IMA FUNgUs · 5(2): 425–438 (2014) doi:10.5598/imafungus.2014.05.02.07 Steven E. Zelski1, Julia A. Balto1, Christine Do1, Huzefa A. Raja1,2, Andrew N. Miller3, and Carol A. Shearer1 ART I CLE Phylogeny and morphology of dematiaceous freshwater microfungi from Perú 1 Department of Plant Biology, University of Illinois at Urbana-Champaign, Room 265 Morrill Hall, 505 South Goodwin Avenue, Urbana, IL 61801, USA; corresponding author e-mail: zelski13@gmail.com 2 Department of Chemistry and Biochemistry, 457 Sullivan Science Building, University of North Carolina, Greensboro, NC 27402-6170, USA 3 Illinois Natural History Survey, University of Illinois at Urbana-Champaign, Champaign, IL 61820, USA Abstract: A survey of freshwater ascomycetes conducted along an elevational gradient in Perú in the Districts of Cusco, Junín, and Madre de Dios yielded specimens of Cancellidium applanatum, Cordana abramovii, Sporoschisma juvenile, S. uniseptatum, and S. saccardoi. With the exception of S. saccardoi, these are new records for Perú. Molecular data was generated for three previously unsequenced species: Cancellidium applanatum, Cordana abramovii and Sporoschisma saccardoi. These taxa are reported herein from the neotropics with an accompanying phylogeny based on partial 28S nuclear ribosomal large-subunit sequence data. The sexual morph of S. saccardoi has previously been linked to Melanochaeta hemipsila through cultural studies. Molecular data from ascospores and conidia of M. hemipsila and S. saccardoi, respectively, were used to demonstrate a genetic connection of the sexual and asexual morphs of these fungi for the irst time, resulting in the new combination Sporoschisma hemipsila being made. Key words: Aquatic fungi Ascomycoyta Cancellidium Cordana Sporoschisma submerged woody debris Article info: Submitted: 20 January 2014; Accepted: 24 November 2014; Published: 10 December 2014. INTRODUCTION During a study of ascomycetes colonizing submerged, decomposing woody and herbaceous debris in freshwater habitats along an elevational gradient in Perú extending from the Peruvian Amazon to the Peruvian Andes (2010–2012), numerous freshwater mitosporic fungi were encountered. Shearer et al. (2007) divided the freshwater mitosporic fungi into three ecological groups: (1) freshwater hyphomycetes; (2) aeroaquatic hyphomycetes; and (3) freshwater miscellaneous mitosporic ascomycetes. This study deals with one aeroaquatic hyphomycete (Cancellidium applanatum), and four species of miscellaneous mitosporic ascomycetes (Cordana abramovii, Sporoschisma saccardoi, S. juvenile, and S. uniseptatum). Cancellidium is typiied by C. applanatum, which was originally collected from submerged wood blocks of Ochroma pyramidale in Kobe, Japan. Cancellidium applanatum has been reported from many Paleotropical localities (Webster & Davey 1980, Shaw 1994, Ho et al. 2001, Sivichai et al. 2002, Fryar et al. 2004, Pinnoi et al. 2006, Pinruan et al. 2007, Zhao et al. 2012). In this study in the Neotropics, multiple collections of Cancellidium applanatum (PE0063) were recovered from low and middle altitudes along the elevational gradient, but not from high altitude aquatic habitats. Yeung et al. (2006) suggested that the congeneric C. pinicola was phylogenetically related to Hypocreales. However, they noted that a connection to Hypocreales was dubious due to the questionable nature of the culture from which the DNA was extracted (Yeung et al. 2006, Zhao et al. 2012). In this study one 28S sequence was generated from a Peruvian specimen and the identity was corroborated with two 28S sequences generated from Thai material. Another dematiaceous fungus, closely resembling Cordana abramovii, was found in 33 of 86 collections from a range of sites. Cordana is typiied by C. pauciseptata. The type is described as acervular, possibly due to the cushiony appearance of the aggregated sporing structures and setae on the substrate (Preuss 1851). The majority of the taxa belonging to the genus are not described as such; rather, conidia simply form on erect conidiophores with surrounding setae. The Peruvian specimens of Cordana abramovii (PE0053) are characterized by pale brown to brown, cylindrical, septate conidiophores with swollen conidiogenous zones; terminal and intercalary polyblastic conidiogenous cells; and golden brown to dark brown, 1-septate, thickwalled, verruculose conidia. Two additional species, C. musae and C. pauciseptata, have previously been reported from Perú (Matsushima 1993). Cordana species are placed in the family Cordanaceae (Cannon & Kirk 2007). Several species of Sporoschisma were collected from multiple sites, including S. uniseptata (15 collections), S. saccardoi (13), S. juvenile (9), and S. parcicuneatum (2). Sporoschisma uniseptata has 1-septate, rarely 2-septate, reddish brown, verruculose conidia; S. saccardoi has brown, 5-septate, doliiform, smooth walled conidia; S. juvenile © 2014 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 speciied 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 http://creativecommons.org/licenses/by-nc-nd/3.0/legalcode. 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 5 · NO. 2 425 ART I CLE Zelski et al. has brown 5-septate, cylindrical, verruculose conidia; and S. parcicuneatum has brown, 1(–3)-septate, cuneiform, verruculose conidia (Goh et al. 1997). Sexual reproductive structures of Melanochaeta hemipsila were found among conidiophores of S. saccardoi from substrates collected in Cusco and Junin. Melanochaeta hemipsila has been connected to S. saccardoi based on studies in which the asexual morph was produced from colonies derived from ascospores (Müller et al. 1969, Nag Raj 1975, Sivichai et al. 2000). The sexual morph of the Peruvian specimens is characterized by: gregarious, supericial, dark brown to black ascomata with short conical beaks; numerous, septate, capitate setae arising from the external ascomal wall; clavate, unitunicate, 8-spored asci with an I- refractive apical apparatus; biseriate, cylindrical to curved, 5-septate ascospores with olivaceous to brown central cells, hyaline end cells, lacking sheaths or appendages. The goals of this study were to: (1) describe, illustrate, and provide voucher specimens and sequences for the foregoing