Introducing Melanoctona Tectonae Gen. Et Sp. Nov. and Minimelanolocus Yunnanensis Sp. Nov. (Herpotrichiellaceae, Chaetothyriales

Home , Capronia, Chaetothyriales, Chaetothyriomycetidae, Cladophialophora, Fonsecaea, Herpotrichiellaceae

Introducing Melanoctona tectonae gen. et sp. nov. and Minimelanolocus yunnanensis sp. nov. (Herpotrichiellaceae,chaetothyriales)

Qing TIAN a,b,c,d,Mingkhuan DOILOM c, d,Zong-Long LUO c,d,e, Putarak CHOMNUNTI c,d,Jayarama D. BHAT f, Jian-Chu XU a,b &Kevin D. HYDE a, b, c, d*

aKey Laboratory for Plant Diversity and Biogeography of East Asia, Kunming Institute of Botany,Chinese Academy of Sciences, Kunming 650201, Yunnan, People’sRepublic of China

bWorld Agroforestry Centre, East and Central Asia, Kunming 650201, Yunnan, People’sRepublic of China

cCenter of Excellence in Fungal Research, Mae Fah Luang University, Chiang Rai, 57100, Thailand

dSchool of Science, Mae Fah Luang University,Chiang Rai, 57100, Thailand

eCollege of Basic Medicine, Dali University,Dali, Yunnan 671000, China

fFormerly at Department of Botany,Goa University,Goa 403 206, India

Abstract – Herpotrichiellaceae is an interesting, but confused family in Chaetothyriales; the latter has been considered to represent anatural and well-defined group. Anew genus Melanoctona was collected on decaying wood of Tectona grandis in Chiang Rai Province, Thailand and is introducedinHerpotrichiellaceae.Ithas aunique combination of morphological and phylogenetic characters. Phylogenetic analyses of combined ITS, LSU and SSU sequence data place Melanoctona in adistinct lineage in Herpotrichiellaceae. Melanoctona is distinguished from other genera in Herpotrichiellaceae by its hyphomycetous formation and muriform, ellipsoidal to ovoid, brown to dark brown conidia. Minimelanolocus yunnanensis sp. nov.isalso introduced. This species inhabits decaying wood in freshwater streams and rivers in Yunnan Province, China. Maximum likelihood, maximum parsimony and Bayesian analyses of combined ITS, LSU and SSU sequence data, as well as adistinct morphology provide evidence for this new species. Comprehensive descriptions and illustrations of Melanoctona tectonae gen. et sp. nov.and Minimelanolocus yunnanensis sp. nov.are provided with notes on their taxonomy and phylogeny. Minimelanolocus yunnanensis is compared with morphologically similar taxa.

Black moulds /Eurotiomycetes /Fresh water /Phylogeny /Taxonomy

*Corresponding author: Kevin D. Hyde, email address: [email protected]

doi/10.7872/crym/v37.iss4.2016.477 478 Q. Tian et al.

IntroductIon

The class Eurotiomycetes is amonophyletic group comprising three major subclasses, Eurotiomycetidae Geiser &Lutzoni, Chaetothyriomycetidae Doweld, and Mycocaliciomycetidae Tibell. Chaetothyriomycetidae accommodates the order Chaetothyriales, which produce ostiolateperithecial ascomata with bitunicate asci, similar to those produced in Dothideomycetes (Geiser et al.,2006). The order Chaetothyriales, presently comprises six families, viz. Chaeto- thyriaceae, Cyphellophoraceae, Epibryaceae, Herpotrichiellaceae, Strelitzianaceae and Trichomeriaceae (Geiser et al.,2006, Kirk et al.,2008, Chomnunti et al.,2012, 2014; Réblová et al.,2013; Feng et al.,2014b; Gueidan et al.,2014; Crous et al., 2015). Most taxa in these families are saprobes, such as species in Herportrichiellaceae which grow on decaying wood and mushrooms(Barr,1987; Untereiner &Naveau, 1999; Untereiner,2000). The asexual morphs of several Herpotrichiellaceae species are black yeasts and are animal or human pathogens or occur on rocks (de Hoog et al., 2000; Prenafeta-Boldú et al.,2006; Réblová et al.,2013; Feng et al.,2014a; Vicente et al.,2014; Zeng et al.,2014; Isola et al.,2016). Genera of Herpotrichiellaceae have been related to species of Coenosphaeria and Trichometasphaeria (Munk, 1953) and latertreated as members of Pleosporales and Dothideales(Müller &von Arx, 1962; Bigelow &Barr,1963; von Arx &Müller, 1975). Although the family Herpotrichiellaceae has been considered to represent a natural and well-definedfamily,the morphological characters used in the delimitation of its genera have been confused (Munk, 1957; Müller &von Arx, 1962; Samuels &Müller,1978). The genus Minimelanolocus was revisited by Liu et al. (2015) who introduced four new species and provided new sequence data and abackbone tree for the order Chaetothyriales and showed the genus is adistinct lineage in Chaetothyriales. In this study,weintroduce Melanoctona tectonae gen. et sp. nov.and Minimelanolocus yunnanensis sp. nov.based on the morphological characteristics and phylogenetic analyses. Descriptions and illustrations are provided for both taxa.

