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The Colletotrichum dracaenophilum,C. magnum and C. orchidearum
species complexes
U. Damm
1*
, T. Sato
2
, A. Alizadeh
3
, J.Z. Groenewald
4
, and P.W. Crous
4
,
5
,
6
1
Senckenberg Museum of Natural History Görlitz, PF 300 154, 02806 Görlitz, Germany;
2
Genetic Resources Center, National Agriculture and Food Research
Organization, Kannondai, Tsukuba, Ibaraki 305-8602, Japan;
3
Department of Plant Protection, Faculty of Agriculture, Azarbaijan Shahid Madani University, Tabriz,
Iran;
4
Westerdijk Fungal Biodiversity Institute, P.O. Box 85167, 3508 AD Utrecht, The Netherlands;
5
Department of Genetics, Biochemistry and Microbiology,
Forestry and Agricultural Biotechnology Institute (FABI), University of Pretoria, Pretoria, South Africa;
6
Wageningen University and Research Centre (WUR),
Laboratory of Phytopathology, Droevendaalsesteeg 1, 6708 PB Wageningen, The Netherlands
*Correspondence: U. Damm,
ulrike.damm@senckenberg.de
Abstract: Although Glomerella glycines,Colletotrichum magnum and C. orchidearum are known as causal agents of anthracnose of soybean, Cucurbitaceae and
Orchidaceae, respectively, their taxonomy remains unresolved. In preliminary analyses based on ITS, strains of these species appear basal in Colletotrichum phy-
logenies, clustering close to C. cliviae,C. brevisporum and other recently described species from tropical or subtropical regions. Phylogenetic analyses (ITS, GAPDH,
CHS-1, HIS3, ACT, TUB2) of 102 strains previously identified as Ga. glycines,C. magnum and C. orchidearum as well as other related strains from different culture
collections and studies placed these taxa in three species complexes, and distinguished at least 24 species, including 11 new species. In this study, C. magnum,
C. orchidearum and C. piperis were epitypified and their taxonomy resolved, while C. cliviicola was proposed as a new name for C. cliviae. Furthermore, a sexual morph
was observed for C. yunnanense, while C. brevisporum,C. cliviicola and C. tropicicola were reported from new hosts or countries. Regarding their conidial morphology,
species in the C. dracaenophilum, C. magnum and C. orchidearum species complexes are reminiscent of C. gloeosporioides or C. boninense s. lat., and were likely to be
confused with them in the past.
Key words: Anthracnose, Ascomycota,Colletotrichum,Gloeosporium,Glomerella, Phylogeny, Systematics.
Taxonomic novelties: New name: Colletotrichum cliviicola Damm & Crous for C. cliviae Yan L. Yang et al.;New species: C. cacao Damm, C. cattleyicola Damm & Toy.
Sato, C. coelogynes Damm, C. lobatum Damm, C. merremiae Damm, C. musicola Damm, C. okinawense Damm & Toy. Sato, C. panamense Damm, C. plurivorum
Damm, Alizadeh & Toy. Sato, C. sojae Damm & Alizadeh, C. vittalense Damm; Epitypifications (basionyms): Glomerella magna S.F. Jenkins & Winstead,
C. orchidearum Allesch., C. piperis Petch.
Available online 7 April 2018; https://doi.org/10.1016/j.simyco.2018.04.001.
INTRODUCTION
During a systematic study of Colletotrichum species, strains were
detected that were reminiscent of C. gloeosporioides regarding
conidial morphology, but did not belong to any of the well-studied
species complexes (Cannon et al. 2012). Several of these strains
were previously identified as Glomerella glycines,Ga. magna
and C. orchidearum.
Glomerella glycines is known as the causal agent of
anthracnose of soybean. It was described by Lehman & Wolf
(1926) from soybean stems as the sexual morph of Colleto-
trichum glycines (Hemmi 1920). The Compendium of Soybean
Diseases (Sinclair 1982) lists two species on soybean, the first
being C. dematium var. truncatum (syn. of C. truncatum,Damm
et al. 2009) represented by the line drawing of C. glycines by
Hemmi (1920), which was apparently regarded as a synonym of
C. truncatum, and the second being Ga. glycines. Further
confusion was caused by connecting C. destructivum to Ga.
glycines (Tiffany & Gilman 1954, Manandhar et al. 1986).
However, a recent molecular study has shown that
C. destructivum belongs to the C. destructivum species complex,
while isolates from the study of Manandhar et al. (1986) were not
closely related to C. destructivum, belonging to a different spe-
cies complex (Damm et al. 2014). In contrast, von Arx & Müller
(1954) treated Ga. glycines as a form of Ga. cingulata with large
ascospores. Based on these records, Ga. glycines was previ-
ously thus connected to at least three different species
complexes.
Glomerella magna (syn. C. magnum) was described on
watermelon (Citrullus lanatus) in the USA and is heterothallic
(Jenkins & Winsteat 1964). According to Jenkins & Winsteat
(1964),Ga. magna is pathogenic to many species of Cucurbi-
taceae, including watermelon, cantaloup (Cucumis melo),
squash and pumpkin (Cucurbita spp.), and sometimes cucumber
(Cucumis sativus). Grand (1985) lists this species as a pathogen
of Citrullus lanatus,Cucumis melo,Cucumis sativus,Cucurbita
pepo,Cucurbita sp. and Trichosanthes anguina in North Car-
olina, USA. Recently, Ga. magna was also reported from
Cucumis sativus,Lagenaria siceraria and Luffa cylindrica
(Cucurbitaceae) in Taiwan (Tsay et al. 2010), as an anthracnose
pathogen of papaya (Carica papaya) in Brazil, Mexico and Costa
Rica (Nascimento et al. 2010, Tapia-Tussel et al. 2016, Molina-
Chaves et al. 2017) and from Lobelia chinensis in China (Li et al.
2013). Wasilwa et al. (1993) found that strains of this species
were less aggressive compared with those of C. orbiculare.
Freeman & Rodriguez (1992, 1993) and Redman et al. (1999)
studied the lifestyle of this fungus, and were able to disrupt
pathogenicity in C. magnum using an ultraviolet radiation treat-
ment, transforming it to a non-pathogenic, endophytic mutualist.
In the ITS and MAT1-2 phylogenies by Du et al. (2005), strain
Peer review under responsibility of Westerdijk Fungal Biodiversity Institute.
© 2018 Westerdijk Fungal Biodiversity Institute. Production and hosting by ELSEVIER B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-
nc-nd/4.0/).
available online at www.studiesinmycology.org STUDIES IN MYCOLOGY 92: 1–46 (2019).
1
Studies in Mycology
L2.5 (CBS 519.07) grouped outside any known species complex.
The sequence of the mating type protein (MAT1-2) gene is also
included in Marcelino et al. (2008), confirming this result. The
species was however never included in any of the recent mul-
tilocus studies of the genus (Cannon et al. 2012, Marin-Felix
et al. 2017), and its close relatives are unknown.
Colletotrichum orchidearum is known as a causal agent of
anthracnose of Orchidaceae. The species was described by
Allescher (1902) with three formae on three different Orchid-
aceae species from the Munich Botanical Garden. Recently,
strains from different Orchidaceae in China were included in a
multilocus analysis of Orchidaceae plants from China and
identified as C. orchidearum (Yang et al. 2011). Farr & Rossman
(2017) list C. orchidearum from numerous Orchidaceae hosts
from Asian, African and Latin American countries. Xu et al.
(2016) also reported this species from Arctium lappa in China.
However, except for Yang et al. (2011) and Xu et al. (2016),
these reports originate from checklists or other reports from the
pre-molecular era. There are sequences of strains called
C. orchidearum from two further studies in GenBank, one of them
displayed a strain as “type strain of C. orchidearum”(Z. Zhang
et al., unpubl. data). However, there are no strains available from
the original publication. The species was lectotypified recently
(Damm et al. 2012a), but still awaits epitypification to fix the
genetic application of the name.
In preliminary analyses based on ITS sequence data, strains
of Ga. glycines,C. magnum and C. orchidearum appear basal in
Colletotrichum phylogenies close to C. cliviae,C. brevisporum
and other recently described species from mainly tropical regions
(Yang et al. 2009, Noireung et al. 2012), and indicating that more
than one species complex was involved. The aim of this study
was therefore to clarify the systematic position of Ga. glycines,
C. magnum and C. orchidearum and related species, resolve the
respective species complexes and characterise the species in
these complexes morphologically and by means of multilocus
sequence analyses.
MATERIALS AND METHODS
Isolates
A total of 102 strains was studied, previously identified as
C. cliviae, C. orchidearum, C. brevisporum, Ga. glycines, C.
magnum and C. dracaenophilum, as well as other related strains
from the culture collections of the Westerdijk Fungal Biodiversity
Institute, Utrecht, The Netherlands (CBS), the Genetic Resources
Center, National Agriculture and Food Research Organization,
Tsukuba, Ibaraki, Japan (MAFF), the University of Tehran, Uni-
versity College of Agriculture and Natural Resources, Tehran, Iran
(UTFC) and CABI Europe UK Centre, Egham, UK (IMI) as well as
from recent studies in the literature. The Iranian strains were part
of recent collections by A. Alizadeh and O. Atghia from cultivated
and wild plants in Iran. Type material (holotypes, lectotypes and
epitypes) of the species studied are located in the fungaria of the
CBS, the US National Fungus Collections, Beltsville, Maryland,
USA (BPI), the fungaria based in the Royal Botanic Gardens,
Kew, UK (IMI and K(M)), the Botanische Staatssammlung Mün-
chen (M), Germany and the Herbarium Hamburgense (HBG),
Germany. All descriptions are based on the ex-holotype or ex-
epitype cultures, if not stated otherwise. Features of other
strains are added if deviant. Subcultures of the ex-holotypes and
ex-epitypes, respectively, as well as all other isolates used for
morphological and sequence analyses are maintained in the
culture collections of CBS, IMI, MAFF and UTFC (Table 1).
Morphological analysis
To enhance sporulation, autoclaved filter paper and double-
autoclaved stems of Anthriscus sylvestris were placed onto the
surface of synthetic nutrient-poor agar medium (SNA; Nirenberg
1976). SNA and OA (oatmeal agar; Crous et al. 2009) cultures
were incubated at 20 °C under near UV light with a 12 h
photoperiod for 10 d. Measurements and photographs of char-
acteristic structures were made according to Damm et al. (2007).
Appressoria on hyphae were observed on the reverse side of
SNA plates or on slide cultures (Damm et al. 2013). Microscopic
preparations were made in clear lactic acid, with 30 measure-
ments per structure and observed with a Nikon SMZ1000 dis-
secting microscope (DM), or with a Nikon Eclipse 80i microscope
using differential interference contrast (DIC) illumination. Colony
characters and pigment production on SNA and OA cultures
incubated at 20 °C under near UV light with 12 h photoperiod
were determined after 10 d. Colony colours were rated according
to Rayner (1970). Growth rates were measured after 7 and 10 d.
Phylogenetic analysis
Genomic DNA of the isolates was extracted using the method of
Damm et al. (2008). The 5.8S nuclear ribosomal RNA gene with
the two flanking internal transcribed spacers (ITS), a 200-bp intron
of the glyceraldehyde-3-phosphate dehydrogenase (GAPDH),
partial sequences of the chitin synthase 1 (CHS-1), histone H3
(HIS3), actin (ACT) and beta-tubulin (TUB2) genes were amplified
and sequenced using the primer pairs ITS-1F (Gardes & Bruns
1993) + ITS-4 (White et al. 1990), GDF1 + GDR1 (Guerber
et al. 2003), CHS-354R + CHS-79F (Carbone & Kohn 1999),
CYLH3F + CYLH3R (Crous et al. 2004b), ACT-512F + ACT-783R
(Carbone & Kohn 1999) and T1 (O'Donnell & Cigelnik 1997) + Bt-
2b (Glass & Donaldson 1995) or T1 + BT4R (Woudenberg et al.
2009), respectively. The PCRs were performed in a 2720 Thermal
Cycler (Applied Biosystems, Foster City, California) in a total
volume of 12.5 μL. The GAPDH,CHS-1,HIS3,ACT and TUB2
PCR mixture contained 1 μL 20× diluted genomic DNA, 0.2 μMof
each primer, 1x PCR buffer (Bioline, Luckenwalde, Germany),
2 mM MgCl
2
,20μM of each dNTP, 0.7 μL DMSO and 0.25 U Taq
DNA polymerase (Bioline). Conditions for PCR of these genes
constituted an initial denaturation step of 5 min at 94 °C, followed
by 40 cycles of 30 s at 94 °C, 30 s at 52 °C and 30 s at 72 °C, and
afinal denaturation step of 7 min at 72 °C, while the ITS PCR was
performed as described by Woudenberg et al. (2009). The DNA
sequences generated with forward and reverse primers were
used to obtain consensus sequences using Bionumerics v. 4.60
(Applied Maths, St-Marthens-Lathem, Belgium), and the align-
ment assembled and manually adjusted using Sequence Align-
ment Editor v. 2.0a11 (Rambaut 2002).
To determine whether the six sequence datasets were
congruent and combinable, tree topologies of 70 % reciprocal
Neighbour-Joining bootstrap with Maximum Likelihood distances
(10 000 replicates) with substitution models determined sepa-
rately for each partition using MrModeltest v. 2.3 (Nylander 2004)
were compared visually (Mason-Gamer & Kellogg 1996).
DAMM ET AL.
2
Table 1. Strains of Colletotrichum spp. studied, with collection details and GenBank accession numbers.
Species Accession no.
1
Host Country GenBank No.
2
ITS GAPDH CHS-1 HIS3 ACT TUB2
C. brevisporum CBS 129957 Anthurium sp. Thailand MG600762 MG600822 MG600869 MG600908 MG600966 MG601029
CBS 129958 Anthurium sp. Thailand MG600763 MG600823 MG600870 MG600909 MG600967 MG601030
CBS 512.75 Carica papaya Australia MG600761 MG600821 MG600868 MG600907 MG600965 MG601028
BCC 38876* Neoregalia sp. Thailand JN050238 JN050227 ——JN050216 JN050244
MFLUCC100182 Pandanus pygmaeus Thailand JN050239 JN050228 ——JN050217 JN050245
MAFF 305751 Passiflora edulis Japan MG600764 MG600824 MG600871 —MG600968 MG601031
C. cacao CBS 119297* Theobroma cacao Costa Rica MG600772 MG600832 MG600878 MG600916 MG600976 MG601039
C. cattleyicola CBS 170.49* Cattleya sp. Belgium MG600758 MG600819 MG600866 MG600905 MG600963 MG601025
MAFF 238321 Cattleya sp. Japan MG600759 ————MG601026
C. cliviicola CBS 125375* Clivia miniata China MG600733 MG600795 MG600850 MG600892 MG600939 MG601000
CSSS2 Clivia miniata China GU109480 GU085868 GU085866 —GU085862 GU085870
CBS 133705 Clivia sp. South Africa MG600732 MG600794 MG600849 MG600891 MG600938 MG600999
C. coelogynes CBS 132504* Coelogyne sp. Germany MG600713 MG600776 MG600836 MG600882 MG600920 MG600980
CBS 132515 Coelogyne sp. Germany MG600714 MG600777 MG600837 MG600883 MG600921 MG600981
C. dracaenophilum CBS 121453 Dracaena sanderana Bulgaria MG600712 MG600775 MG600835 MG600881 MG600919 MG600979
CBS 119360 Dracaena sanderana China MG600711 MG600774 MG600834 MG600880 MG600918 MG600978
CBS 118200 Dracaena sanderana China MG600710 MG600773 MG600833 MG600879 MG600917 MG600977
CBS 118199* Dracaena sanderana China JX519222 JX546707 JX519230 JX546756 JX519238 JX519247
C. excelsum-altitudinum CGMCC 3.15130* Bletilla ochracea China HM751815 KC843502 ——KC843548 JX625211
CGMCC 3.15131 Bletilla ochracea China JX625182 KC843503 ——KC843549 JX625212
C. gloeosporioides CBS 112999* Citrus sinensis Italy JQ005152 JQ005239 JQ005326 JQ005413 JQ005500 JQ005587
C. liaoningense 1 CGMCC 3.17616,
CAUOS2*
Capsicum annuum China KP890104 KP890135 KP890127 —KP890097 KP890111
C. liaoningense 1 CAUOS6 Capsicum annuum China ——KP890131 ——KP890115
C. liaoningense 2 CAUOS3 Capsicum annuum China KP890105 KP890136 KP890128 ——KP890112
C. liaoningense 2 CAUOS4 Capsicum annuum China KP890106 KP890137 KP890129 —KP890099 KP890113
C. lobatum IMI 79736* Piper catalpaefolium Trinidad
and Tobago
MG600768 MG600828 MG600874 MG600912 MG600972 MG601035
C. magnum CBS 519.97* Citrullus lanatus USA MG600769 MG600829 MG600875 MG600913 MG600973 MG601036
IMI 391662 Citrullus lanatus USA MG600771 MG600831 MG600877 MG600915 MG600975 MG601038
CBS 575.97 Citrullus lanatus USA MG600770 MG600830 MG600876 MG600914 MG600974 MG601037
C. merremiae CBS 124955* Merremia umbellata Panama MG600765 MG600825 MG600872 MG600910 MG600969 MG601032
C. musicola CBS 132885* Musa sp. Mexico MG600736 MG600798 MG600853 MG600895 MG600942 MG601003
CBS 127557 Musa sp. Mexico MG600737 MG600799 MG600854 MG600896 MG600943 MG601004
C. okinawense MAFF 240517 Carica papaya Japan MG600767 MG600827 ——MG600971 MG601034
C. orchidearum CORCX6 Cattleya sp. China HM585403 HM585393 HM582027 —HM581997 HM585419
UTFC 262 Cordyline terminalis Iran MG600742 MG600804 ——MG600948 MG601009
CORCG3 Cymbidium hookerianum China HM585402 HM585392 HM582026 —HM581996 HM585416
CBS 135131* Dendrobium nobile Netherlands MG600738 MG600800 MG600855 MG600897 MG600944 MG601005
CBS 136877 Dendrobium nobile Netherlands MG600739 MG600801 MG600856 MG600898 MG600945 MG601006
MAFF 239931 Dendrobium
phalaenopsis
Japan MG600745 MG600807 ——MG600951 MG601012
MAFF 240480 Dendrobium
phalaenopsis
Japan MG600746 MG600808 MG600858 —MG600952 MG601013
UTFC 266 Epipremnum aureum Iran MG600741 MG600803 ——MG600947 MG601008
MFLUCC 12-0531*
(HT of C.
hymenocallidicola)
Hymenocallis sp. Thailand KT290264 KT290263 KT290262 ———
CGMCC 3.14982*
(HT of C. aracearum)
Monstera deliciosa China KX853166 KX893585 ——KX893577 KX893581
CORCX11 Oncidium flexuosum China HM585404 HM585394 HM582028 —HM581998 HM585417
MAFF 238499 Oncidium sp. Japan MG600743 MG600805 ——MG600949 MG601010
MAFF 238779 Oncidium sp. Japan MG600744 MG600806 MG600857 —MG600950 MG601011
MAFF 306084 Phalaenopsis sp. Japan MG600747 ————MG601014
CGMCC 3.14983 Philodendron selloum China KX853167 KX893586 ——KX893578 KX893582
UTFC 265 Philodendron sp. Iran MG600740 MG600802 ——MG600946 MG601007
(continued on next page)
THE COLLETOTRICHUM DRACAENOPHILUM,C.MAGNUM AND C. ORCHIDEARUM SPECIES COMPLEXES
www.studiesinmycology.org 3
Table 1. (Continued).
Species Accession no.
1
Host Country GenBank No.
2
ITS GAPDH CHS-1 HIS3 ACT TUB2
MAFF 240504 Vanda sp. Japan MG600748 MG600809 MG600859 —MG600953 MG601015
C. panamense CBS 125386* Merremia umbellata Panama MG600766 MG600826 MG600873 MG600911 MG600970 MG601033
C. piperis IMI 71397,
CPC 21195*
Piper nigrum Malaysia MG600760 MG600820 MG600867 MG600906 MG600964 MG601027
C. plurivorum MAFF 306008 Abelmoschus esculentus Japan MG600727 MG600790 ——MG600933 MG600994
MAFF 243073 Amorphophallus rivieri Japan MG600730 MG600793 MG600847 —MG600936 MG600997
CORCX9 Arundina graminifolia China HM585398 HM585381 HM582025 —HM581986 HM585423
CGMCC 3.17358 Camellia sinensis China KJ955215 KJ954916 ——KJ954483 KJ955361
LJTJ30*
(HT of
C. sichuanensis)
Capsicum annuum China KP748221 KP823800 ——KP823741 KP823853
LJTJ3 Capsicum annuum China KP748193 KP823773 ——KP823738 KP823850
LJTJ16 Capsicum annuum China KP748207 KP823786 ——KP823739 KP823851
LJTJ22 Capsicum annuum China KP748213 KP823792 ——KP823740 KP823852
MAFF 238697 Carica papaya Japan MG600724 MG600787 ——MG600931 MG600991
CBS 125474* Coffea sp. Vietnam MG600718 MG600781 MG600841 MG600887 MG600925 MG600985
CBS 125473 Coffea sp. Vietnam MG600717 MG600780 MG600840 MG600886 MG600924 MG600984
CORCG2 Cymbidium
hookerianum
China HM585397 HM585380 HM582024 —HM581985 HM585422
MAFF 238875 Glycine max Japan MG600725 MG600788 ———MG600992
LFN0008 Glycine max. Brazil KT696336 KT696289 —KT696311 KT696275 KT696282
CBS 132443 Gossypium sp. Brazil MG600719 MG600782 MG600842 MG600888 MG600926 MG600986
CBS 132444 Gossypium sp. Brazil MG600720 MG600783 MG600843 MG600889 MG600927 MG600987
MAFF 306007 Lycopersicon
esculentum
Japan MG600728 MG600791 MG600846 —MG600934 MG600995
CMM 3742 Mangifera indica Brazil KC702980 KC702941 KC598100 —KC702908 KC992327
CMM 3746 Mangifera indica Brazil KC702981 KC702942 KC598101 —KC702909 KC992328
MAFF 305790 Musa sp. Japan MG600726 MG600789 MG600845 —MG600932 MG600993
MAFF 238315 Oncidium sp. Japan MG600729 MG600792 ——MG600935 MG600996
MAFF 305974 Passiflora edulis Japan MG600731 —MG600848 —MG600937 MG600998
CBS 903.69 Phaseolus lunatus Benin MG600721 MG600784 MG600844 MG600890 MG600928 MG600988
UTFC 261 Phaseolus vulgaris Iran MG600722 MG600785 ——MG600929 MG600989
UTFC 260 Spathiphyllum wallisii Iran MG600723 MG600786 ——MG600930 MG600990
C. sojae SAUCC 1407 Arctium lappa China KT362184 KT362188 KT362187 —KT362189 KT362185
CGMCC 3.15171 Bletilla ochracea China HM751813 KC843501 ——KC843550 KC244161
CAUOS5 Capsicum sp. China KP890107 KP890138 KP890130 ——KP890114
LFN0009 Glycine max Brazil KT696354 KT696295 —KT696318 KT696281 KT696288
UTFC 288 Glycine max Iran MG600755 MG600816 ——MG600960 MG601022
CBS 134.87 Glycine max Italy MG600752 MG600813 MG600863 MG600902 MG600957 MG601019
CBS 181.81 Glycine max Serbia MG600753 MG600814 MG600864 MG600903 MG600958 MG601020
CBS 182.81 Glycine max Serbia MG600754 MG600815 MG600865 MG600904 MG600959 MG601021
ATCC 62257* Glycine max USA MG600749 MG600810 MG600860 MG600899 MG600954 MG601016
IL18A Glycine max USA KC110792 KC110810 —KC110801 KC110828 KC110819
IL26A Glycine max USA KC110793 KC110811 —KC110802 KC110829 KC110820
ATCC 11871 Medicago sativa USA MG600750 MG600811 MG600861 MG600900 MG600955 MG601017
CBS 128510 Medicago sativa USA MG600751 MG600812 MG600862 MG600901 MG600956 MG601018
UTFC 301 Phaseolus vulgaris Iran MG600756 MG600817 ——MG600961 MG601023
UTFC 303 Vigna unguiculata Iran MG600757 MG600818 ——MG600962 MG601024
C. tropicicola 1 BCC 38877,
MFLUCC 110114*
Citrus maxima Thailand JN050240 JN050229 ——JN050218 JN050246
C. tropicicola 1 CBS 127555,
CPC 15927
Citrus sp. Mexico MG600715 MG600778 MG600838 MG600884 MG600922 MG600982
C. tropicicola 1 CBS 133174,
CPC 15924
Citrus sp. Mexico MG600716 MG600779 MG600839 MG600885 MG600923 MG600983
C. tropicicula 2 MFLUCC100167 Paphiopedilum
bellatolum
Thailand JN050241 JN050230 ——JN050219 JN050247
C. vittalense GUFCC 15503 Calamus thwaitesii India JN390935 KC790759 KF451996 —KC790646 KC790892
CBS 126.25 orchid unknown MG600735 MG600797 MG600852 MG600894 MG600941 MG601002
CBS 181.82* Theobroma cacao India MG600734 MG600796 MG600851 MG600893 MG600940 MG601001
C. yunnanense CBS 132135,
AS3.9617*
Buxus sp. China JX546804 JX546706 JX519231 JX546755 JX519239 JX519248
AS3.9616 Buxus sp. China EF369491 —————
DAMM ET AL.
4
Maximum parsimony analyses were performed on the multilocus
alignment (ITS, GAPDH, CHS-1, HIS3, ACT, TUB2) as well as
for each gene separately with PAUP (Phylogenetic Analysis
Using Parsimony) v. 4.0b10 (Swofford 2003) using the heuristic
search option with 100 random sequence additions and tree
bisection and reconstruction (TBR) as the branch-swapping al-
gorithm. Alignment gaps were treated as missing and all char-
acters were unordered and of equal weight. No more than 10
trees of score (length) greater than or equal to 10 were saved in
each replicate. Tree length, consistency index (CI), retention
index (RI), rescaled consistency index (RC) and homoplasy in-
dex (HI) were calculated for the resulting tree. The robustness of
the trees obtained was evaluated by 10 000 bootstrap replica-
tions using the Fast-stepwise addition algorithm (Hillis & Bull
1993). A Markov Chain Monte Carlo (MCMC) algorithm was
used to generate phylogenetic trees with Bayesian probabilities
using MrBayes v. 3.2.6 (Ronquist & Huelsenbeck 2003) for the
combined sequence datasets. Models of nucleotide substitution
for each gene determined by MrModeltest v. 2.3 were included
for each gene partition. The analyses of two MCMC chains were
run from random trees for 10 00 000 generations and sampled
every 100 generations. The likelihood score of the two runs were
2 630 and 2600 and therefore, the first 2 615 (the average of
both) trees were discarded as the burn-in phase of the analysis
and posterior probabilities determined from the remaining trees.
For additional comparison, a Neighbour-Joining analysis was
performed on the multigene alignment using PAUP and 1 000
bootstrap replications. Sequences derived in this study have
been lodged at GenBank, the alignment and trees in TreeBASE
(www.treebase.org/treebase-web/home.html), and taxonomic
novelties in MycoBank (Crous et al. 2004a).
RESULTS
Phylogeny
The six individual datasets did not show any conflicts in tree to-
pology of the 70 % reciprocal bootstrap trees, which allowed us
to combine them. In the multigene analyses (gene boundaries
of ITS: 1–558, GAPDH: 569–900, CHS-1:911–1211, HIS3:
1 222–1 642, ACT: 1 653–1 940, TUB2: 1 951–2 498) of 102
strains previously identified as C. cliviae, C. orchidearum, C.
brevisporum, Ga. glycines, C. magnum,C. dracaenophilum as
well as other related strains, including the outgroup
(C. gloeosporioides strain CBS 112999), 2 500 characters
including the alignment gaps were processed, of which 613
characters were parsimony-informative, 158 parsimony-
uninformative and 1729 constant. After a heuristic search using
PAUP, the maximum of 1000 equally most parsimonious trees
were retained (length = 1470 steps, CI = 0.723, RI = 0.942,
RC = 0.681, HI = 0.277), of which one is shown in Fig. 1. The
topology of the 1 000 trees was similar, which was verified for a
large selection of trees. They differed only in the position of taxa
within the subclades and in the position of some of the subclades
within the main clades. For the Bayesian analyses, a GTR+G
model was selected for ITS, a GTR+I+G model for CHS-1,a
HKY+G model for GAPDH,TUB2 and HIS3 and a HKY+I model for
ACT, and incorporated in the analysis. The consensus tree ob-
tained from Bayesian analyses confirmed the tree topology ob-
tained with parsimony. Bayesian posterior probability values
agreed with bootstrap support values (Fig. 1).