species of freshwater mitosporic fungi for which pure cultures were obtained; (2) compare and contrast these fungi with morphologically similar and genetically related taxa; and (3) construct a molecular phylogeny using 28S large subunit (LSU) nrDNA to elucidate the evolutionary relationships of these fungi with other Ascomycota. MATERIALs AND METHODs Isolates Submerged woody and herbaceous debris was collected from a variety of freshwater habitats that included rivers, streams, backwaters, swamps, and inundated trails. Approximately 30 pieces of debris were put into a sealable plastic bag along with a wet paper towel at each of 86 sampling sites along an altitudinal gradient stretching from 218–3566 m. Samples were shipped to our laboratory at the University of Illinois at Urbana-Champaign. In the laboratory, substrates were placed in moist chambers (sealable plastic boxes lined with moist paper towels) and incubated at room temperature (~25 °C) with 12/12 h light/dark conditions. Samples were examined for reproductive structures within one week of arrival and periodically thereafter for 12 mo with an AO stereomicroscope. Digital images of fruiting structures were taken on an Olympus SZX7 stereomicroscope (Olympus Optical Tokyo) itted with a SPOT RT colour camera using SPOT Advanced software (Diagnostics Instruments, Sterling Hts, MI). Ascomata were removed from the substrate with a dissecting needle and gently teased apart in a drop of distilled water. Conidiophores and conidia were removed in the same manner and gently placed in a drop of distilled water. Fungal tissue was then sandwiched between 25 × 25 and 18 × 18 mm cover slips in distilled water, and placed on a microscope slide for examination. Glycerin was added after examination in preparation for permanent preservation in our herbarium (ILL) according to the protocol of Volkmann-Kohlmeyer & Kohlmeyer (1996). Examination of fungal structures was performed on an Olympus BHS microscope (Olympus Optical, Tokyo) equipped with Nomarski interference and 426 phase optics. Digital micrographs were obtained with the SPOT Insight 12 Mp colour camera and Spot Advanced software. Images were processed with Adobe Photoshop and assembled with Adobe InDesign. For single spore isolation, sterile dissecting needles were used to spread ascospores or conidia on antibiotic water agar (AWA]: 20 g agar (Difco), 0.5 g streptomycin sulfate, 0.5 g penicillin G (Sigma) and 1000 mL deionized H2O. Single germinated ascospores or conidia were transferred to PYG+Ab agar plates: 1.25 g peptone, 1.25 g yeast extract, 18 g agar (Difco), 5 g D-glucose (Acros), 0.5 g streptomycin sulfate, 0.5 g penicillin G (Sigma), and 1000 mL deionized H2O. They were then grown at ambient temperature with 12/12 hr light/dark conditions. DNA isolation, ampliication and analyses DNA extraction was performed on mycelium scraped with a sterile spatula from PYG+Ab agar plates. Mycelium was irst ground into a ine powder in liquid nitrogen with a sterile mortar and pestle and DNA was extracted with a DNeasy Plant Mini Kit (Qiagen Sciences, Valencia, CA) according to the manufacturer’s instructions. PCR of extracted DNA was performed using Illustra Ready-To-Go™ PCR Beads (GE Healthcare) using the primer pair LROR and LR6 (Rehner & Samuels 1994, Vilgalys & Hester 1990) on an MJ Research PTC-200 thermocycler using the following parameters: initial denaturation at 95 °C for 5 min, followed by 40 cycles at 95 °C for 30 s, 50 °C for 15 s, 72 °C for 10 s, with a inal extension step of 72 °C for 10 min. PCR products were puriied using QIAquick PCR Puriication Kit (Qiagen Sciences, Valencia, CA) according to the manufacturer’s instructions. Sequencing reactions (11 µL) using the primers LROR, LR3, LR3R, and LR6 (Rehner & Samuels 1994, Vilgalys &Hester 1990) were carried out using the BigDye® Sequence Terminator kit 3.1 (Applied Biosystems, Foster City, CA). Sanger DNA sequencing was performed on an AB 3730xl DNA Analyzer at the W. M. Keck Center for Comparative and Functional Genomics at the University of Illinois at Urbana-Champaign. In addition to the sequences generated in this study (Table 1), sequences used in a study of Melanochaeta (Mugambi & Huhndorf 2008) were downloaded from GenBank. A taxonomic search of Cordana in GenBank yielded seven LSU sequences as well as a sequence from the sexual morph of Porosphaerella, represented by P. borinquensis. These sequences were added to the gene database. Select Sordariomycetes sequences from Zhang et al. (2006) as well as those of several freshwater ascomycetes were also included. Two members of Magnaporthales and one from Lulworthiales were used as outgroup taxa (Table 2). Sequences were assembled and initially aligned in Sequencher v. 4.9 (Gene Codes, Ann Arbor, MI). Alignment was performed using Muscle v. 3.6 (Edgar 2004) followed by visual correction. Characters at the 5’ and 3’ ends were excluded due to missing data for some taxa, resulting in a inal alignment length of 1062 base pairs. For Maximum Likelihood and Bayesian analyses, jModeltest v. 0.1.1 (Posada 2008) was used to determine the best-it model of nucleotide evolution for the data set. The GTR + I + G model was selected (-lnL 9963.4715). Base pair frequencies were: freqA = 0.2250, freqC = 0.2513, freqG = IMA FUNGUS Dematiaceous freshwater microfungi from Perú Table 1. Sequences generated for this study with voucher specimen location, GenBank number, and CBS strain number. Voucher specimen, Isolate genBank Accession Number CBs no. Cancellidium applanatum ILL 41206, TH0063-1a KF833358 CBS 137654 Cancellidium applanatum ILL 41206, TH0063-1b KF833359 CBS 137655 Cancellidium applanatum ILL 41205, PE0063-1a KF833360 CBS 137653 Cordana abramovii ILL 41204, PE0053-24a KF833361 CBS 137652 Sporoschisma hemipsila ILL 41207, PE0177-21a KF833362 CBS 137656 Sporoschisma hemipsila ILL 41207, PE0177-21b KF833363 ------- Sporoschisma hemipsila ILL 41207, PE0177-21c KF833364 CBS 138600 ART I CLE species Table 2. Sequences retrieved from GenBank for this study. species genBank Accession Number species genBank Accession Number Aniptodera chesapeakensis U46882 Fusoidispora aquatica AY780365 Annulatascus triseptatus AY780049 Gaeumannomyces graminis AF362557 Annulusmagnus triseptatus