probabilities (PP) of each clade being monophyletic. Trees were visualized in FigTree v ( software/). New sequences generated in this study were deposited in NCBI's GenBank nucleotide database (www. ncbi.nlm.nih.gov; Table 1). RESULTS Collection of Diaporthe strains Twenty-one Diaporthe strains including presumed plant pathogens and endophytes were isolated from 11 different host plant species (Table 2) collected from three provinces (Jiangxi, Yunnan, Zhejiang) in the northern part of China. In addition, 28 strains were isolated from the plant samples inspected by Jiangsu Entry-Exit Inspection and Quarantine Bureau. The paraphyly of Diaporthe Phylogenetic analysis was conducted with 224 sequences derived from 76 ingroup taxa from Diaporthaceae with Valsa ambiens as the outgroup (Table 1). The combined alignment comprised characters including gaps (795 for LSU, 558 for ITS, 464 for TEF1). Based on the results of the Mrmodeltest, the following priors were set in MrBayes for the different data partitions: GTR+G models with gamma-distributed rates were implemented for LSU and ITS, HKY+I+G model with invgamma-distributed rates were implemented for TEF1. The Bayesian analysis lasted generations and the consensus tress and posterior probabilities were calculated from the trees left after discarding the first 25 % generations for burn-in (Fig. 1). The generic relationships of Mazzantia, Ophiodiaporthe, Phaeocytostroma, Pustulomyces, and Stenocarpella with Diaporthe from this analysis are shown in Fig. 1. The topology and branching order of the phylogenetic trees inferred from ML and Bayesian methods were essentially similar. Five genera from Diaporthaceae did not form discrete clades from Diaporthe species but are scattered in the latter, although the family remains monophyletic. The paraphyletic nature of Diaporthe, however, is demonstrated (Fig. 1). Ophiodiaporthe formed a well resolved and distinct clade represented by strain YMJ 1364, and clustered together with the ex-type culture of D. sclerotioides (CBS ) (BPP 0.99, MLBS: 90). Stenocarpella, represented by S. maydis and S. macrospora, was well supported (BPP 1, MLBS = 96) and closely related to several species of Phaeocytostroma. Mazzantia, however, was poorly supported for its phylogenetic position in Diaporthaceae (Fig. 1). Phylogenetic analyses of the combined datasets of Diaporthe species In total, 1089 sequences derived from 273 ingroup taxa were combined and Diaporthella corylina was used as outgroup. A total of 2783 characters including gaps (568 for CAL, 554 for HIS, 523 for ITS, 636 for TEF1 and 456 for TUB) were included in the multi-locus dataset, comprising sequences generated from this study and others downloaded from GenBank (Table 2). For the Bayesian inference, GTR+I+G model was selected for CAL, HIS and ITS, HKY+I+G for TEF1 and TUB through the analysis of Mrmodeltest. The maximum likelihood tree conducted by the GTR model confirmed the tree topology and posterior probabilities of the Bayesian consensus tree. The topology and branching order for the phylogenetic trees inferred from ML and Bayesian methods were essentially similar (Fig. 2). Based on the multi-locus phylogeny and morphology, 49 strains were assigned to 13 species, including eight taxa which we describe here as new (Fig. 2). VOLUME 8 NO

225 Gao et al. D. ellipicola LC 0810 D. eres LC 3198 D. biguttusis LC 1106 D. eres CBS D. longicicola LC 1127 D. eres LC /1 D. eres LC /1 D. mahothocarpus LC /1 D. mahothocarpus LC 0732 D. penetriteum LC 3394 D. penetriteum LC 3215 D. penetriteum LC /1 100 D. apiculata LC 3452 D. apiculata LC /0.99 D. oraccinii LC /1 D. oraccinii LC 3166 D. oraccinii LC /1 D. rudis LC 6145 D. rudis LC /1 D. arecae complex LC /0.99 D. arecae complex LC 4155 D. pseudophoenicicola LC / D. pseudophoenicicola LC 6151 D. arecae complex LC 4164 D. hongkongensis LC /0.99 D. hongkongensis LC /0.99 D. xishuangbanica LC 6744 D. xishuangbanica LC /0.99 D. tectonigena LC 6512 D. undulata LC / /1 D. velutina LC 4419 D. velutina LC /0.99 D. velutina LC /1 D. elaeagnicola LC /1 D. elaeagnicola LC D. saccarata CBS D. oncostoma CBS D. decedens CBS D. ampelina FAU /1 D. angelicae CBS /1 D. angelicae AR /0.99 D. neoarctii CBS D. compacta LC /1D. compacta LC /0.99 D. compacta LC /1 99/1 98/1 D. ueckerae LC 3564 D. tectonendophytica LC 6623 Diaporthe sp. LC 6740 Diaporthe sp. LC /0.99 D. sclerotioides CBS / Ophiodiaporthe cyatheae BCRC Diaporthe sp. LC 6232 D. yunnanensis LC / D. discoidispora LC 3503 Diaporthe sp. LC 6170 D. incompleta LC /1Phaeocytostroma ambiguum* CPC /1 Phaeocytostroma ambiguum CPC Phaeocytostroma plurivorum CBS /1 Stenocarpella maydis* CBS Stenocarpella macrospora* CBS /1 Phaeocytostroma megalosporum CBS Pustulomyces_bambusicola MFLUCC 11_0436 Phaeocytostroma sacchari CBS D. detrusa CBS Mazzantia napelli AR3498 D. perjuncta CBS D. ovoicicola LC 1128 D. ternstroemiae LC 0777 D. fusicola LC /0.99 Diaporthe sp. LC D. fusicola LC 0778 D. acaciigena CBS D. pustulata CBS /1 D. pustulata CBS D. pustulata CBS Valsa ambiens CFCC Fig. 1. Phylogenetic tree of the family Diaporthaceae from a maximum likelihood analysis based on the combined multi-locus dataset (ITS, LSU, TEF1). The ML bootstrap values 70 %, bayesian probabilities BPP 0.90 are marked above the branches. The tree is rooted with Valsa ambiens. 158 IMA FUNGUS

226 Diaporthe is paraphyletic Fig. 2. Phylogenetic tree of the genus Diaporthe from a maximum likelihood analysis based on the combined multi-locus dataset (CAL, HIS, ITS, TEF1, TUB). The ML bootstrap values 70 %, bayesian probabilities BPP 0.90 are marked above the branches. The tree is rooted with Diaporthella corylina. The novel species are highlighted. VOLUME 8 NO