MAterIAlS And MethodS

Fungal isolation and specimen examination. – Decaying wood was collected from aquatic habitats in Dali, Yunnan Province, China and decaying teak was collected from terrestrial habitats in Chiang Rai Province, Thailand and returned to the laboratory in Zip lock plastic bags. The specimens were incubatedinamoist chamber for 1-2 days at 25°Cand examined at regular intervals for sporulating resident fungi. The fungi were examined using an Olympus SZH10 stereo microscope, and mounted in water.Micromorphological characters were determined with a Nikon ECLIPSE 80i compound microscope and images were captured with aCanon EOS 600D digital camera. Differential interference contrast microscopy was used to visualize hyaline structures. Measurementswere made with Tarosoft (R) Image FrameWork version 0.9.7. Photographic plates were prepared in Adobe Photoshop version CS6 (Adobe Systems, The United States). After morphological examination, single spore isolations were made following the method described by Chomnunti Melanoctona tectonae gen. et sp. nov.and Minimelanolocus yunnanensis sp. nov.479 et al.(2014). Germinating spores were transferred aseptically to malt extractagar (MEA) or potato dextrose agar (PDA) platesand incubated at 25°Cinnormal daylight. Colony colour and culture characters were observed and recorded after one week and again after three weeks. Specimens are deposited in the Herbarium of Cryptogams Kunming Institute of Botany Academia Sinica (KUN-HKAS), Kunming, China and Mae Fah Luang University (MFLU), Chiang Rai, Thailand. Living cultures are deposited in Mae Fah Luang University Culture Collection (MFLUCC), and duplicated in Kunming Institute of Botany,Chinese Academy of Sciences (KUMCC), Kunming, China. Taxa were registered in Index Fungorum (2016) and Facesoffungi (Jayasiri et al.,2015). Fungal DNA extraction, PCR amplification, sequencing and sequence alignment. – Biospin Fungus Genomic DNA Extraction Kit-BSC14S1 (BioFlux®, P.R. China) was used to extract DNA from fresh mycelium grown on MEA at 25°C for four weeks using the instructions of the manufacturer (Hangzhou, P.R. China). The primers ITS5 and ITS4, NS1 and NS4 (White et al.,1990) and LROR (Rehner &Samuels, 1994) and LR5 (Vilgalys &Hester,1990) were used for the amplification of ITS, SSU and LSU respectively.Polymerase chain reaction (PCR) amplification was carried out following the method of Tian et al. (2015). PCR amplification was confirmed on 1% agarose electrophoresis gels stained with ethidium bromide. The amplified PCR fragments were sequenced by BGI, Ltd Shenzhen, P.R. China). Sequence data are deposited in GenBank (Table 1). The alignment and tree were deposited in TreeBASE under accession number 19479. Phylogenetic analyses. – The closest taxa to our strains were determined with standard nucleotide blast searches in NCBI database (http://www.ncbi.nlm.nih. gov/), and sequences of representative species were selected as in Liu et al. (2015). Sequence data from 35 taxa of Herpotrichiellaceae were selected from Crous et al. (2007), Badali et al. (2008) and