The analyses resulted in detection of three main clades and
24 subclades, presumably representing different Colletotrichum
species. The first clade (orchidearum, 99 % bootstrap support)
consists of eight subclades, of which three subclades include a
large number of strains from several host genera, and five
subclades with one to three strains of either the same host genus
or the same country. The two subclades on the top of the phy-
logeny are short-branched with bootstrap support values of 80 %
and 94 %, respectively. Except for the single-strain clade of
C. piperis, all other subclades are longer-branched with boot-
strap support values ranging between 82 and 100 %. The second
clade (magnum, 99 %) consists of 10 subclades; half of them are
single-strain clades. Two further subclades containing five and
three strains are on short branches. The remaining three sub-
clades consist of strains from three studies, the sequences of
which were downloaded from GenBank; the four strains of
C. liaoningense form two subclades. There are six subclades in
the third clade (dracaenophilum, 86 %); three of them are very
long-branched, while the other three subclades are closely
related. There was only the ITS sequence available of the
second strain of C. yunnanense; the bootstrap support value of
this long branch is therefore only 87 %. One strain previously
identified as C. tropicicola (MFLUCC 10-0167) formed a
Table 1. (Continued).
Species Accession no.
1
Host Country GenBank No.
2
ITS GAPDH CHS-1 HIS3 ACT TUB2
Colletotrichum sp. GZAAS5 09545 Citrus medica China JQ247623 JQ247599 ——JQ247647 JQ247635
COUFAL7300 Sechium edule Brazil KT285378 KT285381 KT285380 —KT285378 KT285383
1
ATCC: American Type Culture Collection, Virginia, USA; BCC: BIOTEC culture collection, Bangkok, Thailand; CBS: Culture collection of the Westerdijk Fungal
Biodiversity Institute, Utrecht, The Netherlands; CGMCC: China General Microbiological Culture Collection Center, Beijing, China; CMM: Culture Collection of Phyto-
pathogenic Fungi Prof. Maria Menezes, Federal Rural University of Pernambuco, Brazil; CPC: Culture collection of Pedro Crous, housed at CBS; COUFAL: Coleç~
ao de
Culturas de Fungos Fitopatog^
enicos da Universidade Federal de Alagoas, Brazil; GUFCC: Goa University, Fungal Culture Collection, Taleigao Plateau, Goa, India;
GZAAS: Guizhou Academy of Agricultural Sciences, Guiyang, China; IMI: Culture collection of CABI Europe UK Centre, Egham, UK; LARS: Culture collection of Long
Ashton Research Station, Bristol, UK (no longer existing); MAFF: MAFF Genebank Project, Ministry of Agriculture, Forestry and Fisheries, Tsukuba, Japan; MFLUCC:
Culture collection of the Mae Fah Luang University, Chiang Rai, Thailand ; SAUCC: Culture collection of the Department of Plant Pathology, College of Plant Protection,
Shenyang Agricultural University, China; STE-U: Culture collection of the Department of Plant Pathology, University of Stellenbosch, South Africa; UTFC: Culture collection
of the University of Tehran, University College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran.
2
ITS: internal transcribed spacers and intervening 5.8S nrDNA; GAPDH: partial glyceraldehyde-3-phosphate dehydrogenase gene; CHS-1: partial chitin synthase-1 gene;
HIS: partial histone H3 gene; ACT: partial actin gene; TUB2: partial beta-tubulin gene. Sequences generated in this study are emphasised in bold face.
* Ex-holotype or ex-epitype cultures.
THE COLLETOTRICHUM DRACAENOPHILUM,C.MAGNUM AND C. ORCHIDEARUM SPECIES COMPLEXES
www.studiesinmycology.org 5
5 changes
CBS 112999 C. gloeosporioides Citrus Italy
AS3.9616 Buxus China
CBS 132135 Buxus China*
CBS 118199 Dracaena China*
CBS 119360 Dracaena China
CBS 121453 Dracaena Bulgaria
CBS 118200 Dracaena China
CBS 132504 Coelogyne Germany*
CBS 132515 Coelogyne Germany
CBS 127555 Citrus Mexico
CBS 133174 Citrus Mexico
BCC 38877 Citrus Thailand*
MFLUCC100167 Paphiopedilum Thailand
CGMCC 3.15131 Bletilla China
CGMCC 3.15130 Bletilla China*
CBS 125473 Coffea Vietnam
CORCX9 Arundina China
UTFC 260 Spathiphyllum Iran
CBS 125474 Coffea Vietnam*
CORCG2 Cymbidium China
CBS 132443 Gossypium Brazil
UTFC 261 Phaseolus Iran
CBS 132444 Gossypium Brazil
CBS 903.69 Phaseolus Benin
LJTJ3 Capsicum China
LJTJ30 Capsicum China(*)
LFN0008 Glycine Brazil
LJTJ22 Capsicum China
CBS 125375 Camellia China
MAFF 238697 Carica Japan
MAFF 305790 Musa Japan
MAFF 306008 Abelmoschus Japan
MAFF 306007 Lycopersicon Japan
MAFF 305974 Passiflora Japan
LJTJ16 Capsicum China
MAFF 238875 Glycine Japan
MAFF 238315 Oncidium Japan
MAFF 243073 Amorphophallus Japan
CMM 3742 Mangifera Brazil
CMM 3746 Mangifera Brazil
CBS 133705 Clivia South Africa
CBS 125375 Clivia China*
CSSS2 Clivia China
CBS 181.82 Theobroma Indi a*
GUFCC 15503 Calamus India
CBS 126.25 orchid unknown
CBS 132885 Musa Mexico*
CBS 127557 Musa Mexico
MFLUCC 12-0531 Hymenocallis Thailand(*)
CGMCC 3.14982 Monstera China(*)
CGMCC 3.14983 Philodendron China
CBS 135131 Dendrobium NL*
MAFF 239931 Dendrobium Japan
MAFF 306084 Phalaenopsis Japan
MAFF 238499 Oncidium Japan
CBS 136877 Dendrobium NL
MAFF 240504 Vanda Japan
CORCG3 Cymbidium China
CORCX6 Cattleya China
CORCX11 Oncidium China
MAFF 240480 Dendrobium Japan
UTFC 266 Epipremnum Iran
UTFC 265 Philodendron Iran
UTFC 262 Cordyline Iran
MAFF 238779 Oncidium Japan
CBS 170.49 Cattleya Belgium*
MAFF 238321 Cattleya Japan
ATCC 62257 Glycine USA*
IL26A Glycine USA
ATCC 11871 Medicago USA
CBS 128510 Medicago USA
CBS 134.87 Glycine Italy
CBS 181.81 Glycine Serbia
CBS 182.81 Glycine Serbia
CGMCC 3.15171 Bletilla China
IL18A Glycine USA
UTFC 288 Glycine Iran
SAUCC 1407 Arctium China
UTFC 303 Vigna Iran
CAUOS5 Capsicum China
UTFC 301 Phaseolus Iran
LFN0009 Glycine Brazil
IMI 71397 Piper Malaysia*
BCC 38876 Neoregelia Thailand*
CBS 512.75 Carica Australia
MFLUCC100182 Pandanus Thailand
CBS 129957 Anthurium Thailand
CBS 129958 Anthurium Thailand
MAFF 305751 Passiflora Japan
IMI 79736 Piper Trin Tobago*
CBS 124955 Merre mia Panama*
CBS 125386 Merremia Panama*
MAFF 240517 Carica Japan*
CBS 519.97 Citrullus USA*
CBS 575.97 Citrullus USA
IMI 391662 Citrullus USA
CAUOS2 Capsicum China*
CAUOS6 Capsicum China
CAUOS3 Capsicum China
CAUOS4 Capsicum China
GZAAS5 09545 Citrus China
COUFAL 7300 Sechium Brazil
CBS 119297 Theobroma Costa Rica*
C. magnum
C. musicola
C. vittalense
C. cliviicola
C. plurivorum
C. piperis
C. sojae
C. cattleyicola
C. orchidearum
C. okinawense
C. panamense
C. merremiae
C. lobatum
C. brevisporum
C. excelsum-altitudinum
C. tropicicola 1
C. coelogynes
C. dracaenophilum
C. yunnanense
C. cacao
Colletotrichum sp.
C. liaoningense 1
C. liaoningense 2
C. tropicicola 2
dracaenophilum orchidearum magnum
86
94
87
92
96
100
100
96
72
72
100
98
99
90
95
80
74
72
84
99
87
94
100
100
92
98
70
82
99
98
79
83 80
86
90
94
100
1.00
1.00
1.00
1.00
1.00
1.00
1.00 1.00
0.95
1.00
1.00
1.00
1.00
1.00
1.00
1.00
0.75
1.00
1.00
1.00
1.00
0.98 1.00
1.00
1.00
0.86
1.00
1.00
1.00
1.00
1.00
1.00
1.00
1.00
1.00
1.00
1.00
0.96
1.00
1.00
1.00
1.00
1.00
0.98
0.96
0.99
1.00
0.99
1.00
Fig. 1. The first of 1000 equally most parsimonious trees obtained from a heuristic search of the combined ITS, GAPDH,CHS-1,HIS3,ACT and TUB2 sequence alignment of
the Colletotrichum dracaenophilum,C. magnum and C. orchidearum species complexes. Bootstrap support values above 70 % (bold) and Bayesian posterior probability values
above 0.90 are shown at the nodes. Colletotrichum gloeosporioides strain CBS 112999 is used as outgroup. Numbers of ex-holotype and ex-epitype isolates are emphasised
with an asterisk. Strain numbers are followed by substrate (host genus) and country of origin, NL = Netherlands, Trin Tobago = Trinidad and Tobago. Species complexes are
indicated by blue lines. Branches that are crossed by diagonal lines are shortened by 50 %.
DAMM ET AL.
6
separate lineage between the C. tropicicola clade containing the
ex-holotype of that species and C. excelsum-altitudinum. All six
subclades contain strains from only one host genus each.
The three subclades, further referred to as C. dracaenophilum,
C. magnum and C. orchidearum species complexes, respectively,
are all well supported in the multilocus phylogeny, however, the
clades are not supported with some of the single-locus phylog-
enies (not shown). Isolates can be best assigned to one of these
species complexes by using TUB2 sequence data.
Taxonomy
Based on DNA sequence data and morphology, the 102 strains
studied (Table 1) are assigned to 24 species, of which 8, 10 and
6 species, respectively, belong to the Colletotrichum dracaeno-
philum,C. magnum and C. orchidearum species complexes,
including 11 species that proved to be new to science and are
described. Five species formed sexual morphs in vitro. All spe-
cies studied in culture are characterised below.
Colletotrichum brevisporum Noireung et al., Cryptog. Mycol.
33: 350. 2012. Fig. 2.
A description of the type specimen is provided by Noireung et al.
(2012). The description below is based on strains from
Anthurium sp. in Thailand and Carica papaya in Australia.
Sexual morph not observed. Asexual morph on SNA (CBS
129957). Vegetative hyphae 2–8μm diam, hyaline to pale brown,
smooth-walled, septate, branched. Chlamydospores not
observed. Conidiomata, conidiophores and setae formed directly
on hyphae. Setae medium brown, verrucose, 60–110 μm long,
2–4-septate, base pale brown, cylindrical, sometimes slightly
inflated, 4–7μm diam, tip rounded to ± acute. Conidiophores
hyaline to pale brown, smooth-walled, septate, branched, to
40 μm long. Conidiogenous cells (hyaline to) pale brown, smooth-
walled, cylindrical to ampulliform, 12–24 × 3–6μm, opening
1–2μm diam, collarette 0.5 μm long, periclinal thickening distinct.
Conidia hyaline, smooth-walled, aseptate, the apex rounded, the
base rounded to truncate, (14–)16.5–19(–19.5) × (4 –)
4.5–5(–5.5) μm, mean ± SD = 17.7 ± 1.3 × 4.8 ± 0.4 μm, L/W
ratio = 3.7, conidia of strain CBS 512.75 shorter, measuring
(10.5–)12.5–14.5(–15.5) × (3.5–)4–5(–6) μm, mean ± SD =
13.4 ± 1.1 × 4.6 ± 0.6 μm, L/W ratio = 2.9. Appressoria not formed
by CBS 129957 after > 2 wk, appressoria of strain CBS 129958
single, dark brown, smooth-walled, irregularly roundish outline,
with an undulate to lobate margin, (5.5–)8–13(–16) × (4–)
6–10(–12.5) μm, mean ± SD = 10.4 ± 2.5 × 8.0 ± 2.2 μm, L/W
ratio = 1.3.
Asexual morph on Anthriscus stem (CBS 129957). Con-
idiomata, conidiophores and setae formed on pale brown,
angular cells, 3–8μm diam. Setae medium brown, verrucose,
Fig. 2. Colletotrichum brevisporum (A–N, U –V. from culture CBS 129957. O–T. from culture CBS 512.75). A–B. Conidiomata. C, I. Tips of setae. D, J. Bases of setae. E–H,
K–N. Conidiophores. O–T. Appressoria. U–V. Conidia. A, C–H, U. from Anthriscus stem. B, I –T, V. from SNA. A–B. Dissecting microscope (DM). C –V. Differential interference
contrast illumination (DIC). Scale bars: A = 100 μm, E = 10 μm. Scale bar of A applies to A –B. Scale bar of E applies to C –V.
THE COLLETOTRICHUM DRACAENOPHILUM,C.MAGNUM AND C. ORCHIDEARUM SPECIES COMPLEXES
www.studiesinmycology.org 7
50–120(–160) μm long, 1–3-septate, base cylindrical to slightly
inflated, 5–6.5 μm diam, tip ± acute to ± rounded. Conidiophores
pale brown, smooth-walled. Conidiogenous cells pale brown,
smooth-walled, ellipsoidal to cylindrical, 8–22 × 4–6μm,
opening 1–2μm diam, conidiogenous cells of strain CBS 512.75
often bent and inflated in the upper part and sometimes
extending to form new conidiogenous loci, collarette 0.5 μm long,
periclinal thickening visible or distinct. Conidia hyaline, smooth-
walled, aseptate, straight, cylindrical, the apex rounded, the
base rounded to truncate, (17–)18–20.5(–22) × (4–)
4.5–5(–5.5) μm, mean ± SD = 19.3 ± 1.2 × 4.6 ± 0.3 μm, L/W
ratio = 4.2, conidia of strain CBS 512.75 shorter and wider,
measuring (11–)12.5–16(–16) × (4.5 –)5 –5.5( –6) μm, mean ±
SD = 13.6 ± 1.3 × 5.3 ± 0.4 μm, L/W ratio = 2.6.
Cultural characteristics (CBS 129957): Colonies on SNA flat with
entire margin, medium hyaline to pale ochreous, filter paper
partly olivaceous grey, Anthriscus stem, filter paper and medium
partly covered with olivaceous grey, iron grey or salmon acervuli
and felty whitish aerial mycelium, reverse same colours;
25–27.5 mm in 7 d (40 mm in 10 d). Colonies on OA flat with
entire margin, surface buff to grey olivaceous, covered with pale
olivaceous grey, iron grey or salmon acervuli and very short, felty
whitish aerial mycelium, reverse buff, pale to olivaceous grey to
olivaceous grey; 26–27 mm in 7 d (40 mm in 10 d). Conidial
mass salmon.
Materials examined:Australia, Victoria, Melbourne, from stem rot of Carica
papaya, collection date and collector unknown (deposited in CBS collection Oct.
1975 by D.G. Parbery), CBS H-21067, CBS 512.75 = INB 13412. Thailand,
Chiang Mai, Mesapok waterfall, from leaf spot of Anthurium sp., 5 Oct. 2010, P.W.
Crous, CBS H-21064, culture CBS 129957; Chiang Mai, Mesapok waterfall, from
leaf spot of Anthurium sp., 5 Oct. 2010, P.W. Crous, culture CBS 129958.
Notes:Colletotrichum brevisporum was described as a causal
agent of leaf anthracnose of Neoregelia sp. and Pandanus
pygmaeus in Thailand (Noireung et al. 2012), and has been
reported as endophytes and pathogens of several host plants in
tropical regions, including Capsicum chinense in Brazil (De
Almeida et al. 2017), Capsicum annuum in China (Liu et al.
2016), Carica papaya in Australia (Shivas et al. 2016) and
Brazil (Vieira et al. 2013), Passiflora edulis in Australia (Shivas
et al. 2016), Citrus medica in China (Peng et al. 2012), Lycium
chinense in Korea (Paul et al. 2014) and Sechium edule in Brazil
(Bezerra et al. 2016).
Although there are only ITS sequences available of the
strains from papaya and passion fruit in the paper of Shivas et al.
(2016), which is not absolute proof of their identity as
C. brevisporum, different strains included in this study (CBS
512.75, MAFF 305751) confirm the occurrence of this species on
these hosts. However, we also included a strain from papaya in
Japan (MAFF 240517), that represents a different species in the
C. magnum species complex (see C. okinawense). We also
report here C. brevisporum on Anthurium in Thailand
(Fig. 2A–N, U–V). A different strain from Anthurium in Thailand
was identified as C. karstii (CBS 129927) by Damm et al.
(2012b), belonging to the C. boninense species complex.
The ITS and ACT sequences of C. brevisporum are the same
as those of C. merremiae and C. lobatum; the ITS sequence is
also identical with that of C. okinawense.Colletotrichum bre-
visporum can be identified based on its GAPDH sequence, but
two groups are resolved. The TUB2 sequences have only one
difference from C. lobatum. There are no CHS-1 and HIS3 se-
quences of the ex-type strain available, but based on the strains
included in this study, there is one additional nucleotide differ-
ence from C. lobatum and C. panamaense, and from C. lobatum
and C. merremiae, respectively.
In a blastn search on NCBI GenBank, the TUB2 sequence of
the ex-type strain BCC 38876 is 100 % identical with the two
strains of the original paper (Noireung et al. 2012), and with the
sequence of C. brevisporum strain CCCM12 from Cucurbita
moschata in China (KY797630, Liu et al. 2018). The only ACT
and GAPDH sequences that are 100 % identical with the ex-
type strain are those from the ex-type strain itself. The ACT
sequence of the strain from Pandanus from the original paper
as well as C. brevisporum strains CCCM12 from Cucurbita
moschata in China (KY797629, Liu et al. 2018), strain IRA93
from Capsicum in Brazil (KU315567, De Almeida et al. 2017),
strains CRI-L1 and CRI-N2 (KT185055, KT185056, L. Huang,
unpubl. data) and four strains from Carica papaya in Brazil
(Vieira et al. 2013) are 99 % identical (1 or 2 nucleotides dif-
ference). The GAPDH sequence of the strain from Pandanus is
99 % identical (2 nucleotides difference). The ITS sequences of
a large number of unidentified strains and strains identified as
C. brevisporum,C. magnum,Ga. magna and Ga. cingulata var.
brevispora are 100 % identical with the ITS sequence of the ex-
type strain of C. brevispora.
Colletotrichum cacao Damm, sp. nov. MycoBank MB824219.
Fig. 3.
Etymology: The species epithet is derived from the host plant,
Theobroma cacao.
Sexual morph not observed. Asexual morph on SNA. Vegetative
hyphae 1.5–5.5 μm diam, hyaline, smooth-walled, septate,
branched. Chlamydospores not observed. Conidiomata, co-
nidiophores and setae formed directly on hyphae. Setae pale to
medium brown, verrucose, 35–85 μm long, 1–2(–4)-septate,
base cylindrical to somewhat inflated, 4–7.5 μm diam,
tip ± acute. Conidiophores pale brown, smooth-walled, septate,
sometimes branched, to 45 μm long. Conidiogenous cells pale
brown, smooth-walled, ellipsoidal, obpyriform to clavate,
9–22 × (3–)4–6μm, opening 1.5–2μm diam, collarette
0.5–1μm long, periclinal thickening visible, sometimes distinct.
Conidia hyaline, smooth-walled, aseptate, straight, cylindrical,
the apex and base rounded, (13–)14–15.5(–17) × 5(–5.5) μm,
mean ± SD = 14.7 ± 0.8 × 5.1 ± 0.2 μm, L/W ratio = 2.9.
Appressoria not formed.
Asexual morph on Anthriscus stem. Conidiomata,co-
nidiophores and setae formed directly on hyphae, no basal cells
observed, but also formed in medium brown, closed conidiomata.
Setae medium brown, smooth-walled, verruculose towards the
tip, 50–130 μm long, 1–5-septate, base cylindrical, conical to
slightly inflated, 4.5–6.5 μm diam, tip ± acute to ± rounded or
functioning as conidiogenous locus. Conidiophores pale brown,
smooth-walled to verrucose, septate, branched, to 60 μm long.
Conidiogenous cells pale brown, smooth-walled to verrucose,
cylindrical to clavate, 9–24 × 3.5–6μm, opening 1.5–2μm diam,
collarette 0.5–1μm long, periclinal thickening visible to distinct.
Conidia hyaline, smooth-walled, aseptate, straight, cylindrical, the
apex and base rounded, (15.5–)16.5–18.5(–20.5) × 4.5–5.5 μm,
mean ± SD = 17.4 ± 1.1 × 5.0 ± 0.3 μm, L/W ratio = 3.5.
Culture characteristics: Colonies on SNA flat with entire margin,
agar medium hyaline to pale cinnamon, filter paper, Anthriscus
stem and agar medium partly covered with isabelline acervuli,
aerial mycelium lacking, reverse same colours; 20–25 mm in 7 d
DAMM ET AL.
8
(30–34 mm in 10 d). Colonies on OA flat with entire margin,
surface buff, almost entirely covered with small iron grey acervuli,
with rosy buff to salmon spore masses towards the centre,
whitish towards the margin, aerial mycelium lacking, reverse
olivaceous buff to olivaceous grey; 22.5–23 mm in 7 d
(34–34.5 mm in 10 d). Conidial mass rosy buff to salmon.
Material examined:Costa Rica, endophyte of Theobroma cacao, collection date
and collector unknown (deposited in CBS collection by A. Rossman, INB 13412)
(CBS H-21068 holotype, culture ex-type CBS 119297 = MCA 2773).
Notes: There are many Colletotrichum species occurring on
Theobroma cacao. For example, C. ignotum,C. theobromicola
and C. tropicale were recognised as endophytes of T. cacao by
Rojas et al. (2010) and described or epitypified in that study; they
belong to the C. gloeosporioides species complex, that was
reviewed recently by Weir et al. (2012). One species from the
C. acutatum species complex, C. sloanei, was described from
Theobroma in Malaysia, belonging to the C. acutatum species
complex (Damm et al. 2012a). A leaf endophyte of T. cacao in
Panama from the study of Rojas et al. (2010) was re-identified as
C. karstii (CBS 124951) by Damm et al. (2012b), belonging to the
C. boninense species complex. In contrast, the species
described here, C. cacao, belongs to the C. magnum species
complex. Rojas et al. (2010) noted several further unidentified
taxa amongst their collections on T. cacao from Panama.
Additional Colletotrichum species that have been described on
T. cacao (see notes under C. sloanei in Damm et al. 2012a)
include C. brachytrichum and C. theobromae from leaves of
T. cacao in Trinidad and fruits of T. cacao in Cameroon that form
conidia that are smaller than those of C. cacao, measuring
10–13.5 × 3–3.7 μm and 9 –12 × 3–5μm, respectively (Saccardo
1906). Colletotrichum cradwickii, described from branches of
T. cacao in Jamaica, forms conidia that are elongate, constricted
in the middle, measuring 14–17 × 5 μm; while C. luxificum was
collected from branches, buds and fruits of T. cacao in Surinam
and Demerara (now Guyana) forming ovoid-oblong conidia with
both ends rounded, that are sometimes slightly constricted in the
centre, measuring 13–19 × 4–5μm(Saccardo & Trotter 1913).
The conidial dimensions are similar to those of C. cacao. How-
ever, in both descriptions a constriction in the centre of the conidia
is mentioned that was not observed in C. cacao.
The formation of setae that function as conidiogenous loci
and conidia in closed conidiomata was not observed in any of the
other species of the three species complexes studied here.
Closed conidiomata are known from the C. boninense and
C. gloeosporioides species complexes (Damm et al. 2012b,B.
Weir, unpubl. data), and setae forming conidia is only known
from C. theobromicola (syn. C. fragariae) in the
C. gloeosporioides species complex (Villanueva-Arce et al.
2005).
Fig. 3. Colletotrichum cacao (from ex-holotype culture CBS 119297). A–B. Conidiomata. C. Tip of a seta. D. Base of a seta. E–F, H –K. Conidiophores. G. Seta. L–M. Conidia.
A, C–F, K –L. from Anthriscus stem. B, G–J, M. from SNA. A–B. DM. C –M. DIC. Scale bars: A = 100 μm, E = 10 μm. Scale bar of A applies to A–B. Scale bar of E applies to
C–M.
THE COLLETOTRICHUM DRACAENOPHILUM,C.MAGNUM AND C. ORCHIDEARUM SPECIES COMPLEXES
www.studiesinmycology.org 9
Colletotrichum cacao can be identified with all loci studied.
There is no TUB2 sequence in GenBank that is > 96 % identical,
no ACT sequence > 97 % identical, no HIS3 sequence > 91 %
identical, no CHS-1 sequence > 98 % identical and no GAPDH
sequence > 96 % identical to the respective sequences of
C. cacao. The ITS sequence of strain CBS 119297 is 100 %
identical to that of Colletotrichum sp. MCA 2773 (DQ286217), the
same strain sequenced by Farr et al. (2006),Colletotrichum sp.
FH2 (FJ919388) from bitter gourd, probably from India (V.
Jayakumar et al., unpubl. data), fungal endophyte STRI:ICBG-
Panama:TK766 from a tropical woody plant (KF436361,
Higginbotham et al. 2013) and C. gloeosporioides isolates S166,
S170, S183 and S193 from tissue of Aristolochia triangularis in
Brazil (MF076612–MF076615, A.K. Stuart et al., unpubl. data).
Colletotrichum cattleyicola Damm & Toy. Sato, sp. nov.
MycoBank MB824220. Fig. 4.
Etymology: The species epithet is derived from the host plant,
Cattleya.
Sexual morph not observed. Asexual morph on SNA. Vegetative
hyphae 1–8μm diam, hyaline, smooth-walled, septate,
branched. Chlamydospores not observed. Conidiomata, co-
nidiophores and setae formed directly on hyphae. Setae medium
brown, verruculose to verrucose, 50–80 μm long, 2–3-septate,
base cylindrical, slightly inflated to conical, 4.5–6μm diam,
tip ± acute to ± rounded, setae of strain MAFF 238321 up to
280 μm long. Conidiophores hyaline to pale brown, smooth-
walled, septate, branched, to 35 μm long. Conidiogenous cells
hyaline to pale brown, smooth-walled, cylindrical to ellipsoidal,
9–35 × 3.5–5.5 μm, opening 1.5–2μm diam, collarette
0.5–1μm long, periclinal thickening visible. Conidia hyaline,
smooth-walled, aseptate, straight, sometimes very slightly
curved, cylindrical, the apex and base rounded, (10.5–)
14.5–19(–20) × (4–)5–5.5(–6) μm, mean ± SD = 16.9 ± 2.1 ×
5.3 ± 0.4 μm, L/W ratio = 3.2, conidia of strain MAFF 238321
longer, measuring (14–)16.5 –20,5( –22) × 5 –6( –6.5) μm,
mean ± SD = 18,7 ± 2.0 × 5.5 ± 0.3 μm, L/W ratio = 3.4.
Appressoria single, pale to dark brown, smooth-walled, elongate
rectangular, clavate to irregular in outline, with an undulate to
lobate margin, (6–)8.5–13.5(–16) × (3.5–)4–6.5(–8) μm,
mean ± SD = 11.0 ± 2.5 × 5.1 ± 1.3 μm, L/W ratio = 2.2,
appressoria of strain MAFF 238321 wider and more variable in
size, measuring (5–)6–16.5(–29) × (4.5–)5.5–11.5(–16.5) μm,
mean ± SD = 11.3 ± 5.3 × 8.4 ± 2.9 μm, L/W ratio = 1.3.
Asexual morph on Anthriscus stem. Conidiomata, co-
nidiophores and setae formed on pale brown, angular cells,
3–7.5 μm diam. Setae pale to medium brown, verrucose,
50–100 μm long, 1–3-septate, base conical, to ± inflated,
4.5–7.5 μm diam, tip ± rounded, setae of strain MAFF 238321 up
to 330 μm long. Conidiophores pale brown, smooth-walled to
verruculose, simple or septate and branched, to 50 μm long.
Conidiogenous cells pale brown, smooth-walled to verruculose,
Fig. 4. Colletotrichum cattleyicola (from ex-holotype culture CBS 170.49). A–B. Conidiomata. C, H. Tips of setae. D, I. Bases of setae. E–G, J–M. Conidiophores. N–S.