227 Gao et al. Fig. 2. (Continued). 160 IMA FUNGUS

228 Diaporthe is paraphyletic Fig. 2. (Continued). VOLUME 8 NO

229 Gao et al. Fig. 2. (Continued). 162 IMA FUNGUS

230 Diaporthe is paraphyletic Table 2. Sources of isolates and GenBank accession numbers used in the phylogenetic analyses of Diaporthe. Newly sequenced material is indicated in bold type. Species names * Culture collection no. Isolation sources Host family GenBank Accession Numbers References ITS TEF1 TUB HIS CAL D. acaciigena CBS (ex-type) Acacia retinodes Mimosaceae KC KC KC KC KC Gomes et al. (2013) D. acerina CBS Acer saccharum Aceraceae KC KC KC KC KC Gomes et al. (2013) D. acutispora CGMCC = LC 6161 Coffea sp., endophyte Rubiaceae KX KX KX KX KX This study LC 6142 Camellia sasanqua, endophyte Theaceae KX KX KX KX KX This study LC 6160 Camellia sasanqua, endophyte Theaceae KX KX KX KX KX This study D. alleghaniensis CBS (ex-type) Betula alleghaniensis, branches Betulaceae KC KC KC KC KC Gomes et al. (2013) D. alnea CBS (ex-type) Alnus sp. Betulaceae KC KC KC KC KC Gomes et al. (2013) CBS Alnus sp. Betulaceae KC KC KC KC KC Gomes et al. (2013) D. ambigua CBS Pyrus communis Rosaceae KC KC KC KC KC Gomes et al. (2013) CBS Aspalathus linearis, crown Fabaceae KC KC KC KC KC Gomes et al. (2013) D. ampelina CBS Vitis vinifera Vitaceae AF AY JX AY Gomes et al. (2013) CBS Vitis vinifera Vitaceae KC KC KC KC KC Gomes et al. (2013) D. amygdali CBS (ex-type) Prunus dulcis Rosaceae KC KC KC KC KC Gomes et al. (2013) CBS Vitis vinifera Vitaceae KC KC KC KC KC Gomes et al. (2013) D. anacardii CBS (ex-epitype) Anacardium occidentale Anacardiaceae KC KC KC KC KC Gomes et al. (2013) D. angelicae CBS (ex-epitype) Heracleum sphondylium Apiaceae KC KC KC KC KC Gomes et al. (2013) CBS Foeniculum vulgare Apiaceae KC KC KC KC KC Gomes et al. (2013) D. apiculata LC 4152 Camellia, leaf Theaceae KP KP KP KP Gao et al. (2016) LC 3418, (ex-type) Camellia sinensis, leaf, endophyte Theaceae KP KP KP KP Gao et al. (2016) D. arctii CBS Arctium sp. Arecaceae KC KC KC KC KC Gomes et al. (2013) D. arecae CBS Citrus sp., fruit Rutaceae KC KC KC KC KC Gomes et al. (2013) CBS (ex-isotype) Areca catechu, fruit Arecaceae KC KC KC KC KC Gomes et al. (2013) D. arengae CBS (ex-type) Arenga engleri Arecaceae KC KC KC KC KC Gomes et al. (2013) D. asheiola CBS , CPC 16508, Vaccinium ashei Ericaceae KJ KJ KJ KJ Lombard et al. (2014) (ex-type) CBS , CPC Vaccinium ashei Ericaceae KJ KJ KJ KJ Lombard et al. (2014) D. aspalathi CBS Aspalathus linearis Fabaceae KC KC KC KC KC Gomes et al. (2013) CBS , (ex-type) Aspalathus linearis Fabaceae KC KC KC KC KC Gomes et al. (2013) D. australafricana CBS Vitis vinifera Vitaceae KC KC KC KC KC Gomes et al. (2013) CBS Vitis vinifera Vitaceae KC KC KC KC KC Gomes et al. (2013) D. baccae CBS Vaccinium corymbosum Ericaceae KJ KJ Lombard et al. (2014) CBS (ex-type) Vaccinium corymbosum Ericaceae KJ KJ Lombard et al. (2014) VOLUME 8 NO

231 Gao et al. Table 2. (Continued). Species names * Culture collection no. Isolation sources Host family GenBank Accession Numbers References ITS TEF1 TUB HIS CAL D. batatas CBS Ipomoea batatas Convolvulaceae KC KC KC KC KC Gomes et al. (2013) D. beckhausii CBS Viburnum sp. Caprifoliaceae KC KC KC KC KC Gomes et al. (2013) D. beilharziae BRIP (ex-type) Indigofera australis Papilionaceae JX JX KF Thompson et al. (2015) D. benedicti CFCC (ex-type) Juglans mandshurica Juglandaceae KP KP KP KP KP Fan et al. (2015) CFCC Juglans mandshurica Juglandaceae KP KP KP KP KP Fan et al. (2015) D. betulae CFCC (ex-type) Betula platyphylla Betulaceae KT KT KT KT KT Du et al. (2016) CFCC Betula platyphylla Betulaceae KT KT KT KT KT Du et al. (2016) D. betulicola CFCC (ex-type) Betula albosinensis Betulaceae KX KX KX KX KX Du et al. (2016) CFCC Betula albosinensis Betulaceae KX KX KX KX KX Du et al. (2016) D. bicincta DP0659, CBS Juglans sp., dead wood Juglandaceae KC KC KC KC Udayanga et al. (2014a) D. biconispora ZJUD 60, CGMCC Citrus sinensis Rutaceae KJ KJ KJ KJ Huang et al. (2015) ZJUD 61, CGMCC Fortunella margarita Rutaceae KJ KJ KJ KJ Huang et al. (2015) ZJUD 62, CGMCC Citrus grandis Rutaceae KJ KJ KJ KJ Huang et al. (2015) D. biguttulata ZJUD 47, CGMCC Citrus limon Rutaceae KJ KJ KJ KJ Huang et al. (2015) (ex-type) ZJUD 48, CGMCC Citrus limon Rutaceae KJ KJ KJ KJ Huang et al. (2015) D. biguttusis CGMCC (ex-type) Lithocarpus glabra Fagaceae KF KF KF Gao et al. (2015) D. brasiliensis CBS (ex-type) Aspidosperma tomentosus Apocynaceae KC KC KC KC KC Gomes et al., 2013 LGMF 926 Aspidosperma tomentosus Apocynaceae KC KC KC KC KC Gomes et al., 2013 D. canthii CBS (ex-type) Canthium inerme Rubiaceae JX KC KC KC Du et al. (2016) D. carpini CBS Carpinus betulus Corylaceae KC KC KC KC KC Gomes et al. (2013) D. caulivora CBS (ex-neotype) Glycine max Fabaceae KC KC KC KC KC Gomes et al. (2013) CBS Glycine soja Fabaceae KC KC KC KC KC Gomes et al. (2013) D. celastrina CBS Celastrus scandens Celastraceae KC KC KC KC Gomes et al. (2013) D. cf. heveae 1 CBS Hevea brasiliensis Euphorbiaceae KC KC KC KC KC Gomes et al. (2013) D. cf. heveae 2 CBS Hevea brasilliensis, leaf Euphorbiaceae KC KC KC KC KC Gomes et al. (2013) D. chamaeropis CBS Chamaerops humilis, dead part Arecaceae KC KC KC KC KC Gomes et al. (2013) of leaf CBS Spartium junceum, dead branch Fabaceae KC KC KC KC KC Gomes et al. (2013) D. charlesworthii BRIP 4884m (ex-type) Rapistrum rugostrum Brassicaceae KJ KJ KJ Thompson et al. (2015) D. cinerascens CBS Ficus carica Moraceae KC KC KC KC KC Gomes et al. (2013) D. citri CBS Citrus sinensis Rutaceae KC KC KC KC KC Gomes et al. (2013) CBS KC KC KC KC KC Gomes et al. (2013) AR 3405 Citrus sp. Rutaceae KC KC KC KJ Udayanga et al. (2014b) 164 IMA FUNGUS