Liu et al. (2015) (Table 1). Multiple alignments were made by MAFFTv.7.036 (Katoh &Standley, 2013), and adjusted manually using BioEdit v. 7.2 (Hall, 1999) and ClustalX v. 1.83 (Thompson et al.,1997). The tree was rooted using sequence data from Cyphellophora laciniata (CBS 190.61). MODELTEST v. 2.0 (Nylander,2004), following Akaike Information Criterion, was used to determine the best-fitmodel of evolution for each data set for Maximum Likelihood analyses and MrModel test for Bayesian analyses. Maximum-likelihood (ML) analysis was performed in RAxML (Stamatakis, 2008) implemented in raxmlGUI v.0.9b2 (Silvestro &Michalak, 2012). One thousand non-parametric bootstrap iterations were employed with the available models of generalized time reversible (GTR +Gsubstitution model.) and adiscrete gamma distribution (Stamatakis et al.,2008; Liu et al.,2011). The number of replicates was automatically inferred using the stopping criterion (Pattengale et al.,2009). Maximum likelihood bootstrap values equal to or greater than 50% are given as the first set of numbers above the nodes (Fig. 1). Maximum parsimony (MP) analysis was conducted with PAUP v. 4.0b10 (Swofford, 2002) using the heuristic search option with 1,000 random taxa addition and tree bisection reconnection (TBR) as branch-swapping algorithm. Maxtrees were setup unlimited and azero of maximum branch length collapsed, all multiple, equally parsimonious trees were saved. Gaps were treated as missing data. Tree length (TL), consistency index (CI), retention index (RI), rescaled consistency index (RC) and homoplasy index (HI) were calculated. The robustness of the best parsimonious tree was evaluated by abootstrap (BT) value using 1,000 replications 480 Q. Tian et al. bold / / / / / / / / 3 in 554285 SSU FJ358302 FJ358301 FJ358303 KF155195 KF155192 KF155194 AY numbers shown are // // // // // // LSU FJ358234 FJ358233 FJ358235 JQ342182 KF928518 KF928514 EU035403 EU035402 EU035405 EU035409 KC809994 KC809995 KC809992 KC809989 KC809990 accession sequences GenBank ITS 251087 U103995 FJ385273 JQ342180 EU035402 EU035403 EU137322 EU137266 EU137292 EU103997 EU103989 EU137291 EU103988 EU103984 EU103985 EU103986 EU137318 EU035403 EU035409 EU035405 AY DQ008141 generated ewly /E / nezuela nezuela nezuela Japan Japan China China U.S.A. Brazil Brazil .N U.S.A. U.S.A. U.S.A. Locality Portugal Australia Australia Germany Denmark Ve Ve Ve Netherlands study cat male male plant male child male human lesion, forearm, male phylogenetic abies male human skin limb, wood the right imbricatula lesion, wood in ce juice drink drink bambusoides bambusoides asymptomatic tea infection, in on Picea abscess, toe ce Sour abscess, of pple ycosis ports ports lesion otting lesion phaeohyphomycosis, ecaying griseus lesion, rain treated kin racheal Roots eus kin isseminated nterdigital Hypotrachyna Phyllostachys Phyllostachys Chromoblastomycosis, species ubcutaneous hromoblastom ubcutaneous Stenocer of 2 / /S / /S /S /R / / /S / pe TS TB TI TT TC TI TD TD TS TA LT Ty numbers 1 4747 2793 4405 8724 4747 2222 5144 ollection/ 640.96 160.54 260.83 