Appressoria. T–U. Conidia. A, C–G, T. from Anthriscus stem. B, H–S, U. from SNA. A–B. DM. C–U. DIC. Scale bars: A = 100 μm, E = 10 μm. Scale bar of A applies to A –B.
Scale bar of E applies to C–U.
DAMM ET AL.
10
doliiform to cylindrical, 7 –18 × 4.5–5μm, opening 1 –1.5 μm diam,
collarette 0.5–1μm long, periclinal thickening visible, sometimes
distinct. Conidia hyaline, smooth-walled, aseptate, straight, cy-
lindrical, the apex and base rounded, (13–)16–19(–20.5) × (5–)
5.5–6(–6.5) μm, mean ± SD = 17.6 ± 1.4 × 5.9 ± 0.3 μm, L/W
ratio = 3.0, conidia of strain MAFF 238321 longer, measuring
(16–)18.5–21.5(–24.5) × (4.5–)5–6(–6.5) μm, mean ± SD =
20.0 ± 1.7 × 5.6 ± 0.4 μm, L/W ratio = 3.6.
Culture characteristics: Colonies on SNA flat with entire margin,
hyaline to pale honey, agar medium partly covered with very
short whitish aerial mycelium, reverse same colours; growth
30–32.5 mm in 7 d (40 mm in 10 d). Colonies on OA flat with
entire margin; olivaceous buff to primrose, covered with very
short whitish aerial mycelium, reverse same colours, growth
31.5–34 mm in 7 d (40 mm in 10 d). Conidial mass whitish to
very pale salmon.
Materials examined:Belgium, from a root of Cattleya sp., collection date and
collector unknown (isolated by J. van Holder, identified by A.C. Stolk, deposited in
the CBS collection Feb. 1949) (CBS H-21502 holotype, culture ex-holotype CBS
170.49). Japan, Mie Prefecture, from a lesion on a stem sheath of Cattleya sp.,
unknown collection date, T. Kobayashi (isolated by T. Kobayashi, Jul. 2000),
GLM-F111629, culture MAFF 238321 = CBS 143245 = GLMC 1836.
Notes:Colletotrichum cattleyicola belongs to the C. orchidearum
species complex. Based on our study, C. cattleyicola is only
known from a root of Cattleya in Belgium and from lesions on a
stem sheath of Cattleya sp. in Japan. The strain from Belgium
was originally identified as C. orchidearum, which is however
epitypified in this study and belongs to the C. dracaenophilum
species complex. The strain from Japan was originally identified
as C. gloeosporioides (Sato et al. 2012) and later re-identified as
C. orchidearum as well (T. Sato, unpubl. data).
Numerous Colletotrichum/Gloeosporium species were
described on Orchidaceae, among them four species that were
described on Cattleya. Colletotrichum cattleyae Verpl. was
described from dead leaves of Brasso-Cattleya
(Brassavola ×Cattleya) hybrid 'Woluwe'in Belgium with oblong
to ovoid conidia that measure 7–10 × 3.5–7μm,
mean ± SD = 9.56 ± 0.18 × 6.22 ± 0.12 μm, L/W ratio = 3.2
(Verplancke 1935b); they are shorter and have a different shape
than conidia of C. cattleyicola that are cylindrical. Gloeosporium
cattleyae Henn., Hedwigia 48: 16 (1908) [Nom. illegit., Art. 53.1]
was described from Cattleya leopoldii in Sao Paulo, Brazil, with
ellipsoidal conidia with both ends rounded, that are larger than
those of C. cattleyicola, measuring 15–22 × 7–11 μm, while Gl.
cattleyae Henn. var. macrospora Verpl. forms conidia that are
even larger, measuring 21–31 × 7 –10 μm(Verplancke 1935a).
The earlier homonym Gl. cattleyae Sacc. & D. Sacc. (Saccardo
1906) was described from dead leaves of Cattleya mossia in
Paris, France, with elongate conidia with both ends rounded,
measuring 15–20 × 4–6μm. They regard this fungus as the
asexual morph of Physalospora cattleyae Maubl. & Lasnier
(Saccardo 1905) that forms ascospores that measure
20–25 × 5–7μm. The conidial size is similar to that of
C. cattleyicola; however, we were not able to locate the type
specimen in order to verify the morphology of this species.
Another species was described fom Cattleya sp. in Italy,
C. servazzii (Gallucci-Rangone 1955); conidia are larger than
those of C. cattleyicola, measuring 26 × 7 μm.
Appressoria of C. cattleyicola are narrow and very different
in shape (mostly clavate or elongate cylindrical) compared to
other species in the C. orchidearum complex. Colletotrichum
cattleyicola can be identified based on its unique ITS, HIS3
and TUB2 sequences, while the ACT sequence is the same
as that of C. vittalense and the GAPDH sequence is the same
as those of C. sojae and C. orchidearum.TheITSofstrain
CBS 170.49 is identical to that of Colletotrichum strains ITCC
5213 from Cattleya in India (JN390844, Sharma et al. 2013b)
and Colletotrichum sp. strain GLB3 from damaged roots of
Vanilla planifolia in Mexico (KX953436, M.C.C. Gonzalez-
Chavez et al., unpubl. data), indicating possible further oc-
currences of this species. The closest match with the TUB2
sequence of strain CBS 170.49 was with 99 % identity (5
nucleotides difference) Colletotrichum sp. MST 6-3 from
leaves of Coffea arabica in Puerto Rico (KJ883603, M.C.C.
Gonzalez-Chavez et al., unpubl. data). There is only one
nucleotide difference between the GAPDH sequence of strain
CBS 170.49 and those of three unidentified Colletotrichum
isolates, C08116, C07004 and C07010 (GU935864–
GU935866), probably from Korea (Choi et al. 2011). The ACT
sequence of C. cattleyicola is 100 % identical with that of
C. cliviae strain GUFCC15503 from Calamus thwaitesii in
India (KC790646, Sharma et al. 2013a) that is re-identified as
C. vittalense in this study.
Colletotrichum cliviicola Damm & Crous, nom. nov. Myco-
Bank MB824221. Fig. 5.
Basionym:Colletotrichum cliviae Yan L. Yang et al., Fungal Di-
versity 39: 133. 2009, nom. illeg. [ICN (Melbourne) Art. 53.1], non
Chaetostroma cliviae Oudem., Verslagen van de Gewone Ver-
gaderingen der Wis- en Natuurkundige Afdeeling: 226. 1896.
Colletotrichum cliviae (Oudem.) Arnaud, Bulletin de la Soci
et
ede
pathologie v
eg
etale de France I: 37. 1914. Colletotrichum cliviae
(Oudem.) Petr., Sydowia 1 (1–3): 82 (1947), nom. illeg. [ICN
(Melbourne) Art. 53.1].
Etymology: The species epithet is derived from the host plant,
Clivia.
Sexual morph not observed. Asexual morph on SNA. Vegetative
hyphae 1–10 μm diam, hyaline, smooth-walled, septate,
branched. Chlamydospores not observed. Conidiomata, co-
nidiophores formed directly on hyphae. Setae (only formed after
> 2 wk) pale to dark brown (opaque), sometimes with a pale
brown to white tip, smooth-walled to verruculose, 50–130 μm
long, 2–4-septate, base cylindrical or slightly inflated,
4.5–5.5 μm diam, tip ± acute. Conidiophores hyaline, smooth-
walled, septate, branched, to 30 μm long, after 3 wk turning
pale brown and elongating up to 70 μm. Conidiogenous cells
hyaline, smooth-walled, cylindrical to doliiform, often ± flexuous,
upper part sometimes surrounded by a mucous sheath,
7.5–23 × 4.5–7.5 μm, opening 1.5–2μm diam, collarette
0.5–1μm long, periclinal thickening distinct. Conidia hyaline,
smooth-walled, aseptate, straight, cylindrical, the apex and base
rounded, (11–)15.5–20.5(–26.5) × (4 –)5.5 –6.5( –7) μm,
mean ± SD = 17.9 ± 2.5 × 5.9 ± 0.6 μm, L/W ratio = 3.0, after
3 wk often 1-septate, forming anastomosis or secondary conidia
(microcyclic conidiation) from short phialides formed from conidia
or anastomosis tubes. Appressoria single, medium to dark
brown, smooth-walled, elliptical, subcircular or irregular in
outline, with an undulate to lobate margin, (7–)
8.5–11.5(–12.5) × (4.5–)6.5–8.5( –9.5) μm, mean ± SD =
10.0 ± 1.7 × 7.4 ± 1.1 μm, L/W ratio = 1.3., appressoria of strain
CBS 133705 differ in shape and are longer, clavate to navicular
in outline, with an undulate to lobate margin, (5.5–)
THE COLLETOTRICHUM DRACAENOPHILUM,C.MAGNUM AND C. ORCHIDEARUM SPECIES COMPLEXES
www.studiesinmycology.org 11
12–21.5(–24) × (4.5–)5–8(–10.5) μm, mean ± SD = 16.6 ±
4.8 × 6.4 ± 1.5 μm, L/W ratio = 2.6.
Asexual morph on Anthriscus stem. Conidiomata, co-
nidiophores formed on pale brown, angular cells, 3.5–8.5 μm
diam. Setae (only formed after >2 wk) pale to dark brown
(opaque), smooth-walled to verruculose, 110–220 μm long, 2 –7-
septate, base cylindrical or slightly inflated, 3.5–7.5 μm diam,
tip ± acute. Conidiophores hyaline to pale brown, smooth-walled,
septate, to 30 μm long, after 3 wk elongating up to 80 μm.
Conidiogenous cells hyaline to pale brown, smooth-walled, cy-
lindrical to doliiform, often ± flexuous, occasionally extending to
form new conidiogenous loci, 7–19 × 4.5–7.5 μm, after 3 wk
elongating up to 40 μm, opening 1.5–2μm diam, collarette
0.5–1μm long, periclinal thickening distinct. Conidia hyaline,
smooth-walled, aseptate, straight, cylindrical, the apex and base
rounded, (13–)16.5–20.5(–22.5) × (5–)6–6.5(–7) μm,
mean ± SD = 18.4 ± 1.9 × 6.2 ± 0.4 μm, L/W ratio = 3.0, after
3 wk often 1–3-septate, forming anastomosis or secondary
conidia (microcyclic conidiation) from short phialides formed from
conidia or anastomosis tubes.
Culture characteristics: Colonies on SNA flat with entire margin,
hyaline to pale saffron, agar medium and Anthriscus stems partly
covered with whitish to pale grey aerial mycelium, Anthriscus
stems also covered with grey to black conidiomata, reverse hyaline
to pale saffron, partial smoke grey to greyish sepia; growth
35–36 mm in 7 d (40 mm in 10 d). Colonies on OA flat with
entire margin; grey olivaceous to olivaceous black, partly covered
with floccose whitish to pale grey aerial mycelium, reverse pale
olivaceous grey to olivaceous grey, growth 38–40 mm in 7 d (
40 mm in 10 d). Conidial mass whitish to very pale salmon.
Materials examined:China, Yunnan Province, Kunming, on leaf of Clivia miniata,
10 Aug. 2008, Y.L. Yang (GZAAS 080005 holotype [not seen], GLM-F112434
isotype, culture ex-holotype CBS 125375 = CSSK4 = GLMC 1861). South Af-
rica, Western Cape Province, Stellenbosch, Botanical Garden, on Clivia sp., 18.
Jul. 2012, P.W. Crous, culture CBS 133705 = CPC 21079.
Notes:Oudemans (1896) described a fungus on leaves of Clivia
nobilis (Amaryllidaceae) in the Netherlands as Chaetostroma
cliviae.Arnaud (1914) combined this species in the genus Col-
letotrichum;Petrak (1947) did the same, apparently unaware of
the publication by Arnaud (1914). Both combinations of Chae-
tostroma cliviae are not listed in Index Fungorum and MycoBank,
which was apparently the reason for Yang et al. (2009) to
overlook them and describe C. cliviae as a new species from
leaves of Clivia miniata in China. Colletotrichum cliviae (Oud.)
Arnaud has priority over the two later homonyms C. cliviae
(Oud.) Petr. and C. cliviae Yan L. Yang et al.; both are therefore
illegitimate names (Art. 53.1). In order to replace the latter name,
the new name C. cliviicola nom. nov. is provided.
Fig. 5. Colletotrichum cliviicola (A–O, W–X. from ex-holotype culture CBS 125375). A–B. Conidiomata. C, J. Tips of setae. D, K. Bases of setae. E–G, L –N. Conidiophores.
H. Two 1-septate conidia forming anastomoses. I, O. Aging conidia forming conidiogenous loci (arrow heads). Q–V. Appressoria. W–X. Conidia. A, C–I, W. from Anthriscus
stem. B, J–V, X. from SNA. A–B. Dissecting microscope (DM). C –X. Differential interference contrast illumination (DIC). Scale bars: A = 200 μm, B = 100 μm, E = 10 μm. Scale
bar of E applies to C–X.
DAMM ET AL.
12
Conidia of Chaetostroma cliviae are cylindrical with both ends
rounded, measuring 23–28 × 5–7μm(Saccardo and Sydow,
1899), that means they are longer than those of C. cliviicola,
measuring (11 –)15.5–20, 5( –26.5) × (4–)5.5 –6.5(–7) μmonSNA
and (13–)16.5–20.5(–22.5) × (5–)6–6.5(–7) μmonAnthriscus
stems. Another species described on Clivia miniata in greenhouses
in Turnau (today Turnov, Czech Republik), C. himantophylli Bub
ak
&Kab
at 1907, was regarded as a synonym of C. cliviae (Oud.)
Arnaud by Arnaud (1914). However, its conidia are narrower than
this species, and C. cliviicola, measuring 14 –24 × 4–4.5 μm
(Bub
ak & Kab
at 1907). A later homonym, C. himantophylli Verpl. &
Claess. 1934 (Nom. illegit., Art. 53.1), described from Clivia nobilis
in Belgium, forms cylindrical but narrower conidia, measuring
15–23 × 4 –4.5 μm(Trotter & Cash 1972).
Colletotrichum cliviae Yan L. Yang et al. was reported as an
anthracnose pathogen of Arundina graminifolia,Capsicum sp.,
Clivia miniata,Cymbidium hookerianum and Zamioculas zamii-
folia in China (Diao et al. 2017, Yang et al. 2009, 2011, Zhou & Li
2017) and of soybean, lima bean and grapevine in Brazil (Santos
et al. 2018, Sousa et al. 2018, Barbieri et al. 2017), on Cattleya
sp., Calamus thwaitesii,Phaseolus sp. and Saccharum sp. in
India (Sharma et al. 2013b, Chowdappa et al. 2014), Myrianthus
arboreus in Cameroon, Citrus limon in Vietnam (Douanla-Meli
et al. 2018) and as an endophyte on Camellia sinensis and
Mangifera indica in Brazil and China, respectively (Vieira et al.
2014, Liu et al. 2015). However, in this study, only strains from
Clivia grouped with the ex-holotype strain of C. cliviicola, while all
“C. cliviae”strains from hosts other than Clivia (Yang et al. 2011,
Vieira et al. 2014, Liu et al. 2015, Barbieri et al. 2017), including
strains from the MAFF culture collection that were included in our
study, were revealed to be mostly C. plurivorum, a species
closely related to C. cliviicola, C. sojae or C. vittalense.
In a study of Douanla-Meli et al. (2018),C. plurivorum (as C.
sichuanensis) was regarded as a synonym of C. cliviicola (as
C. cliviae), although both species formed well supported clades
in the phylogeny based on a multilocus data set and there was
no indication of disconcordance between the gene trees. Based
on this study, C. cliviicola is a distinct species. The C. cliviae
clade in Douanla-Meli et al. (2018) also included strains from an
undetermined ornamental plant in India that could also be Clivia.
Strains from the studies of Douanla-Meli et al. (2018), Liu
et al. (2015) and Yang et al. (2011) previously identified as
C. cliviae that were re-identified as C. plurivorum in this study,
formed a sexual morph. However, no sexual morph was
observed in strains from Clivia, identified as C. cliviicola in this
study (Yang et al. 2009, 2011, this study). Moreover, microcyclic
conidiation and the formation of anastomoses were observed in
the ex-type strain of C. cliviicola, but not in C. plurivorum. This
species is also the fastest growing species compared to all
species treated in this study.
Colletotrichum cliviicola belongs to the C. orchidearum spe-
cies complex and differs from the closely related C. plurivorum in
its TUB2,HIS and GAPDH sequences, while their CHS-1 se-
quences are identical. In the ITS and ACT trees (not shown),
C. cliviicola was also separated from C. plurivorum, but forms a
subgroup within the C. plurivorum clade.
In a pathogenicity test by Yang et al. (2011), this species
caused symptoms on Clivia spp. and Bletilla striata (Orchid-
aceae), but on none of the nine other test plants belonging to
Amaryllidaceae and other plant families.
Colletotrichum coelogynes Damm, sp. nov. MycoBank
MB824222. Fig. 6.
Etymology: Named after the host plant, Coelogyne.
Sexual morph not observed. Asexual morph on SNA. Vegetative
hyphae 1.5–11.5 μm diam, hyaline, smooth-walled, septate,
branched. Chlamydospores not observed. Conidiomata,co-
nidiophores and setae formed directly on hyphae. Setae medium
brown, smooth-walled, often verruculose towards the tip,
50–190 μm long, 2–8-septate, base cylindrical, 3.5–5.5 μm diam,
tip ± acute. Conidiophores pale to medium brown, smooth-walled,
septate, branched, to 80 μm long. Conidiogenous cells pale to
medium brown, smooth-walled, cylindrical, 12–24 × 3.5–5.5 μm,
the upper part often surrounded by a gelatinous sheath, opening
1.5–2.5 μm diam, collarette 0.5 μm long, periclinal thickening
sometimes visible. Conidia hyaline, smooth-walled, aseptate,
straight, cylindrical, the apex and base rounded, often with a
prominent scar and slightly constricted in the middle, (15–)
15.5–18( –20) × (4.5–)5–6(–6.5) μm, mean ± SD = 16.7 ± 1.2 ×
5.5 ± 0.4 μm, L/W ratio = 3.0. Appressoria single or in loose
groups, (pale) medium to dark brown, smooth-walled,
navicular, ± circular or irregular in outline, with an undulate or
entire margin, (8–)10.5–16.5(–20.5) × (6.5–)8–12( –16) μm,
mean ± SD = 13.4 ± 3.1 × 9.8 ± 2.1 μm, L/W ratio = 1.4.
Asexual morph on Anthriscus stem. Conidiomata, co-
nidiophores and setae formed on pale to medium brown, angular
cells, 4–9.5 μm diam. Setae medium brown, verruculose to
verrucose towards the tip, 70–210 μm long, 2–7-septate, base
cylindrical to conical, 5–10 μm diam, tip ± acute, often with a
constriction close to the tip. Conidiophores pale to medium
brown, smooth-walled, septate, branched, to 60 μm long. Con-
idiogenous cells pale to medium brown, smooth-walled, cylin-
drical, 16–25 × 3–5μm, opening 1–1.5 μm diam, collarette
0.5 μm long, periclinal thickening visible, sometimes distinct.
Conidia hyaline, smooth-walled, aseptate, straight, cylindrical,
the apex and base rounded, often with a prominent scar and
slightly constricted in the middle, 16–19(–23.5) × 5–6μm,
mean ± SD = 17.5 ± 1.6 × 5.5 ± 0.3 μm, L/W ratio = 3.2.
Culture characteristics: Colonies on SNA flat with entire margin,
medium buff to pale honey, chervil stem, filter paper and SNA
medium partly covered with greyish acervuli and short whitish
aerial mycelium, reverse buff to pale honey with the grey acervuli
shining through; 26–27.5 mm in 7 d (32.5–35 mm in 10 d).
Colonies on OA flat with entire margin, surface salmon, partly
covered with dark grey to orange acervuli and floccose whitish to
pale grey aerial mycelium, reverse vinaceous buff, rosy buff to
purplish grey; 27.5–32.5 mm in 7 d (40 mm in 10 d). Conidial
mass orange.
Materials examined:Germany, Munich, glasshouse, from leaves of Coelogyne
sp., 20 Nov. 2010, U. Damm (CBS H-21069 holotype, culture ex-holotype CBS
132504); Munich, glasshouse, from leaves of Coelogyne sp., 20 Nov. 2010, U.
Damm, culture CBS 132515.
Notes: Typical for C. coelogynes are the large conidiomata and
the comparatively high growth rate; C. coelogynes is the fastest
growing species in the C. dracaenophilum complex. In contrast
to C. orchidearum,C. coelogynes forms conidia that often have a
prominent scar and are slightly constricted in the middle, as well
as have longer setae. See C. orchidacearum for other species
described and reported from Orchidaceae.
THE COLLETOTRICHUM DRACAENOPHILUM,C.MAGNUM AND C. ORCHIDEARUM SPECIES COMPLEXES
www.studiesinmycology.org 13
Colletotrichum coelogynes can be identified with all loci
studied. Closest matches with the ITS sequence of
C. coelogynes strain CBS 132504 in GenBank were with 99 %
identity (1 and 4 nucleotides difference) endophytic Colleto-
trichum isolates, probably both from Dendrobium spp. from
China (FJ042517, Yuan et al. 2009 and FJ544250, C. Gao &
S.X. Guo, unpubl. data). Closest matches with the TUB2 and the
GAPDH sequences of strain CBS 132504 were with 92 % and
84 % identity, respectively, the two C. tropicicola strains from
Noireung et al. (2012). Closest matches with the ACT and HIS3
sequences were, both with 92 % identity, C. tropicicola strain
MFLUCC 11-0114 and the two C. excelsum-altitudinum strains
(Noireung et al. 2012, Tao et al. 2013) and C. pseudomajus strain
CBS 571.88 and C. radicis strain CBS 529.93 (KF687864,
KF687847, C. gigasporum complex, Liu et al. 2014), respec-
tively. Closest match with the CHS-1 sequence of strain CBS
132504 with 95 % identity were C. yunnanense strain CBS
132135 (JX519231, Cannon et al. 2012) as well as sequences of
several species belonging to the C. gloeosporioides und
C. gigasporum complexes.
Colletotrichum dracaenophilum D.F. Farr & M.E. Palm, Mycol.
Res. 110: 1401. 2006. Fig. 7.
Sexual morph not observed. Asexual morph on SNA. Vegetative
hyphae 1.5–5.5 μm diam, hyaline, smooth-walled, septate,
branched. Chlamydospores not observed. Conidiomata,co-
nidiophores and setae formed directly on hyphae. Setae medium
brown, smooth-walled, verruculose towards the tip, 50–130 μm
long, 2–5-septate, base cylindrical, sometimes slightly inflated,
5–8μm diam, tip ± acute, often ending in a conidiogenous
opening. Conidiophores hyaline (setae ending in a conidiogenous
opening medium brown), smooth-walled, septate, branched.
Conidiogenous cells hyaline (on setae ending in a conidiogenous
opening medium brown), smooth-walled, cylindrical, the upper part
sometimes surrounded by a gelatinous sheath, 10–16 ×
5.5–6.5 μm, opening 1.5–2μm diam, collarette 0.5 μm long,
periclinal thickening distinct. Conidia hyaline, smooth-walled,
aseptate, straight, cylindrical, the apex rounded, the base
rounded or truncate, sometimes tapering to the base and some-
times slightly curved, (15.5–)20–24.5(–28.5) × (5.5–)6.5–7μm,
mean ± SD = 22.2 ± 2.3 × 6.6 ± 0.3 μm, L/W ratio = 3.4, conidia of
strain CBS 121453 shorter, measuring (15.5–)17–21.5(–25.5) ×
6–7(–8) μm, mean ± SD = 19.3 ± 2.4 × 6.7 ± 0.4 μm, L/W ra-
tio = 2.9. Appressoria single or in loose groups, dark brown,
smooth-walled, navicular, cigar- to bullet-shaped in outline, with an
lobate, crenate or undulate margin, (7–)10–18(–22.5) × (5.5–)
6–8.5(–10.5) μm, mean ± SD = 13.9 ± 4.0 × 7.4 ± 1.3 μm, L/W
ratio = 1.9, appressoria of strain CBS 121453 wider, measuring
(5–)8–19(–27) × (4.5–)6.5–11.5(–15.5) μm, mean ± SD =
13.5 ± 5.5 × 9.1 ± 2.6 μm, L/W ratio = 1.5.
Fig. 6. Colletotrichum coelogynes (from ex-holotype culture CBS 132504). A–B. Conidiomata. C, H. Tips of setae. D, I. Bases of setae. E–G, J–K. Conidiophores. L–Q.
Appressoria. R–S. Conidia. A, C–G, R. from Anthriscus stem. B, H –Q, S. from SNA. A–B. DM. C–S. DIC. Scale bars: A = 200 μm, E = 10 μm. Scale bar of A applies to A –B.
Scale bar of E applies to C–S.
DAMM ET AL.
14
Asexual morph on Anthriscus stem. Conidiomata, no basal
cells were found, on which conidiophores and setae are formed.
Setae pale to medium brown, smooth-walled, verrucose towards
the tip, 60–190 μm long, 4–6-septate, base cylindrical
to ± inflated, 4–7μm diam, very thin towards the tip,
tip ± rounded. Conidiophores pale brown, smooth-walled. Con-
idiogenous cells pale brown, smooth-walled, cylindrical to
ampulliform, 10–16 × 5–7μm, opening 1–1.5 μm diam, col-
larette 0.5 μm long, rarely observed, periclinal thickening distinct.
Conidia hyaline, smooth-walled, aseptate, straight, cylindrical,
the apex rounded, the base rounded or truncate, sometimes with
a prominent scar, sometimes tapering towards the base, (17.5–)
20–25(–28) × 5.5–6.5 μm, mean ± SD = 22.4 ± 2.6 ×
6.1 ± 0.4 μm, L/W ratio = 3.7.
Culture characteristics: Colonies on SNA flat with entire margin,
hyaline to pale ochreous, filter paper partly pale ochreous, chervil
stem, filter paper and SNA medium partly covered with short
white aerial mycelium, reverse same colours; 16.5–17 mm in 7 d
(24–25.5 mm in 10 d). Colonies on OA flat with entire margin,
surface moist, saffron to orange due to sporulation, or with some
buff sectors with grey spots, aerial mycelium lacking, reverse
saffron to buff; 14.5–15.5 mm in 7 d (25–26.5 mm in 10 d).
Conidial mass saffron to orange.
Materials examined:Bulgaria, Plovdiv, from plants of Dracaena sanderiana,
bought from the market, collection date unknown, S.G. Bobev, CBS 121453 = AR
4406 = No. 1688/1. China, intercepted in San Francisco, California, USA, from
dying stems of Dracaena sanderiana, 25 Sep. 2002, J.R. Nelson (BPI 871498
holotype [not seen], culture ex-holotype CBS 118199 = MEP 1532).
Notes:Colletotrichum dracaenophilum was described as a stem
pathogen of Dracaena sanderiana; it is based on Dracaena
stems that were intercepted in California, USA, but originated
from China (Farr et al. 2006). The species is only known from
Dracaena, from D. sanderiana in Australia, Bulgaria, China,
Egypt and Florida (USA), and on D. braunii in Brazil (Farr et al.
2006, Bobev et al. 2008, Sharma et al. 2014, Macedo & Barreto
2016, Morsy & Elshahawy 2016, Shivas et al. 2016). The reports
from Bulgaria and Florida (USA) could be debated, as the plants
were imported from other countries, probably from Asia (Bobev
et al. 2008) and could have been infected by the fungus prior to
import. The strain from Bobev et al. (2008) is included in this
study.
Another species on Dracaena, C. petchii, was found on
D. marginata and Dracaena sp. in Italy and the Netherlands,
respectively, and on D. sanderiana in China and Australia, and
belongs to the C. boninense species complex (Damm et al.
2012b, Shivas et al. 2016). Both species, C. dracaenophilum
and C. petchii, are only known from Dracaena spp.
Fig. 7. C. dracaenophilum (from ex-holotype culture CBS 118199). A–B. Conidiomata. C, H. Tips of setae. D. Base of a setae and conidiophores. I. Base of a setae. E–G,
J–M. Conidiophores. M. Seta ending in a conidiogenous opening. N–S. Appressoria. T–U. Conidia. A, C–G, T. from Anthriscus stem. B, H–S, U. from SNA. A–B. DM. C–U.
DIC. Scale bars: A = 100 μm, D = 10 μm. Scale bar of A applies to A–B. Scale bar of D applies to C–U.
THE COLLETOTRICHUM DRACAENOPHILUM,C.MAGNUM AND C. ORCHIDEARUM SPECIES COMPLEXES
www.studiesinmycology.org 15
Colletotrichum dracaenophilum can be identified with all loci
studied. The closest matches with the sequences of all loci of the
ex-type strain CBS 118199 in GenBank are all those of the
C. dracaenophilum strains included in this study or mentioned
above. This species belongs to the C. dracaenophilum species
complex. Conidia of C. dracaenophilum are larger than those of
all other species treated in this study.