232 Diaporthe is paraphyletic Table 2. (Continued). Species names * Culture collection no. Isolation sources Host family GenBank Accession Numbers References ITS TEF1 TUB HIS CAL D. citriasiana ZJUD 30 (ex-type) Citrus unshiu, dead wood Rutaceae JQ JQ KC KC Huang et al. (2015) ZJUD 33 Citrus paradise, stem-end rot Rutaceae JQ JQ KC KC Huang et al. (2015) fruit D. citrichinensis ZJUD 34 Citrus sp. Rutaceae JQ JQ KC Huang et al. (2015) ZJUD 35 Citrus unshiu, dead wood Rutaceae JQ JQ KC KC Huang et al. (2015) ZJUD 36 Citrus unshiu, dead wood Rutaceae KC KC KC KC Huang et al. (2015) D. compacta LC3083 (ex-type) Camellia sinensis, leaf, Theaceae KP KP KP KP Gao et al. (2016) endophyte LC3084 Camellia sinensis, leaf, Theaceae KP KP KP KP Gao et al. (2016) endophyte D. convolvuli CBS Convolvulus arvensis Convolvulaceae KC KC KC KC KC Huang et al. (2015) D. crataegi CBS Crataegus oxyacantha Rosaceae KC KC KC KC KC Gomes et al. (2013) D. crotalariae CBS (ex-type) Crotalaria spectabilis Fabaceae KC KC KC KC KC Gomes et al. (2013) D. cuppatae CBS Aspalathus linearis Fabaceae KC KC KC KC KC Gomes et al. (2013) D. cynaroidis CBS Protea cynaroides Proteaceae KC KC KC KC KC Gomes et al. (2013) D. cytosporella AR 5149 Citrus sinensis Rutaceae KC KC KC KC Udayanga et al. (2014b) D. decedens CBS Corylus avellana Corylaceae KC KC KC KC KC Gomes et al. (2013) CBS Corylus avellana Corylaceae KC KC KC KC KC Gomes et al. (2013) D. detrusa CBS Berberis vulgaris Berberidaceae KC KC KC KC KC Gomes et al. (2013) CBS Berberis vulgaris Berberidaceae KC KC KC KC KC Gomes et al. (2013) D. discoidspora ZJUD 87, CGMCC Citrus sinensis Rutaceae KJ KJ KJ KJ Huang et al. (2015) ZJUD 89, CGMCC Citrus unshiu Rutaceae KJ KJ KJ KJ Huang et al. (2015) D. elaeagni CBS Elaeagnus sp., twig Elaeagnaceae KC KC KC KC KC Gomes et al. (2013) D. elaeagni-glabrae CGMCC = LC 4802 Elaeagnus glabra, pathogen Elaeagnaceae KX KX KX KX KX This study LC 4806 Elaeagnus glabra, pathogen Elaeagnaceae KX KX KX KX KX This study D. ellipicola CGMCC (ex-type) Lithocarpus glabra, diseased Fagaceae KF KF KF Gao et al. (2015) leaves D. endophytica CBS (ex-type) Schinus terebinthifolius Anacardiaceae KC KC KC KC KC Gomes et al. (2013) LGMF 911 Schinus terebinthifolius Anacardiaceae KC KC KC KC KC Gomes et al. (2013) D. eres AR5193, CBS (exepitype) Ulmus laevis Ulmaceae KJ KJ KJ KJ Udayanga et al. (2014a) CBS Castanea sativa Fagaceae KC KC KC KC Udayanga et al. (2014a) D. eugeniae CBS Eugenia aromatica, leaf Mrytaceae KC KC KC KC KC Gomes et al. (2013) D. fibrosa CBS Rhamnus cathartica Rhamnaceae KC KC KC KC KC Gomes et al. (2013) CBS Rhamnus cathartica Rhamnaceae KC KC KC KC KC Gomes et al. (2013) VOLUME 8 NO

233 Gao et al. Table 2. (Continued). Species names * Culture collection no. Isolation sources Host family GenBank Accession Numbers References ITS TEF1 TUB HIS CAL D. foeniculina CBS Pyrus pyrifolia Rosaceae KC KC KC KC KC Gomes et al. (2013) Theaceae KC KC KC KC KC Gomes et al. (2013) CBS (ex-type of P. theicola) Camellia sinensis, leaves and branches CBS Foeniculum vulgare Apiaceae KC KC KC KC KC Gomes et al. (2013) D. fraxiniangustifolia BRIP (ex-epitype) Fraxinus-angustifolia subsp. Oleaceae JX JX KF Tan et al. (2013) oxycapa D. ganjae CBS (ex-type) Cannabis sativa, dead leaf Cannabaceae KC KC KC KC KC Gomes et al. (2013) D. gardeniae CBS Gardenia florida, stem Rubiaceae KC KC KC KC KC Gomes et al. (2013) D. goulteri BRIP 55657a (ex-type) Helianthus annuus Asteraceae KJ KJ KJ Thompson et al. (2015) D. gulyae BRIP (ex-type) Helianthus annuus Asteraceae JF JN KJ Thompson et al. (2015) D. helianthi CBS Helianthus annuus Asteraceae KC KC KC KC KC Gomes et al. (2013) CBS (ex-type) Helianthus annuus Asteraceae KC KC KC KC KC Gomes et al. (2013) D. helicis AR 5211 Hedera helix Araliaceae KJ KJ KJ KJ KJ Udayanga et al. (2014a) D. hickoriae CBS (ex-epitype) Carya glabra Juglandaceae KC KC KC KC KC Gomes et al. (2013) D. hongkongensis CBS (ex-type) Dichroa febrifuga, fruit Hydrangeaceae KC KC KC KC KC Gomes et al. (2013) D. hordei CBS Hordeum vulgare Poaceae KC KC KC KC KC Gomes et al. (2013) D. impulsa CBS Sorbus aucuparia Rosaceae KC KC KC KC KC Gomes et al. (2013) CBS Sorbus americana Rosaceae KC KC KC KC KC Gomes et al. (2013) D. incompleta CGMCC = LC 6754 Camellia sinensis, pathogen Theaceae KX KX KX KX KX This study LC 6706 Camellia sinensis, pathogen Theaceae KX KX KX KX This study D. inconspicua CBS (ex-type) Maytenus ilicifolia, endophytic in Celastraceae KC KC KC KC KC Gomes et al. (2013) petiole D. infecunda CBS (ex-type) Schinus terebinthifolius Anacardiaceae KC KC KC KC KC Gomes et al. (2013) LGMF 908 Schinus terebinthifolius Anacardiaceae KC KC KC KC KC Gomes et al. (2013) D. kongii BRIP (ex-type) Helianthus annuus Asteraceae JF JN KJ Thompson et al. (2011) D. lichicola BRIP (ex-type) Litchi chinensis Sapindaceae JX JX KF Tan et al. (2013) D. longicicola CGMCC (ex-type) Lithocarpus glabra Fagaceae KF KF KF Gao et al. (2015) D. longicolla FAU 599 Glycine max Fabaceae KJ KJ KJ KJ Udayanga et al. (2015) D. longispora CBS (ex-type) Ribes sp. Grossulariaceae KC KC KC KC KC Gomes et al. (2013) D. lusitanicae CBS (ex-type) Foeniculum vulgare Apiaceae KC KC KC KC Gomes et al. (2013) CBS Foeniculum vulgare Apiaceae KC KC KC KC KC Gomes et al. (2013) D. macintoshii BRIP 55064a Rapistrum rugostrum Brassicaceae KJ KJ KJ Thompson et al. (2015) D. mahothocarpus CGMCC Lithocarpus glabra Fagaceae KC KC Gao et al. (2014) D. manihotia CBS Manihot utilissima, leaves Euphorbiaceae KC KC KC KC KC Gomes et al. (2013) 166 IMA FUNGUS