126.86 173.52 306.94 259.83 147.84 987.96 491.70 556.83 834.96 11 11 11 accession 11 11 11 11 122642 129337 ec Isolate CBS CBS CBS CBS CBS CBS CBS CBS CBS CBS CBS CBS CBS CBS CBS CBS CBS CBS CBS CBS CBS Cultur collection culture esii and chaetospira australiensis emmonsii carrion ii carrionii boppii yegr bantiana saturnica devriesii minourae arxii samo ë ns is subtilis chaetospira minourae immunda chaetosp ira parmeliae potulenturum potulenturum xon Ta GenBank 1. ble Cladophialophora Cladophialophora Cladophialophora Cladophialophora Ta Cladophialophora Cladophialophora Cladophialophora Cladophialophora Cladophialophora Cladophialophora Cladophialophora Cladophialophora Cladophialophora Cladophialophora Cladophialophora Cladophialophora Cladophialophora Cladophialophora Cladophialophora Cladophialophora Cladophialophora Melanoctona tectonae gen. et sp. nov.and Minimelanolocus yunnanensis sp. nov.481 ae in / / 3 SSU R215616 Y554289 shown FJ358310 FJ358314 FJ358307 KR215614 KR215618 KR215615 KR215617 DQ823108 DQ823106 KX258780 KX258783 MFlucc :M are numbers 1K CBS, // /A at LSU FJ358242 FJ358246 FJ358239 AF050281 EU041862 KR21561 KR215613 KR215609 KR215610 KR215612 DQ823101 DQ823099 KX258779 KX258782 accession gene. sequences housed nrRNA Crous GenBank ITS the 366914 156963 366925 857539 of generated AF050281 EU035416 EU041805 Pedro AY AY AY AY KR215606 KR215608 KR215605 KR215607 KR215604 DQ826739 DQ826737 KX258778 KX258781 of ewly subunit Belgium. .N – collection America small USA China China China China China Brazil china Canada 8S Locality Uruguay Germany thailand study Switzerland Netherlands orking South SSu :1 Louvain-la-Neuve, cPc :W gene, grandis phylogenetic Louvain, male nrRNA de the Netherlands, the ctona in wood wood wood wood wood wood of Te ce The male punctatus ale ale oil of aspiration-biopsy catholique Sour rowncoal kin, subunit ecaying ecaying ecaying ecaying ecaying treated ecaying Utrecht, wood ge Ictalurus lar hromoblastomycosis, 8S ymphnode, l’Universite species ecaying de of lSu :2 Schimmelcultures, 2 gene, ycoth è que /M /D /M /B // pe T TL TC TD TD TS TS TD TD td td voor Ty numbers nrDNA Mucl :M 5.8S 669 657 1 12-0389 16-0764 entraalbureau 5831 15-0416 15-0415 15-0259 15-0414 15-0237 accession ollection/ 507.90 289.93 271.37 181.65 190.61 11 ec ncluding OL-ID OL-ID Thailand, Isolate cBS :C 2i MUCL9760 CBS CBS CBS CBS CBS CBS AFT AFT Cultur and MFLUCC MFLUCC MFLUCC MFLUCC MFLUCC Life, MFlucc MFlucc collection of culture. ChiangRai, ee Tr region1 culture spacer Fungal x-lectotype Collection, and :e the lt yunnanensi s obscurus melanicus asiaticus curvatus aquaticus xon osoi laciniata Culture anceps Ta tectonae transcribed verrucosa monophora pedr culture, ssembling jeanselmei pisciphil a xenobiotica GenBank pilosella 1. nternal (continued) ol :A x-type onia ble ItS :i t :e AFt Exophiala Fonsecaea Fonsecaea Phialophora Minimelanolocus Exophiala 3 Fah. LuangUniversity 2 Ta 1 bold Rhinocladiella Minimelanolocus Exophiala Capr Melanoctona Cyphellophora Minimelanolocus Minimelanolocus Minimelanolocus Minimelanolocus 482 Q. Tian et al.