GQ996540 Gnomonia gnomon AF408361 Apiognomonia errabunda AF408334 Halosphaeria appendiculata U46885 Ascitendus austriacus GQ996539 Lasiosphaeria ovina AF064643 Bellojisia rhynchostoma EU999217 Lentomitella cirrhosa AY761085 Bullimyces aurisporus JF775590 Lentomitella crinigera AY761086 Bullimyces communis JF775585 Lindra thalassiae DQ470947 Bullimyces cosaricensis JF775591 Melanochaeta aotearoae AF466082 Calosphaeria barbirostris EF577059 Melanochaeta aotearoae AF466081 Ceratolenta caudata JX066705 Melanochaeta hemipsila EU583218 Ceratostomella cuspidata FJ617558 Melanochaeta hemipsila EU583217 Ceratostomella pyrenaica DQ076323 Melanochaeta hemipsila AF466083 Chaetomidium arxii FJ666359 Melanochaeta hemipsila AF466084 Chaetosphaeria innumera AY017375 Melanopsamella vermiculariodes AF064644 Chaetosphaeria ovoidea AF064641 Neurosopra crassa AF286411 Chaetosphaeria pulviscula AF466091 Nohea umiumi U46893 Chaetosphaeria spinosa AFF466079 Ohiostoma stenoceras DQ836904 Chaetosphaeria tropicalis AF466080 Ophioceras tenuisporum AY346295 Chatosphaeria capitata AFF466061 Ophiostoma pilferum DQ470955 Conlarium duplumascospora JN936993 Papulosa amerospora DQ470950 Cordana ellipsoidea HE672156 Porosphaerella borinquensis EF063573 Cordana ellipsoidea HE672166 Rhamphoria delicatula AF261068 Cordana inaequalis HE672157 Rhodoveronaea varioseptata FJ617560 Cordana pauciseptata HE672158 Riomyces rotundus JF775589 Cordana pauciseptata HE672159 Sordaria imicola AY780079 Cordana pauciseptata HE672160 Tainosphaeria crassipes AF466089 Cordana solitaria HE672161 Thielavia subthermophila HM448442 Cryptadelphia groenendalensis EU528007 Thyridium vestitum AY544671 Cryptadelphia polyseptata AY281102 Valsa ambiens AF362564 Diaporthe eres AF408350 Xylomelasma sordida AY761087 Fragosphaeria purpurea AF096191 0.3204, and freqT = 0.2033. The analysis estimated a rate matrix of transitions and transversions in which r[AC] = 0.8185, r[AG] = 2.3648, r[AT] = 1.8097, r[CG] = 0.5711, r[CT] = 7.3857, and r[GT] = 1. Invariable sites comprised 0.416 of the data set and the gamma shape parameter was 0.427. Maximum likelihood analysis was performed with RAxML VOLUME 5 · NO. 2 v. 7.0.4 (Stamakis et al. 2008) on the LSU dataset on the CIPRES Portal v. 2.0 (Miller et al. 2010) using default settings and GTR with 1000 fast bootstrap searches. Bayesian analysis was conducted using MrBayes v. 3.1.2 with two runs and four chains under default settings (Huelsenbeck & Ronquist 2001, Ronquist & Huelsenbeck 427 ART I CLE Zelski et al. 428 Table 3. Collection locations of specimens examined in this study. All collections are of submerged woody debris. Taxa present at each site are abbreviated as follows: Ca = Cancellidium applanatum, Co = Cordana abramovii, Sh = Sporoschisma hemipsila, Sj = Sporoschisma juvenile, and Su = Sporoschisma uniseptata. All Perú collections are made by S.E. Zelski and H. A. Raja, except for C-1797 collected by S.E. Zelski and J. A. Balto. The Thai collections were made by S. E. Zelski. Collection Country state site details Taxa C-1696 Madre de Dios Palm swamp off the Interoceanic Highway near Puerto Maldonado, 12˚42’48.0954”S, 69˚28’11.28”W, 239m, water 23.3 C, pH 5.9, 20 May 2010 Su C-1697 Madre de Dios Semi-aquatic habitat on Trail 1, 12˚34’06.52”S, 70˚06’04.57”W, 263m, 22 May 2010 Ca C-1698 Madre de Dios Stream at Trail 10, 12˚37’48.95”S, 70˚05’23.69”W, 287m, water 22.3 C, pH 5.6, 22 May Ca 2010 C-1699 Madre de Dios Creek at Trail 23, 12˚33’31.03”S, 70˚05’56.96”W, 280 m, water 22.2 C, pH 6.4, 22 May Ca 2010 C-1700 Madre de Dios Stream at Trail 28, 12˚34’02.81”S, 70˚05’42.96”W, 272 m, water 22.7 C, pH 5.9, 22 May 2010 Perú Ca C-1702 Madre de Dios Rio Amigos, 12˚34’02.86”S, 70˚04’56.26”W, 218m, water 25.3 C, pH 7.9, 22 May 2010 Su C-1703 Madre de Dios Pozo Don Pedro, palm swamp at end of Trail 17, 12˚33’34.27”S, 70˚06’38”W, 243m, 2 May 2010 C-1704 Madre de Dios Oxbow lake at Trail 14, 12˚34’14.74”S, 70˚05’23.69”W, 241m, water 23.0 C, pH 6.7, 23 Su May 2010 C-1705 Madre de Dios Seasonal lake at Trail 29, 12˚34’16.98”S, 70˚05’06.70”W, 244m, water 23.2 C, pH 6.4, 23 May 2010 Ca, Sj, Su C-1708 Madre de Dios Rio Amigos, 12˚33’46.476”S, 70˚04’41.808”W, 218 m, water 25.3 C, pH 7.9, 23 May 2010 Co C-1709 Cusco River at Quincemil Trail 1, trailhead 13˚14’22.5594”S, 70˚46’12.6114”W, 688m, water 21.0 C, pH 6.3, 26 May 2010 Ca C-1710 Cusco Stream at Quincemil Trail 1, 13˚13’58.25945”S, 70˚46’37.7754”W, 675m, water 22.2C, Sh pH 7.2, 26 May 2010 C-1711 Cusco Stream at Quincemil Trail 1, trailhead 13˚14’22.5594”S, 70˚46’12.6114”W, 688m, water Co 21.2 C, pH 7.1, 26 May 2010 C-1712 Cusco Stream at Quincemil Trail 1, trailhead 13˚14’22.5594”S, 70˚46’12.6114”W, 688m, water Ca, Co 21.2 C, pH 6.8, 26 May 2010 C-1713 Cusco Stream at Quincemil Trail 1, trailhead 13˚14’22.5594”S, 70˚46’12.6114”W, 688m, water Ca, Co 21.0 C, pH 6.0, 26 May 2010 C-1714 Cusco Stream at Quincemil Trail 1, trailhead 13˚14’22.5594”S, 70˚46’12.6114”W, 688m, water Ca, Co 21.2 C, pH 5.5, 26 May 2010 C-1715 Cusco Semi-aquatic habitat along Quincemil Trail 1, trailhead 13˚14’22.5594”S, 70˚46’12.6114”W, 688m, water 21.2 C, pH 6.8, 26 May 2010 Ca, Su C-1716 Cusco Stagnant ditch along Quincemil Trail 2, 13˚13’40.404”S, 70˚45’14.184”W, 659m, water 22.8 C, pH 5.3, 26 May 2010 Ca, Co C-1717 Cusco Stream at Quincemil Trail 2, trailhead 13˚13’ 34.07”S, 70˚45’12.67”W, 653m, water 21.8 C, pH 6.2, 26 May 2010 Ca C-1719 Cusco Stream at Quincemil Trail 2, trailhead 13˚13’ 34.00”S, 70˚45’10.62”W, 653m, water 21.9 C, pH 6.5, 26 May 2010 Ca, Co C-1720 Cusco Stream at Quincemil Trail 3, 13˚18’27.756”S, 70˚48’44.9274”W, 757m, water 20.7 C, pH 6.0, 27 May 2010 Co, Sj C-1722 Cusco Stream at Quincemil Trail 3, 13˚18’27.756”S, 70˚48’44.9274”W, 757m, water 21.3 C, pH 7.5, 27 May 2010 Co, Su C-1723 Cusco Stream at Quincemil Trail 3, 13˚18’27.76”S, 70˚48’44.93”W, 757m, water 22.3 C, pH 7.5, 27 May 2010 Ca C-1725 Cusco River at Quincemil Trail 3, 13˚18’53.128”S, 70˚48’44.8194”W, 817m, water 20.3 C, pH 7.6, 27 May 2010 Sj C-1726 Cusco Stream crossing Interoceanic Highway, 13˚17’7.008”S, 70˚47’13.632”W, 653m, water 21.7 C, pH 7.6, 27 May 2010 Sh, Sj C-1727 Cusco Stream crossing Interoceanic Highway, 13˚27’4.3914”S, 70˚54’11.3754”W, 1372m, water 15.0 C, pH 7.6, 28 May 2010 Sh, Sj