Colletotrichum excelsum-altitudinum G. Tao et al. [as
‘excelsum-altitudum’], Fungal Diversity 61: 152. 2013.
Description: See Tao et al. (2013).
Notes:Colletotrichum excelsum-altitudinum was described from
healthy leaves of Bletilla ochracea (Orchidaceae) in Guizhou,
China, and reported from this host together with 16 other
endophytic species, including a further six new Colletotrichum
species from the same host (Tao et al. 2013).
Colletotrichum excelsum-altitudinum belongs to the
C. dracaenophilum species complex. This species forms shorter
conidia than the other species in this complex and can be
identified with sequences of all loci available (ITS, GAPDH,ACT,
TUB2), best with GAPDH and TUB2. The ITS sequence of the
ex-type strain, CGMCC 3.15130, is 100 % identical with the two
sequences of C. excelsum-altitudinum (Tao et al. 2013) and with
that from C. excelsum-altitudinum isolate OBitC1 from
Momordica charantia (Cucurbitaceae) in India (KU239167, P.
Chowdappa et al., unpubl. data). The GAPDH sequence of the
ex-type strain, CGMCC 3.15130, is 100 % identical with the two
sequences of C. excelsum-altitudinum (Tao et al. 2013); the
sequences of all other species are 96 % identical. The ACT
sequence of strain CGMCC 3.15130 is 100 % identical with the
two sequences of C. excelsum-altitudinum (Tao et al. 2013) and
99 % identical (1 and 2 nucleotides difference) with those of the
C. tropicicola strains from Citrus and Paphiopedilum (Noireung
et al. 2012), while the TUB2 sequence is 100 % and 99 %
identical (1 nucleotides difference) with those of the C. excelsum-
altitudinum strains and 99 % identical (10 nucleotides difference)
with those of both C. tropicicola strains.
Colletotrichum liaoningense Y.Z. Diao et al., Persoonia 38: 34.
2017.
Description: See Diao et al. (2017).
Notes: This species belongs to the C. magnum species complex
and is so far only known from Capsicum in China (Diao et al.
2017).
There are five strains from Capsicum sp. in China cited in
Diao et al. (2017) that belong to the species complexes treated in
this paper; four strains were described as C. liaoningense, while
one strain was identified as C. cliviae. The C. cliviae strain
CAUOS5 was re-identified as C. sojae (C. orchidearum species
complex) in this study. However, there are several irregularities
related to the sequence data from Diao et al. (2017). We suspect,
for example, that the ACT sequence KP890098 is actually from
CAUOS5 as well and not from CAUOS3, as it is identical with
that of several strains of C. sojae; there is no ACT sequence of
CAUOS5 listed in the paper. The number of this strain in the
strain table is given as CAUOS6. The sequences of the four
C. liaoningense strains deposited by Diao et al. (2017) also all
differ from one another. For example, the ITS sequence of strain
CAUOS2, the ex-type strain of C. liaoningense differs in
numerous positions from those of the other three strains of this
species, while the TUB2 sequences are identical with
C. magnum (see notes of this species).
The four strains of C. liaoningense form two clades in the
phylogeny of this study, suggesting that their identification re-
quires verification.
Colletotrichum lobatum Damm, sp. nov. MycoBank
MB824223. Fig. 8.
Etymology: The species epithet is derived from the lobate edge
of the appressoria.
Sexual morph not observed.
Asexual morph on SNA. Vegetative hyphae 1–6.5 μm diam,
hyaline, smooth-walled, septate, branched. Chlamydospores not
observed. Conidiomata, conidiophores and setae formed directly
on hyphae. Setae pale to medium brown, verrucose, 35–90 μm
long, 1–3-septate, base cylindrical to slightly inflated, 4.5–7μm
diam, tip ± rounded to ± acute. Conidiophores hyaline to pale
brown, smooth-walled, septate, branched, to 45 μm long. Con-
idiogenous cells hyaline to pale brown, smooth-walled, cylindri-
cal, 5–30 × 3–5μm, opening 1.5–2μm diam, collarette
0.5–1μm long, distinct, periclinal thickening distinct. Conidia
hyaline, smooth-walled, aseptate, straight, cylindrical, the apex
and base rounded, (10.5–)12.5–14.5(–16) × (4–)4.5–5μm,
mean ± SD = 13.7 ± 1.0 × 4.7 ± 0.3 μm, L/W ratio = 2.9.
Appressoria not formed on the backside of the SNA plate, but in
slide culture, single, medium brown, smooth-walled, subglobose,
elliptical to irregular in outline, with an lobate or undulate margin,
(7–)7.5–15.5(–25) × (5.5–)6.5–11 ( –15) μm, mean ± SD =
11.6 ± 4.0 × 8.9 ± 2.3 μm, L/W ratio = 1.3.
Asexual morph on Anthriscus stem. Conidiomata, co-
nidiophores and setae formed on pale brown, angular cells,
3.5–7μm diam. Setae medium brown, verrucose, 45–90 μm
long, 1–3-septate, base cylindrical, sometimes slightly inflated,
4–6.5 μm diam, tip ± acute to ± rounded. Conidiophores hyaline
to pale brown, smooth-walled to verruculose, simple or septate,
branched, to 30 μm long. Conidiogenous cells pale brown,
smooth-walled to verruculose, cylindrical to doliiform,
7.5–15 × 3.5–5.5 μm, opening 1.5–2μm diam, collarette 0.5 μm
long, periclinal thickening distinct. Conidia hyaline, smooth-
walled, aseptate, straight, cylindrical, the apex and base
rounded, (13.5–)14.5–17(–18) × (4–)4.5–5(–5.5) μm,
mean ± SD = 15.6 ± 1.2 × 4.9 ± 0.3 μm, L/W ratio = 3.2.
Culture characteristics: Colonies on SNA flat with entire margin,
hyaline, honey to pale cinnamon, agar medium, filter paper and
Anthriscus stem partly covered with tiny grey to salmon acervuli,
aerial mycelium lacking, reverse same colours, growth
23–24.5 mm in 7 d (33.5–35 mm in 10 d). Colonies on OA flat
with entire margin; cinnamon, entirely covered by tiny grey dots
formed by conidiomata, aerial mycelium lacking, reverse buff to
vinaceous buff, growth 25.5–27 mm in 7 d (35.5 –37.5 mm in
10 d). Conidial mass salmon.
Material examined:Trinidad and Tobago, from Piper catalpaefolium, collection
date and collector unknown (IMI 79736 holotype, CBS H-21506 isotype, culture
ex-holotype IMI 79736 = CPC 21196).
Notes: There are two species on Piper spp. treated in this study,
C. lobatum and C. piperis.Colletotrichum lobatum belongs to the
C. magnum species complex, while C. piperis belongs to the
C. orchidearum complex. The conidial sizes and shapes are very
similar. In contrast to C. piperis,C. lobatum forms lobate
appressoria and setae that are unbranched, and colonies that
DAMM ET AL.
16
grow faster. Moreover, C. lobatum strain IMI 79736 originates
from Latin America, while C. piperis is only known from Asia
(Malaysia, Sri Lanka). See C. piperis for other species described
and reported from Piper spp.
The mostly regularly lobed roundish appressoria are also
different from the irregular appressoria of the closely related
C. brevisporum that also has faster growing colonies.
The species can be identified best by its unique GAPDH
sequence. There is respectively one nucleotide difference in the
TUB2,CHS-1 and HIS3 sequences to those of C. brevisporum,
while ITS and ACT sequences of the two species are identical.
The closest matches in a blastn search with the GAPDH
sequence in GenBank were with 97 % C. liaoningense strains
CAUOS3 and CAUOS4 (KP890136, KP890137, Diao et al.
2017), while the closest matches with the TUB2 sequence
were with 99 % identity (1 nucleotide difference) C. brevisporum
strains L57/LC0600 (ex-holotype strain) and BTL23/LC0870
(JN050244 and JN050245, Noireung et al. 2012) and strain
CCCM12 from Cucurbita moschata (KY797630, Liu et al. 2018).
The result of the blastn search with the ITS sequence was the
same as that for C. brevisporum.
Colletotrichum magnum (S.F. Jenkins & Winstead) Rossman &
W.C. Allen, IMA Fungus 7: 4. 2016. Fig. 9.
Basionym:Glomerella magna S.F. Jenkins & Winstead, Phyto-
pathology 54: 453. 1964.
Sexual morph on filter paper (only observed on the type spec-
imen BPI 596678). Ascomata perithecia, solitary, superficial or
immersed, non-stromatic, medium to dark brown, subglobose to
pyriform, 355–500 × 200–355 μm, ostiolate, with a neck.
Peridium composed of medium brown flattened textura angularis
with cells 8–17 μm diam. Ascogenous hyphae and interascal
tissue not observed. Asci unitunicate, probably 8-spored, but the
number of spores per ascus could not be seen, cylindrical,
tapering to apex and base, smooth-walled, 71–122 ×
12–15.5 μm. Ascospores initially hyaline and aseptate but can
become pale brown and septate with age, smooth-walled,
allantoid, curved most in the middle, with rounded ends, (23 –)
27–37(–43.5) × (4.5–)5–6.5(–7) μm, mean ± SD =
32.1 ± 5.0 × 5.6 ± 0.7 μm, L/W ratio = 5.7.
Asexual morph on filter paper (type specimen BPI 596678).
Conidiomata, conidiophores (disintegrated) and setae formed on
pale brown, angular cells. Setae medium brown, verruculose to
verrucose, 50–70 μm long, 1–3-septate, base cylindrical,
3–4μm diam, tip rounded. Conidiophores and conidiogenous
cells not observed. Conidia hyaline, smooth-walled, aseptate,
straight, cylindrical, the apex and base rounded, (17–)
Fig. 8. Colletotrichum lobatum (from ex-holotype culture IMI 79736). A–B. Conidiomata. C, G. Tips of setae. D, H. Bases of setae. E–F, I –K. Conidiophores. L–Q.
Appressoria. R–S. Conidia. A, C–F, R. from Anthriscus stem. B, G–Q, S. from SNA. A –B. DM. C–S. DIC. Scale bars: A = 100 μm, E = 10 μm. Scale bar of A applies to A–B.
Scale bar of E applies to C–S.
THE COLLETOTRICHUM DRACAENOPHILUM,C.MAGNUM AND C. ORCHIDEARUM SPECIES COMPLEXES
www.studiesinmycology.org 17
18–20(–20.5) × 4.5–5(–5.5) μm, mean ± SD = 19.1 ±
1.0 × 4.9 ± 0.3 μm, L/W ratio = 3.9.
Asexual morph on SNA (ex-epitype strain CBS 519.97).
Vegetative hyphae 1.5–11 μm diam, hyaline, smooth-walled,
septate, branched. Chlamydospores not observed. Con-
idiomata, conidiophores formed directly on hyphae. Setae not
observed. Conidiophores hyaline, smooth-walled, septate, not
branched, to 20 μm long, conidiophores of strain CBS 575.97
pale to medium brown, smooth-walled to verruculose, septate,
branched, to 50 μm long. Conidiogenous cells hyaline, smooth-
walled, cylindrical to ellipsoidal, often ± flexuous,
13–20 × 4–5μm, conidiogenous cells of strain CBS 575.97 pale
to medium brown, smooth-walled to verruculose, cylindrical to
clavate, opening 1.5–2μm diam, collarette 0.5 μm long, peri-
clinal thickening visible. Conidia hyaline, smooth-walled, asep-
tate, straight, sometimes very slightly curved, cylindrical, the
apex and base rounded, (11–)15.5–19( –20.5) × 4 –4.5 μm,
mean ± SD = 17.4 ± 1.8 × 4.2 ± 0.2 μm, L/W ratio = 4.1.
Appressoria single or in loose groups, medium to dark brown,
smooth-walled, ± circular, elliptical, clavate, spathulate or irreg-
ular in outline, with an entire or undulate margin, (5–)
6.5–12.5(–16) × (3.5–)4.5–7.5(–10) μm, mean ± SD = 9.4 ±
2.9 × 5.9 ± 1.5 μm, L/W ratio = 1.6.
Asexual morph on Anthriscus stem (ex-epitype strain CBS
519.97). Conidiomata, conidiophores formed directly on hyphae
or on pale brown, angular cells, 3–8.5 μm diam. Setae not
observed. Conidiophores pale brown, smooth-walled, sometimes
septate, rarely branched, to 20 μm long. Conidiogenous cells
pale brown, smooth-walled, subsphaerical to broadly ellipsoidal,
sometimes ampulliform, 5.5–12 × 4–7μm, opening 1.5–2μm
diam, collarette not observed, periclinal thickening visible. Con-
idia hyaline, smooth-walled, aseptate, sometimes very slightly
curved, cylindrical, the apex and base rounded, (17–)
18–20(–21.5) × 4.5–5(–5.5) μm, mean ± SD = 19.2 ± 1.1 ×
4.8 ± 0.3 μm, L/W ratio = 4.0, conidia of strain CBS 575.97
narrower, measuring (17–)19 –22( –24) × (3.5 –)4–4.5(–5) μm,
mean ± SD = 20.5 ± 1.4 × 4.3 ± 0.3 μm, L/W ratio = 4.8.
Culture characteristics: Colonies on SNA flat with entire margin,
hyaline to pale cinnamon, aerial mycelium lacking, in strain CBS
575.97 Anthriscus stem and medium partly covered with felty
white aerial mycelium, reverse same colours; 24.5–25.5 mm in
7 d (36–36.5 mm in 10 d), strain CBS 575.97 faster growing,
28.5–31.5 mm in 7 d (40 mm in 10 d). Colonies on OA flat
with entire margin, surface isabelline to citrine, towards the
margin straw, aerial mycelium lacking, reverse olivaceous buff to
olivaceous, 24.5–26 mm in 7 d (36.5 mm in 10 d), strain CBS
575.97 differing in colour, mycelium formation and growth rate:
dark olivaceous, olivaceous to olivaceous black, partly covered
with short felty whitish aerial mycelium, reverse olivaceous buff,
olivaceous grey to iron grey, 25.5–27.5 mm in 7 d (36 –40 mm
in 10 d). Conidial mass saffron.
Materials examined:USA, from Citrullus lanatus, collection date and collector
unknown, [BPI 596678 (S120 A × S120 F, dried culture on filter paper) holotype];
from Citrullus lanatus, collection date and collector unknown (CBS H-21063
epitype, here designated MBT380418, culture ex-epitype CBS
519.97 = L2.5 = LARS 688); from Citrullus lanatus, collection date and collector
unknown, culture CBS 575.97 = DXD = LARS 687; from Citrullus lanatus,
collection date and collector unknown (deposited in IMI collection by D.S.
Freeman), culture IMI 391662 = L2 5 = CPC 19388.
Notes: An anthracnose disease of cucurbits caused by Ga.
magna was reported from the USA (Jenkins & Winstead 1964).
The causal agent was isolated from Citrullus lanatus and
described as a heterothallic species with very large asci and
ascospores that were formed by strains crossed in the labora-
tory. The species was combined into Colletotrichum by Rossman
et al. (2016).
Compatible strains 120A (=ATCC 15015 = S120A) and 120F
(=ATCC 15016 = S120F) that were crossed in the laboratory to
produce the sexual morph were deposited in ATCC by Jenkins &
Winstead (1964). In the resulting publication, the authors state
that they deposited herbarium material in BPI as S 120A and S
120F, apparently two specimens and not indicating it as type
material. That would mean the species is not validly published,
as it was published after 1 Jan. 1958 without designating a type
(Art. 40.1). However, the label on BPI 596678 says “possible
type”; further information on the specimen: “S.F.J. & N.N.W. No.
S120 A × S120 F, Comm. N.N. Winstead May 10, 1963”clearly
reveals that the authors deposited only one specimen, the result
of the crossing of the two strains and not two, and that the details
of this specimen agree with the information in the original
description. BPI 596678 is therefore regarded as the holotype.
The two strains 120A and 120F that were crossed in order to
form the sexual morph have no type status, and there is no ex-
holotype strain retained. In a subsequent PhD thesis at Cornell
University, Ithaca, NY, USA, that was not available to us, S.
Brown conducted additional crosses of the two original strains.
Strains CBS 519.97 (=LARS 688 = L2.5) and CBS 575.97
(=LARS 687 = DXD) are derived from them (B.G. Turgeon, in
litt.). One of these strains, CBS 519.97, was selected as the
basis of the epitype.
Conidia of the holotype specimen BPI 596678 measured in
this study are smaller than those listed in the original description
by Jenkins & Winstead [1964; (24–)28(–40) × (4–)4.5(–6) μm],
measuring (17–)18–20(–20.5) × 4.5–5(–5.5) μm]. Conidial
measurements from the ex-epitype strain CBS 519.97 agree with
those from the holotype. Other features observed on the holotype
do not diverge from the original description.
Two further species were described on Citrullus in Russia and
the USA, respectively. They are possible Colletotrichum spp.
However, none of them seems to be conspecific with
C. magnum:Gloeosporium lagenaria var. citrulli forms shorter
conidia (14 × 5 μm, Potebnia 1907) than C. magnum, while
conidia of Volutella citrulli are elliptical or clavate, sometimes
slightly curved, measuring 15–20 × 3–4μm(Stoneman 1898).
Conidia of C. magnum are wider and not clavate.
Although Jenkins & Winstead (1964) proved the pathogenicity
of Ga. magna to many species of Cucurbitaceae, especially
watermelon, squash, pumpkin and cantaloup, strains and
specimens from that study are all from watermelon. Glomerella
magna is reported from other Cucurbitaceae in the USA, China
Fig. 9. Colletotrichum magnum (A–J, Q–R. from ex-epitype culture CBS 519.97. K –P. from culture CBS 575.97. S–AA. from holotype BPI 596678). A–B, W. Conidiomata.
C–J. Conidiophores. K–P. Appressoria. Q–R, T. Conidia. S. Ascoma. V. Outer surface of peridium. X–Y. Ascospores. Z. Seta. U, AA. Asci. A, C–F, Q. from Anthriscus stem;
B, G–P, R. from SNA. S–AA. from filter paper. A–B, S, W. DM. C–R, T –V. X –AA. DIC. Scale bars: A, S = 100 μm, C = 10 μm. A applies to A–B. S applies to S, W. C applies
to C–R, T –V, X –AA.
THE COLLETOTRICHUM DRACAENOPHILUM,C.MAGNUM AND C. ORCHIDEARUM SPECIES COMPLEXES
www.studiesinmycology.org 19
and Taiwan (Grand 1985, Tsay et al. 2010), from Carica papaya
in Brazil, Costa Rica and Mexico (Nascimento et al. 2010, Tapia-
Tussel et al. 2016, Molina-Chaves et al. 2017) and Lobelia
chinensis in China (Li et al. 2013), however, most of them either
lack molecular data or have ITS data only. By means of
sequence comparison, most of the respective strains could be
confirmed as belonging to the C. magnum species complex.
However, there is no species identification possible in this
complex based on ITS data.
In contrast, further reports of Colletotrichum species on Cit-
rullus lanatus include C. truncatum (as C. capsici) in India,
C. gloeosporioides and C. gloeosporioides f. sp. cucurbitae in
Brazil, C. lagenarium in the USA, Libya and Zimbabwe as well as
C. orbiculare in several countries worldwide (Farr & Rossman
2017). Probably none of these reports was based on molecu-
lar data and some could refer to C. magnum. The only strain of a
different species from Citrullus identified based on molecular
data is strain CBS 128524 from Citrullus lanatus in New Zealand
that was identified as C. karstii, belonging to the C. boninense
species complex (Damm et al. 2012b).
Colletotrichum magnum is difficult to differentiate based on
sequence data. Of the loci included, only the GAPDH and
possibly also the HIS3 sequences are unique. However, HIS3
sequences are not available of all species that are included in the
multilocus phylogeny in this study. The other loci are identical
with strains of other species, mainly C. liaoningense (Diao et al.
2017). Conidia of C. magnum are sometimes slightly curved on
both media, having a different shape than those of the closely
related C. okinawense, and are longer than those of the also
closely related C. liaoningense (71–122 × 12–15.5 μm, Diao
et al. 2017). The ascospores are curved and larger than those
of any other Colletotrichum species treated in this study and only
exceeded in length by C. gigasporum (Rakotoniriana et al. 2013)
and C. taiwanense (Sivanesan & Hsieh 1993), the former
belonging to the C. gigasporum complex, while the systematic
position of the latter is dubious (Liu et al. 2014). Both species
have filiform, 0–1- or 3–8-septate, respectively, ascospores that
are less strongly curved than those of C. magnum.
Closest matches with the GAPDH sequence of strain CBS
519.97 were with 99 % identity (1 nucleotide difference) Ga.
magna strain AK7 probably from watermelon in the USA
(DQ792850, Liu et al. 2007a) and C. brevisporum isolate LJTJ59
(KP943513) from Capsicum in China (F. Liu & G. Gong, unpubl.
data). Based on blastn searches, the ITS sequence of the ex-
type strain CBS 519.97 is 100 % identical e.g. to those of Ga.
magna and Colletotrichum sp. strains from Carica papaya in
Brazil (Nascimento et al. 2010), Mexico (E.T. Arechiga-Carvajal
et al., unpubl. data) and Malaysia (J.H. Sim et al., unpubl. data).
However, the ITS of C. magnum is identical with several other
species in the C. magnum complex.
Colletotrichum magnum was the basis for a number of mo-
lecular, morphological and pathogenicity studies on appresso-
rium formation, pathogenic and symbiotic lifestyles of fungi in
plants (Bhairi et al. 1990, Freeman & Rodriguez 1993, Redman
Fig. 10. Colletotrichum merremiae (from ex-holotype culture CBS 124955). A–B. Conidiomata. C. Tip of a seta. D. Base of a seta. H. Seta. E–G, I–L. Conidiophores. M–R.
Appressoria. S–T. Conidia. A, C–G, S. from Anthriscus stem. B, H–R, T. from SNA. A–B. DM. C –T. DIC. Scale bars: A = 100 μm, E = 10 μm. Scale bar of A applies to A–B.
Scale bar of E applies to C–T.
DAMM ET AL.
20
et al. 1999, Rodriguez et al. 2004). For example, mutation of
pathogenic C. magnum strains resulted in the loss of a virulence
factor and transformation into an endophytic fungus (Freeman &
Rodriguez 1993).
Colletotrichum merremiae Damm, sp. nov. MycoBank
MB824224. Fig. 10.
Etymology: The species epithet is derived from the host plant,
Merremia.
Sexual morph not observed. Asexual morph on SNA. Vegetative
hyphae 1.5–7μm diam, hyaline to pale brown, smooth-walled,
septate, branched. Chlamydospores not observed. Con-
idiomata, conidiophores and setae formed directly on hyphae.
Setae (pale to) medium brown, smooth-walled, verrucose to-
wards the tip, 30–65 μm long, 1–2-septate, base cylindrical
or ± inflated, 4–5.5 μm diam, tip ± acute. Conidiophores pale
brown, smooth-walled to verruculose, septate, branched, to
40 μm long. Conidiogenous cells pale brown, smooth-walled to
verruculose, cylindrical, sometimes flexuous, 10–21 × 3–5μm,
opening 1.5–2μm diam, collarette 0.5 μm long, periclinal
thickening visible, sometimes distinct. Conidia hyaline, smooth-
walled, aseptate, straight, cylindrical, the apex rounded, the
base rounded to truncate, (12.5–)14–15.5(–16.5) × (4–)
4.5–5(–5.5) μm, mean ± SD = 14.8 ± 0.9 × 4.6 ± 0.3 μm, L/W
ratio = 3.2. Appressoria single, medium brown, smooth-
walled, ± circular to elliptical in outline, with an entire or undulate
margin, (4.5–)5–7.5(–10) × (3–)3.5–5(–5.5) μm, mean ± SD =
6.4 ± 1.3 × 4.3 ± 0.7 μm, L/W ratio = 1.5.
Asexual morph on Anthriscus stem. Conidiomata, co-
nidiophores and setae formed directly on hyphae, no basal cells
observed. Setae medium brown, verruculose, 45–80 μm long,
1–2-septate, base cylindrical, conical to ± inflated, 4–7.5 μm
diam, tip ± acute. Conidiophores pale brown, smooth-walled,
septate, rarely branched. Conidiogenous cells pale brown,
smooth-walled, cylindrical to ellipsoidal, flexuous and with con-
strictions, 8–12 × 4.5–6.5 μm, opening 1.5–2μm diam, collarette
0.5 μm long, periclinal thickening visible. Conidia hyaline, smooth-
walled, aseptate, straight, cylindrical, the apex rounded, the base
rounded to truncate, (14–)15–16.5(–19) × (4–)4.5–5μm,
mean ± SD = 15.8 ± 0.9 × 4.6 ± 0.2 μm, L/W ratio = 3.4.
Culture characteristics: Colonies on SNA flat with entire margin,
hyaline to pale cinnamon, aerial mycelium lacking, reverse same
colours, 21.5–22.5 mm in 7 d (31.5–35 mm in 10 d). Colonies on
OA flat with entire margin, olivaceous, towards the margin honey
to buff, partly covered with short felty whitish aerial mycelium,
reverse olivaceous buff to olivaceous grey, 24–24.5 mm in 7 d
(35–35.5 mm in 10 d). Conidial mass rosy buff to saffron.
Material examined:Panama, Gamboa, wet lowland forest, leaf endophyte of
Merremia umbellata, Nov. 2004, S. VanBael & Z. Maynard, D3-1 (CBS H-21065
holotype, culture ex-type CBS 124955 = Q855).
Notes:Merremia is a genus of flowering plants in the Con-
volvulaceae;M. umbellata is commonly known as hogvine (www.
wikipedia.org). No Colletotrichum species was previously
described from Merremia and no strain from this host genus was
included in any of the recently studied species complexes. The
USDA fungal databases (Farr & Rossman 2017) only list Rojas
et al. (2010), the publication where the two strains, CBS 124955
and CBS 125386, described as two different species in this
study, C. merremiae and C. panamense, were derived from. Both
species belong to the C. magnum species complex.
The conidiogenous cells of C. merremiae are usually cylin-
drical and often flexuous or with constrictions, while those of
C. panamense are mostly subglobose to ellipsoidal. Conidia of
C. merremiae are on both media on average at least 2 μm
shorter than those of C. panamense. Typical are also the small
roundish appressoria, that are very different from the compact
appressoria of the closely related C. brevisporum and
C. lobatum. According to the measurements in Diao et al. (2017),
the appressoria of C. liaoningense are even smaller
(3.5–5 × 2.5–4.5 μm).
Colletotrichum merremiae can be identified based on its
unique GAPDH,HIS3 and TUB2 sequences. HIS3 differs in one,
GAPDH and TUB2 each in four nucleotides from C. brevisporum.
The ITS and ACT sequences are identical with those of
C. brevisporum and C. lobatum, that of ITS also with
C. okinawense. The ITS, GAPDH,CHS-1,HIS3,ACT and TUB2
sequences of the two species from Merremia differ in 3, 8, 5, 12,
9 and 5 nucleotides, respectively.
Closest match in a blastn search with the TUB2 sequence of
C. merremiae strain CBS 124955 is with 99 % identity (3 nucle-
otides difference) C. brevisporum strain CCCM12 from Cucurbita
moschata in China (KY797630, Liu et al. 2018) and with the
GAPDH sequence with 98 % identity the two C. brevisporum
strains from the paper of Noireung et al. (2012).NoHIS3 sequence
in GenBank is more than 91 % identical to that of C. merremiae.A
blastn search with the ITS sequence of C. merremiae resulted in
more than 50 identical sequences from strains that were identified
as C. brevisporum/Ga. cingulata var. brevisporum,C. magnum/
Ga. magna and unidentified Colletotrichum strains including that of
the same species sequenced by Rojas et al. (2010, GU994392).
Colletotrichum musicola Damm, sp. nov. MycoBank
MB824225. Fig. 11.
Etymology: The species epithet is derived from the host plant,
Musa.
Sexual morph on Anthriscus stem (only observed in strain CBS
132885). Ascomata perithecia, formed after 4 wk, solitary, su-
perficial, non-stromatic, pyriform, ostiolate, glabrous, medium to
dark brown, 200–320 × 130–220 μm diam, ostiolate, glabrous;
Peridium 5–10 μm thick, composed of 2–4 layers of medium to
dark brown flattened textura angularis with cells 5–16.5 μm
diam. Ascogenous hyphae hyaline, smooth-walled, delicate,
rarely visible. Interascal tissue formed of paraphyses, hyaline,
smooth-walled, cylindrical, with a round tip, disintegrating quickly,
septate, apically free, 40–55 μm long, at the basis 2–6μm wide.