234 Diaporthe is paraphyletic Table 2. (Continued). Species names * Culture collection no. Isolation sources Host family GenBank Accession Numbers References ITS TEF1 TUB HIS CAL D. maritima NB 382-2E Picea rubens needle Pinaceae KU KU KU Tanney et al. (2016) NB 463-3A Picea rubens needle Pinaceae KU KU KU Tanney et al. (2016) NB I (ex-type) Picea rubens needle Pinaceae KU KU KU Tanney et al. (2016) D. masirevicii BRIP Chrysanthemoides monilifera Rosaceae KJ KJ KJ Huang et al. (2015) subsp. rotundata BRIP 57892a (ex-type) Helianthus annuus Asteraceae KJ KJ KJ Huang et al. (2015) D. mayteni CBS (ex-type) Maytenus ilicicolia Celastraceae KC KC KC KC KC Gomes et al. (2013) D. megalospora CBS Sambucus canadensis Caprifoliaceae KC KC KC KC KC Gomes et al. (2013) D. melonis CBS Glycine soja Fabaceae KC KC KC KC KC Gomes et al. (2013) CBS (ex-isotype) Cucumis melo Cucurbitaceae KC KC KC KC KC Gomes et al. (2013) D. middletonii BRIP Chrysanthemoides monilifera Rosaceae KJ KJ KJ Thompson et al. (2015) subsp. rotundata BRIP 54884e (ex-type) Rapistrum rugostrum Brassicaceae KJ KJ KJ Thompson et al. (2015) D. miriciae BRIP 55662c Glycine max Fabaceae KJ KJ KJ Thompson et al. (2015) BRIP 54736j (ex-type) Helianthus annuus Asteraceae KJ KJ KJ Thompson et al. (2015) BRIP 56918a Vigna radiata Papilionaceae KJ KJ KJ Thompson et al. (2015) D. multigutullata ZJUD 98 Citrus grandis Rosaceae KJ KJ KJ KJ Huang et al. (2015) D. musigena CBS ; CPC Musa sp., leaves Musaceae KC KC KC KC KC Gomes et al. (2013) (ex-type) D. neilliae CBS Spiraea sp. Rosaceae KC KC KC KC KC Udayanga et al. (2014a) D. neoarctii CBS (ex-type) Ambrosia trifida Asteraceae KC KC KC KC KC Gomes et al. (2013) D. neoraonikayaporum MFLUCC Tectona grandis Verbenaceae KU KU KU KU Doilom et al. (2017) MFLUCC Tectona grandis Verbenaceae KU KU KU KU Doilom et al. (2017) MFLUCC Tectona grandis Verbenaceae KU KU KU KU Doilom et al. (2017) D. nobilis CBS Laurus nobilis, stem Lauraceae KC KC KC KC KC Gomes et al. (2013) D. nomurai CBS Morus sp. Moraceae KC KC KC KC KC Gomes et al. (2013) D. nothofagi BRIP (ex-type) Nothofagus cunninghamii Fagaceae JX JX KF Tan et al. (2013) D. novem CBS Glycine max Fabaceae KC KC KC KC KC Gomes et al. (2013) CBS (ex-type) Glycine max Fabaceae KC KC KC KC KC Gomes et al. (2013) D. oncostoma CBS Robinia pseudoacacia, leaf spot Fabaceae KC KC KC KC KC Gomes et al. (2013) CBS Robinia pseudoacacia Fabaceae KC KC KC KC KC Gomes et al. (2013) D. oraccinii LC 3166 (ex-type) Camellia sinensis, leaf, Theaceae KP KP KP KP Gao et al. (2016) endophyte LC 3296 Camellia sinensis, leaf, Theaceae KP KP KP KP Gao et al. (2016) endophyte VOLUME 8 NO

235 Gao et al. Table 2. (Continued). Species names * Culture collection no. Isolation sources Host family GenBank Accession Numbers References ITS TEF1 TUB HIS CAL D. ovalispora ZJUD 93, CGMCC Citrus limon Rosaceae KJ KJ KJ KJ Huang et al. (2015) D. oxe CBS (ex-type) Maytenus ilicifolia Celastraceae KC KC KC KC KC Gomes et al. (2013) CBS Maytenus ilicifolia Celastraceae KC KC KC KC KC Gomes et al. (2013) D. padi var. padi CBS Prunus padus Rosaceae KC KC KC KC KC Gomes et al. (2013) CBS Alnus glutinosa Betulaceae KC KC KC KC KC Gomes et al. (2013) D. paranensis CBS (ex-type) Maytenus ilicifolia Celastraceae KC KC Kc KC KC Gomes et al. (2013) D. pascoei BRIP (ex-type) Persea americana Lauraceae JX JX KF Tan et al. (2013) D. penetriteum LC 3353 Camellia sinensis, leaf Theaceae KP KP KP KP Gao et al. (2016) LC 3394 Camellia sinensis, leaf Theaceae KP KP KP KP Gao et al. (2016) D. perjuncta CBS (ex-type) Ulmus glabra Ulmaceae KC KC KC KC KC Gomes et al. (2013) D. perniciosa CBS Malus pumila, bark Rosaceae KC KC KC KC KC Gomes et al. (2013) D. perseae CBS Perseae gratissima, young fruit Lauraceae KC KC KC KC KC Gomes et al. (2013) D. phaseolorum AR 4203, CBS Phaseolus vulgaris Fabaceae KJ KJ KJ KJ Huang et al. (2015) CBS Caperonia palustris Euphorbiaceae KC KC KC KC KC Gomes et al. (2013) CBS Aster exilis Asteraceae KC KC KC KC KC Gomes et al. (2013) Podocarpaceae KX KX KX KX KX This study D. podocarpimacrophylli D. pseudomangiferae CGMCC = LC 6155 Podocarpus macrophyllus, endophyte LC 6144 Podocarpus macrophyllus, endophyte LC 6194 Podocarpus macrophyllus, endophyte LC 6197 Podocarpus macrophyllus, endophyte LC 6200 Podocarpus macrophyllus, endophyte Podocarpaceae KX KX KX KX This study Podocarpaceae KX KX KX KX KX This study Podocarpaceae KX KX KX KX KX This study Podocarpaceae KX KX KX KX KX This study LC 6229 Olea europaea, endophytes Oleaceae KX KX KX KX KX This study CBS (ex-type) Mangifera indica Anacardiaceae KC KC KC KC KC Gomes et al. (2013) CBS Mangifera indica, peel of fruit Anacardiaceae KC KC KC KC KC Gomes et al. (2013) D. pseudophoenicicola CBS (ex-type) Phoenix dactylifera, dead tops of Anacardiaceae KC KC KC KC KC Gomes et al. (2013) green leaves CBS Mangifera indica, showing Anacardiaceae KC KC KC KC KC Gomes et al. (2013) dieback D. pterocarpi MFLUCC Pterocarous indicus Papilionaceae JQ JX JX JX Udayanga et al. (2012) MFLUCC Pterocarous indicus Papilionaceae JQ JX JX JX Udayanga et al. (2012) 168 IMA FUNGUS

236 Diaporthe is paraphyletic Table 2. (Continued). Species names * Culture collection no. Isolation sources Host family GenBank Accession Numbers References ITS TEF1 TUB HIS CAL D. pterocarpicola MFLUCC a (ex-type) Piterocarpus indicus Papilionaceae JQ JX JX JX Udayanga et al. (2012) MFLUCC b Piterocarpus indicus Papilionaceae JQ JX JX JX Udayanga et al. (2012) D. pulla CBS Hedera helix Araliaceae KC KC KC KC Udayanga et al. (2014a) D. pustulata CBS Acer pseudoplatanus Aceraceae KC KC KC KC KC Gomes et al. (2013) CBS Acer pseudoplatanus Aceraceae KC KC KC KC KC Gomes et al. (2013) D. raonikayaporum CBS (ex-type) Spondias mombin Anacardiaceae KC KC KC KC KC Gomes et al. (2013) D. rhoina CBS Rhus toxicodendron Anacardiaceae KC KC KC KC KC Gomes et al. (2013) D. rudis CBS (ex-type) Vitis vinifera Vitaceae KC KC KC KC KC Machingambi et al. (2015) CBS Vitis Vinifera Vitaceae KC KC KC KC KC Machingambi et al. (2015) D. saccarata CBS (ex-type) Protea repens, cankers Proteceae KC KC KC KC KC Gomes et al. (2013) D. sackstonii BRIP 54669b (ex-type) Helianthus annuus Asteraceae KJ KJ KJ Gomes et al. (2013) D. salicicola BRIP (ex-type) Salix purpurea Salicaceae JX JX KF Gomes et al. (2013) D. schini LGMF 910, CPC Schinus terebinthifolius, Anacardiaceae KC KC KC KC KC Thompson et al. (2015) endophytic in leaf CBS (ex-type) Schinus terebinthifolius, Anacardiaceae KC KC KC KC KC Tan et al. (2013) endophytic in leaf D. sclerotioides CBS (ex-type) Cucumis sativus Cucurbitaceae KC KC KC KC KC Gomes et al. (2013) CBS Cucumis sativus Cucurbitaceae KC KC KC KC KC Gomes et al. (2013) D. scobina CBS Fraxinus Excelsior, living and Oleaceae KC KC KC KC KC Gomes et al. (2013) dead twig D. serafiniae BRIP 55665a (ex-type) Helianthus annuus Asteraceae KJ KJ KJ Gomes et al. (2013) BRIP Lupinus albus Rosetta Fabaceae KJ KJ KJ Gomes et al. (2013) D. siamensis MFLUCC 10_0573a Dasymaschalon sp. Annonaceae JQ JX JX Thompson et al. (2015) MFLUCC 10_0573b Dasymaschalon sp. Annonaceae JQ JX JX Thompson et al. (2015) D. sojae CBS Glycine soja Fabaceae KC KC KC KC KC Udayanga et al. (2012) CBS Euphorbia nutans Euphorbiaceae KC KC KC KC KC Udayanga et al. (2012) FAU 635 Glycine max Fabaceae KJ KJ KJ KJ Gomes et al. (2013) D. sterilis CBS (ex-type) Vaccinium corymbosum Ericaceae KJ KJ KJ KJ Gomes et al. (2013) CBS Vaccinium corymbosum Ericaceae KJ KJ KJ KJ Huang et al. (2015) D. stewartii CBS FJ GQ Lombard et al. (2014) D. stictica CBS Buxus sampervirens, dead twig Buxaceae KC KC KC KC KC Lombard et al. (2014) D. subclavata ZJUD 83, CGMCC Citrus grandis cv. Shatianyou Rosaceae KJ KJ KJ KJ Udayanga et al. (2011) VOLUME 8 NO