C-1728 Cusco Stream crossing Interoceanic Highway, 13˚35’23.3154”S, 70˚57’21.888”W, 2562m, water 9.7 C, pH 8.3, 28 May 2010 Ca, Sj C-1730 Madre de Dios Stream at Trail 14, 12˚34’14.7”S, 70˚05’23.69”W, 241m, water 25.1 C, pH 7.3, 30 Sep 2010 Ca, Co C-1733 Madre de Dios Stream at Trail 28, 12˚34’02.81”S, 70˚05’42.96”W, 272 m, water 23.3 C, pH 6.8, 30 Sep 2010 Ca, Co C-1735 Madre de Dios Stream at Trail 23, 12˚33’31.03”S, 70˚05’56.96”W, 280m, water 23.6 C, pH 6.8, 30 Sep Ca, Co, Su 2010 Ca IMA FUNGUS Dematiaceous freshwater microfungi from Perú Table 3. (Continued). state site details Taxa C-1736 Madre de Dios Rio Amigos, 12˚33’25.22”S, 70˚05’59.89”W, 288 m, water 31.4 C, pH 8.0, 1 Oct 2010 Ca, Co, Su C-1737 Madre de Dios CICRA. Rio Amigos, 12˚34’13.008”S, 70˚41’14.7714”W, 218 m, water 31.4 C, pH 8.0, 1 Oct 2010 Ca C-1739 Cusco River at end of Quincemil Trail 1, trailhead 13˚14’22.5594”S, 70˚46’12.6114”W, 688m, water 19.6 C, pH 8.3, 3 Oct 2010 Ca, Co, Sh C-1740 Cusco Stream Quincemil Trail 1, trailhead 13˚14’22.5594”S, 70˚46’12.6114”W, 688m, water 19.0 C, pH 8.3, 3 Oct 2010 Ca, Sh, Su C-1741 Cusco Stream at Quincemil Trail 1, trailhead 13˚14’22.5594”S, 70˚46’12.6114”W, 688m, water Ca, Co 19.2 C, pH 7.7, 3 Oct 2010 C-1742 Cusco Stream at Quincemil Trail 1, trailhead 13˚14’22.5594”S, 70˚46’12.6114”W, 688m, water Ca 19.1 C, pH 6.7, Oct 2010 C-1743 Cusco Semi-aquatic habitat along Quincemil Trail 1, trailhead 13˚14’22.56”S, 70˚46’12.61”W, 688m, 26 May 2010 C-1745 Cusco Stream at Quincemil Trail 1, trailhead 13˚14’22.5594”S, 70˚46’12.6114”W, 688m, water Ca, Co 19.7 C, pH 5.8, 3 Oct 2010 C-1746 Cusco Rio Caliente, 1km south of Quincemil, 13˚13’20.87”S, 70˚44’30.07”w, 626m, water 25.0 Co, Su C, pH 7.2, 3 Oct 2010 C-1747 Cusco Quincemil. Stream at Quincemil Trail 2, 13˚13’31.0434”S, 70˚45’10.6194”W, 653 m, water 24.0 C, pH 7.4, 4 Oct 2010 Ca C-1748 Cusco Stream at Quincemil Trail 2, 13˚13’31.04”S, 70˚45’10.62”W, 653m, water 25.0 C, pH 7.3, 4 Oct 2010 Ca, Co C-1749 Cusco Stream at Quincemil Trail 3, trailhead 13˚18’22.756”S, 70˚48’44.9274”W, 757 m, water Ca 20.5 C, pH 7.18, 4 Oct 2010 C-1750 Cusco Stream at Quincemil Trail 3, trailhead 13˚18’22.756”S, 70˚48’44.9274”W, 757 m, water Co, Sh, Su 21.6 C, pH 7.1, 4 Oct 2010 C-1751 Cusco Stream at Quincemil Trail 3, trailhead 13˚18’22.756”S, 70˚48’44.9274”W, 757 m, water Ca, Sj 21.4 C, pH 7.5, 4 Oct 2010 C-1752 Cusco Stream at Quincemil Trail 3, trailhead 13˚18’27.76”S, 70˚48’44.93”W, 757m, water 21.5 Ca C, pH 7.5, 4 Oct 2010 C-1753 Cusco Stream with red algae along Quincemil trail 3, 13˚18’27.756”S, 70˚48’44.9274”W, 757m, water 21.8 C, pH 7.2, 4 Oct 2010 Co C-1754 Cusco River at end of Quincemil Trail 3, 13˚18’23.65”S, 70˚48’47.02”W, 772m, water 21.0 C, pH 7.7, 4 Oct 2010 Co C-1755 Cusco Stream crossing the Interoceanic Highway, 13˚17’7.008”s, 70˚47’13.632”W, 737m, water 22.0 C, pH 7.7, 4 Oct 2010 Ca, Co, Sh, Sj, Su C-1756 Cusco Stream crossing Interoceanic Highway, 13˚27’52.1994”S, 70˚53’52.44”W, 1463m, water 15.33 C, pH 8.2, 5 Oct 2010 Sh, Sj C-1757 Cusco Stream crossing Interoceanic Highway, 13˚32’37.95”S, 70˚53’18.95”W, 3421m, water 17.9 C, pH 8.3, 5 Oct 2010 Sh C-1758 Cusco Stream along Interoceanic Highway, 13˚37’40.3674”S, 71˚24’23.9394”W, 3566m, water Su 17.4 C, pH 8.4, o5 Oct 2010 C-1768 Madre de Dios Pozo Don Pedro, palm swamp at end of Trail 17, 12˚33’34.27”S, 70˚06’38”W, 243 m, water 25.4 C, pH 7.9, 9 Apr 2011 Ca C-1769 Madre de Dios Stream at Trail 20, 12˚33’25.22”S, 70˚05’59.89”W, 238m, water 23.1 C, pH 8.3, 9 Apr 2011 Ca, Co C-1770 Madre de Dios Stream at Trail 23, 12˚33’31.03”S, 70˚05’56.96”W, 280m, water 23.3 C, pH 7.8, 9 Apr 2011 Ca, Co C-1772 Madre de Dios Stream at Trail 19, 12˚34’01.04”S, 70˚05’43.24”W, 275 m, water 23.7 C, pH 5.1, 9 Apr 2011 Ca, Co C-1773 Madre de Dios Stream at Trail 28, 12˚34’02.81”S, 70˚05’42.96”W, 272 m, water 23.7 C, pH 5.1, 9 Apr 2011 Ca Ca C-1774 Cusco Rio Frio, 13 ˚13’20.87”S, 70 ˚44’30.07”W, 626m, water 25.3 C, pH 8.0, 12 Apr 2011 Ca, Co, Sh C-1775 Cusco River at end of Quincemil Trail 1, trailhead 13˚14’22.5594”S, 70˚46’12.6114”W, 688m, water 21.5 C, pH 7.7, 12 Apr 2011 Ca, Sh C-1776 Cusco Stream at Quincemil Trail 1, trailhead 13˚14’22.5594”S, 70˚46’12.6114”W, 688m, water Ca 21.4 C, pH 7.8, 12 Apr 2011 C-1777 Cusco Stream at Quincemil Trail 1, trailhead 13˚14’22.5594”S, 70˚46’12.6114”W, 688m, water Ca, Co 21.3 C, pH 6.0, 12 Apr 2011 C-1778 Cusco Stream at Quincemil Trail 1, trailhead 13˚14’22.5594”S, 70˚46’12.6114”W, 688m, water Ca, Co 21.3 C, pH 6.0, 12 Apr 2011 VOLUME 5 · NO. 2 ART I CLE Collection Country 429 Zelski et al. ART I CLE Table 3. (Continued). Collection Country state site details C-1779 Cusco Stream at Quincemil Trail 1, trailhead 13˚14’22.5594”S, 70˚46’12.6114”W, 688m, water Ca, Co 21.7 C, pH 6.8, 12 Apr 2011 C-1780 Cusco Stream at Quincemil Trail 3, trailhead 13˚18’27.756”S, 70˚48’44.9279”W, 757m, water 21.7 C, pH 6.8, 13 Apr 2011 Co, Su C-1781 Cusco Stream at Quincemil Trail 3, trailhead 13˚18’27.756”S, 70˚48’44.9279”W, 757m, water 21.5 C, pH 7.0, 13 Apr 2011 Co C-1782 Cusco Stream at Quincemil Trail 3, 13˚18’27.756”S, 70˚48’44.9279”W, 757m, water 21.2 C, pH 7.8, 13 Apr 2011 Ca, Co C-1783 Cusco River backwater at Quincemil Trail 3, 13˚18’27.756”S, 70˚48’44.9279”W, 757m, water 22.0 C, pH 7.1, 13 Apr 2011 Ca C-1784 Cusco Stream with red algae at Quincemil trail 3, 13˚18’27.756”S, 70˚48’44.9274”W, 757m, water 21.8 C, pH 7.15, 3 Apr 2011 Su C-1787 Cusco Stream crossing Interoceanic Highway, 13°21’2.4114”S, 71°39’21.9954”W, 3327m, water 11.6 C, pH 8.3, 27 May 2010 Sh C-1797 Junin River near Satipo, 11˚20’1.7154”S, 74˚37’36.192”W, 891m, water 22.0 C, pH 9.0, 21 May 2012 Sh Chiang Mai Mushroom Research Center, 19˚7’4.512”N, 98˚44’2.2194”E, 904m, water 23.5 C, pH 7.6, 9 Jul 2012 Ca C-1832 Chiang Mai Tham Rusee Nature Trail, 18˚40’24.4794”N, 90˚54’38.3754”E. 1149m, water 22.4 C, pH 7.2, 18 Jun 2012 Ca C-1833 Chiang Mai Sri Lanna National Park. Boa Tong Waterfall. 