Asci unitunicate, 8-spored, cylindrical to clavate, tapering to apex
and base, smooth-walled, 76–91.5 × 9.5–12.5 μm. Ascospores
uni- or biseriately arranged, aseptate, hyaline, sometimes pale
brown, smooth-walled, lunate to fusiform (12.5–)
16–21(–25.5) × (4.5–)5.5–6.5(–7) μm, mean ± SD = 18.4 ±
2.4 × 6.0 ± 0.5 μm, L/W ratio = 3.1.
Sexual morph on SNA (only observed in strain CBS 132885).
Ascomata pyriform to subspherical, 100–370 × 90–300 μm.
Peridium 5–10 μm thick, composed of 2–4 layers of pale to
medium brown flattened textura angularis with cells 4.5–15.5 μm
diam. Ascogenous hyphae hyaline, smooth, delicate, rarely
visible. Interascal tissue formed of paraphyses, hyaline, smooth-
walled, cylindrical, disintegrating quickly, rarely observed. Asci
unitunicate, 8-spored, narrowly clavate, cylindrical to flask-
shaped, fasciculate, 35–90.5 × 10–13.5 μm. Ascospores uni-
or biseriately arranged, aseptate, hyaline, sometimes pale brown,
THE COLLETOTRICHUM DRACAENOPHILUM,C.MAGNUM AND C. ORCHIDEARUM SPECIES COMPLEXES
www.studiesinmycology.org 21
smooth-walled, lunate to fusiform, (15–)17.5–21.5(–24.5) × (5 –)
5.5–6.5(–8) μm, mean ± SD = 19.5 ± 2.2 × 6.0 ± 0.6 μm, L/W
ratio = 3.2.
Asexual morph on SNA. Vegetative hyphae 1–10.5 μm diam,
hyaline, smooth-walled, septate, branched. Chlamydospores not
observed. Conidiomata, conidiophores and setae formed directly
on hyphae. Setae pale to medium brown, verruculose to verru-
cose, 65–95 μm long, 1–2-septate, base cylindrical to ± inflated,
4.5–6.5 μm diam, tip ± rounded. Conidiophores hyaline to pale
brown, smooth-walled, single or septate and branched, to 30 μm
long. Conidiogenous cells hyaline to pale brown, smooth-walled,
cylindrical to conical, 7.5–14.5 × 4–6μm, opening 1–1.5 μm
diam, collarette 0.5 μm long, periclinal thickening observed,
collarette of strain CBS 127557 0.5–1μm long and periclinal
thickening distinct. Conidia hyaline, smooth-walled, aseptate,
straight, cylindrical to ellipsoidal, the apex and base rounded,
(10.5–)12.5–16.5(–19) × 4.5–5.5(–6) μm, mean ± SD = 14.5 ±
2.1 × 5.2 ± 0.5 μm, L/W ratio = 2.8, conidia of strain CBS
127557 slightly larger, measuring (13.5–)14.5 –17( –19) ×
5–5.5(–6) μm, mean ± SD = 15.6 ± 1.2 × 5.3 ± 0.3 μm, L/W
ratio = 2.9. Appressoria single or in loose groups, medium to dark
brown, smooth-walled, elliptical, navicular, bullet-shaped or
irregular outline, with undulate or lobate margin, (7.5–)
9.5–15(–18.5) × (4.5–)5–7.5(–10) μm, mean ± SD = 12.3 ±
2.8 × 6.4 ± 1.3 μm, L/W ratio = 1.9, appressoria of strain CBS
127557 slightly larger, measuring (9–)11–16.5(–20) × (5–)
6–11( –13) μm, mean ± SD = 13.7 ± 2.8 × 8.3 ± 2.5 μm, L/W
ratio = 1.6.
Asexual morph on Anthriscus stem not observed in strain
CBS 132885, but few conidia and one seta observed in strain
CBS 127557. Seta medium brown, verruculose, 62 μm long, 2-
septate, base slightly inflated, 5.5 μm diam, tip ± acute. Co-
nidiophores and conidiogenous cells not observed. Conidia hy-
aline, smooth-walled, aseptate, straight, cylindrical to ellipsoidal,
the apex and base rounded, (14–)14.5–17(–17.5) × (5–)
5.5–6μm, mean ± SD = 15.5 ± 1.2 × 5.6 ± 0.3 μm, L/W
ratio = 2.8.
Culture characteristics: Colonies on SNA flat with entire margin,
hyaline to honey, agar medium, filter paper and Anthriscus stem
partly covered with felty whitish aerial mycelium and grey to black
conidiomata/ascomata, reverse same colours; growth
27.5–29.5 mm in 7 d (40 mm in 10 d). Colonies on OA flat
with entire margin; olivaceous buff, grey olivaceous to oliva-
ceous, partly covered with short felty whitish aerial mycelium and
grey conidiomata/ascomata, reverse olivaceous buff to oliva-
ceous grey, growth 28–30 mm in 7 d (40 mm in 10 d). Conidial
mass pale salmon.
Materials examined:Mexico, from Musa sp., 16 Dec. 2008, M. de J. Yanez
Morales (CBS H-21500 holotype, culture ex-holotype CBS 132885 = CPC
16328); Tapachula, Chiapas, from Musa sp., 16 Dec. 2008, M. de J. Yanez
Morales, CBS H-21501, culture CBS 127557 = CPC 16329.
Notes:Colletotrichum musicola can be identified with all loci
studied. Closest matches with the ITS sequence of isolate CBS
132885 were with 99 % identity (1 nucleotide difference)
“C. gloeosporioides”isolates CG0305 from Glycine max in
Taiwan (FJ172224, Chen et al. 2006) and AC4-M-Mexico from
post-harvest fruit of Musa acuminata in Mexico (M. Espinoza-
Ortega et al., unpubl. data), possibly a similar strain or even
one of the strains studied here. Closest matches to the TUB2
sequence of CBS 132885 are with seven nucleotides difference
(99 % identity) C. orchidearum strains CORCG3 and CORCX6
from Cymbidium and Cattleya, respectively, in China (HM585418
and HM585416, Yang et al. 2011). Closest match with the
GAPDH sequence of isolate CBS 132885 was with 99 % identity
(2 nucleotides difference) “C. cliviae”strains LC0551 and
LC1238, isolated as endophytes from Pennisetum purpureum in
Thailand (KC835389, KC835390, Manamgoda et al. 2013).
There is no ACT and no CHS-1 sequence with > 98 % identity
and no HIS3 sequence with > 96 % identity to those of
C. musicola available in GenBank.
This species belongs to the C. orchidearum species complex.
Several Colletotrichum species are known from Musa, for
example C. aotearoa,C. chrysophilum,C. musae,C. siamense,
C. theobromicola and C. tropicale belonging to the
C. gloeosporioides complex (Weir et al. 2012, Sharma et al.
2015, Vieira et al. 2018), C. karstii, belonging to the
C. boninense complex (Damm et al. 2012b), C. paxtonii and
C. scovillei belonging to the C. acutatum complex (Damm et al.
2012a, Zhou et al. 2017) and C. gigasporum belonging to the
C. gigasporum complex (Liu et al. 2014). Further Colletotrichum
species were described on Musa spp. without sequence data
(see notes under C. paxtonii in Damm et al. 2012a). We could
not locate the type material of these species to confirm their
taxonomic positions.
One sexual morph, Ga. musarum, was described by Petch
(1917) on Musa paradisiaca in Ceylon (Sri Lanka) as associ-
ated with Gloeosporium musarum Cooke & Massee; its asco-
spores are hyaline, aseptate, cymbiform (boat-shaped,
navicular), straight or curved, obtuse and measure
14–18 × 3.5–4μm. Ascospores of C. musicola have a different
shape and are larger, measuring on average 19.5 × 6 μm (SNA)
or 18.4 × 6 μm(Anthriscus stem). Ascospore shape and size of
strains called Ga. musae by Rodrigues & Owen (1992) were
again different and cannot be linked either. The sexual morph of
the latter species was obtained by means of laboratory cross-
ings, while C. musicola formed a sexual morph in pure culture; it
is apparently homothallic. Compared to other sexual morphs
developed in the C. orchidearum complex, C. musicola forms
ascospores that are similar in length to those of C. sojae,
however much wider in the middle and with a different shape
(fusiform).
Colletotrichum okinawense Damm & Toy. Sato, sp. nov.
MycoBank MB824226. Fig. 12.
Etymology: The species epithet is derived from the Japanese
island Okinawa, where the species was collected.
Sexual morph not observed. Asexual morph on SNA. Vegetative
hyphae 2–8μm diam, hyaline, smooth-walled, septate,
branched. Chlamydospores not observed. Conidiomata, co-
nidiophores formed directly on hyphae. Setae not observed.
Conidiophores hyaline, smooth-walled, septate, branched.
Conidiogenous cells hyaline, smooth-walled, cylindrical to
Fig. 11. Colletotrichum musicola (A –C, I–N, V–AF from ex-holotype culture CBS 132885. D–H, O–U. from culture CBS 127557). A–C. Conidiomata. D, M. Tips of setae. E,
N. Bases of setae. F–L. Conidiophores. O–T. Appressoria. U–V. Conidia. W, Z, AA. Ascomata. X. Outer surface of peridium. Y. Peridium in cross section. AB. Ascospores. AC.
Paraphyses. AD–AF. Asci. D–E, U, Y –AC, AF. from Anthriscus stem. A–C, F –T, V, W –X, AD. from SNA. AE. from OA. A–C, W, Z, AA. DM. D–V, X –Y, AB –AF. DIC. Scale
bars: A = 100 μm, E = 10 μm. A applies to A–C, W, Z–AA. E applies to D –V, X –Y, A B –AF.
THE COLLETOTRICHUM DRACAENOPHILUM,C.MAGNUM AND C. ORCHIDEARUM SPECIES COMPLEXES
www.studiesinmycology.org 23
doliiform, sometimes lacking a basal septum, 8–25 × 3–6μm,
opening 1–2μm diam, collarette 0.5–1μm long, periclinal
thickening observed. Conidia hyaline, smooth-walled, aseptate,
straight, clavate to cylindrical, the apex rounded, the base
rounded or truncate, (10.5–)12–14.5(–16.5) × (3.5–)
4.5–5.5 μm, mean ± SD = 13.4 ± 1.3 × 5.0 ± 0.4 μm, L/W ra-
tio = 2.7, few conidia up to 22 μm long were observed.
Appressoria single or in loose groups, pale to medium brown,
smooth-walled, elliptical, navicular, bullet-shaped or irregular in
outline, with undulate or lobate margin, (5.5–)
7–15(–23) × (2.5–)5–10.5(–13) μm, mean ± SD = 11.1 ± 3.9 ×
7.7 ± 2.6 μm, L/W ratio = 1.4.
Asexual morph on Anthriscus stem. Conidiomata, co-
nidiophores formed on pale to medium brown, angular cells,
3–7μm diam. Setae not observed. Conidiophores hyaline to
very pale brown, smooth-walled, septate, branched, to 25 μm
long. Conidiogenous cells hyaline, smooth-walled, cylindrical to
conical, 4.5–21 × 3.5–5.5 μm, opening 1–2μm diam, collarette
0.5 μm long, periclinal thickening observed. Microcyclic con-
idiation observed, sometimes from conidia still attached to the
conidiogenous cell. Conidia hyaline, smooth-walled, aseptate,
straight, clavate to cylindrical, the apex rounded, the base
rounded or truncate, (12–)12.5–15(–16.5) × (4.5–)5–5.5 μm,
mean ± SD = 13.9 ± 1.3 × 5.1 ± 0.3 μm, L/W ratio = 2.7.
Culture characteristics: Colonies on SNA flat with entire margin,
medium hyaline, aerial mycelium lacking, reverse same colours;
31.5–34 mm in 7 d (40 mm in 10 d). Colonies on OA flat with
entire margin, surface saffron to salmon, aerial mycelium lacking,
reverse salmon; 31.5–34 mm in 7 d (40 mm in 10 d). Conidial
mass saffron.
Materials examined:Brazil, Minas Gerais, Uberaba, on stems of Carica papaya,
Jun. 1892, E. Ule n. 1947 (H holotype of Gloeosporium papayae). Japan,
Okinawa prefecture, Miyakojima Island, from a petiole of Carica papaya, 28 Sep.
2007, S. Sato (isolated Sep. 2007 by T. Sato) (GLM-F 111630 holotype of
C. okinawense, culture ex-holotype MAFF 240517 = NBRC 104626 = CBS
143246 = GLMC 1837).
Notes: This species belongs to the C. magnum species complex.
Strain MAFF 240517 was originally identified as
C. gloeosporioides by J. Moriwaki and recently re-identified as
C. brevisporum by T. Sato. Based on the multilocus phylogeny in
this study this species is closely related to C. brevisporum, but
represents a distinct species.
Many Colletotrichum species have been reported or were
described on papaya (Carica papaya)(Farr & Rossman 2017); a
number of them were studied based on molecular data. For
example, C. acutatum and C. simmondsii were described on
Carica papaya in Australia. They belong to the C. acutatum
species complex; both species occur on many other host plants
as well (Damm et al. 2012a). A strain from Carica papaya in
Brazil was identified as C. karstii (CBS 106.91); this species
belongs to the C. boninense species complex and also occurs on
many host plants (Damm et al. 2012b). Colletotrichum
Fig. 12. Colletotrichum okinawense (from ex-holotype culture MAFF 240517). A–B. Conidiomata. C–E, J–M. Conidiophores. F–I. Microcyclic conidiation. N–S. Appressoria.
T–U. Conidia. A, C–I, T. from Anthriscus stem. B, J –S, U. from SNA. A–B. DM. C–U. DIC. Scale bars: A = 100 μm, C = 10 μm. Scale bar of A applies to A–B. Scale bar of C
applies to C–U.
DAMM ET AL.
24
queenslandicum, originally described as C. gloeosporioides var.
minus from Carica papaya in Australia, and C. siamense strain
ICMP 18739 from Carica papaya in South Africa belong to the
C. gloeosporioides species complex (Weir et al. 2012). There are
also two other strains from papaya included in this study, one
from Japan (MAFF 238697) that is re-identified as C. plurivorum
(C. orchidearum complex), and one from Australia (CBS 512.75)
that was re-identified as C. brevisporum that belongs to the
C. magnum complex, too.
Two species were described by Hennings (1895, 1908) from
papaya in Brazil: C. papayae Henn. 1908 and Gloeosporium
papayae Henn. 1895. They are not synonyms as listed in
Saccardo et al. (1931); Petrak (1953) combined the latter in
Colletotrichum as C. papayae (P. Henn.) Petr. 1953, apparently
not being aware of the older homonym (nom. illegit., Art. 53.1).
Conidia of C. papayae Henn. are cylindrical, straight to curved,
measuring 12–20 × 5–7μm, while those of Gl. papayae Henn.
are cylindrical to subclavate, obtuse, straight, measuring
11 –14 × 5–6μm according to Hennings (1895) and (12.5–)
13–15(–15.5) × 4.5–6μm, mean ± SD = 14 ± 1.2 ×
5.3 ± 0.5 μm, L/W ratio = 2.7 according to our measurements.
The conidia size of strain MAFF 240517 is similar to that of Gl.
papayae. However, the shape of the conidia observed on the
type specimen of Gl. papayae was cylindrical with parallel walls
or even a constriction at the middle, not distinctly attenuated to
the base as those of C. okinawense.
GAPDH,ACT and TUB2 sequences of C. okinawense differ
from all other species treated in this study (no HIS3 sequence
available), while the ITS is identical to that of C. brevisporum,
C. merremiae and C. lobatum, and the CHS-1 sequence to that
of C. brevisporum. The ACT sequence is 100 % identical to those
of C. brevisporum isolates CMM1672, CMM1702, CMM1822 and
CMM2005 from Carica papaya in Brazil (KC702903–KC702906,
Vieira et al. 2013) that possibly belong to the same species. The
closest matches with the TUB2 sequence are with 99 % identity
(6 nucleotides difference) to the four C. liaoningense strains from
Capsicum in China (KP890111 –KP890113, KP890115, Diao
et al. 2017). The closest matches with the GAPDH sequence
are with 97 % identity to the C. liaoningense strains CAUOS 3
and CAUOS 4 (KP890136, KP890137, Diao et al. 2017). The ITS
sequence is 100 % identical to several unidentified isolates and
isolates identified as C. brevisporum,C. magnum,Ga. magna
and C. cingulata var. brevispora.
In contrast to all other species in the C. magnum complex,
conidia of C. okinawense are predominantly clavate; it is also the
fastest growing species in this complex. Microcyclic conidiation
has only rarely been observed in the genus Colletotrichum. Only
one other species with this feature is known to us, C. cliviicola,
that belongs to the C. orchidearum complex. Secondary conidia
of C. okinawense are formed directly from conidiogenous
openings on conidia, and sometimes already when the mother
cell itself is still attached to its conidiogenous cell, while sec-
ondary conidia of C. cliviicola are usually formed on short pegs or
phialides that appear on older conidia that are sometimes
already septate.
Colletotrichum orchidearum Allesch., Rabenh. Krypt.-Fl., Edn
2 (Leipzig) 1(7): 563. 1902 (1903). Fig. 13.
Synonyms:Colletotrichum hymenocallidicola Chethana et al.,
Fungal Diversity 75: 160. 2015.
Colletotrichum aracearum L.W. Hou & L. Cai, Mycosphere 7:
1115. 2016.
Sexual morph on SNA. Ascomata perithecia, formed after 4 wk,
solitary, non-stromatic, subglobose, ostiolate, 200–300 ×
175–250 μm. Peridium 6–15 μm thick, composed of 2–3 layers
of medium brown flattened textura angularis with cells 5–22 μm
diam. Ascogenous hyphae hyaline, smooth, delicate. Interascal
tissue formed of paraphyses, hyaline, smooth-walled, mostly
cylindrical but tapering towards the round tip, disintegrating
quickly, septate, constricted at the septa, branched, apically free,
40–70 μm long and up to 6–8.5 μm diam. Asci unitunicate, 8-
spored, fasciculate, smooth-walled, cylindrical to slightly
clavate, tapering to apex and base, round tip, no apical appa-
ratus observed, 70–100 × 11–15 μm. Ascospores uni- or
biseriately arranged, aseptate, hyaline, smooth-walled, allantoid,
with round ends, (13.5–)16–20(–22) × 5–5.5(–6) μm,
mean ± SD = 18.0 ± 1.9 × 5.2 ± 0.3 μm, L/W ratio = 3.5.
Sexual morph on Anthriscus stem.Ascomata perithecia,
formed after 4 wk, solitary, non-stromatic, subglobose to ovoidal,
dark brown, ostiolate; outer wall composed of medium brown
flattened textura angularis with cells 17–22 μm diam. Asci 8-
spored, cylindrical to clavate, unitunicate, fasciculate, thin-
walled, 56–77 × 9–11.5 μm. Ascospores uni- or biseriately ar-
ranged, aseptate, hyaline, smooth-walled, allantoid, with round
ends, (13–)15.5–19(–21) × 4.5–5.5(–6) μm, mean ± SD =
17.4 ± 1.8 × 5.1 ± 0.4 μm, L/W ratio = 3.4.
Asexual morph on SNA. Vegetative hyphae 1–11 μm diam,
hyaline to pale brown, smooth-walled to verruculose, septate,
branched. Chlamydospores not observed. Conidiomata, co-
nidiophores and setae formed directly on hyphae. Setae medium
brown, verruculose to verrucose, 55–110 μm long, 1 –2(–4)-
septate, base ± inflated, 5.5–6μm diam, tip rounded. Co-
nidiophores hyaline to pale brown, smooth-walled to verruculose,
septate, branched. Conidiogenous cells hyaline to pale brown,
smooth-walled to verruculose, cylindrical to doliiform,
8–40 × 4–5,5 μm, opening 0.5–2μm diam, collarette 0.5–1μm
long, periclinal thickening sometimes observed. Conidia hyaline,
smooth-walled, aseptate, straight, cylindrical, the apex rounded,
the base truncate or rounded, (13–)14–20(–28) ×
4.5–5.5(–6.5) μm, mean ± SD = 17.0 ± 3.2 × 5.1 ± 0.5 μm, L/W
ratio = 3.4, CBS 136877 forms shorter and wider conidia,
measuring (12.5–)13–15.5(–17) × 5–6.5(–7) μm,
mean ± SD = 14.2 ± 1.2 × 5.7 ± 0.7 μm, L/W ratio = 2.5.
Appressoria single, medium to dark brown, smooth-walled,
clavate, navicular, elliptical or circular in outline, with a lobate
or undulate margin, (9–)10.5–17.5(–22.7) × (5–)
6–10.5(–13) μm, mean ± SD = 14.0 ± 3.3 × 8.4 ± 2.3 μm, L/W
ratio = 1.7.
Asexual morph on Anthriscus stem. Conidiomata, co-
nidiophores and setae formed on pale brown, angular cells,
4.5–8μm diam. Setae (few observed) medium to dark brown,
verruculose, 75–130 μm long, 1–5-septate, base cylindrical,
conical to ± inflated, 4–8μm diam, tip acute to ± rounded.
Conidiophores hyaline to pale brown, smooth-walled. Con-
idiogenous cells hyaline to pale brown, smooth-walled, cylindrical
to doliiform, 7–10 × 4.5–6μm, opening 1 μm diam, collarette
0.1–1μm long, periclinal thickening not observed. Conidia hy-
aline, smooth-walled, aseptate, straight, cylindrical, the apex
rounded, the base truncate or rounded, (14.5–)
15.5–17(–17.5) × (4.5–)5–5.5(–6) μm, mean ± SD = 16.2 ±
0.9 × 5.2 ± 0.3 μm, L/W ratio = 3.1.
Culture characteristics: Colonies on SNA flat with entire margin,
hyaline to pale honey, agar medium, filter paper and Anthriscus
THE COLLETOTRICHUM DRACAENOPHILUM,C.MAGNUM AND C. ORCHIDEARUM SPECIES COMPLEXES
www.studiesinmycology.org 25
stem partly covered with grey to black fruiting bodies and sparse
aerial mycelium, reverse same colours; growth 30–31.5 mm in
7d(40 mm in 10 d). Colonies on OA flat with entire margin;
buff, honey, grey olivaceous to olivaceous grey, partly covered
with short floccose white aerial mycelium, reverse buff to oliva-
ceous grey, growth 30–32 mm in 7 d (40 mm in 10 d). Conidial
mass whitish to pale yellow.
Materials examined:Germany, Munich, glasshouses of Botanical Garden, on
dead and dying leaves of Eria javanica (syn. E. stellata), Apr. 1895, J.E. Weiss
[M-0140831 lectotype of C. orchidearum (named as forma eriae)]; Iran, Gole-
stan province, Gorgan, from leaves of Epipremnum aureum (syn. Scindapsus
aureus), Oct. 2013, A. Alizadeh, strain UTFC 266 = 249C. Netherlands, Utrecht
(indoor plant), from anthracnose on leaf of Dendrobium nobile, Apr. 2013, I.
Benoit-Gelber (CBS H-21910 epitype, here designated MBT380419, culture ex-
epitype CBS 135131); ibid., culture CBS 136877.
Notes: The type specimens of the three forms of C. orchidearum,
C. orchidearum f. cymbidii,C. orchidearum f. eriae and
C. orchidearum f. physosiphonis (Allescher 1902) were recently
investigated by Yang et al. (2011), and strains from different
orchid genera from China clustering in the multilocus phylogeny
of that study were identified as C. orchidearum based on
morphological resemblance. As C. orchidearum was described
in Europe, the authors were however reluctant to select an
epitype from their Asian collection. Damm et al. (2012a) studied
the specimens as well and lectotypified C. orchidearum with the
specimen M-0140831 from leaves of Eria javanica.
Different collections of Colletotrichum species from Orchid-
aceae in Europe were treated in this study, including strains of
the species studied by Yang et al. (2011), that were isolated from
a leaf of Dendrobium nobile in the Netherlands cultivated as an
indoor plant, confirming the occurrence of this species in Europe.
Conidia from the lectotype specimen were measured in Damm
et al. (2012a): (13.5–)15.5–19.5 × 5–6μm, mean ± SD =
17.2 ± 1.6 × 5.6 ± 0.3 μm, L/W ratio = 3.1. Conidia from one of
the strains from the Netherlands, CBS 135131, agree in conidial
dimensions, measuring on average (13–)14–20(–28) ×
4.5–5.5(–6.5) μm on SNA and (14.5–)15.5–17(–17.5) × (4.5 –)
5–5.5(–6) μmonAnthriscus stems. Moreover, conidiomata were
minute as observed on the lectotype specimen and the length of
the setae was similar (setae from lectotype: 56–120 ×
4.5–6μm). Strain CBS 135131 was therefore used as basis of
the epitype.
Our collection from Munich, Germany, had similar conidial
dimensions as C. orchidearum; however, conidia often develop a
prominent scar and are slightly constricted in the middle.
Moreover, the conidiomata of this collection were very large,
while those of C. orchidearum were small and longer setae were
formed. The species collected in Munich belongs to the
C. dracaenophilum complex and was described as
C. coelogynes in this study. Another species from orchids
described in this study is C. cattleyicola that also belongs to the
C. orchidearum species complex.
There are numerous other Colletotrichum species that were
described on Orchidaceae, including C. orchidophilum,
belonging to the C. acutatum species complex (Damm et al.
2012a), C. cymbidiicola,C. oncidii and C. karstii, belonging to
the C. boninense species complex (Yang et al. 2011, Damm et al.
2012b), C. arxii belonging to the C. gigasporum complex (Liu
et al. 2014), C. ochracea,C. caudasporum,C. duyunensis,
C. endophytum,C. guizhounensis and C. bletillum, belonging to
the C. caudatum,C. graminicola and C. spaethianum species
complexes, respectively (Tao et al. 2013), C. coelogynes and
C. excelsum-altitudinum in the C. dracaenophilum species
complex (Tao et al. 2013, this study).
There are sequences of strains called C. orchidearum from
three studies in GenBank. Strains from different Orchidaceae
hosts from the study of Yang et al. (2011) and isolate SAUCC
1407 from Arctium lappa (Xu et al. 2016) both from China were
included in our phylogeny. The strains from Orchidaceae were
confirmed as C. orchidearum, while the isolate from Arctium
lappa was re-identified as C. sojae. One of two ITS sequences of
“C. orchidearum”isolate NW248 from an unpublished and un-
named study was displayed as “type strain of C. orchidearum”
(EU520211, EU732727; Z. Zhang et al., unpubl. data); it was
revealed to be no Colletotrichum sp. at all but a species of
Cytospora (Diaporthales).
The description of C. hymenocallidicola (Ariyawansa et al.
2015) is based on one strain of which five loci were
sequenced (ITS, GAPDH,CHS-1,ACT,TUB2). Blastn searches
on NCBI GenBank with all sequences of this species revealed
that the ACT and TUB2 sequences do not conform with the
placement of the species based on ITS, GAPDH and CHS-1
sequences. While the closest matches of the ITS, GAPDH and
CHS-1 sequences were with 98–99 % identity strains identified
as C. cliviae, placing the species in the C. orchidearum complex,
the ACT sequence (KT290260) was 93 % identical with those of
C. pseudomajus and C. vietnamense (both C. gigasporum
complex) and only 83 % identical with strains identified as
C. cliviae, and the TUB2 sequence (KT290261) was 100 %
identical with the ex-epitype strain of C. truncatum (Damm et al.
2009). The ACT and TUB2 sequences were possibly mixed-up
with those of other strains studied by the authors. Further, the
beginning part of the GAPDH sequence apparently includes
artefacts as it is completely different from all related species.
Therefore, C. hymenocallidicola and C. aracearum were not in
the same clade in a recent phylogeny of the genus Colleto-
trichum (Marin-Felix et al. 2017). The ACT and TUB2 sequences
of C. hymenocallidicola as well as the first 22 nucleotides of
GAPDH were excluded from the alignment in this study. It is
highly recommended to re-sequence the ex-type strain of this
species, in order to confirm our results with the other loci and
establish a solid basis of this apparently common species.
Colletotrichum hymenocallidicola and C. aracearum were
described recently as pathogens of Hymenocallis sp. in Thailand
(Ariyawansa et al. 2015) and Monstera deliciosa and Philoden-
dron selloum in China (Hou et al. 2016), respectively, and shown
to be synonyms of C. orchidearum in this study. Based on this
study, C. orchidearum is known from monocots such as Hyme-
nocallis (Amaryllidaceae), Monstera,Philodendron,Scindapus
(Araceae), Cordyline (Asparagaceae) as well as many Orchid-
aceae, including Cattleya,Cymbidium,Dendrobium,Oncidium,
Phalaenopsis and Vanda in Asia (China, Iran, Japan, Thailand)
and Europe (as indoor plant in the Netherlands).
Fig. 13. C. orchidearum (A–W, Y –Z. from ex-epitype culture CBS 133131, X, AA. from culture CBS 136877). A–B. Conidiomata, arrow head in A: Conidioma with seta. C, G.