237 Gao et al. Table 2. (Continued). Species names * Culture collection no. Isolation sources Host family GenBank Accession Numbers References ITS TEF1 TUB HIS CAL ZJUD 95, CGMCC Citrus unshiu Rosaceae KJ KJ KJ KJ Gomes et al. (2013) D. subordinaria CBS Plantago lanceolata Plantaginaceae KC KC KC KC KC Huang et al. (2015) CBS Plantago lanceolata Plantaginaceae Kc KC KC KC KC Huang et al. (2015) D. tecomae CBS Tabebuia sp. Bignoniaceae KC KC KC KC KC Gomes et al. (2013) D. tectonae MFLUCC Tectona grandis Verbenaceae KU KU KU KU Gomes et al. (2013) MFLUCC Tectona grandis Verbenaceae KU KU KU KU Gomes et al. (2013) D. tectonendophytica MFLUCC Tectona grandis Verbenaceae KU KU KU KU Doilom et al. (2017) D. tectonigena MFLUCC Tectona grandis Verbenaceae KU KU KU KU Doilom et al. (2017) D. terebinthifolii CBS Schinus terebinthifolius Anacardiaceae KC KC KC KC KC Doilom et al. (2017) LGMF 907 Schinus terebinthifolius Anacardiaceae KC KC KC KC KC Doilom et al. (2017) D. thunbergii MFLUCC 10_0756a Thunbergia laurifolia Acanthaceae JQ JX JX JX Doilom et al. (2017) MFLUCC 10_0756b Thunbergia laurifolia Acanthaceae JQ JX JX JX Doilom et al. (2017) D. toxica CBS (ex-type) Lupinus angustifolius, stem Fabaceae KC KC KC KC KC Udayanga et al. (2012) CBS Lupinus sp. Fabaceae KC KC KC KC KC Udayanga et al. (2012) D. tulliensis BRIP 62248a Theobroma cacao Sterculiaceae KR KR KR Gomes et al. (2013) D. ueckerae FAU 656 Cucumis melo Cucurbitaceae KJ KJ KJ KJ Gomes et al. (2013) FAU 658 Cucumis melo Cucurbitaceae KJ KJ KJ KJ Crous et al. (2015) D. undulata CGMCC = LC 6624 Unknown host, pathogen - KX KX KX KX Huang et al. (2015) LC 8110 Unknown host, pathogen - KY KY KY Huang et al. (2015) LC 8111 Unknown host, pathogen - KY KY KY This study D. unshiuensis ZJUD 51, CGMCC Fortunella margarita Rutaceae KJ KJ KJ KJ This study ZJUD 52, CGMCC Citrus unshiu Rosaceae KJ KJ KJ KJ This study D. vaccinii CBS (ex-type) Oxycoccus macrocarpos Ericaceae KC KC KC KC KC Huang et al. (2015) CBS Vaccinium corymbosum Ericaceae KC KC KC KC KC Huang et al. (2015) D. vawdreyi BRIP 57887a Psidium guajava Sterculiaceae KR KR KR Gomes et al. (2013) D. velutina CGMCC = LC 4421 Neolitsea sp., pathogen Lauraceae KX KX KX KX Gomes et al. (2013) LC 4419 Neolitsea sp., pathogen Lauraceae KX KX KX KX KX Crous et al. (2015) LC 4641 Callerya cinerea, pathogen Fabaceae KX KX KX KX KX This study LC 4788 Unknown host, pathogen - KX KX KX KX KX This study LC 6708 Camellia sinensis, pathogen Theaceae KX KX KX KX This study D. vexans CBS Solanum melongena Solanaceae KC KC KC KC KC This study D. virgilia CMW (ex-type) Virgilia oroboides Unknown KP KP This study 170 IMA FUNGUS

238 Diaporthe is paraphyletic Table 2. (Continued). Species names * Culture collection no. Isolation sources Host family GenBank Accession Numbers References ITS TEF1 TUB HIS CAL CMW Virgilia oroboides Unknown KP KP Gomes et al. (2013) D. woodii CBS Lupinus sp. Fabaceae KC KC KC KC KC Gomes et al. (2013) D. woolworthii CBS Ulmus americana Ulmaceae KC KC KC KC KC Gomes et al. (2013) D. xishuangbanica CGMCC = LC 6707 Camellia sinensis, pathogen Theaceae KX KX KX KX This study LC 6744 Camellia sinensis, pathogen Theaceae KX KX KX This study D. yunnanensis CGMCC = LC6168 Coffea sp., endophytes Rubiaceae KX KX KX KX KX This study LC 8106 Coffea sp., endophytes Rubiaceae KY KY KY KY This study LC 8107 Coffea sp., endophytes Rubiaceae KY KY KY KY This study Diaporthe sp. LC 6496 Camellia sinensis, endophytes Theaceae KX KX KX KX KX This study LC 6512 Camellia sinensis, endophyte Theaceae KX KX KX KX KX This study LC 6232 Theobroma cacao, endophyte Sterculiaceae KX KX KX KX KX This study LC 8108 Theobroma cacao, endophyte Sterculiaceae KY KY KY KY This study LC 8109 Theobroma cacao, endophyte Sterculiaceae KY KY KY KY This study LC 6623 Unknown host, pathogen - KX KX KX KX This study LC 8114 Unknown host, pathogen - KY KY KY This study LC 8115 Unknown host, pathogen - KY KY KY This study LGMF 947 Glycine max, seed Fabaceae KC KC KC KC KC Gomes et al. (2013) CBS Man, abscess - KC KC KC KC KC Gomes et al. (2013) Diaporthe sp. 1 CGMCC = LC 0771 Alnus sp., pathogen Betulaceae KX KX KX KX KX This study Diaporthe sp. 2 CGMCC = LC 6140 Acer sp., endophyte Aceraceae KX KX KX KX KX This study LC8112 Acer sp., endophyte Aceraceae KY KY KY KY This study LC8113 Acer sp., endophyte Aceraceae KY KY KY KY This study Diaporthella corylina CBS Corylus sp., dying stems Corylaceae KC KC KC KC KC Gomes et al. (2013) P. conorum CBS Penus pentaphylla Pinaceae KC KC KC KC KC Gomes et al. (2013) P. emicis BRIP 45089a (ex-type) Emex australis Polygonaceae JF JX JX JX Udayanga et al. (2012) P. fukushii CBS Pyrus pyrifolia Roseceae KC KC KC KC KC Gomes et al. (2013) BRIP 45089b Emex australis Polygonaceae JQ JX JX JX Udayanga et al. (2012) -: not provided in literatures. VOLUME 8 NO