19˚4’10.848”N, 99˚4’46.8834”E, 508m, water 22.6 C, pH 7.2, 23 Jun 2012 Co C-1827 Thailand 2003). A total of 10 000 000 generations were run with trees sampled every 1 000 generations, resulting in a total of 10 000 trees. The irst 1 000 trees were discarded as burnin, and the remaining 9 000 trees were used to calculate posterior probabilities (PP). The consensus of the trees was viewed in Dendroscope v. 2.7.4 (Huson et al. 2007). RAxML analyses of the dataset produced a single most likely tree (ln -9231.511787) on which bootstrap support (> 75) and PP values (> 95) are indicated on the tree. Sequences generated in this study and the alignment used for phylogenetic analysis were deposited respectively in GenBank and in TreeBASE (www.treebase.org, submission 15251). REsULTs Field collections The entire results of ield collections will be reported in a separate paper on elevational distribution patterns of freshwater ascomycetes. For this study, ive species of dematiaceous hyphomycetes were selected for morphological and molecular phylogenetic study, as noted above (p. 425). Cancellidium applanatum, Cordana abramovii, S. juvenile, and S. uniseptatum are reported here as new records for Perú. Specimens examined are listed in the taxonomy portion of this paper with collection numbers whose details are given in Table 3. Phylogenetic analyses A single most likely tree from RAxML analysis (Fig. 1) indicated that Cancellidium applanatum groups with other freshwater Sordariomycetidae, its closest sequenced relative being Thyridium vestitum. The three sequences used in this analysis form a strongly supported monophyletic clade, with the Peruvian specimen separated from a clade containing 430 Taxa two specimens from Thailand. Inclusion of the C. pinicola sequence from GenBank (DQ144048) places that sequence irmly in Hypocreales (results not shown) as Yeung et al. (2006) reported. A BLAST search using that sequence produces a 100 % match to Trichoderma koningiopsis, suggesting contamination of the C. pinicola isolate. The results of this analysis indicate that the taxonomic placement of C. applanatum is in Sordariomycetes incertae sedis at this time. Cordana abramovii clusters with other Cordana species in a well-supported monophyletic clade (Fig. 1). Cordana has been linked to Porosphaerella via Porosphaerella cordanophora and was irst placed in Trichosphaeriaceae (Müller & Samuels 1982) and later Chaetosphaeriaceae (Réblová et al. 1999). Réblová & Winka (2000) provided molecular evidence that did not support the inclusion of Cordana in Chaetosphaeriaceae, and this study supports their conclusion. Cordanaceae is a separate lineage, widely separated from Chaetosphaeriaceae in our phylogenetic analysis. Porosphaerella borinquensis is closely related, but basal to, Cordanaceae in this analysis, not nesting within the clade. Porosphaerella borinquensis has a Pseudobotrytis terrestris asexual morph, and it has been suggested that the mitosporic morph may be a compound form of basic Cordana features (Fernández & Huhndorf 2004). Sporoschisma saccardoi has long been linked via cultural studies to Melanochaeta hemipsila and our study supports the sexual-asexual morph connection using LSU sequences from both states. Multiple attempts to sequence the 28S 5′ and 3′ ends of M. hemipsila (KF833362) were made without success. This missing data may account for the long branch for that sequence. The Peruvian specimen is placed in a wellsupported clade with M. hemipsila and M. aotearoae within Chaetosphaeriaceae, agreeing with prior molecular studies (Fernandez et al. 2006, Mugambi & Huhndorf 2008). IMA FUNGUS Dematiaceous freshwater microfungi from Perú ART I CLE Chaetosphaeriaceae 100 Melanochaeta aotearoae AF466082 Melanochaeta aotearoae AF466081 96 Sporoschisma hemipsila EU583218 Sporoschisma hemipsila EU583217 100 Sporoschisma hemipsila AF466083 Sporoschisma hemipsila AF466084 82 Sporoschisma hemipsila KF833364 87 Sporoschisma hemipsila KF833363 Sporoschisma hemipsila KF833362 99 Melanopsammella vermicularioides AF064644 100 Chaetosphaeria ovoidea AF064641 87 Chaetosphaeria pulviscula AF466091 Tainosphaeria crassiparies AF466089 Chaetosphaeria innumera AY017375 100 Chaetosphaeria tropicalis AF466080 99 Chaetosphaeria spinosa AF466079 Chaetosphaeria capitata AF466061 Thielavia subthermophila HM448442 100 Sordaria fimicola AY780079 Neurospora crassa AF286411 Sordariales 82 Lasiosphaeria ovina AF064643 Bellojisia rhynchostoma EU999217 Chaetomidium arxii FJ666359 Cordana pauciseptata HE672158 99 Cordana pauciseptata HE672159 92 Cordana pauciseptata HE672160 Cordana ellipsoidea HE672156 82 93 Cordana ellipsoidea HE672166 Cordanaceae Cordana solitaria HE672161 97 Cordana abramovii KF833361 77 Cordana inaequalis HE672157 Porosphaerella borinquensis EF063573 Bullimyces aurisporus JF775590 100 Bullimyces communis JF775585 Bullimyces costaricensis JF775591 100 Rhamphoria delicatula AF261068 Rhodoveronaea varioseptata FJ617560 96 Ceratostomella pyrenaica DQ076323 Ceratostomella cuspidata FJ617558 Xylomelasma sordida AY761087 100 Cryptadelphia groenendalensis EU528007 Cryptadelphia polyseptata AY281102 Fragosphaeria purpurea AF096191 100 Ophiostoma piliferum DQ470955 Ophiostoma stenoceras DQ836904 100 Annulusmagnus triseptatus GQ996540 100 Annulatascus trisepatatus AY780049 Annulatascaceae Ascitendus austriacus GQ996539 98 Lentomitella cirrhosa AY761085 Lentomitella crinigera AY761086 Fusoidispora aquatica AY780365 Calosphaeria barbirostris EF577059 Ceratolenta caudata JX066705 100 Riomyces rotundus JF775589 Conlarium duplumascosporaa JN936993 100 Cancellidium applanatum KF833359 99 Cancellidium applanatum KF833358 Cancellidium applanatum KF833360 Thyridium vestitum AY544671 Papulosa amerospora DQ470950 100 Nohea umiumi U46893 100 Microascales Halosphaeria appendiculata U46885 Aniptodera chesapeakensis U46882 Diaporthe eres AF408350 100 Valsa ambiens AF362564 Diapothales 100 Gnomonia gnomon AF408361 Apiognomonia errabunda AF408334 Gaeumannomyces graminis AF362557 Magnaporthales Ophioceras tenuisporum AY346295 Lindra thalassiae DQ470947 Lulworthiales Fig. 1. Most likely tree (ln -9231.511787) from LSU nrDNA analysis obtained with RAxML. ML bootstrap support values > 75 are indicated at nodes, BPP support values > 95 indicated by thickened branches. VOLUME 5 · NO. 2 431 ART I CLE Zelski et al. Fig. 2. Cancellidium applanatum (PE0063-1). A. Habit view. B–D. Conidia. E. Base of conidium. F, g. Strings of monilioid cells. Bars: A = 200 µm, B–G = 20 µm. TAXONOMY Cancellidium applanatum Tubaki, Trans. Mycol. Soc. Japan 16: 358 (1975). (Fig. 2) Description: Colonies on PYG+Ab agar 2 cm diam at 30 days, white to pale yellow, becoming dark grey at the center as conidia are formed, mycelium immersed with scant aerial hyphae, margin entire, discrete, reverse whitish to buff to 432 pale yellow. Conidiophores micronematous, mononematous, arising terminally or laterally from the hyphae, simple, erect, hyaline, smooth walled. Conidia bulbils formed as inlated ends of conidiophores, 160–220 × 51–98 (x̅ = 183.4 × 74.9 µm, n = 30), shiny, silver to black when young, brown with age, obovate to obcordate, composed of parallel rows of septate rectangular cells radiating from point of attachment with conidiophore, outer cells surrounding strings of monilioid cells. IMA FUNGUS Dematiaceous freshwater microfungi from Perú Distribution: Known from Australia, Brazil, China, Hong Kong, Japan, Malaysia, Perú, and Thailand. Notes: This fungus was recovered from a variety of habitats representing a range of environmental conditions. It is saprobic on submerged woody and palm debris in lentic and lotic habitats. The specimens examined in this study are characterized by the production of bulbils on the surface of the substrate that appear silver, brown, or black depending on age, and are composed of parallel rows of cells encapsulating strings of monilioid cells. Surprisingly, this fungus was not reported by Matsushima (1993, 1995), who studied the fungi colonizing decomposing plant debris along the same river system we sampled. It occurred at water temperatures ranging from 18.7–31.7 °C and pH 5.1–8.3. It was recovered from altitudes ranging from 218–817 m. As the fungus was not recovered from higher elevations and its distribution appears to be mainly tropical (with the exception of the type locality, which has a subtropical climate), it may be that C. applanatum is adapted to warmer habitats. Cordana abramovii Seman & Davydk., Novosti Sist. Nizsh. Rast. 20: 115 (1983). (Fig. 3) Description: Conidiophores gregarious, erect, straight or lexuous, to 6-septate, smooth, brown, paler towards the apex, 620–990 µm long × 5–6.5 µm wide (between conidiogenous swellings), base to 18 µm diam. Conidiogenous cells polyblastic (to 8), terminal and intercalary, one swelling per cell (8.5–13 µm wide), denticulate. Conidia enteroblastic, verruculose, tan to reddish brown, pyriform to obovate, thick walled (to 3.0 µm), transversely uniseptate with a septal pore, and tapered base bearing the scar of schizolytic abscission, 21–29 µm long × 11.5–16 µm wide (x̅ = 24.6 × 14.4, n = 30). Specimens examined: C-1714, PE0053-1; C-1741, PE0053-3; C-1750, PE0053-4; C-1746, PE0053-5; C-1719, PE0053-9; C-1713, PE0053-11; C-1720, PE0053-12; C-1716, PE0053-13; C-1711, PE0053-14; C-1722, PE0053-15; C-1708, PE0053-16; C-1712, PE0053-17; C-1755, PE0053-18; C-1736, PE0053-20; C-1735, VOLUME 5 · NO. 2 PE0053-21; C-1753, PE0053-22; C-1739, PE0053-23; C-1782, PE0053-24; C-1779, PE0053-25; C-1748, PE0053-26; C-1770, PE0053-27; C-1730, PE0053-28; PE0053-30; C-1754, PE005334; C-1733, PE0053-40; C-1745, PE0053-42; C-1744, PE0053-43; C-1777, PE0053-44; C-1833, TH0053-1. ART I CLE Specimens examined: C-1709, PE0063-1; C-1715, PE0063-3; C-1753, PE0063-4; C-1714, PE0063-5; C-1719, PE0063-6; C-1742, PE0063-7; C-1742, PE0063-8; C-1705, PE0063-10; C-1715, PE0063-12; C-1698, PE0063-13; C-1717, PE0063-14; C-1723, PE0063-15; C-1713, PE0063-16; C-1716, PE0063-18; C-1700, PE0063-19; C-1699, PE0063-20; C-1712, PE0063-21; C-1752, PE0063-23; C-1745, PE0063-26; C-1734, PE0063-27; C-1744, PE0063-28; C-1732, PE0063-29; C-1730, PE0063-30; C-1729, PE0063-31; C-1697, PE0063-36; C-1755, PE0063-38; C-1751, PE0063-42; C-1736, PE0063-44; C-1735, PE0063-45; C-1747, PE0063-46; C-1749, PE0063-47; C-1739, PE0063-48; C-1737, PE0063-50; C-1733, PE0063-52; C-1748, PE0063-56; C-1740, PE0063-63; C-1731, PE0063-68; C-1741, PE0063-70; C-1777, PE0063-81; C-1769, PE0063-82; C-1772, PE0063-83; C-1832, TH0063-1; C-1827, TH0063-2. Distribution: Known from Brunei, Perú, Russia, Seychelles, and Thailand. Notes: Morphologically, the Peruvian specimens reported and described herein most closely match the description of C. abramovii. The conidiophores in the Peruvian specimens are thinner than the type (5–6.5 vs. (8–)10–12.5 µm), as are the swellings of the conidiogenous zones (8.5–13 µm vs. 18 µm). Conidia are thick walled and approximately the same size (21–29 × 11.5–16 µm vs. 27–31 × 15–15.5 µm) as the type. The Peruvian specimens, however, have verruculose wall ornamentation, a feature not noted by Seman & Davydkina (1983). These morphological differences, as well as the geographic distance between the collection localities, suggest that the Peruvian specimens may represent a variation of C. abramovii s. str. or even a new species. Hyde & Goh (1998) provide evidence of a similar situation in their reports of C. abramovii var. seychellensis, an anatomically similar taxon possessing conidia with a purple, pitted episporium, and C. abramovii var. abramovii, possessing brown conidia and lacking an episporium. These variants were collected in the Old World tropics, while the type was reported from northern Ossetia. The specimens of C. abromovii in this study are restricted to Perú. Further molecular evidence should be gathered to increase our understanding of the phylogenetic afinities of these highly similar taxa as well as other members of Cordanaceae. Information from additional geographically separated specimens as well as additional molecular data, especially ITS, would shed light on whether C. abramovii represents a species complex with geographical variation, or whether these are distinct species. This fungus was recovered from a variety of habitats with a range of environmental conditions. Its habit is thus far known to be saprobic on submerged woody and palm debris in lentic and lotic habitats. Water temperature ranges from 18.7–31.7 °C and pH ranges from 5.1–8.3. Its altitudinal range is from 218–772 m. sporoschisma hemipsila (Berk. & Broome) Zelski, A.N. Mill., & Shearer, comb. nov. MycoBank MB807636 (Fig. 4) Basionym: Sphaeria hemipsila Berk. & Broome, Bot. J. Linn. Soc. 14: 126 (1873). Synonyms: Lasiosphaeria hemipsila (Berk. & Broome) Sacc., Syll. Fung. 2: 198 (1883). Chaetosphaeria hemipsila (Berk. & Broome) Petch., Ann. Roy. Bot. Gard. Peradenija 6: 336 (1917). Melanochaeta hemipsila (Berk. & Broome) E. Müll. et al., Revue Mycol. 