Tips of setae. D, F, H. Bases of setae. E, I–L. Conidiophores. M–R. Appressoria. S–T. Conidia. U. Ascoma. V. Peridium in cross section. W. Outer surface of peridium. X.
Paraphyses. Y. Ascospores. Z–AA. Asci. A, C–F, S. from Anthriscus stem. B, G–R, T –AA. from SNA. A –B, U. DM. C –T, V –AA. DIC. Scale bars: A = 100 μm, E = 10 μm. A
applies to A–B. E applies to C–T, V –AA.
THE COLLETOTRICHUM DRACAENOPHILUM,C.MAGNUM AND C. ORCHIDEARUM SPECIES COMPLEXES
www.studiesinmycology.org 27
Colletotrichum orchidearum can be differentiated with se-
quences of all loci included, except for GAPDH; sequences of
CHS-1 and HIS3 are only available of part of the strains included.
GAPDH sequences of C. orchidearum and C. sojae are identical.
In a blastn search in GenBank, the TUB2 sequence of strain
CBS 135131 was 100 % identical with those of the strains from
Orchidaceae in China and 99 % identical (one nucleotide dif-
ference) with those of the two C. aracearum strains, while the two
C. cliviicola strains CSSS1 and CSSS1 (Yang et al. 2009) differ
in eight nucleotides; all are included in this study. In a blastn
search with the ACT sequence, the two C. aracearum strains
were identical and the strains from Orchidaceae in China differed
in one nucleotide; while the C. cliviicola strains differed in four
nucleotides. The ITS sequence of strain CBS 135131 was
identical with those of the two C. aracearum (strains,
“C. gloeosporioides”isolate C10 from Cymbidium in China (Yao
et al. 2013), Colletotrichum sp. strain AR3750 and Glomerella sp.
strain AR3749 from Dendrobium and Cattleya, respectively, in
Thailand (Farr et al. 2006) and differed in one nucleotide from
“C. gloeosporioides”isolate C14 from Cymbidium in China (?)
(J.A. Yao et al., unpubl. data) and the ex-type strain of
C. hymenocallidicola, while the C. cliviicola strains differed in
seven nucleotides.
No sexual morph was observed in the strain from Hymeno-
callis (Ariyawansa et al. 2015), but in those from Monstera
deliciosa (Hou et al. 2016) and from Orchidaceae (Yang et al.
2011, this study). In pathogenicity tests by Yang et al. (2011),
strain CORCX6 from Cattleya sp. caused lesions on fruits of
peppers, apple and tomato by wound/drop inoculation, but none
on unwounded fruits.
Colletotrichum panamense Damm, sp. nov. MycoBank
MB824227. Fig. 14.
Etymology: The species epithet is derived from the country
where the species was collected, Panama.
Sexual morph not observed. Asexual morph on SNA. Vegetative
hyphae 1.5–7.5 μm diam, hyaline, smooth-walled, septate,
branched. Chlamydospores not observed. Conidiomata, co-
nidiophores and setae formed directly on hyphae. Setae medium
brown, smooth-walled, verrucose towards the tip, 40–100 μm
long, 2(–3)-septate, base inflated, 6–7μm diam, tip ± acute.
Conidiophores pale brown, smooth-walled, septate or aseptate.
Conidiogenous cells pale brown, smooth-walled, broadly ellip-
soidal, doliiform to cylindrical, 7–16 × 5.5–7μm, often interca-
lary, opening 1.5–2μm diam, collarette 0.5 –1μm long, periclinal
thickening distinct. Conidia hyaline, smooth-walled, aseptate,
straight, cylindrical, the apex and base rounded, (14.5–)
15.5–18(–19.5) × (4.5–)5–5.5 μm, mean ± SD = 16.9 ± 1.2 ×
5.0 ± 0.3 μm, L/W ratio = 3.4. Appressoria not formed.
Fig. 14. Colletotrichum panamense (from ex-holotype culture CBS 125386). A–B. Conidiomata. C, H. Tips of setae. D, I. Bases of setae. E–G, J–N. Conidiophores. O–P.
Conidia. A, C–G, O. from Anthriscus stem. B, H –N, P. from SNA. A–B. DM. C –P. DIC. Scale bars: A = 100 μm, G = 10 μm. Scale bar of A applies to A–B. Scale bar of G
applies to C–P.
DAMM ET AL.
28
Asexual morph on Anthriscus stem. Conidiomata, co-
nidiophores and setae formed directly on hyphae, no basal cells
observed. Setae medium brown, smooth-walled, verrucose to-
wards the tip, 60–100 μm long, 2–3-septate, base inflated,
5.5–7.5 μm diam, tip ± acute to ± rounded. Conidiophores pale
brown, smooth-walled, septate, sometimes branched, to 30 μm
long. Conidiogenous cells pale brown, smooth-walled, ovoidal,
broadly ellipsoidal to subsphaerical, 6–11.5 × 4.5–7.5 μm,
opening 1.5–2μm diam, collarette not observed, periclinal
thickening sometimes visible or distinct. Conidia hyaline, smooth-
walled, aseptate, straight, cylindrical, the apex and base
rounded, (14–)16–20(–22) × 5–5.5 μm, mean ± SD = 18 ±
2.1 × 5.3 ± 0.2 μm, L/W ratio = 3.4.
Culture characteristics: Colonies on SNA flat with entire margin,
medium hyaline to pale ochreous to pale luteous, filter paper
partly pale luteous, aerial mycelium lacking, reverse same col-
ours; 18.5–20 mm in 7 d (27.5–30 mm in 10 d). Colonies on OA
flat with entire margin, surface rosy buff to saffron with small grey
dots due to sporulation, aerial mycelium lacking, reverse rosy
buff to buff; 22.5–25.5 mm in 7 d (34–33.5 mm in 10 d). Conidial
mass saffron.
Material examined:Panama, Gamboa, wet lowland forest, leaf endophyte of
Merremia umbellata, Nov. 2004, S. Van Bael & Z. Maynard, D2-13 (CBS H-21066
holotype, culture ex-type CBS 125386 = Q882).
Notes:Colletotrichum panamense belongs to the C. magnum
species complex like C. merremiae that is also described from
Merremia umbellata in Panama in this study. In contrast to
C. merremiae, the conidiogenous cells of C. panamense are
often subglobose to ellipsoidal, on SNA often also intercalary.
Conidia of C. panamense are on both media on average at least
2μm longer.
Colletotrichum panamense differs with all loci studied from all
other species of the genus. The ITS sequence of strain CBS
125386 is 100 % identical to Colletotrichum sp. Q882
(GU994392, Rojas et al. 2010), which is the same strain. Other
sequences have 3 bp differences. Closest matches with the
TUB2 sequence of the ex-type strain of C. panamense are with
99 % identity (5 nucleotides difference) the four C. liaoningense
strains that are included in our study (Diao et al. 2017). Closest
matches with the CHS-1 sequence are with 99 % identity (1
nucleotide difference) to those of the ex-type strain of
C. brevisporum sequenced by Liu et al. (2014, KF687760) and
C. brevisporum strain CCCM12 from Cucurbita moschata in
China (KY797631, Liu et al. 2018). No ACT sequence in Gen-
Bank is closer than 97 % identical and no HIS3 sequence closer
than 90 % identical to those of C. panamense.
Colletotrichum piperis Petch, Ann. R. bot. Gdns Peradeniya
6(3): 239. 1917. Fig. 15.
Fig. 15. Colletotrichum piperis (from ex-epitype culture IMI 71397). A–B. Conidiomata. C, H. Tips of setae. D, I. Bases of setae. E–G, J–K. Conidiophores. L–Q. Appressoria.
R–S. Conidia. A, C–G, R. from Anthriscus stem. B, H –Q, S. from SNA. A–B. DM. C–S. DIC. Scale bars: A = 100 μm, E = 10 μm. Scale bar of A applies to A–B. Scale bar of
E applies to C–S.
THE COLLETOTRICHUM DRACAENOPHILUM,C.MAGNUM AND C. ORCHIDEARUM SPECIES COMPLEXES
www.studiesinmycology.org 29
Sexual morph not observed. Asexual morph on SNA. Vegetative
hyphae 1–8μm diam, hyaline, smooth-walled, septate,
branched. Chlamydospores not observed. Conidiomata, co-
nidiophores and setae formed directly on hyphae. Setae pale to
medium brown, verruculose to verrucose, 30–95 μm long, 1–3-
septate, base cylindrical, 4.5–7μm diam, tip ± acute
to ± rounded. Conidiophores hyaline to pale brown, smooth-
walled to verruculose, septate, branched, to 65 μm long. Con-
idiogenous cells hyaline to pale brown, smooth-walled to verru-
culose, cylindrical, 10–25 × 3.5–5.5 μm, opening 2–2.5 μm
diam, collarette 1–1.5 μm long, periclinal thickening distinct.
Conidia hyaline, smooth-walled, aseptate, straight, cylindrical to
obclavate, the apex rounded, the base rounded to truncate,
(11.5–)13–18.5(–28) × 4 –5( –6) μm, mean 15.7 ± 2.7 ×
4.7 ± 0.4 μm, L/W ratio = 3.4. Appressoria not formed at the
reverse side of SNA plates, but few observed on slide culture
after 1 month, single, medium brown, smooth-walled, elliptical,
bullet-shaped to rectagular in outline, with an entire margin,
(3.5–)6–10(–13) × (3–)4–6(–8.5) μm, mean ± SD =
7.9 ± 2.1 × 5.0 ± 1.1 μm, L/W ratio = 1.6.
Asexual morph on Anthriscus stem. Conidiomata,co-
nidiophores and setae formed on pale brown, angular cells. Setae
pale to medium brown, verruculose to verrucose, often branched,
50–120 μm long, 1–3-septate, base cylindrical, sometimes
slightly inflated, 4.5–7μm diam, tip ± acute to ± rounded. Co-
nidiophores pale brown, smooth-walled, septate, sometimes
branched, to 35 μm long. Conidiogenous cells pale brown,
smooth-walled, cylindrical to doliiform, 8–22 × 4 –6.5 μm, opening
1–1.5 μm diam, collarette 0.5–1μm long, periclinal thickening
observed. Conidia hyaline, smooth-walled, aseptate, straight,
cylindrical, the apex rounded, the base rounded to truncate, (14–)
15.5–17.5(–19.5) × (4–)4.5–5(–5.5) μm, mean ± SD = 16.6 ±
1.0 × 4.7 ± 0.3 μm, L/W ratio = 3.6.
Culture characteristics: Colonies on SNA flat with entire margin,
hyaline, cinnamon to isabelline in the centre, agar medium, filter
paper and Anthriscus stem partly covered with grey to salmon
acervuli, aerial mycelium lacking, reverse same colours; growth
15.5–16.5 mm in 7 d (26–27.5 mm in 10 d). Colonies on OA flat
with entire margin; grey olivaceous, with a buff margin, covered
with translucent salmon conidia masses and partly covered with
very short aerial mycelium, reverse olivaceous grey to iron-grey,
growth 17.5–19 mm in 7 d (26–29 mm in 10 d). Conidial mass
salmon.
Materials examined:Malaysia,fromPiper nigrum, unknown collection date and
collector (IMI history: 1957 P. Holliday HS 134, 1958 CABI) (IMI 71397 epitype
of C. piperis Petch, here designated, MBT380420, CBS H-21503 isoepitype,
culture ex-epitype IMI 71397). Puerto Rico, Caguas, from Piper umbellatum,9
Feb. 1913, FL Stevens, BPI 399521 possible paratype of C. piperis Stevens. Sri
Lanka, Medulsima, from leaf of Piper betle,T.Petch4544,17Mar.1915(K(M)
235609 B, part of holotype of C. piperis Petch). Ta iwan,Tainan,fromPiper
betle, 29 Oct. 1907, K. Sawada, BPI 397531 possible holotype of
C. geniculatum.
Notes: There were several species described on Piper spp.
including C. dasturii,C. geniculatum,C. necator,C. piperis Petch
and C. piperis Stevens. The two species named C. piperis were
described in the same year (1917), C. piperis Petch from leaves
of Piper betle and P. nigrum in Sri Lanka (Ceylon) and C. piperis
Stevens from Piper umbellatum in Puerto Rico, respectively
(Petch 1917, Stevens 1917). According to Roy (1948), who
investigated the publication dates, the name C. piperis Stevens is
illegitimate (nom. illegit., Art. 53.1) as it was published between
10
th
July and 18
th
August 1917 and a homonym of the earlier
C. piperis Petch (published 4
th
July 1917) and was therefore
replaced by C. stevensii.
Conidia of C. piperis Petch measure 12–19 × 3.5–4.5 μm
according to Petch (1917) and 10–16 × 4–6μm and (13.5–)
14–16(–17.5) × (4.5–)4.5–5.5(–6) μm, mean ± SD = 15.1 ±
1.0 × 5.1 ± 0.4 μm, L/W ratio = 3.0 according to measurements
from the type specimen by Roy (1948) and the present study.
The conidial size and shape of strain IMI 71397 agree with these
measurements and our observations (not shown), respectively.
This strain originated from P. nigrum in Malaysia, the same re-
gion of Asia than the holotype of C. piperis Petch (Sri Lanka).
Strain IMI 71397 is therefore used as the basis for the epitype of
this species. In contrast, conidia of C. stevensii (syn. C. piperis
Stevens) are larger than C. piperis Petch, measuring
17–27 × 7 μm(Stevens 1917); which was confirmed by mea-
surements of conidia of a possible paratype specimen of
C. piperis Stevens in this study: (16.5–)19–23.5(–25.5) × (5 –)
5.5–7.5(–8.5) μm, mean ± SD = 21.2 ±
2.3 × 6.4 ± 0.9 μm, L/W ratio = 3.3. Colletotrichum necator that
was described from fruit of Piper sp. in Singapore by Massee
(1912) also forms larger conidia (20–23 × 5–7μm) than
C. piperis Petch. Sawada (1959) described C. geniculatum from
P. betle in Taiwan. Conidia from the type specimen measure
12.5–14(–15) × (5–)5.5–6.5(–7) μm, mean ± SD = 13.3 ±
0.8 × 6.2 ± 0.6 μm, L/W ratio = 2.2; conidiophores were not
observed, setae measure 35–57 × 4–5.5 μm. The conidia size
of C. geniculatum is similar to that of C. piperis Petch. However,
the conidia are shorter and wider, resulting in a smaller L/W ratio
and a different shape. Roy (1948) described C. dasturii from
P. betle in India, Bengal; its conidia are curved, while C. piperis
Petch has straight conidia.
Sequence comparison of Colletotrichum spp. from Piper spp.
in GenBank suggests that species from several species com-
plexes can be found on this host genus. For example, strains
from a study of leaf endophytes in Piper hispidum in Brazil
identified by ITS belong to the C. boninense and
C. gloeosporioides species complexes (Orlandelli et al. 2012). A
strain from Piper nigrum (IMI 324991, unknown location) was
recently identified as C. fioriniae belonging to the C. acutatum
species complex (Damm et al. 2012a), while a C. siamense
strain from a leaf lesion of P. nigrum in Australia (James et al.
2014) and two C. truncatum strains (CBS 127.57, IMI 63597)
from Peperomia magnoliifolia (Piperaceae) in India (Damm et al.
2009) belong to the C. gloeosporioides and C. truncatum species
complexes, respectively. Farr & Rossman (2017) further list
C. capsici (syn. of C. truncatum), C. dematium and
C. gloeosporioides. However, the respective sources are from
the pre-sequence era and therefore need confirmation.
Another species on Piper,C. lobatum, is described in this
study from Piper catalpaefolium in Trinidad and Tobago with very
similar conidial shape and size, but very different appressoria
that are more complex than those of C. piperis Petch. Colleto-
trichum lobatum belongs to the C. magnum species complex,
while C. piperis belongs to the C. orchidearum complex. Apart
from the rather simple appressoria with entire edge, C. piperis
sometimes forms branched setae, a feature rarely observed in
the genus Colletotrichum. Moreover, C. piperis is the slowest
growing species in the C. orchidearum complex.
Colletotrichum piperis can be identified with all loci included in
this study. Closest matches in a blastn search with the ITS
sequence of C. piperis strain IMI 71397 on GenBank were with
99 % identity (5 nucleotides difference) several strains identified
DAMM ET AL.
30
as C. gloeosporioides, fungal sp., C. magnum,C. orchidearum,
fungal endophyte, C. trifolii,Glomerella glycines,Colletotrichum
sp., C. aracearum and Glomerella sp. There is no GAPDH
sequence in GenBank with 96 % identity and no TUB2
sequence with 97 % identity to that of strain IMI 71397. The
closest match with the ACT sequence was with 99 % identity (3
nucleotides difference) C. cliviae strain GUFCC15503 from
Calamus thwaitesii in India (KC790646, Sharma et al. 2013a).
Colletotrichum plurivorum Damm, Alizadeh & Toy. Sato, sp.
nov. MycoBank MB824228. Fig. 16.
Synonym:Colletotrichum sichuanensis G.S. Gong & F.L. Liu,
Scientific Reports 6(32761): 6. 2016. nom. inval., Art. 40.1
(Melbourne).
Etymology: The species epithet is based on the large host range
of this species.
Sexual morph on Anthriscus stem (observed in strains CBS
125474 and UTFC 260). Ascomata perithecia, formed after 4 wk,
solitary, superficial or immersed, non-stromatic, globose to
obpyriform, ostiolate, glabrous or covered by sparse white aerial
mycelium, medium to dark brown, 100–230 × 95–160 μm,
surrounded by pale brown, smooth-walled to verruculouse hy-
phae. Peridium 12–18 μm thick, composed of 3–5 layers of pale
brown flattened textura angularis with cells 5–18 μm diam.
Ascogenous hyphae hyaline, smooth, delicate, rarely visible.
Interascal tissue formed of paraphyses, hyaline, smooth-walled,
cylindrical, with a rounded tip, disintegrating quickly, septate,
apically free, 50–70 μm long, base 4.5–7.5 μm wide, branched
at the base. Asci unitunicate, 8-spored, cylindrical, smooth-
walled, 50–65.5 × 10.5–12.5 μm, the base broadly truncate,
asci of strain UTFC 260 larger, measuring 70–90 × 9 –13.5 μm.
Ascospores uni- or biseriately arranged, aseptate, in strain UTFC
260 also septate ascospores observed, initially hyaline, turning
pale brown with age, smooth-walled, allantoid to fusiform, with
both ends rounded, (13–)14.5–18(–22) × (4–)5–6(–7) μm,
mean ± SD = 16.4 ± 1.8 × 5.5 ± 0.5 μm, L/W ratio = 3.0.
Sexual morph on SNA (observed in strains CBS 125474,
UTFC 260 and UTFC 261). Ascomata perithecia, globose to
obpyriform, 180–340 × 180–230 μm, ostiolate, medium to dark
brown, glabrous. Peridium 12–18 μm thick, composed of 4–6
layers of pale brown flattened textura angularis with cells
5–19 μm diam. Interascal tissue formed of paraphyses, hyaline,
smooth-walled, cylindrical with a rounded tip, disintegrating
quickly, septate, apically free, 40–70 μm long, base 4.5 –6μm
wide, branched at the base. Asci unitunicate, 8-spored, cylin-
drical to clavate, fasciculate, 80.5–93 × 8.5–12 μm. Ascospores
uni- or biseriately arranged, aseptate, initially hyaline, turning
pale brown with age, smooth-walled, allantoid to fusiform, with
both ends rounded, (13–)15–19(–21) × (5–)5.5–6(–7) μm,
mean ± SD = 17.0 ± 2.1 × 5.7 ± 0.4 μm, L/W ratio = 3.0.
Asexual morph on SNA (mostly based on strain CBS 903.69,
UTFC 260 and UTFC 261, only few conidia observed in strain
CBS 125474). Vegetative hyphae 1–8μm diam, hyaline to pale
brown, smooth-walled, septate, branched. Chlamydospores not
observed. Conidiomata, conidiophores and setae formed directly
on hyphae. Setae not observed in strain CBS 125474, setae of
strain CBS 903.69 medium brown, verruculose to verrucose,
40–130 μm long, 1–(2)-septate, base cylindrical to slightly
inflated, 3–6μm diam, tip ± rounded to ± acute, setae of strain
UTFC 260 2–3-septate. Conidiophores and conidiogenous cells
not observed in strain CBS 125474, conidiophores of strain CBS
903.69 hyaline to pale brown, smooth-walled, septate, branched,
to 40 μm long. Conidiogenous cells of strain CBS 903.69 hyaline
to pale brown, smooth-walled, cylindrical clavate to doliiform,
6.5–19 × 3.5–4.5 μm, often integrated, some polyphialides
observed, opening 1–2μm diam, collarette 0.5–1μm long,
periclinal thickening distinct. Conidia (only three observed in
strain CBS 125474) hyaline, smooth-walled, aseptate, straight,
cylindrical, the apex and base rounded, 15–17 × 5.5 μm,
mean ± SD = 16.0 ± 0.9 × 5.6 ± 0.1 μm, L/W ratio = 2.9, conidia
of strain CBS 903.69 shorter, measuring (9.5–)
11.5–14(–15) × (4.5–)5–6μm, mean ± SD = 12.7 ± 1.3 ×
5.4 ± 0.4 μm, L/W ratio = 2.4, conidia of strain UTFC 261 shorter
and narrower, measuring (10–)10.5 –11.5(–12.5) × (3.5–)
4–4.5(–5) μm, mean ± SD = 11 ± 1.1 × 4.5 ± 0.5 μm, L/W
ratio = 2.6, conidia of strains UTFC 260 and UTFC 261 some-
times slightly curved. Appressoria single, pale, medium to dark
brown, smooth-walled, navicular to bullet-shaped or irregular in
outline, with an undulate, crenate to strongly lobate margin, (9–)
12.5–18.5(–22.5) × (4.5–)6.5–11.5(–15.5) μm, mean ± SD =
15.4 ± 3.0 × 8.8 ± 2.6 μm, L/W ratio = 1.7, appressoria of strain
CBS 903.69 smaller, measuring (5–)8.5 –15.5( –24.5) × (5–)
6.5–9(–10) μm, mean ± SD = 11.9 ± 3.4 × 7.6 ± 1.3 μm, L/W
ratio = 1.6.
Asexual morph on Anthriscus stem. Conidiomata, co-
nidiophores and setae formed on pale brown, angular cells,
2.5–6.5 μm diam. Setae medium brown, verruculose to verru-
cose, 40–95 μm long, 1–2-septate, base conical, slightly infla-
ted, 4.5–8μm diam, tip ± acute. Conidiophores pale brown,
smooth-walled, simple or septate and branched, to 30 μm long.
Conidiogenous cells pale brown, smooth-walled, cylindrical,
clavate to doliiform, 7–19 × 4–5.5 μm, opening 1–1.5 μm diam,
collarette 0.5–1μm long, periclinal thickening visible, sometimes
distinct. Conidia hyaline, smooth-walled, aseptate, straight, cy-
lindrical, sometimes slightly clavate, the apex and base rounded,
(15–)16.5–20(–22.5) × (5–)5.5–6.5(–8) μm, mean ± SD =
18.4 ± 1.8 × 5.9 ± 0.5 μm, L/W ratio = 3.1, conidia of strain CBS
903.69 shorter, measuring (12.5–)13.5 –16.5(–16.5) × (5–)
5.5–6μm, mean ± SD = 14.6 ± 1.1 × 5.6 ± 0.2 μm, L/W
ratio = 2.6.
Culture characteristics: Colonies on SNA flat with entire margin,
hyaline to pale olivaceous grey, agar medium, filter paper and
Anthriscus stem partly covered with pale olivaceous grey aerial
mycelium and grey conidiomata/ascomata, reverse same col-
ours; growth 27.5–31 mm in 7 d (40 mm in 10 d). Colonies on
OA flat with entire margin; olivaceous buff, grey olivaceous to
olivaceous, partly covered with short felty whitish aerial mycelium
and pale purplish grey to pale olivaceous grey conidiomata/
ascomata, reverse pale olivaceous grey to olivaceous grey,
growth 28.5–31.5 mm in 7 d (40 mm in 10 d). Conidial mass
greyish white to very pale salmon, in strain UTFC 260 pale buff to
orange.
Materials examined:Benin, Dahomey, Porto Novo, from leaf spot of Phaseolus
lunatus, collection date and collector unknown (isolated by G. Weststeijn and
deposited in CBS collection Oct. 1969 by G. Weststeijn), CBS H-21497,culture CBS
903.69. Brazil,fromGossypium sp., collection date and collector unknown, culture
CBS 132443 = CPC 18211; from Gossypium sp., collection date and collector
unknown, cultureCBS 132444 = CPC 18212. Iran, Golestan province, Gorgan,from
leaves of Spathiphyllum wallisii, May 2013, A. Alizadeh, strain UTFC 260 = 176C;
Guilan province, Koochesfehan area, from pods of Phaseolus vulgaris, Oct. 2013,
O. Atghia & A. Alizadeh, strain UTFC261 = 513C. Vietnam,Da Lat-Lam Dong, from
anthracnose on leaf of Coffea sp., collection date unknown, P. Nguyen & E. Liljeroth
(CBS H-21496 holotype, culture ex-holotype CBS 125474 = LD11(L3)); Da Lat-
THE COLLETOTRICHUM DRACAENOPHILUM,C.MAGNUM AND C. ORCHIDEARUM SPECIES COMPLEXES
www.studiesinmycology.org 31
Lam Dong, from anthracnose on leaf of Coffea sp., collection date unknown, P.
Nguyen & E. Liljeroth, culture CBS 125473 = LD 33(L1).
Notes:Colletotrichum plurivorum was first described as
C. sichuanensis from Capsicum annuum in the Sichuan Province
of China (Liu et al. 2016). The species is based on four “holotype
living cultures”, one of which was designated as holotype; no
type specimen has been deposited in a fungarium, and the name
is therefore invalid (Art. 40.1). This species is described as a new
species based on a strain from Coffea in Vietnam (Nguyen et al.
2010). In a study of Douanla-Meli et al. (2018),C. sichuanensis
was regarded as a synonym of C. cliviicola (as C. cliviae),
although both species formed well supported clades in the
phylogeny of that study based on a multilocus data set and there
was no indication of disconcordance between the gene trees.
Based on the data of our study confirming these clades,
C. cliviicola and C. plurivorum are regarded as distinct species.
Colletotrichum plurivorum belongs to the C. orchidearum
species complex. It has a large host range, including Anacardia-
ceae (Mangifera), Araceae (Amorphophallus, Spathiphyllum),
Caricaceae (Carica), Fabaceae (Glycine, Phaseolus), Malvaceae
(Abelmoschus, Gossypium), Musaceae (Musa), Orchidaceae
(Arundina, Cymbidium, Oncidium), Passifloraceae (Passiflora),
Rubiaceae (Coffea), Solanaceae (Capsicum, Lycopersicon) and
Theaceae (Camellia). Based on DNA sequence comparisons,
strains from Vitis spp. (Vitaceae)inBrazil(Santos et al. 2018) and
China (Lei et al. 2016), Dioscorea (Dioscoreaceae) in Puerto Rico
(S.E. Fuentes Aponte et al., unpubl. data), Myrianthus arboreus
(Urticaceae) in Cameroon and Citrus limon (Rutaceae) in Vietnam
(Douanla-Meli et al. 2018), previously identified as C. cliviae or
Colletrichum sp. were revealed to be C. plurivorum as well. Strains
from studies on lima beans (Phaseolus lunatus) in Brazil (Sousa
et al. 2018, Cavalcante et al. 2018) probably also belong to this
species, based on sequence comparisons of GAPDH and ACT
sequences (TUB2 was mixed-up with a C. truncatum strain) or a
multilocus phylogeny, respectively. Many species have been re-
ported or were described from these hosts (Farr & Rossman 2017,
this study). It is likely that this species has been described previ-
ously in the pre-molecular era based on morphology. Due to the
small differences in morphology (e.g. conidial size and shape)
between the species of the C. orchidearum complex on the one
hand and the large interspecific variation on the other, it is hardly
possible to link such old species.
Colletotrichum plurivorum formed a sexual morph in culture.
However, some strains predominantly formed a sexual morph,
while the asexual morph was hardly or not observed; other
strains predominantly formed an asexual morph, while the sexual
morph was lacking or both morphs were formed. Similar ob-
servations were made in other studies. For example, strain
CGMCC 3.17358 from Camellia in China (Liu et al. 2015) was
sterile on SNA and PDA, but developed the sexual morph on
Anthriscus stems; no asexual morph was observed. In contrast,
strains from Orchidaceae in Yang et al. (2011) formed both a
sexual and an asexual morph. In the study of Douanla-Meli
et al.(2018) isolates of C. plurivorum (as C. cliviae) from Citrus
limon in Vietnam only produced the sexual morph, while isolates
from Myrianthus arboreus in Cameroon produced both the
asexual and the sexual morph.