239 Gao et al. Fig. 3. Diaporthe acutispora (CGMCC ). A B. 30-d-old culture on PNA medium. C. Conidiomata. D E. Conidiophores. F G. Alpha conidia. Bars: C = 100 µm; D G = 10 µm. TAXONOMY Diaporthe acutispora Y.H. Gao & L. Cai, sp. nov. MycoBank MB (Fig. 3) Etymology: Named after the acute spores. Diagnosis: Diaporthe acutispora is phylogenetically distinct and morphologically differs from species reported from the host genera Coffea and Camellia in the larger conidiophores and alpha conidia (Table 3). Type: China: Yunnan Province: Aini Farm, on healthy leaves of Coffea sp., 20 Sep. 2014, W.J. Duan (HMAS holotype, dried culture; CGMCC = LC 6161 ex-type culture). Description: On PNA: Conidiomata pycnidial, globose, brownish, embedded in tissue, erumpent at maturity, µm diam, often with a yellowish conidial cirrus exuding from the ostioles. Conidiophores µm, cylindrical, hyaline, septate, branched, straight or slightly curved, tapering towards the apex. Alpha conidia abundant in culture, µm (x = 8.4 ± ± 0.2, n = 30), aseptate, hyaline, ellipsoidal to fusoid, multi-guttulate. Beta conidia not observed. Culture characters: Cultures incubated on PDA at 25 C in darkness, growth rate 7.5 mm diam/d. Colony entirely white at surface, reverse with pale brown pigmentation, white, fluffy aerial mycelium. Additional material examined: China: Yunnan Province: Xishuangbanna, on healthy leaves of Camellia sasanqua, 20 Sep. 2014, W.J. Duan, culture LC 6142; ibid. culture LC Diaporthe elaeagni-glabrae Y.H. Gao & L. Cai, sp. nov. MycoBank MB (Fig. 4) Etymology: Named after the host species Elaeagnus glabra. Diagnosis: Diaporthe elaeagni-glabrae can be distinguished from the closely related species D. elaeagni (96 % in ITS, 93 % in TEF1, 94 % in TUB, 96 % in HIS, and 94 % in CAL) and D. stictica (96 % in ITS, 95 % in TEF, 97 % in TUB, 96 % in HIS, and 96 % in CAL) (Fig. 2). Diaporthe elaeagni-glabrae differs from other species recorded from Elaeagnus in the significantly longer alpha conidia (Table 3). Type: China: Jiangxi Province: on diseased leaves of Elaeagnus glabra, 5 Sep. 2013, Y.H. Gao (HMAS holotype, dried culture; CGMCC = LC 4802 ex-type culture). 172 IMA FUNGUS

240 Diaporthe is paraphyletic Table 3. Synoptic characters of Diaporthe spp. referred to in this study. Host genera Species Conidiomata (μm) Conidiophores (μm) Alpha conidia (μm) Beta conidia (μm) References Coffea P. coffeae Uecker (1988) Camellia D. acutispora This study D. amygdali (4.18 ) ( 9.64) (1.63 ) ( 3.31) - Diogo et al. (2010) D. apiculata ( 416) (20.0 ) Gao et al. (2016) D. compacta Gao et al. (2016) D. discoidispora Huang et al. (2015) D. eres (6 ) ( 9) 3 4 (18 )22 28(29) Udayanga et al. (2014b) D. foeniculacea (5.4 )6.8 7( 9) (2 ) ( 3.1) (16.8 ) ( 24.2) (1.1 ) ( 1.7) Phillips (2003) D. foeniculina ( 18) 1 2 (7.5 )8.5 9( 9.2) (2 ) ( 2.7) (20 )22 28( 29) (1.1 ) ( 2) Udayanga et al. (2014c) D. hongkongensis to (5 )6 7( 8) (2 )2.5( 3) Gomes et al. (2013) D. oraccinii Gao et al. (2016) D. penetriteum ( 27) Gao et al. (2016) D. ueckerae (9 )12 28( 30) (6 ) ( 8.6) (2 ) Udayanga et al. (2014a) D. xishuangbanica This study D. yunnanensis This study P. acaciicola Diedicke (1911) P. theae Petch (1925) Elaeagnus P. arnoldiae Uecker (1988) P. elaeagni Uecker (1988) P. elaeagnicola Chang et al. (2005) D. elaeagni-glabrae This study D. incompleta This study Neolitsea D. velutina This study AR, DP, FAU: Isolates in culture collection of Systematic Mycology and Microbiology Laboratory, USDA-ARS, Beltsville, Maryland, USA; BCRC: Bioresource Collection and Research Center, Taiwan; BRIP: Australian plant pathogen culture collection, Queensland, Australia; CBS: Westerdijk Fungal Biodiversity Institute, Utrecht, The Netherlands; CFCC: China Forestry Culture Collection Center, China. CGMCC: China General Microbiological Culture Collection; CMW: culture collection (CMW) of the Forestry and Agricultural Biotechnology Institute; CPC: working collection of Pedro Crous maintained at the Westerdijk Institute; LGMF: Culture collection of Laboratory of Genetics of Microorganisms, Federal University of Parana, Curitiba, Brazil; LC: Working collection of Lei Cai, housed at Institute of Microbiology, CAS, China; MFLUCC: Mae Fah Luang University Culture Collection; ZJUD: Zhe Jiang University, China. VOLUME 8 NO

241 Gao et al. Fig. 4. Diaporthe elaeagni-glabrae (CGMCC ). A B. 14-d-old culture on PDA; C. Conidiomata; D H. Conidiophores; I. Alpha conidia; J. Beta conidia. Bars: C = 100 µm; D J = 10 µm. Description: On PDA: Conidiomata globose, to µm, erumpent, with slightly elongated black necks, yellowish or dirty white, spiral conidial cirri extruding from ostioles. Conidiophores µm, cylindrical, phialidic, septate, branched, sometimes inflated. Alpha conidia µm (x = 8.3 ± ± 0.3, n = 30), hyaline, fusiform or oval, usually biguttulate. Beta conidia µm ( x = 15.1 ± ± 0.2, n = 40), hyaline, filiform, smooth, curved, base truncate. Culture characters: Cultures incubated on PDA at 25 C in darkness, growth rate 7 mm diam/d. Colony pale yellowish, greenish to brownish at the centre, reverse pale yellowish and brownish at the centre with age. Aerial mycelium white, sparse, fluffy, with irregular margin and visible conidiomata at maturity. Additional material examined: China: Jiangxi Province: on diseased leaves of Elaeagnus glabra, 5 Sep. 2013, Y.H. Gao, culture LC Diaporthe helianthi Munt.-Cvetk. et al., Nova Hedwigia 34: 433 (1981). (Fig. 5) Description: Sexual morph not produced. Conidiomata pycnidial globose to subglobose, dark brownish to black, erumpent or immersed in medium, translucent conidia exuded from the ostioles, µm diam. Conidiophores cylindrical, straight or sinuous, apical or base sometimes swelling, µm (x = 16 ± ± 0.5, n = 30). Beta conidia filiform, hamate or slightly curved, base truncate, tapering towards one apex, µm (x = 20 ± ± 0.4, n = 20). Alpha conidia not observed. Culture characters: Cultures on PDA at 25 C in dark, with 12/12 h alternation between daylight and darkness pure white (surface) and pale yellow to cream (reverse). Colony pellicular, forming less pigmented sectors, with concentric rings of gummy mycelium. Growth rate was 10.5 mm diam/d. Material examined: Ukraine: from seeds of Helianthus annuus, 30 Oct. 2015, W.J. Duan culture LC Japan: Lagerstroemia indica, 30 Oct. 2015, W.J. Duan, culture LC Notes: Diaporthe helianthi, responsible for stem canker and grey spot disease of sunflower (Helianthus annuus) (Muntanola-Cvetkovic et al. 1981), has been listed in the Chinese quarantine directory. There is increasing evidence that this serious sunflower pathogen is being quickly and globally disseminated with international trade. The cases reported here were intercepted from imported sunflower seeds from Ukraine and Lagerstroemia indica from Japan. 174 IMA FUNGUS