33: 377 (1969). Chaetosphaeria coelestina Höhn., Sitzungsber. Akad. Wiss. Wein, Math.-Naturwiss. Kl, 1 Abt. 118: 324 (1909). 433 ART I CLE Zelski et al. Fig. 3. Cordana abramovii (PE0053-24). A. Habit view. B. Conidiophore and conidia. C–H. Conidia. Bars: A = 100, B = 20 µm, C–H = 10 µm. Sporoschisma saccardoi E. W. Mason & S. Hughes, Mycol. Pap. 31: 20 (1949). Description: Colonies on PYG + Ab 2 cm diam at 30 d, effuse, velutinous, with mixed tufts of conidiophores and sterile capitate setae. Mycelium immersed, composed of pale to dark brown hyphae. Capitate setae arising from a bulbous stroma 45–60 µm diam or from ascoma, pale brown, becoming paler towards the apex, straight or slightly lexuous, 5–6 septate, 434 150–200 µm long, 5–6.5 µm with subhyaline terminal swelling 10–12 µm wide. Ascomata supericial, 284–400 µm high × 280–370 µm wide (x̅ = 325 µm × 325 µm, n = 10), globose to subglobose, dark brown to black, gregarious, with capitate setae. Paraphyses to 7 µm wide at base, tapering to a rounded apex ~ 3.5 µm wide, as long as asci, free at apices, hyaline, septate, constricted at septa, unbranched. Asci 165– 230 × 13.5–22 µm (x̅ = 186.6 × 16.8, n = 10), cylindrical to cylindro-clavate, 8-spored, biseriate, pedicellate, with an I- IMA FUNGUS Dematiaceous freshwater microfungi from Perú ART I CLE Fig. 4. Sporoschisma saccardoi (PE0349-1). A. Habit view of sexual and asexual states. B. Capitate setae arising from ascoma. C. Asci. D. Young asci and paraphyses. E. Ascus apical rings. F, g. Ascospores. H. Conidiophore. I–K. Conidia. Bars: A = 100 µm, B–K = 20 µm. VOLUME 5 · NO. 2 435 ART I CLE Zelski et al. Fig. 5. A–E. Sporoschisma juvenile (PE0127-7). A. Conidiophore and capitate hypha. B. Young conidiophore. C–E. Conidia. F–M. Sporoschisma uniseptatum (PE0172-8). F. Conidiophore. g. Conidiophore and chains of conidia. H. Conidiophores and capitose hyphae. I–M. Conidia. Bars = 20 µm. 436 IMA FUNGUS Dematiaceous freshwater microfungi from Perú Specimens examined: C-1727, PE0177-1; C-1726, PE0177-2; C-1710, PE0177-3; C-1750, PE0177-4; C-1756, PE0177-5; C-1739, PE0177-6; C-1755, PE0177-7; C-1740, PE0177-10; C-1775, PE0177-15; C-1757, PE0177-12; C-1797, PE0177-21. Distribution: Known from Australia, Brunei Darussalam, Ecuador, Europe, Hong Kong, Indonesia, Kenya, Malaysia, Perú, South Africa, Taiwan, and Thailand. Notes: This fungus was recovered from a variety of habitats with a range of environmental conditions. Water temperature ranged from 11.6–22.2 °C and pH ranged from 7.1–9.0. It was recovered from altitudes ranging from 626–3566 m. The new combination is required as the epithet hemipsila takes precedence over saccardoi. As neither name is widely used, we see no case for not following the rule of priority under the ICN. sporoschisma juvenile Boud., Icones Mycol.1: 12 (1904). (Fig. 5A–E) Description: Setae interspersed among conidiophores, erect, straight or lexuous, to 6-septate, smooth, brown, paler towards apex, 100–150 × 4–6 µm, apex 5–7 µm wide, apex hyaline, capitate, coated with mucilage. Conidiophores scattered to gregarious, arising from dark interwoven hyphae, straight or lexuous, cylindrical, 110–280 µm long, 7–10 µm wide just above substrate, dark brown, smooth. Conidiogenous cells monophialidic, terminal, integrated, lageniform, consisting of a slightly swollen venter 14–20 µm wide and a tubular collarette 80–110 × 9.5–12 µm. Conidia produced in basipetal chains, cylindrical, ends rounded, 34–44 × 10.5–14.5 µm (x̅ = 38 × 12.93, n = 30), 3-septate, pale brown, verruculose. Specimens examined: C-1705, PE0127-1; C-1720, C-1727, PE0127-2; C-1728, PE0127-3; C-1725, PE0127-4; C-1720, PE01275; C-1756, PE0127-6; 1751, PE0127-7; C-1726, PE0127-12. Distribution: Known from Australia, Czechoslovakia, France, Hong Kong, Perú, Seychelles, and the UK. Notes: This fungus was recovered from a variety of habitats with a range of environmental conditions, and at altitudes VOLUME 5 · NO. 2 ranging from 244–2562 m. Water temperature ranged from 9.7–22 °C and pH ranged from 6-8.3. sporoschisma uniseptatum Bhat & W.B. Kendr., Mycotaxon 49: 71 (1993). (Fig. 5F–M) ART I CLE refractive apical apparatus 2–2.5 µm high × 4.5–5.5 µm wide (x̅ = 2.3 × 5.2, n = 10). Ascospores 44–57 × 7–9.5 µm (x̅ = 51 × 8 µm, n = 30), cylindrical, bent, 5-septate, not constricted at septa, smooth walled, with lipid droplets in each cell, apices rounded, central cells olivaceous to brown, end cells hyaline, without sheaths or appendages. Conidiophores scattered to gregarious, arising from substrate or directly from ascomata, up to 190 µm long. Conidiogenous cells monophialidic, 9–13 µm wide below venter and 17–20 µm wide above, venter to 22 µm wide, dark brown, paler at the torn apex, simple, erect, dark brown, smooth walled. Conidia formed enteroblastically inside the tubular collarette of the conidiogenous cell and emerging in a chain, doliiform, 48–60 × 11–13.5 µm (x̅ = 55.5 × 12.5 µm, n = 30), 5-septate, occasionally constricted at septa, central cells brown, end cells hyaline. Synonym: Melanochaeta garethjonesii Sivichai & HywelJones, Mycol. Res. 104: 481 (2000). Description: Conidiophores dark brown, erect, straight or lexuous, septate, cylindrical, terminating with phialidic conidiogenous cells, 125–190 µm long × 9–11 µm wide, to 22 µm wide at the swollen venter. Capitate setae present among conidiophores, erect, straight or lexuous, 3–6 septate, smooth, pale brown, paler towards the sub-hyaline apex, 120–175 x 8–10 µm, swollen apex 6–13 µm wide, surrounded by mucilage. Conidia 25.5–32.5 × 11–14 µm (x̅ = 30.8 × 12.6 µm, n = 30), formed in chains, cylindrical, truncate at both ends, slightly verruculose, 1-septate, pale brown, uniform in colour. Specimens examined: C-1704, PE0172-1; C-1696, PE0172-2; C-1702, PE0172-3; C-1722, PE0172-4; C-1715, PE0172-5; C-1705, PE0172-6; C-1746, PE0172-7; C-1755, PE0172-8; C-1735, PE01729; C-1758, PE0172-12; C-1750, PE0172-10; C-1740, PE0172-14; C-1736, PE0172-16; C-1784, PE0172-20. Distribution: Known from Australia, Brunei Darrusalam, Canada, China, Ecuador, French Guiana, Hong Kong, India, Indonesia, Italy, Malaysia, Perú, Seychelles, South