In pathogenicity tests by Liu et al. (2016) strains described as
C. sichuanensis from Capsicum annuum in China were able to
infect fruits of Capsicum annuum and Pyrus pyrifolia. The
pathogenicity of strain UTFC 261 from Phaseolus vulgaris in Iran
was confirmed by Atghia (2015).
Colletotrichum plurivorum can be identified by GAPDH,HIS3,
and TUB2 sequences with few nucleotides difference in each
gene to the closely related C. cliviicola. The asexual morph of the
two species is similar. However, C. plurivorum is slower growing
on SNA and OA than C. cliviicola. Morover, microcyclic con-
idiation and the formation of anastomoses were observed in the
ex-type strain of C. cliviicola, but not in C. plurivorum, while no
sexual morph is so far known in C. cliviicola.
In a blastn search in GenBank with the TUB2 sequence of the
ex-type strain CBS 125474 the sequences of several strains
were found to be 100 % identical: C. cliviae strains AH1B6,
AH1B5 (Wang et al. 2016b) and LF774 (Liu et al. 2015) from tea
plants on China, of which the latter was included in this study (as
CBS 125375), Colletotrichum sp. strain FJ074 from grapevine in
China (Lei et al. 2016) as well as Colletotrichum isolates LJTJ3,
LJTJ22 and LJTJ30 that were described by Liu et al. (2016) as
C. sichuanensis and included in this study. The closest matches
with the HIS3 sequence of the ex-type strain are with 99 %
identity (1 nucleotide difference) six C. cliviae strains from soy-
beans in Brazil (Barbieri et al. 2017), of which one, strain
LFN0008, is included in this study. The GAPDH sequence of the
ex-type strain of C. plurivorum is 100 % identical to several
strains that were previously identified as C. cliviae, including
G61B, G14B, F85241C from Dioscorea alata in Puerto Rico
(S.E. Fuentes Aponte et al., unpubl. data), JQS from Zamio-
culcas zamiifolia in China (Zhou & Li 2017), AV1 from grapevine
in Brazil (Santos et al. 2018), AH1A2 and AH1B5 from tea plants
on China (Wang et al. 2016b), HP157 from papaya in Mexico (I.
Marquez-Zequera, unpubl. data), CORCG2 from Cymbidium in
China (Yang et al. 2011) that is included in this study, and the six
strains from soybeans in Brazil (Barbieri et al. 2017). The blastn
search with the ITS sequence of strain CBS 125474 resulted in
40 sequences with 100 % identity. Most, if not all of these
strains probably represent C. plurivorum as well, but this needs
to be confirmed based on GAPDH,HIS3,orTUB2 sequences.
The clades representing C. plurivorum and C. sojae in the
phylogeny in this study include strains from Phaseolus spp. in
Iran and Benin. The well-known anthracnose pathogen of com-
mon beans, C. lindemuthianum had been reported to form sexual
morphs in culture that were called Ga. lindemuthiana (Shear &
Wood 1913)orGa. cingulata f. sp. phaseoli (Kimati & Galli
1970). However, in an extensive study on C. lindemuthianum
that included the epitypification of this species, several methods
were unsuccessfully applied to induce the formation of a sexual
morph of this species (Liu et al. 2013). Liu et al. (2013) assumed
the sexual morphs not to be conspecific with C. lindemuthianum.
The species studied by Shear & Wood (1913) formed conidia
that measure 10.5–16.5 × 4.5–5μm on corn meal agar and
12–16.5 × 4.5–6μm on host tissue and curved ascospores that
measure 15–22.5 × 4.5–6μm; it could well be a species of the
C. orchidearum complex. The ascospores of C. sojae studied by
us are narrower, while the ascospores of C. plurivorum observed
Fig. 16. Colletotrichum plurivorum (A, C–H, O–T, V –AD. from ex-holotype culture CBS 125474. B, I–N, U. from culture CBS 903.69). A–B. Conidiomata. C, I. Tips of setae.
D, J. Bases of setae. E–H, K–N. Conidiophores. O–S. Appressoria. T–U. Conidia. V–W. Ascomata. X. Paraphyses. Y. Outer surface of peridium. Z. Peridium in cross section.
AA–AC. Asci. AD. Ascospores. A, C –H,T, V, X, AA, AD. from Anthriscus stem. B, I–S, U, W, Y–Z, AB–AC. from SNA. A–B, V –W. DM. C–U, X–AD. DIC. Scale bars:
A = 100 μm, T = 10 μm. A applies to A–B, V–W. T applies to C –U, X–AD.
THE COLLETOTRICHUM DRACAENOPHILUM,C.MAGNUM AND C. ORCHIDEARUM SPECIES COMPLEXES
www.studiesinmycology.org 33
in this study and the studies of Liu et al. (2015) and Yang et al.
(2011) have a different shape, being only very slightly curved.
However, ascospores reported in Liu et al. (2016) from Capsicum
in China have a similar shape, and are similarly arranged in the
ascus. It is therefore possible that C. plurivorum is the same
fungus as “Ga. lindemuthiana”. In contrast, ascospores of Ga.
lindemuthiana shown by Rodríguez-Guerra et al. (2005) are
broader than both species, measuring 15.5–29 × 4.5–7μm; the
size differs from both. Kimati & Galli (1970) observed two types
of asci and ascospores of Ga. cingulata f. sp. phaseoli:(1–)
4(–8)-spored asci with mean ascospores dimensions,
20 × 6.5 μm, and 8-spored asci, with ellipsoidal ascospores,
measuring on average 10 × 4 μm, that also do not match with
any of the species treated in this study.
While no sequence data were included in these papers, few
studies showed that strains from Phaseolus vulgaris with sexual
morphs and those forming asexual morphs belong to different
RAPD groups (Talamini et al. 2006) or clades in phylogenies
based on ITS and HMG sequence data (Barcelos et al. 2011,
2014), one of which clustered with Ga. magna strains
(Barcelos et al. 2014). A blast search with a sequence of strain
UFLAG07-3 (KF604738) from that study was 99 % identical (4
nucleotides difference) with Ga. glycines strain LFN0009 from
soybean in Brazil (Barbieri et al. 2017), that is included in the
C. orchidearum subclade in this study; based on that it is
probably none of the species treated in this study. More loci need
to be sequenced to resolve the position of this species.
There are also two strains from Gossypium in Brazil that were
identified as C. plurivorum. However, the common causal agents
of cotton anthracnose and ramulose, C. gossypii var. gossypii
and C. gossypii var. cephalosporioides, respectively, belong to
the C. gloeosporioides complex (Salustiano et al. 2014,U.
Damm, unpubl. data).
Colletotrichum sojae Damm & Alizadeh sp. nov. MycoBank
MB824229. Fig. 17.
Non Vermicularia truncata Schwein., Trans. Am. phil. Soc. 4(2):
230. 1832. Glomerella glycines Lehman & F.A. Wolf, J. Agric.
Res., Washington 33(4): 381. 1926. [=Colletotrichum truncatum
(Schwein.) Andrus & W.D. Moore].
Etymology: The species epithet is derived from the host plant
Glycine max, soybean.
Sexual morph on Anthriscus stem (observed in strains ATCC
62257, CBS 134.87, UTFC 288, UTFC 301 and UTFC 303).
Ascomata perithecia, solitary, superficial, non-stromatic, globose
to subglobose, ostiolate, pale to medium brown, 80–150 μm
diam, glabrous or covered with few stiff hairs, 10–70 μm long.
Peridium 10–15 μm thick, composed of 3 layers of pale brown
flattened textura angularis.Ascogenous hyphae hyaline, smooth-
walled, delicate, rarely visible. Interascal tissue not observed.
Asci mostly immature asci of strain ATCC 62257 observed,
mature asci apparently disintegrating quickly, unitunicate, 8-
spored, cylindrical to clavate, smooth-walled, the base broadly
truncate, mature asci of strain UTFC 288 thin-walled, clavate,
measuring 77–80 × 11–11.5 μm. Ascospores aseptate, hyaline,
smooth-walled, fusiform, curved, sometimes additionally slightly
flexed in the middle, with rounded ends, (14–)
15.5–23(–33.5) × (3.5–)4–5μm, mean ± SD = 19.3 ± 3.7 ×
4.5 ± 0.3 μm, L/W ratio = 4.3, ascospores of strain UTFC 288
wider, measuring 14.5–20 × 5 –6.5 μm, mean ± SD = 17 ±
1.49 × 5.5 ± 0.4, L/W ratio = 3.1.
Sexual morph on SNA (only observed in strain CBS 134.87).
Ascomata perithecia, solitary or in clusters, superficial or
immersed in the agar medium, non-stromatic, ± globose, ostio-
late, pale to medium brown, glabrous. Asci not observed. As-
cospores aseptate, hyaline, smooth-walled, fusiform, curved,
with rounded ends, (18.5–)20–27(–37) × (4–)4.5–5μm,
mean ± SD = 27.7 ± 3.5 × 4.6 ± 0.3 μm, L/W ratio = 5.2.
Asexual morph on SNA. Vegetative hyphae 1–7.5 μm diam,
hyaline to pale brown, smooth-walled, septate, branched. Chla-
mydospores not observed. Conidiomata, conidiophores and
setae formed directly on hyphae. Setae medium brown, smooth-
walled to verruculose, 30–75 μm long, 2–3-septate, base cy-
lindrical, conical to slightly inflated, 3.5–6μm diam, tip ± rounded
to slightly acute. Conidiophores hyaline, smooth-walled, simple,
sometimes septate, to 25 μm long. Conidiogenous cells hyaline,
smooth-walled, cylindrical to ovoidal, often integrated and conical
or clavate, 7–22.5 × 3–6.5(–8) μm, opening 1–2μm diam,
collarette 0.5–1μm long, periclinal thickening distinct. Conidia
hyaline, smooth-walled, initially aseptate, becoming septate with
age, straight, the apex and base rounded, (11–)
14–17(–17.5) × (4.5–)5–6(–6.5) μm, 15.7 ± 1.4 × 5.4 ± 0.4 μm,
L/W ratio = 2.9, conidia of strain ATCC 11871 narrower,
measuring (14–)15–18(–19.5) × (4–)4.5–5(–5.5) μm,
mean ± SD = 16.5 ± 1.5 × 4.9 ± 0.3 μm, L/W ratio = 3.4.
Appressoria (only few observed) single, medium brown, smooth-
walled, navicular, bullet-shaped or irregular in outline, with an
undulate to lobate margin, (9–)12–19(–24) × (6–)
8–11.5(–13) μm, mean ± SD = 15.3 ± 3.6 × 9.7 ± 1.8 μm, L/W
ratio = 1.6, appressoria of strain UTFC 288 smaller, measuring
11–21 × 5 –9.5 μm, mean ± SD = 14.5 ± 3.1 × 7.5 ± 1.4 μm, L/W
ratio = 1.9.
Asexual morph on Anthriscus stem. Sporulation sparse.
Conidiomata not observed in strain ATCC 62257, conidiophores
and setae formed by strain ATCC 11871 on pale brown angular
cells, 3–8.5 μm diam. Setae (few observed) medium to dark
brown, smooth-walled or verruculose, 50–130 μm long, 1–12-
septate, base cylindrical to conical, or ± inflated, 3–8.5 μm
diam, tip ± acute to ± rounded. Conidiophores and con-
idiogenous cells not observed in strain ATCC 62257. Co-
nidiophores in strain ATCC 11871 hyaline to pale brown, smooth-
walled, septate, branched, to 40 μm long. Conidiogenous cells in
strain ATCC 11871 hyaline to pale brown, smooth-walled, cylin-
drical, 8–25 × 4–5.5 μm, opening 1–2μm diam, collarette 0.5 μm
long, rarely seen, periclinal thickening observed. Conidia hyaline,
smooth-walled, aseptate, straight, the apex and base rounded,
(12.5–)14–17.5(–18.5) × 5–6(–6.5) μm, mean ± SD =
15.8 ± 1.5 × 5.5 ± 0.4 μm, L/W ratio = 2.9.
Culture characteristics: Colonies on SNA flat, with entire margin,
hyaline to honey, covered by thin, floccose, whitish to pale grey,
aerial mycelium, reverse same colours; 28–29 mm in 7 d
(38.5–40 mm in 10 d). Colonies on OA flat with entire margin,
Fig. 17. Colletotrichum sojae (B–D, I–V, Y –AE. from ex-holotype culture ATCC 62257. A, E–H. from culture ATCC 11871. W–X. from culture CBS 134.87). A–B. Con-
idiomata. C. Tip of a seta. D. Base of a seta. I. Seta. E–H, J –M. Conidiophores. N–S. Appressoria. T–U. Conidia. V. Peridium in cross section. W–X. Ascomata. Y. Outer
surface of peridium. Z. Ascospores. AA–AD. Immature asci. AE. Disintegrating apex of an ascus apparently after ascospore release. A, C –H, T, V, X–AE. from Anthriscus
stem. B, I–S, U, W. from SNA. A–B, W–X. DM. C–V, Y –AE. DIC. Scale bars: A = 100 μm, T = 10 μm, Y = 40 μm. A applies to A –B, W–X. T applies to C –U, V, Y –AE.
THE COLLETOTRICHUM DRACAENOPHILUM,C.MAGNUM AND C. ORCHIDEARUM SPECIES COMPLEXES
www.studiesinmycology.org 35
buff, olivaceous buff to olivaceous, partly covered with thin,
floccose, whitish aerial mycelium, reverse buff to olivaceous
grey, 30–32.5 mm in 7 d (40 mm in 10 d). Conidial mass not
observed in strain ATCC 62257, those of strain UTFC 288 pale
yellow.
Materials examined:Iran, Golestan province, Tooskestan area, isolated from
leaves of Glycine max, Oct. 2013, A. Alizadeh & O. Atghia, strain UTFC
288 = 442C; Guilan province, Roodsar area, from leaves of Phaseolus vulgaris,
Oct. 2013, A. Alizadeh & O. Atghia, strain UTFC 301 = 507C; Guilan province,
Loolman-Astaneh ashrafie road, from leaves of Vigna unguiculata, Oct. 2013, A.
Alizadeh & O. Atghia, strain UTFC 303 = 524C. Italy, Bologna, from Glycine max,
unknown collection date and collector (sent to CBS Jan. 1987 for identification by
P. Giunchi, Osservatorio Malattie Delle Piante, Bologna, Italy, identification by
H.A. van der Aa, No. 9944), CBS H-12958, culture CBS 134.87. Serbia and
Montenegro, Novi Sad, from Glycine max, unknown collection date, Z. Klocokar-
Smit (isolated 1980 by Z. Klocokar-Smit and deposited in CBS collection Jan.
1981), CBS H-12907, culture CBS 181.81; Novi Sad, from Glycine max, unknown
collection date and collector (isolated 1980 by Z. Klocokar-Smit and deposited in
CBS collection Jan. 1981), culture CBS 182.81. USA, Illinois, from Glycine max,
unknown collection date, J.B. Manandhar (received from R. O'Connell as Ga.
glycines, before from J.B. Sinclair) (CBS H-21495 holotype of C. sojae, culture
ex-holotype ATCC 62257 = CPC 19367); Iowa, from stem of Medicago sativa,
unknown collection date and collector (received from R. O'Connell as Ga. gly-
cines, before from L.H. Tiffany), culture ATCC 11871 = CPC 18948; Iowa, from
stem of Medicago sativa unknown collection date and collector (received from R.
O'Connell as C. destructivum, before from A. Alfaro, originally from L.H. Tiffany),
culture CBS 128510 = LARS 204 = CECT 2873 = Alfaro 212 = ATCC
11871 = DSM 1167 (apparently the same strain as above); North Carolina,
Raleigh, from anthracnose of Glycine max, unknown collection date, S.G. Leh-
man & F.A. Wolf [BPI 596658 (dried culture) holotype of Ga. glycines]; North
Carolina, Raleigh, from anthracnose of Glycine soja (=G. max), unknown
collection date and collector (isolated by S.G. Lehman as strain 1936, deposited
in CBS collection as Ga. glycines in May 1932 by J.A. Stevenson), (strain CBS
195.32 ex-holotype strain of Ga. glycines).
Notes: Soybean (Glycine max) anthracnose was first reported in
1917 from Korea, where it was attributed to C. glycines (Hemmi
1920). Anthracnose occurs in all soybean producing areas and
can cause yield losses of up to 100 % (Sinclair 1982). Lehman &
Wolf (1926) studied soybean anthracnose in the USA and
noticed an asexual morph they identified as C. glycines and a
sexual morph they regarded as the sexual morph of this species,
hence naming it Ga. glycines.
The only specimen located that could directly be linked to
Lehman & Wolf is BPI 596658. A handwritten note with that
specimen reads “ATCC #1936, G. glycines (Hori) n.n. (n.n. was
later crossed out and replaced by “Lehman & Wolf”), S.G.
Lehmann and Frederick A. Wolf, Jour. Agric. Research 33:
381–390, Aug. 15, 1926”, referring to the publication of that
species. We therefore regard this specimen as the holotype of
Ga. glycines.
Specimen BPI 596658 contains two dried cultures of Ga.
glycines strain ATCC 1936 (Label on tubes: Glomerella glycines,
A.T.C.C. # 1936, 11-19-1929, Vaxter agar on soybean). No
conidia or ascospores were observed on these cultures. The
same number (1936) appeared with the strain information of
strain CBS 195.32 that was isolated by S.G. Lehman from Glycine
max in Raleigh, North Carolina, USA, originally identified as Ga.
glycines and deposited to the CBS collection in May 1932 by J.A.
Stevenson. This means CBS 195.32 is apparently the same strain
as ATCC 1936 and therefore regarded as the ex-holotype strain of
Ga. glycines. The ATCC collection lists a strain ATCC 1936, but
recorded as “unknown”. According to a request at ATCC (A.
Khashnobish, in lit.), no strain ATCC 1936 exists anymore.
However, strain CBS 195.32 was identified as C. truncatum
by Damm et al. (2009). Drawings and description of the asexual
morph of Ga. glycines observed by Lehman & Wolf (1926) agree
with this. Colletotrichum truncatum is a common pathogen of
soybean (Ramos et al. 2013, Yang et al. 2014, Rog
erio et al.
2016). However, it is dubious that this soybean pathogen
formed a sexual morph in the study of Lehmann & Wolf, as a
sexual morph was never observed for any strain of C. truncatum,
neither in the extensive study of Damm et al. (2009) nor in any
other study on this pathogen (e.g. Shenoy et al. 2007, Than et al.
2008, Rog
erio et al. 2016).
Lehman & Wolf (1926) stated that they made isolations “from
diseased stems and pods bearing the conidial stage, from
infected seed, and from old, decaying stems bearing the
ascogenous stage”and further: “No efforts have been made to
make single-spore cultures from either conidia or ascospores...”
There is no indication that the authors would have found conidia
and ascospores on the same plant parts. They transferred blocks
of agar containing several spores each. Although the authors did
not observe cultural differences, which was obviously the reason
for them to conclude they were studying one species, it is likely
that two species were isolated and even mixed cultures were
prepared: “Some strains from conidia have born conidia alone,
even after repeated transfer and cultivation on a variety of media;
other strains from ascospores, however, have consistently yiel-
ded both the conidial and ascospore stages when cultivated
upon the same kind of media”(Lehman & Wolf 1926). So, it is
likely that the sexual morph and the asexual morph observed by
them are different species. Yang et al. (2014) reported two
species to be common pathogens of soybean in the USA,
namely “Ga. glycines”and C. truncatum. Apparently, Lehman &
Wolf never noticed the asexual morph that really belongs to the
observed sexual morph.
According to Tiffany & Gilman (1954) and Lin & Wu (1966),
the asexual morph of Ga. glycines is not C. glycines, but a
species similar to C. destructivum.Manandhar et al. (1986)
claimed C. destructivum to be the asexual morph of Ga. gly-
cines by morphological comparison of strains from Glycine
forming a sexual morph as described for Ga. glycines (Lehman &
Wolf 1926) that were therefore considered to be Ga. glycines,
and strains from Medicago that were considered to be
C. destructivum and gave a detailed description of both morphs.
Strains from Manandhar et al. (1986) from both hosts (ATCC
62257 from Glycine max and ATCC 11871 from Medicago), were
included in this study and confirmed to be conspecific. However,
these strains are not closely related to C. destructivum that was
epitypified in a recent study and belongs to the C. destructivum
complex (Damm et al. 2014).
One of the two strains from Manandhar et al. (1986) easily
formed the very typical sexual morph in culture that agrees with
the pictures and the description of Lehman & Wolf (1926):
perithecia 220–340 μm diam, asci 70–106 × 9.5–13.5 μm,
mean = 80 × 12 μm; ascospores in culture were (13.12–)
18.75–28.12(–43.35) μm long. The ascospores of C. plurivorum,
also occurring on soybeans and also described in this study,
have a different shape (see Fig. 16).
The name Ga. glycines was applied by Lehman & Wolf
(1926) based on the asexual morph that was identified by
them as C. glycines and the simultaneously observed sexual
morph that was apparently wrongly connected to the asexual
morph. However, the type specimen of Ga. glycines only in-
cludes the asexual morph that was re-identified as C. truncatum
by Damm et al. (2009).Colletotrichum truncatum is based on
Vermicularia truncata (Schweinitz 1832), which has priority over
DAMM ET AL.
36
Ga. glycines. There is no name available for the second species
that forms the sexual morph. Consequently, this species is
described here as a new species, C. sojae.
According to von Arx (1957),C. glycines Hori ex Hemmi is a
synonym of C. dematium f. truncata, which is a synonym of
C. truncatum (Damm et al. 2009). However, von Arx neither saw
the type specimen nor a strain of C. glycines. In the Compendium
of Soybean Diseases, the two species on soybean are treated as
C. truncatum (= C. glycines) and Ga. glycines (Sinclair 1982). It
is possible that C. glycines and C. truncatum are synonyms as
both types represent species with curved conidia. However, Yang
et al. (2014) collected three Colletotrichum species with curved
conidia on soybeans in the USA, C. incanum,C. chlorophyti and
C. truncatum.Colletotrichum glycines Hori ex Hemmi could be a
synonym of any of these species, or even a different species. We
did not study the type of C. glycines and there is no ex-type strain
of C. glycines available for molecular analysis. In order to clarify
its systematic position, C. glycines would need to be epitypified.
Another species named C. glycines,C. glycines Gonz. Frag.
(Gonz
alez Fragoso 1924) was not described from soybean, but
from Wisteria sinensis (syn.: Glycine sinensis) in Portugal. The
name is illegitimate, according to Art. 53.1.
Within the strains of this species there is considerable vari-
ation in the formation of sexual and asexual morphs. There are
strains that form the asexual morph only and strains that pre-
dominantly form the sexual morph. It is very likely the latter case
is the reason for Lehman & Wolf (1926) to overlook the asexual
morph of this species.
Colletotrichum sojae occurs mainly on Fabaceae (Glycine,
Medicago,Phaseolus, Vigna), but also on Amaranthaceae
(Amaranthus), Asteraceae (Arctium), Solanaceae (Capsicum)
and Orchidaceae (Bletilla). There is another species occurring on
Fabaceae (including Glycine and Phaseolus) treated in this study
(see C. plurivorum).
Colletotrichum sojae can be identified by HIS3,ACTand TUB2
sequences, while the ITS is identical with that of C. vittalense.
Closest matches in blastn searches with the TUB2 sequence of
the ex-type strain of C. sojae, ATCC 62257, resulted in 100 %
matches with Ga. glycines strain ATCC 62257 (Yang et al. 2014),
Colletotrichum strain CGMCC 3.15171 from Bletilla in China (Tao
et al. 2013), C. cliviae strain CAUOS5 from Capsicum in China
(Diao et al. 2017) and with 99 % identity (1 nucleotide difference)
Ga. glycines strains IL18A and IL26A (Yang et al. 2014) and
C. orchidearum isolate SAUCC 1407 (Xu et al. 2016), both
included in this study. The ACT sequence of strain ATCC 62257
resulted in 100 % identity with the three Ga. glycines strains of
Yang et al. (2014),Colletotrichum strains CGMCC 3.15171 from
Bletilla (Tao et al. 2013) and CAUGOS3 from Capsicum in China
(Diao et al. 2017), all included in this study, as well as Colleto-
trichum sp. SKH-2010 isolates C07010 and C08116 (GU944760,
GU935805, Choi et al. 2011) and with 99 % identity (1 and 2
nucleotides difference), C. orchidearum isolate SAUCC 1407 (Xu
et al. 2016) and Ga. glycines strain LFN0009 (Barbieri et al. 2017),
both included in this study. The GAPDH of strain ATCC 62257 is
100 % identical with Ga. glycines strains IL26A, ATCC 62257,
IL18A (Yang et al. 2014) and LFN0009 (Barbieri et al. 2017) and
C. aracearum strains LC1033 and LC1041 (Hou et al. 2016), all
included in this paper, as well as Colletotrichum isolates C08116,
C07004 and C07010 (GU935864, GU935865, GU935866, Choi
et al. 2011). Closest matches in blastn searches with the ITS
sequence of the ex-type strain resulted in a large number of
identical sequences from several host plants in different countries.
Colletotrichum orchidearum isolate SAUCC 1407 from Arc-
tium lappa in China (Xu et al. 2016), C. cliviae strains LFN0009
(as Ga. glycines in GenBank) from Glycine max in Brazil
(Barbieri et al. 2017) and CAUOS5 from Capsicum in China
(Diao et al. 2017), Colletotrichum sp. from Bletilla in China
(CGMCC 3.15171, Tao et al. 2013) as well as Ga. glycines
isolates IL18A, IL26A from soybean in the USA (Yang et al.
2014) could be re-identified as C. sojae based on a sequence
comparison in this study. We suspect the ACT sequence
KP890098 is actually from CAUOS5 as well, and not from
CAUOS3. There is no ACT sequence of CAUOS5 listed in the
paper (Diao et al. 2017). Colletotrichum sp. SKH-2010 isolates
C07010, C07004 and C08116 were also identified as C. sojae.
These strains were cited from Choi et al. (2011) in GenBank, but
not included in that paper. As there is no strain information, we
also did not include the strains in our phylogeny. In contrast,
strain MAFF 238875 from Glycine max in Japan previously
identified as C. cliviae was revealed to be C. plurivorum.
Moreover, the ITS of “Ga. glycines”isolate IFO 7384 from an
unknown host (GenBank AB057435, Moriwaki et al. 2002)is
100 % identical with that of CBS 127604, the ex-holotype strain
of C. lentis (Damm et al. 2014).
Colletotrichum tropicicola Phouliv. et al., Cryptog. Mycol.
33(3): 353. 2012. Fig. 18.
A description of the type specimen is provided by Noireung et al.
(2012). The description below is based on strains from Citrus sp.
collected in Mexico.
Sexual morph not observed.
Asexual morph on SNA (CBS 127555). Vegetative hyphae
1.5–10.5 μm diam, hyaline, smooth-walled, septate, branched.
Chlamydospores not observed. Conidiomata, conidiophores and
setae formed directly on hyphae. Seta (only one observed)
medium brown, verruculose, 81 μm long, 3-septate, base cylin-
drical, 5.5 μm diam, tip ± rounded. Conidiophores hyaline,
smooth-walled, septate, branched, to 40 μm long. Conidiogenous
cells hyaline, smooth-walled, cylindrical, (3–)5–18 × (2–)
3–4.5 μm, often extending to form new conidiogenous loci,
opening 1.5–2μm diam, collarette 1 μm long, periclinal thick-
ening distinct. Conidia hyaline, smooth-walled, aseptate, straight,
cylindrical, the apex rounded, sometimes tapering to the rounded
or truncate base, (13.5–)14–17.5(–22) × 4.5–5(–5.5) μm,
mean ± SD = 15.7 ± 1.7 × 4.8 ± 0.3 μm, L/W ratio = 3.3, conidia
of strain CBS 133174 wider, measuring (14–)
15–17(–18.5) × (5–)5.5–6μm, mean ± SD = 16.2 ±
1.0 × 5.5 ± 0.3 μm, L/W ratio = 2.9. Appressoria single or in loose
groups, medium to dark brown, smooth-walled, ellipsoidal to
bullet-shaped in outline, with an entire or undulate margin, (7–)
8–11( –14.5) × (4 –)5.5 –7( –7.5) μm, mean ± SD = 9.5 ±
1.5 × 6.2 ± 0.7 μm, L/W ratio = 1.6, appressoria of strain CBS
133174 larger, only few observed, measuring
11.5–20(–24.5) × 5–8(–10) μm, mean ± SD = 15.8 ± 4.1 ×
6.7 ± 1.6 μm, L/W ratio = 2.4.