242 Diaporthe is paraphyletic Fig. 5. Diaporthe helianthi (LC 6185). A B. 7-d-old culture on PDA; C. Conidiomata; D F. Conidiophores; G H. Beta conidia. Bars: C = 100 µm; D H = 10 µm. Diaporthe incompleta Y.H. Gao & L. Cai, sp. nov. MycoBank MB (Fig. 6) Etymology: Named after the absence of alpha conidia. Diagnosis: Diaporthe incompleta is phylogenetically distinct and differs morphologically from other species recorded from Elaeagnus and Camellia in the much longer beta conidia (Table 3). Type: China: Yunnan Province: Xishuangbanna, on diseased of Elaeagnus glabra, 19 Apr. 2015, F. Liu (HMAS holotype, dried culture; CGMCC = LC 6754 ex-type culture). Description: Conidiomata pycnidial, subglobose to globose, brownish to black, µm diam, cream to pale luteous conidial droplets exuding from the central ostioles. Conidiophores µm, cylindrical, hyaline, septate, unbranched, smooth, slightly curved, tapering towards apex. Alpha conidia not observed. Beta conidia µm (x = 30.5 ± ± 0.4, n = 30), smooth, hyaline, filiform, base subtruncate, straight or curved. Culture characters: Cultures incubated on PDA at 25 C in darkness, growth rate 16.5 mm diam/d. Colony entirely white, flat, reverse pale yellowish, becoming brownish zoned at the centre with age. Aerial mycelium white, cottony, margin lobate, conidiomata visible at maturity. Additional material examined: China: Yunnan Province: Xishuangbanna, on diseased leaves of Camellia sinensis, 19 Apr. 2015, F. Liu, culture LC VOLUME 8 NO

243 Gao et al. Fig. 6. Diaporthe incompleta (CGMCC ). A. Leaves of host plant; B C. 7-d-old culture; D. Conidiomata; E F. Conidiophores; G. Beta conidia. Bars: D = 100 µm; E G = 10 µm. Diaporthe podocarpi-macrophylli Y.H. Gao & L. Cai, sp. nov. MycoBank MB (Fig. 7) Etymology: Named after the host plant Podocarpus macrophyllus. Diagnosis: Diaporthe podocarpi-macrophylli can be distinguished from the phylogenetically closely related species D. pseudophoenicicola (97 % identity in ITS, 90 % in TEF1, 98 % in TUB, 97 % in HIS, and 97 % in CAL). Morphologically, D. podocarpi-macrophylli differs from other species occurring on the host genera Podocarpus and Olea, i.e. D. cinerascens and Phomopsis podocarpi in its wider and shorter alpha co- 176 IMA FUNGUS

244 Diaporthe is paraphyletic Fig. 7. Diaporthe podocarpi-macrophylli (CGMCC ). A B. 30-d-old culture on PDA; C. Conidiomata; D F. Conidiophores; G I. Alpha and beta conidia. Bars: C = 100 µm; D I = 10 µm. nidia and the presence of beta conidia (Chang et al. 2005, Gomes et al. 2013; Type: Japan: on healthy leaves of Podocarpus macrophyllus, 20 Sep. 2014, W.J. Duan (HMAS holotype, dried culture; CGMCC = LC 6155 ex-type culture). Description: Conidiomata pycnidial in culture on PDA, solitary or aggregated, deeply embedded in the PDA, erumpent, dark brown to black, µm diam, yellowish translucent conidial drops exuding from the ostioles. Alpha conidiophores µm (x = 12.3 ± ± 0.3, n = 30), hyaline, septate, branched, cylindrical, straight to sinuous, sometimes inflated, occurring in dense clusters. Beta conidiophores µm (x = 15.3 ± ± 0.3, n = 30), cylindrical to clavate, hyaline, septate, branched, smooth, straight. Alpha conidia µm (x = 6.3 ± ± 0.7, n = 50), unicellular, aseptate, fusiform, hyaline, usually biguttulate and acute at both ends. Beta conidia µm (x = 19.5 ± ± 0.4, n = 30), hyaline, aseptate, eguttulate, filiform, curved, tapering towards both ends, base truncate. Culture characters: Cultures incubated on PDA at 25 C in darkness, growth rate 12.5 mm diam/d. Colony at first white, becoming cream to yellowish, flat, with dense and felted mycelium, reverse pale brown with brownish dots with age, with visible solitary or aggregated conidiomata at maturity. VOLUME 8 NO

245 Gao et al. Fig. 8. Diaporthe undulata (CGMCC ). A. Leaves of host plant; B C. 30-d-old culture on PNA medium; D. Conidiomata; E. Conidiophores; F G. Alpha conidia. Bars: D = 100 µm; E G = 10 µm. Additional material examined: Japan: on healthy leaves of Podocarpus macrophyllus, 20 Sep. 2014, W.J. Duan, culture LC 6141; ibid. culture LC 6144; ibid. culture LC 6156; ibid. culture LC China: Zhejiang Province: on healthy leaves of P. macrophyllus, 10 Jul. 2015, W.J. Duan, culture LC 6194; ibid. culture LC 6195; ibid. culture LC 6196; ibid. culture LC 6197; ibid. culture LC 6198; ibid. culture LC 6199; ibid. culture LC 6200; ibid. culture LC 6201; ibid. culture LC 6202; ibid. culture LC Italy: on healthy leaves of Olea europaea, 20 Sep. 2014, W.J. Duan, culture LC Diaporthe undulata Y.H. Gao & L. Cai, sp. nov. MycoBank MB (Fig. 8) Description: Conidiomata pycnidial, irregular, embedded in the needle, erumpent, necks, hairy, µm long, coated with short hyphae, one to several necks forming from a single pycnidium. Conidiophores obpyriform, hyaline, phiailidic, septate, branched, µm (x = 9.7 ± ± 0.5, n = 20). Alpha conidia ellipsoid, hyaline, biguttulate, rounded at both ends, (x = 5.8 ± ± 0.3, n = 50). Beta conidia not observed. Culture characters: Cultures incubated on PDA at 25 C in darkness, growth rate 10.5 mm diam/d. Colony entirely white, reverse pale yellowish and dark brownish at the centre with age. Aerial mycelium white, cottony, dense, with undulate margin and visible conidiomata at maturity. Etymology: Named after the colony s undulate margin. Diagnosis: Diaporthe undulata differs from the most closely related species, D. biconispora, in several loci (94 % in ITS, 84 % in TEF1, and 93 % in TUB), and from other Diaporthe species in the obpyriform conidiophores and shorter and wider alpha conidia (Table 3). Type: China-Laos border: on diseased leaves of unknown host, 19 Apr. 2014, F. Liu (HMAS holotype, dried culture; CGMCC = LC 6624 ex-type culture). Additional material examined: China-Laos border: unknown host, 19 Apr. 2014, F. Liu, culture LC 8110; ibid. culture LC Diaporthe velutina Y.H. Gao & L. Cai, sp. nov. MycoBank MB (Fig. 9) Etymology: Named after the felted colony. Diagnosis: Diaporthe velutina is distinguished from D. anacardii in the ITS, TEF1, TUB and HIS loci (99 % in ITS, 95 % in TEF1, 99 % in TUB, and 98 % in HIS), and from 178 IMA FUNGUS