Africa, Sri Lanka, Taiwan, and Thailand. Notes: The fungus was recovered from a variety of habitats with a range of environmental conditions, and altitudes ranging from 218–757 m. Water temperature ranged from 19–31.4 °C and pH ranged from 5.9–8.0. ACKNOWLEDgEMENTs John Paul Janovec, Antonio Quijano, and Janet Quijano provided logistical support and aided in collecting. Renán Clodomiro Valega Rosas provided assistance in obtaining collecting permits, export permits, and shipping samples. We thank the staff of the Los Amigos Biological Station / Centro de Investigación y Capacitación Rio Los Amigos (CICRA). We could not have conducted this research without the sanction of the Peruvian governmental agencies DGFFS, MINAG and MINAM. We also thank Zack Weber for his enormous contribution to the worklow of the laboratory. Financial support for this research was provided by the National Science Foundation (NSF Grants DEB 08-44722 and DEB-1214369). REFERENCEs Cannon PF, Kirk PM (2007) Fungal Families of the World. Wallingford: CAB International. Edgar RC (2004) MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Research 32: 1792–1797. 437 ART I CLE Zelski et al. 438 Fernández FA, Huhndorf SM (2004) Neotropical pyrenomycetes: Porosphaerella borinquensis sp. nov. and its Pseudobotrytis terrestris anamorph. Fungal Diversity 17: 11–16. Fernández FA, Miller AN, Huhndorf SM, Lutzoni FM, Zoller S (2006) Systematics of the genus Chaetosphaeria and its allied genera: morphological and phylogenetic diversity in north temperate and neotropical taxa. Mycologia 98: 121–130. Fryar SC, Booth W, Davies J, Hodgkiss IJ, Hyde KD (2004) Distribution of fungi on wood in the Tutong River, Brunei. Fungal Diversity 17: 17–38. Ho WH, Hyde KD, Hodgkiss IJ, Yanna (2001) Fungal communities on submerged wood from streams in Brunei, Hong Kong, and Malaysia. Mycological Research 105: 1492–1501. Huelsenbeck JP, Ronquist FR (2001) Mr. Bayes: Bayesian inference of phylogenetic trees. Bioinformatics 17: 754–755. Huhndorf SM, Greif M, Mugambi GK, Miller AN (2008) Two new genera in the Magnaporthaceae, a new addition to Ceratosphaeria and two new species of Lentomitella. Mycologia 100: 940–955. Huson DH, Richter DC, Rausch C, Dezulian T, Franz M, et al. (2007) Dendroscope: an interactive viewer for large phylogenetic trees. BMC Bioinformatics 8: 460. Hyde KD, Goh TK (1998) Fungi on submerged wood in the Riviere St. Marie-Louis, The Seychelles. South African Journal of Botany 64: 330–336. Matsushima T (1993) Matsushima Mycological Memoires 7: 1–75. Published by the author, Kobe, Japan. Matsushima T (1995) Matsushima Mycological Memoires 8: 1–54. Published by the author, Kobe, Japan. Miller MA, Pfeiffer W, Schwartz T (2010) Creating the CIPRES Science Gateway for inference of large phylogenetic trees. Proceedings of the Gateway Computing Environments workshop (GCE), 14 Nov. 2010: 1–8. New Orleans. Mugambi GK, Huhndorf SM (2008) A new species of Melanochaeta from Kenya. Sydowia 60: 261–266. Müller E, Harr J, Sulmont P (1969) Deux Ascomycètes dont le stade conidian présent des conidies phaeophragmiées endogènes. Revue de Mycologie 33: 369–378. Müller E, Samuels GJ (1982) Anamorphs of pyrenomycetous ascomycetes II. Porosphaerella gen. nov. and its Cordana anamorph. Sydowia 35: 150–154. Nag Raj TR, Kendrick B (1975) A Monograph of Chalara and Allied Genera. Waterloo, ON: Wilfrid Laurier University Press. Pinnoi A, Lumyong S, Hyde KD, Jones EBG (2006) Biodiversity of fungi on the palm Eleiodoxa conferta in Sirindhorn peat swamp forest, Narathiwat, Thailand. Fungal Diversity 22: 205–218. Pinruan U, Hyde KD, Lumyong S, Mckenzie EHC, Jones EBG (2007) Occurrence of fungi on tissue of the peat swamp palm Licuala longicalyata. Fungal Diversity 25: 157–173. Posada D (2008) jModelTest: phylogenetic model averaging. Molecular Biology and Evolution 25: 1253–1256. Preuss CGT (1851) Uebersicht untersuchter Pilze, besonders aus der Umgegend von Hoyerswerda. Linnaea 24: 99–153. Réblová M, Barr ME, Samuels GJ (1999) Chaetosphaeriaceae, a new family for Chaetosphaeria and its relatives. Sydowia 51: 49–70. Réblová M, Winka K (2000) Phylogeny of Chaetosphaeria and its anamorphs based on morphological and molecular data. Mycologia 92: 939–954. Rehner SA, Samuels GJ (1994) Taxonomy and phylogeny of Gliocladium analyzed from nuclear large subunit DNA sequences. Mycological Research 98: 625–634. Ronquist F, Huelsenbeck JP (2003) MRBAYES 3.1.2: Bayesian phylogenetic inference under mixed models. Bioinformatics 19: 1572–1574. Seman EO, Davydkina TA (1983) De genere Cordana Preuss in URSS. Novosti Sistematiki Nizshikh Rastenii 20: 114–118. Shaw DE (1994) The aero-aquatic fungus Cancellidium applanatum K. Tubaki in Queensland. Mycologist 8: 162–163. Shearer CA, Descals E, Kohlmeyer B, Kohlmeyer J, Marvanová L, et al. (2007) Fungal biodiversity in aquatic habitats. Biodiversity and Conservation 16: 49–67. Sivichai S, Hywel-Jones NL, Somrithipol S (2000) Lignicolous freshwater Ascomycota from Thailand: Melanochaeta and Sporoschisma anamorphs. Mycological Research 104: 478–485. Sivichai S, Jones EBG, Hywel-Jones N (2002) Fungal colonisation of wood in a freshwater stream at Tad Phu, Khao Yai National Park, Thailand. Fungal Diversity 10: 13–129. Stamakis A, Hoover P, Rougemont J (2008) A rapid bootstrap algorithm for the RAxML web servers. Systematic Biology 57: 758–771. Vilgalys R, Hester M (1990) Rapid identiication and mapping of enzymatically ampliied ribosomal DNA from several Cryptococcus species. Journal of Bacteriology 172: 4238–4246. Volkmann-Kohlmeyer B, Kohlmeyer J (1996) How to prepare truly permanent microscope slides. Mycologist 10: 107–108. Webster J, Davey A (1980) Two aero-aquatic hyphomycetes from Malaysia. Transactions of the British Mycological Society 75: 341–345. Yeung QSY, Jeewon R, Hyde KD (2006) Cancellidium pinicola sp. nov. from Pinus massoniana and its phylogeny. Cryptogamie Mycologie 27: 295–304. Zhang N, Castlebury LA, Miller AN, Huhndorf SM, Schoch CL, et al. (2006) An overview of the systematics of the Sordariomycetes based on a four-gene phylogeny. Mycologia 98: 1076–1087. Zhao G, Yu P, Liu X (2012) Cancellidium and Canalisporium (hyphomycetes) from China. Nova Hedwigia 96: 221–236. IMA FUNGUS