Asexual morph on Anthriscus stem (CBS 127555). Con-
idiomata, conidiophores formed directly on hyphae. Setae not
observed. Conidiophores hyaline, smooth-walled, septate,
branched, to 30 μm long. Conidiogenous cells hyaline, smooth-
walled, cylindrical to ± inflated, 8–25 × 3.5–4.5(–6) μm, opening
1.5–2μm diam, collarette 0.5 μm long, periclinal thickening
distinct. Conidia hyaline, smooth-walled, aseptate, straight, cy-
lindrical, the apex rounded, sometimes tapering to the rounded or
THE COLLETOTRICHUM DRACAENOPHILUM,C.MAGNUM AND C. ORCHIDEARUM SPECIES COMPLEXES
www.studiesinmycology.org 37
truncate base, (13.5–)15–17(–18) × 4.5–5(–5.5) μm,
mean ± SD = 15.9 ± 1.0 × 5.0 ± 0.2 μm, L/W ratio = 3.2, conidia
of strain CBS 133174 larger, measuring (15.5–)17–19.5
(–20) × 5–5.5 μm, mean ± SD = 18.2 ± 1.0 × 5.4 ± 0.2 μm, L/W
ratio = 3.4.
Cultural characteristics (CBS 127555). Colonies on SNA flat with
entire margin, medium buff to pale honey, Anthriscus stem, filter
paper and SNA medium partly covered with greyish to orange
acervuli and whitish to grey aerial mycelium, reverse buff to
honey with grey to orange acervuli shining through; 15 –18 mm in
7 d (28.5–32 mm in 10 d). Colonies on OA flat with entire margin,
surface buff, pale olivaceous grey to olivaceous grey, partly
covered with floccose to felty pale olivaceous grey to olivaceous
grey aerial mycelium and grey to orange acervuli, reverse buff,
rosy buff, pale olivaceous grey to olivaceous grey; 16–17.5 mm
in 7 d (30–32.5 mm in 10 d). Conidial mass orange.
Materials examined:Mexico, Tamaulipas, from Citrus sp., 16 Oct. 2008, M. de
Jesus Yanez-Morales, CBS H-21070, culture CBS 127555 = CPC 15927;
Tamaulipas, from Citrus sp., 16 Oct. 2008, M. de Jesus Yanez-Morales, culture
CBS 133174 = CPC 15924.
Notes: This species belongs to the C. dracaenophilum species
complex. Noireung et al. (2012) described C. tropicicola from
leaves of Citrus maxima and Paphiopedilum bellatolum in
Thailand. Further strains from Citrus sp. in Mexico could be
identified as C. tropicicola in this study based on the multilocus
phylogeny. While these two strains formed a well-supported clade
with the ex-type strain of C. tropicicola in our phylogeny, the strain
from Paphiopedilum formed a sister clade to C. excelsum-altitu-
dinum and could represent a further species (referred to as
“C. tropicicola 2”in the phylogeny). Thus C. tropicicola s. str. only
occurs on Citrus spp., according to our data.
Colletotrichum tropicicola can be identified with sequences of
all loci available (ITS, GAPDH,ACT,TUB2), but best with GAPDH
and TUB2. The ITS sequence of the ex-type strain, MFLUCC 11-
0114, is 99 % identical (2, 7 and 6 nucleotides difference) to those
of the apparently wrongly identified C. cordylinicola strain LC0886
(KC790944, Sharma et al. 2013a), the two C. excelsum-altitudi-
num isolates (Tao et al. 2013) and MFLUCC 10-0167, the
C. tropicicola strain from Paphiopedilum, respectively. The
GAPDH sequence of the ex-type strain is 99 % identical (4 nu-
cleotides difference) to that of the C. tropicicola strain from
Paphiopedilum and 96 % identical with the two C. excelsum-alti-
tudinum isolates. The ACT sequence of the ex-type strain is 99 %
identical (3 and 11 nucleotides difference) to that of the
C. tropicicola strain from Paphiopedilum and the two C. excelsum-
altitudinum isolates, respectively.The TUB2 sequence of the ex-
type strain is 98 % identical (10 and 8 nucleotides difference) with
the two original C. excelsum-altitudinum isolates and that of the
C. tropicicola strain from Paphiopedilum, respectively.
Colletotrichum vittalense Damm, sp. nov. MycoBank
MB824230. Fig. 19.
Fig. 18. Colletotrichum tropicicola (A–B, E–R. from culture CBS 127555. C–D from culture CBS 133174). A–B. Conidiomata. C, G. Tips of setae. D–H. Bases of setae. E–F,
I–J. Conidiophores. K–P. Appressoria. Q–R. Conidia. A, C–F, Q. from Anthriscus stem. B, G–P, R. from SNA. A–B. DM. C –R. DIC. Scale bars: A = 100 μm, E = 10 μm. A
applies to A–B. E applies to C–R.
DAMM ET AL.
38
Etymology: The species epithet is derived from Vittal, the place in
Karnataka, India, where the type was collected.
Sexual morph on SNA (only observed in strain CBS 181.82).
Ascomata perithecia, formed after 4 wk, solitary, superficial or
immersed in the agar medium, non-stromatic, globose to obpyri-
form, ostiolate, glabrous, dark brown, 250–300 × 200–250 μm.
Peridium 10–15 μm thick, composed of 3 –5 layers of medium
brown flattened textura angularis with cells 5.5–15 μmdiam.
Ascogenous hyphae hyaline, smooth, delicate, rarely visible. Inter-
ascal tissue formed of paraphyses, hyaline, smooth-walled, cylin-
drical, disintegrating quickly, septate, apically free. Asci unitunicate,
8-spored, cylindrical to clavate, tapering to apex and base, smooth-
walled, 53–83 × 9–11 μm, the base broadly truncate. Ascospores
uni- or biseriately arranged, aseptate, hyaline to pale brown,
smooth-walled, allantoid, fusiform to ellipsoidal, with both ends
rounded, curved or straight, (11.5–)14.5–19(–23.5) × (4–)
4.5–5.5(–6) μm, mean ± SD = 16.8 ± 2.3 × 5.1 ± 0.4 μm, L/W
ratio = 3.3.
Sexual morph on Anthriscus stem (only observed in strain
CBS 181.82). Ascomata perithecia, formed after 4 wk, solitary,
superficial, non-stromatic, globose to obpyriform, ostiolate,
glabrous, dark brown, 150–320 × 130–200 μm diam. Peridium
11 –13 μm thick, composed of 4–6 layers of medium brown
flattened textura angularis with cells 5–15.5 μm diam. Paraph-
yses not observed. Asci 8-spored, unitunicate, cylindrical to
clavate, 53–83 × 9–11 μm. Ascospores uni- or biseriately ar-
ranged, aseptate, hyaline to pale brown, smooth-walled, allan-
toid, fusiform to ellipsoidal, with both ends rounded, curved or
straight, (12–)14–16.5(–17.5) × (4.5–)5–5.5(–6) μm,
mean ± SD = 15.1 ± 1.3 × 5.3 ± 0.4 μm, L/W ratio = 2.8.
Asexual morph on SNA. Vegetative hyphae 1.5–6.5 μm diam,
hyaline to pale brown, smooth-walled, septate, branched. Chla-
mydospores not observed. Conidiomata, conidiophores and
setae formed directly on hyphae. Setae medium to dark brown,
verruculose to verrucose, 50–120 μm long, 1–3-septate, base
cylindrical to conical, 3.5–5.5 μm diam, tip ± acute. Co-
nidiophores pale brown, smooth-walled to verruculose, septate,
branched, to 60 μm long. Conidiogenous cells pale brown,
smooth-walled to verruculose, cylindrical to clavate,
7–22.5 × 2.5–6.5 μm, opening 1.5–2μm diam, collarette
0.5–1μm long, periclinal thickening distinct, conidiogenous cells
of strain CBS 126.25 sometimes integrated. Conidia hyaline,
smooth-walled, aseptate, straight, cylindrical, sometimes slightly
clavate with the widest part at the base, the apex and base
rounded, (11.5–)13.5–16(–18.5) × 5–5.5(–7) μm,
mean ± SD = 14.8 ± 1.4 × 5.4 ± 0.4 μm, L/W ratio = 2.8, conidia
of strain CBS 126.25 slightly longer, measuring (11–)
12.5–15(–18) × 5–5.5 μm, mean ± SD = 13.6 ±
1.2 × 5.2 ± 0.2 μm, L/W ratio = 2.6. Appressoria single or in loose
groups, medium to dark brown, smooth-walled, navicular, bullet-
shaped, elliptical or irregular in outline, with a lobate or undulate
margin, (6–)9.5–16.5(–22.5) × (5–)7.5–11 ( –14.5) μm,
mean ± SD = 13.1 ± 3.5 × 9.1 ± 1.7 μm, L/W ratio = 1.4,
appressoria of strain CBS 126.25 smaller, measuring (7–)
8.5–14.5(–18.5) × (5–)5.5–8(–9.5) μm, mean ± SD = 11.3 ±
3.0 × 6.8 ± 1.1 μm, L/W ratio = 1.7.
Asexual morph on Anthriscus stem. Conidiomata, co-
nidiophores and setae formed on medium brown, angular cells,
3–7μm diam. Setae medium brown, verruculose to verrucose,
55–120 μm long, 1–2(–3)-septate, base cylindrical to conical,
4.5–7.5 μm diam, tip ± acute. Conidiophores pale brown,
smooth-walled. Conidiogenous cells pale brown, smooth-walled,
cylindrical to doliiform, 7.5–10.5 × 3.5–5.5 μm, opening 1–2μm
diam, collarette ± 0.5 μm long, periclinal thickening distinct.
Conidia hyaline, smooth-walled, aseptate, straight, cylindrical,
the apex and base rounded, 15–17(–18.5) × 5–5.5(–6) μm,
mean ± SD = 16.1 ± 0.9 × 5.3 ± 0.3 μm, L/W ratio = 3.0, conidia
of strain CBS 126.25 shorter, measuring (12.5–)
13.5–15.5(–17.5) × 5–5.5 μm, mean ± SD = 14.5 ±
1.0 × 5.2 ± 0.2 μm, L/W ratio = 2.8.
Culture characteristics: Colonies on SNA flat with entire margin,
hyaline to pale olivaceous grey, agar medium, filter paper and
Anthriscus stem partly covered with felty white to pale olivaceous
grey aerial mycelium and grey to black conidiomata/ascomata,
reverse same colours; growth 29–30.5 mm in 7 d (40 mm in
10 d), strain CBS 126.25 grows more slowly: 22.5–24 mm in 7
d (31–32.5 mm in 10 d). Colonies on OA flat with entire margin;
olivaceous buff, grey olivaceous to olivaceous, partly covered
with short felty whitish aerial mycelium and pale grey con-
idiomata/ascomata, reverse olivaceous buff, pale olivaceous
grey to olivaceous grey, growth 28.5–30 mm in 7 d (40 mm in
10 d), strain CBS 126.25 grows more slowly: 24–26 mm in 7 d
(28.5–34 mm in 10 d). Conidial mass greyish white.
Materials examined:India, Karnataka, Vittal, from Theobroma cacao (patho-
genic), collection date and collector unknown (isolated and deposited in CBS
collection by F.H. Beyma, May 1928) (CBS H-21498 holotype, culture ex-
holotype CBS 181.82). Unknown, from an unknown Orchidaceae plant, collec-
tion date and collector unknown (isolated and deposited in CBS collection by K.N.
Murthy, Mar. 1928), CBS H-21499, culture CBS 126.25.
Notes:Colletotrichum vittalense belongs to the C. orchidearum
species complex and is only known as pathogen of Theobroma
cacao in India and from an unknown Orchidaceae plant.
Several species were described on Theobroma cacao,
including C. cacao in the C. magnum complex that is also
described in this study. Numerous species were described on
Orchidaceae hosts, including C. cattleyicola, also belonging to
the C. orchidearum species complex and C. orchidearum
belonging to the C. orchidearum species complex described and
epitypified, respectively, in this study. See notes under C. cacao,
C. cattleyicola and C. orchidearum for further species and re-
ports on these hosts. The two C. vittalense strains from the CBS
collection were previously identified as Ga. cingulata var. cin-
gulata and C. gloeosporioides,Ga. lagenaria and Gl. affine, the
latter is a species that was described from Hoya (Apocynaceae)
and Vanilla (Orchidaceae) from the botanical garden in Padua,
Italy, with similar conidial shape (cylindrical with both ends
rounded) and size (14–20 × 4–6μm) as C. vittalense (Saccardo
1878). However, we could not locate the type material of this
species to confirm its taxonomic position.
A typical feature of C. vittalense are the very short asco-
spores with round ends that vary between straight to strongly
curved. This species can be identified by its unique GAPDH,
CHS-1,HIS3 and TUB2 sequences. In a blastn search on
GenBank with the GAPDH sequence of the ex-type strain of
C. vittalense, CBS 181.82, the only 100 % identical sequence
was C. cliviae strain GUFCC15503 from Calamus thwaitesii in
India (KC790759, Sharma et al. 2013a) that was included in the
phylogeny of this study and re-identified as C. vittalense. Strain
GUFCC15503 originates from Netravali in Goa, also in the south-
west of India like Vittal. The GAPDH sequence of the ex-type
strain of C. cliviicola differs in 2 nucleotides from that of the
ex-type strain of C. vittalense. Strain GUFCC15503 (KF451996,
THE COLLETOTRICHUM DRACAENOPHILUM,C.MAGNUM AND C. ORCHIDEARUM SPECIES COMPLEXES
www.studiesinmycology.org 39
Fig. 19. Colletotrichum vittalense (from ex-holotype culture CBS 181.82). A–B. Conidiomata. C, G. Tips of setae. D–H. Bases of setae. E–F, I –K. Conidiophores. L–Q.
Appressoria. R–S. Conidia. T. Outer surface of peridium. U. Peridium in cross section. V. Paraphyses. W. Ascoma. X. Ascospores. Y–Z. Asci. A, C–F, R, U, W–Z. from
Anthriscus stem. B, G–Q, S –T, V. from SNA. A–B, W. DM. C–V, X –Z. DIC. Scale bars: A = 100 μm, E = 10 μm. A applies to A–B, W. E applies to C–V, X –Z.
DAMM ET AL.
40
Fig. 20. Colletotrichum yunnanense (from ex-holotype culture CBS 132135). A–B. Conidiomata. C, G. Tips of setae. D–H. Bases of setae. E–F, I –K. Conidiophores. L–Q.
Appressoria. R–S. Conidia. T. Ascoma. U. Peridium in cross section. V. Outer surface of peridium. W–X. Asci. Y. Tip of an ascus. Z. Paraphyses. AA. Ascospores. A, C–F, R.
from Anthriscus stem. B, G–Q, S–AA. from SNA. A –B, T. DM. C–S, U –AA. DIC. Scale bars: A = 100 μm, F = 10 μm. A applies to A–B, T. F applies to C–S, U–AA.
THE COLLETOTRICHUM DRACAENOPHILUM,C.MAGNUM AND C. ORCHIDEARUM SPECIES COMPLEXES
www.studiesinmycology.org 41
Sharma et al. 2015) was also the closest match in a blastn
search with the ACT,CHS-1 and TUB2 sequences of strain CBS
181.82 with none, one or two nucleotides difference, respectively.
The ITS sequence of strain CBS 181.82 is 100 % identical with
that of strains from various hosts in different countries from
different studies, including C. sojae strains from Glycine max in
the USA (Yang et al. 2014), that were included in this study. The
ITS of the ex-type strain of C. cliviicola differs in 3 nucleotides.
Colletotrichum yunnanense Xiao Ying Liu & W.P. Wu, Myco-
taxon 100: 139. 2007. Fig. 20.
SexualmorphonSNA.Ascomata perithecia, solitary, superficial or
immersed in the agar medium, non-stromatic, subglobose, ostiolate,
medium to dark brown, 150–200 × 140 –260 μm. Peridium
10–13 μm thick, composed of 4–6 layers of pale brown textura
angularis.Ascogenous hyphae hyaline, smooth-walled, delicate.
Interascal tissue formed of paraphyses, hyaline, smooth-walled,
cylindrical, apically free, tapering towards the round tip, dis-
integrating quickly, septate, branched, 50–65 μm long,
base 2–2.5 μmdiam.Asci unitunicate, 8-spored, cylindrical to
clavate, tapering to apex and base, smooth-walled,
57–65.5 × 10.5–11.5 μm, the base broadly truncate. Ascospores
uni- or biseriately arranged, aseptate, hyaline, smooth-walled,
fusiform with acute ends, slightly curved, (13.5–)
14.5–17.5(–19.5) × 5–5.5(–6) μm, mean ± SD = 16.1 ±
1.7 × 5.4 ± 0.3 μm, L/W ratio = 3.0.
Asexual morph on SNA. Vegetative hyphae 1–9μm diam,
hyaline, smooth-walled, septate, branched. Chlamydospores not
observed. Conidiomata, conidiophores and setae formed directly
on hyphae. Setae medium brown, verrucose, warts up to 2.5 μm
diam, 70–130 μm long, 2–5-septate, base slightly inflated,
4–6.5 μm diam, tip ± acute. Conidiophores pale brown, smooth-
walled, septate, branched, to 40 μm long. Conidiogenous cells
pale brown, smooth-walled, clavate, 8–28 × 5–7μm, the upper
part often surrounded by a gelatinous sheath, opening 1.5 –2μm
diam, collarette 0.5 μm long, periclinal thickening observed.
Conidia hyaline, smooth-walled, aseptate, straight, cylindrical,
the apex and base rounded, with a prominent scar, (13–)
15.5–18(–19.5) × 4.5–5(–5.5) μm, rarely up to 29 μm long,
mean ± SD = 16.7 ± 1.2 × 5.0 ± 0.2 μm, L/W ratio = 3.4.
Appressoria in loose groups, medium to dark brown, smooth-
walled, subglobose to ellipsoidal in outline, with an entire or
lobate margin, (3.5–)5.5–8(–10) × (3–)4.5–7(–7.5) μm,
mean ± SD = 6.8 ± 1.3 × 5.8 ± 1.1 μm, L/W ratio = 1.2.
Asexual morph on Anthriscus stem. Conidiomata, co-
nidiophores and setae formed on pale brown, angular cells,
3.5–7.5 μm diam. Setae medium brown, verrucose, 75–120 μm
long, 2–3-septate, base cylindrical, conical to slightly inflated,
5–8μm diam, tip ± rounded. Conidiophores (hyaline to) pale
brown, smooth-walled, septate, branched, to 35 μm long. Con-
idiogenous cells (hyaline to) pale brown, smooth-walled, cylin-
drical, ellipsoidal to doliiform, 9.5–26 × 3.5–6μm, the upper part
often surrounded by a gelatinous sheath, opening 1–1.5 μm
diam, collarette 0.5 μm long, periclinal thickening visible,
sometimes distinct. Conidia hyaline, smooth-walled, aseptate,
straight, cylindrical, the apex and base rounded, with a prominent
scar, (16–)17–18.5( –20.5) × 5 –5.5 μm,
mean ± SD = 17.7 ± 0.9 × 5.2 ± 0.2 μm, L/W ratio = 3.4.
Culture characteristics: Colonies on SNA flat with entire margin,
hyaline, filter paper partly straw, agar medium, filter paper and
Anthriscus stem partly covered with grey acervuli and thin whitish
aerial mycelium, reverse same colours; growth 10–11.5 mm in
7 d (17–17.5 mm in 10 d). Colonies on OA flat with entire margin;
buff to grey olivaceous, partly covered with short whitish to grey
aerial mycelium and salmon to grey acervuli, reverse buff, honey,
olivaceous to olivaceous-grey, growth 11–12.5 mm in 7 d
(18–19.5 mm in 10 d). Conidial mass salmon.
Material examined:China, Yunnan, Kunming Botanical Garden, from healthy
leaves of Buxus sp., 5 Nov. 2004, W.P. Wu (WU47182 holotype [not seen],
culture ex-holotype CBS 132135 = AS 3.9617 = LC1526).
Notes:Colletotrichum yunnanense was described from healthy
leaves of Buxus sp. in Yunnan (China); the species was isolated
as an endophyte; no sexual morph was observed by incubating it
on PDA at 25 °C in the dark (Holotype WUWP 47182, Liu et al.
2007b). The ex-holotype strain was re-examined on SNA and
OA in this study, and the formation of a sexual morph was
observed on SNA. Colletotrichum yunnanense is the only spe-
cies in the C. dracaenophilum complex for which a sexual morph
is known. It is the species in this complex with the smallest
appressoria; because of their often roundish outline, these
appressoria also have the lowest L/W ratio. The growth rate of
C. yunnanense is the lowest of all species treated in this study.
There are no reports of this species other than that published
by Liu et al. (2007b). Moreover, only one other Colletotrichum
species was reported on Buxus:C. theobromicola on Buxus
microphylla var. japonica in the USA (Singh et al. 2015).
Colletotrichum yunnanense can be identified with all loci
studied. The closest matches with the sequences of all loci of the
ex-type strain CBS 132135 in GenBank are all those of this strain
itself and of the ITS sequence of strain AS 3.9616, respectively.
One further ITS sequence, LN552210, of the endophytic Colle-
totrichum sp. BS4 from leaves of Buxus sinica in Guangzhou,
China (Wang et al. 2016a) was 99 % identical (6 nucleotides
difference) to that of strain CBS 132135. The differences in the
sequences are likely to be artefacts, as they are in the beginning
of the sequence, where there are several Ns. We assume that
this strain is also representative of C. yunnanense. The authors
discovered three new antibacterial azaphilones, colletotrichones
A−C(1−3) and one known compound, chermesinone B (4a).
This isolate tested positive against environmental and human
pathogenic clinical bacterial strains (Wang et al. 2016a). The
boxwood plant, Buxus sinica, is used in traditional Chinese
medicine.
Colletotrichum sp. strains GZAAS5.09545 and COUFAL7300
Notes: Based on our multilocus phylogeny, two strains,
GZAAS5.09545 from citron leaves (Citrus medica,Rutaceae)in
Puer City, Yunnan, China (Peng et al. 2012) and COUFAL7300
from anthracnose of chayote fruits (Sechium edule, Cucurbita-
ceae) in Antonio Calos, Santa Catarina State, Brazil (Bezerra
et al. 2016) formed a clade with high bootstrap support. This
clade could represent a further unnamed species in the
C. magnum species complex. Both strains were originally
identified as C. brevisporum. The latter strain caused lesions on
fruits of Sechium edule and Cucumis melo (Bezerra et al. 2016).
DISCUSSION
All three species complexes treated in this study, namely of
C. dracaenophilum, C. magnum and C. orchidearum, form
DAMM ET AL.
42
straight conidia, similar to those in the C. gloeosporioides spe-
cies complex (Weir et al. 2012). They could have been regarded
as C. gloeosporioides in the past. This is corroborated by records
in Farr & Rossman (2017), and previous identifications linked to
sequences deposited in GenBank.
Except for C. dracaenophilum that forms larger conidia, the
conidial sizes in all three species complexes differ more within
the species (provided that more than one strain was available/
measured) than between them. Differences were mainly
observed among appressoria and ascospore sizes and shapes.
But these structures were not always available as they were not
formed by all species and cultures. In this study, nine previously
described species were assigned to one of the three species
complexes treated in this study. Together with the C. orbiculare
complex, these species representing the three species com-
plexes constitute the most basal lineages of the genus Colleto-
trichum (Marin-Felix et al. 2017).
Many species in the three species complexes are possibly
host specific—apparently mainly specialised to monocots,
while others occur on many hosts. Most of the species were
predominately isolated from plants in tropical or subtropical re-
gions of the world, mainly from Asia and Latin America. The rare
collections from the temperate climate in Central-Europe are
from ornamental plants.
One of the plant families often found as host plants in the
three species complexes is the Orchidaceae. This family is often
colonised by Colletotrichum species. Farr & Rossman (2017) list
34 Colletotrichum species on Orchidaceae. A number of Colle-
totrichum species were described on Orchidaceae, including
C. orchidophilum in the C. acutatum complex (Damm et al.
2012a), C. cymbidiicola and C. oncidii in the C. boninense
complex (Damm et al. 2012b) and C. arxii in the C. gigasporum
complex (Liu et al. 2014). There are also several species on
Orchidaceae treated in this study: C. excelsum-altitudinum (Tao
et al. 2013) and C. coelogynes and a possible further Colleto-
trichum species on Paphiopedilum in Thailand in the
C. dracaenophilum species complex, and C. cattleyicola,
C. orchidearum,C. plurivorum,C. sojae and C. vittalense in the
C. orchidearum species complex.
The Colletotrichum dracaenophilum species complex con-
tains a few apparently host-specific species. Based on the few
strains available, these species seem to be uncommon. Most of
the species are rather distantly related; it is not a species
complex in the strict sense. All species form cylindrical conidia
with round ends, some with sometimes truncate bases or with a
basal scar. In some species, we also observed conidiogenous
cells that are extending to form new conidiogenous loci or mu-
cous layers in the upper parts of the conidiogenous cells, similar
to the C. boninense complex (Damm et al. 2012b). A sexual
morph is only known in C.yunnanense.
The Colletotrichum magnum species complex consists of
nine closely related species. Except for C. brevisporum and an
undescribed species from two independent publications, they are
known only from one host species each. However, for some of
these species only a single strain is available. A sexual morph is
only known from Ga. magna that is heterothallic; the sexual
morph was the result of a laboratory crossing (Jenkins &
Winstead 1964). The ascospores are curved and very large,
larger than those of any other Colletotrichum species treated in
this study and probably only exceeded in length by those of
C. gigasporum (Rakotoniriana et al. 2013) and C. taiwanense
(Sivanesan & Hsieh 1993), the former belonging to the
C. gigasporum complex, while the systematic position of the
latter is dubious (Liu et al. 2014).
The Colletotrichum orchidearum species complex currently
consists of eight closely related species, including three species
(C. orchidearum,C. plurivorum,C. sojae) that are very common and
occur on manyhost species and a number of less common species
that seem to be either host-specific(C. cliviicola,C. musicola,
C. cattleyicola,C. piperis) or restricted to a specific country and
region (C. vittalense). Sexual morphs were observed in cultures of
many of the species of the C. orchidearum complex; we assume
them to be homothallic. However, this needs to be confirmed.
In some species in the C. orchidearum complex we observed
different strains that were only/predominantly producing the
sexual or the asexual morph, respectively. A dominating sexual
morph of one Colletotrichum species and another Colletotrichum
species forming only the asexual morph co-occurring on the
same host, is one of the reasons for wrong connections of sexual
and asexual morphs of Colletotrichum species. This was clearly
the case with Ga. glycines by Lehman & Wolf (1926). A similar
scenario is possible with the sexual morphs ascribed to
C. lindemuthianum that were called Ga. lindemuthiana or Ga.
cingulata f. sp. phaseoli (Shear & Wood 1913, Kimati & Galli
1970). It is also possible that the asexual morphs were just
assumed to be C. lindemuthianum based on the host plants
(common beans) or C. gloeosporioides based on conidial
morphology, respectively. Epitypifications of the respective spe-
cies would clarify this situation.
Nothing is known about the infection strategies of the indi-
vidual species in the three species complexes treated here, and
little is known about their distribution, host specificity and their
impact on the host plants. The high number of sequences from
unidentified or basically only tentatively identified strains in
GenBank suggests a larger distribution and probably more
species in these complexes than included in the present study.
ACKNOWLEDGEMENTS
We thank the curatorsof the CBS, MAFF,UTFC and CABI culture collectionsas well
as Dr Richard O'Connell, UMR1290 BIOGER-CPP, INRA-AgroParisTech, 78850
Thiverval-Grignon, France, Dr Isabelle Benoit-Gelber, Department of Biology,
Concordia University, Montreal, Canada, and Omid Atghita, Department of Plant
Protection, College of Agriculture and Natural Resources, University of Tehran,
Karaj, Iran, for kindly supplying isolates or collectingplant material for this study. We
kindly thank the curators of the fungaria at the Westerdijk Fungal Biodiversity
Institute, Utrecht, The Netherlands, the Herbarium Hamburgense, Hamburg, Ger-
many, the Botanische Staatssammlung München, Germany, the Royal Botanic
Gardens in Kew, UK, and the US NationalFungus Collections, Beltsville, Maryland,
USA, for providing access to important herbarium specimens. Prof. Dr Eva H.
Stukenbrock, Department of Environmental Genomics, Botanical Institute,
Christian-Albrechts University of Kiel, Germany, is gratefully acknowledged for
financial support in sequencing the UTFC strains. Prof. Dr Uwe Braun, Institut für
Geobotanik und Botanischer Garten, Martin-Luther-Universit€
at Halle-Wittenberg,
Germany, is thanked for verifying the Latin names and Dr Jens Wesenberg,
Department of Botany, Senckenberg Museum of Natural History Görlitz, Germany,
for the identificationof an orchid host plant. This research was also supportedby the
Dutch Ministry of Agriculture, Nature and Food Qualitythrough an endowment of the
FES programme “Versterking infrastructuur Plantgezondheid”.
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