246 Diaporthe is paraphyletic Fig. 9. Diaporthe velutina (CGMCC ). A. Diseased leaves; B C. 30-d-old culture on PDA; D. Conidiomata; E. Conidiophores; E. Alpha and beta conidia. Bars: D = 100 µm; E F = 10 µm. other Diaporthe species reported from Camellia sinensis in the more variable size of the alpha conidia (Table 3). Type: China: Jiangxi Province: on diseased leaves of Neolitsea sp., 5 Sep. 2013, Y.H. Gao (HMAS holotype, dried culture; CGMCC = LC 4421 ex-type culture). Description: Conidiomata pycnidial, globose, black, embedded in PDA, aggregated in clusters, µm diam, cream translucent drop of conidia exuded from the central ostioles. Conidiophores µm, cylindrical, hyaline, branched, densely aggregated, slightly tapering towards the apex, sometimes slightly curved. Alpha conidia µm (x = 6.9 ± ± 0.2, n = 50), unicellular, aseptate, hyaline, fusoid to ellipsoid or clavate, bi-guttulate or multi-guttulate. Beta conidia µm (x = 16.1 ± ± 0.4, n = 30), smooth, hyaline, apex acutely rounded, curved. Culture characters: Cultures incubated on PDA at 25 C in darkness, growth rate mm diam/d. Colony entirely white, surface mycelium greyish to brownish at the centre, dense, felted, conidiomata erumpent at maturity, reverse centre yellowish to brownish. Additional material examined: China: Jiangxi Province: Yangling, on diseased leaves of Neolitsea sp., 5 Sep. 2013, Y.H. Gao, culture LC 4419; ibid. culture LC 4422; Gannan Normal University, unknown host, 23 Apr. 2013, Q. Chen, culture LC 4788; Fengshan, on diseased leaves of Callerya cinerea, 5 Sep. 2013, Y.H. Gao, culture LC Yunnan Province: Xishuangbanna, on diseased leaves of Camellia sinensis, 19 Apr. 2015, F. Liu, culture LC 6708; loc. cit., on healthy leaves of C. sinensis, 21 Apr. 2015, F. Liu, culture LC Diaporthe xishuangbanica Y.H. Gao & L. Cai, sp. nov. MycoBank MB (Fig. 10) Etymology: Named after the locality, Xishuangbanna. Diagnosis: Diaporthe xishuangbanica can be distinguished from the phylogenetically closely related D. tectonigena in several loci (98 % in ITS, 90 % in TEF1, and 96 % in TUB) VOLUME 8 NO

247 Gao et al. Fig. 10. Diaporthe xishuangbanica (CGMCC ). A B. 7-d-old culture on PDA; C D. 30-d-old culture on PNA medium; E. Conidiomata; F K. Conidiophores; L N. Alpha conidia. Bars: E = 100 µm; F N = 10 µm. (Fig. 2), and from other Diaporthe species reported from Camellia in the longer conidiophores and alpha conidia (Table 3). Type: China: Yunnan Province: Xishuangbanna, on diseased leaves of Camellia sinensis, 19 Apr. 2015, F. Liu (HMAS holotype, dried culture; CGMCC = LC 6744 ex-type culture). Description: Conidiomata pycnidial, globose, µm diam, scattered on the pine needle. Conidiophores cylindrical, µm (x = 20.9 ± ± 0.3, n = 40), branched, septate, straight, sometimes sinuous or lateral. Alpha conidia µm (x = 8.3 ± ± 0.3, n = 30), fusiform, hyaline, multi-guttulate. Beta conidia not observed. Culture characters: Cultures incubated on PDA at 25 C in darkness, growth rate 17.5 mm diam/d. Colony entirely white, reverse pale yellowish to greenish. Aerial mycelium white, velvety, margin well defined, with visible conidiomata at maturity. Additional material examined: China: Yunnan Province: Xishuangbanna, on diseased leaves of Camellia sinensis, 19 Apr. 2015, F. Liu, culture LC 6707 (CGMCC ). Diaporthe yunnanensis Y.H. Gao & L. Cai, sp. nov. MycoBank MB (Fig. 11) Etymology: Named after the location where the fungus was collected, Yunnan Province. Diagnosis: Diaporthe yunnanensis can be distinguished from the phylogenetically closely related D. siamensis (96 % in ITS, 91 % in TEF1, and 94 % in TUB) (Fig. 2), and from other Diaporthe species reported on the genus Camellia in the smaller alpha conidia (Table 3). Type: China: Yunnan Province: Xishuangbanna, on healthy leaves of Coffea sp., 20 Sep. 2014, W.J. Duan (HMAS holotype, dried culture; CGMCC = LC 6168 ex-type culture). Description: Conidiomata pycnidial, µm diam, globose or irregular, erumpent, solitary or aggregated together, dark brown to black. Conidia exuding from the pycnidia in white to cream drops. Conidiophores cylindrical, straight or slightly curved. Alpha conidia µm ( x = 5.5 ± 1 2 ± 0.5, n = 30), fusiform, hyaline, biguttulate, with one end obtuse and the other acute. Beta conidia µm (x = 27.5 ± ± 0.3, n = 30), hyaline, 180 IMA FUNGUS

248 Diaporthe is paraphyletic Fig. 11. Diaporthe yunnanensis (fcgmcc ). A B. 7-d-old culture on PDA; C. Conidiomata; D. Conidiophores; E. Alpha and beta conidia; F. Beta conidia. Bars: C = 100 µm; D F = 10 µm. aseptate, hamate or curved, base truncate. Culture characters: Colonies on PDA flat, with a moderate growth rate of 5.5 mm diam/d, with abundant dirty white and yellowish pigmented mycelium, dry, felted, extensive thin, and in reverse the centre cream, with zone rings of pale to dark brownish pigmentation. Additional material examined: China: Yunnan Province: Xishuangbanna, on healthy leaves of Coffea sp., 20 Sep. 2014, W.J. Duan, culture LC 8106; ibid. culture LC Diaporthe sp. 1 (Fig. 12) Description: Conidiomata pycnidial, subglobose to globose, dark brown to black, deeply embedded in the substrate, scattered on the substrate surface, embedded in PDA, clusters in group of 2 7 pycnidia, µm, yellowish drop of conidia diffusing from the central ostioles. Conidiophores µm, cylindrical, hyaline, septate, branched, straight to sinuous, base inflated, slightly tapering towards the apex. Alpha conidia µm (x = 9.9 ± ± 0.4, n = 30), unicellular, hyaline, fusoid to ellipsoid or clavate, two or several large guttulae observed, base subtruncate. Beta conidia µm (x = 26.0 ± ± 0.5, n = 30), smooth, hyaline, curved, base subtruncate, tapering towards one apex. Culture characters: Cultures incubated on PDA at 25 C in darkness, growth rate 7.83 mm diam/day. Colony entire, white to dirty pink, cottony, sparse, brownish to black conidiomata erumpent at maturity, coated with white hypha, granular at margin, reverse pale brown, with brownish dots when maturity. Material examined: China: Zhejiang Province: Gutianshan Nature Reserve (29º20 N 18º14 E), on leaves of Alnus mill, Jan. 2010, Y.Y. Su (culture CGMCC = LC 0771). Notes: The present culture belongs to the Diaporthe eres complex, which is reported from a very wide range of host plants and includes mostly opportunistic pathogens or secondary invaders on saprobic host substrata (Udayanga et al. 2014a, Gao et al. 2016). Species delimitation in this complex is currently unclear. Udayanga et al. (2015) accepted nine phylogenetic species in the D. eres complex, including D. alleghaniensis, D. alnea, D. bicincta, D. celastrina, D. eres, D. helicis, D. neilliae, D. pulla, and D. vaccinia. Gao et al. (2016) examined 17 isolates belonging to the D. eres VOLUME 8 NO

249 Gao et al. Fig. 12. Diaporthe sp. 1 (CGMCC ). A. Leaves of host plant; B C. 30-d-old culture on PDA; D. Conidiomata; E F. Conidiophores; G. Beta conidia; H I. Alpha conidia. Bars: D = 100 µm; E I = 10 µm. complex, and reported that many presented intermediate morphology among species and the phylogenetic analyses often resulted in ambiguous clades with short branch and moderate statistical support. The identification of taxa in this group remains unresolved. 182 IMA FUNGUS