(Hillis &Bull, 1993). The best scoring phylogram was drawn in Treeview v. 1.6.6 (Page, 1996). Maximum parsimony bootstrap values equal to or greater than 50% are given as the second set of numbers above the nodes (Fig. 1). ABayesian analysis was conducted with MrBayes v. 3.1.2 (Ronqvist & Huelsenbeck, 2003) to evaluate posterior probabilities (PP) (Rannala &Yang, 1996; Zhaxybayeva &Gogarten, 2002) by Markov Chain Monte Carlo sampling (BMCMC). Four simultaneous Markov chains were run for 5,000,000 generations and trees were sampled every 500th generation and 50,000 trees were obtained. The first 5,000 trees, representing the burn-in phase of the analyses, were discarded, while the remaining

Fig. 1. Phylogenetic tree generated by Maximum likelihood (RAxML) analysis of acombined ITS, LSU and SSU sequence dataset. Bootstrap support values for maximum likelihood (ML, red) and maximum parsimony (MP,black) equal to or greater than 50% are given above the nodes. Bayesian posterior probabilities (BPP,blue) equal to or higher than 0.90 are given below the nodes. Hyphen (“–”)indicates avalue lower than 50% for either MP or BPP.Ex-type strains and reference strains are in bold. Newly generated sequences are in blue. The tree is rooted with Cyphellophora laciniata CBS 190.61. Melanoctona tectonae gen. et sp. nov.and Minimelanolocus yunnanensis sp. nov.483

45,000 trees were used for calculating posterior probabilities in the majority rule consensus tree (critical value for the topological convergence diagnostic is 0.01). Bayesian Posterior Probabilities (BYPP) equal to or greater than 0.90 are given below each node (Fig. 1).

reSultS

Phylogenetic analyses Most of the core genera of Herpotrichiellaceae (Badali et al.,2008; Liu et al., 2015) are included in our phylogenetic analysis (Fig. 1). Forty-five strains are included in the combined ITS, LSU and SSU sequence analyses which comprise 2483 characters with gaps (627 characters for ITS, 891 for LSU and 965 for SSU). Cyphellophora laciniata (Cyphellophora, Cyphellophoraceae,Chaetothyriales) is used as the outgroup taxon. Tree topology of the maximum-likelihood analysis is similartothe maximum parsimony analysis and the Bayesian analysis. The best scoring RAxML tree with a final likelihood value of -11477.832085 is presented (Fig. 1). Most strains are from previous studies for which there is no morphological data and they must be regarded as putatively named strains (Badali et al.,2008; Liu et al.,2015). Single gene analyses are carried out and compared with each species, topology of the tree and clade stability.The two strains generated in the present study cluster within the family Herpotrichiellaceae. The genus Cladophialophora in this study comprises 20 strains which cluster in three strongly supported clades and include the major human-opportunistic species (Fig. 1). Clade Iincludes the pathogenic species C. carrionii, C. boppii and Phialophora verrucosa;Clade II includesthe pathogenic species C. bantiana, C. arxii, C. immunda, C. devriesii, C. saturnica, C. emmonsii,aswell as Fonsecaea monophora;Clade III includes C. potulentorum and C. australiensis which was isolated from sugared drinks. Due to the similarities of morphological characters of these black yeast-like taxa, further research is needed on this genus. In the present study,the phylogenetic analyses show that Melanoctona tectonae sp. nov.(MFLUCC 12-0389) represents as lineage distinct from any other genera in Herpotrichiellaceae (Fig. 1), and forms asingle clade basal to Capronia pilosella (0.94 BPP). Thus, we introduce Melanoctona gen. nov.toaccommodate the species Melanoctona tectonae. Minimelanolocus is awell-defined genus in Herpotrichiellaceae.Liu et al. (2015) indicated that multi-gene phylogenetic analyses for this genus are required to determine the species limits as there is few molecular data available. Minimelanolocus yunnanensis sp. nov.(MFLUCC 16-0764) was collected on submerged wood in streams in Yunnan Province, China. The combined analysis of the LSU and SSU dataset confirms its placement in Minimelanolocus. In the present study,the phylogenetic analyses show that Melanoctona tectonae sp. nov.(MFLUCC 12-0389) represents alineage distinct from other genera in Herpotrichiellaceae (Fig. 1), thus, we introduce Melanoctona gen. nov.to accommodate the species Melanoctona tectonae. 484 Q. Tian et al.

tAXonoMY

Minimelanolocus R.F.Castañeda &Heredia, Cryptog. Mycol. 22(1): 7(2001) Saprobic on decaying wood in aquatic habitat. Colonies superficial on the host, effuse, scattered, hairy,brightly coloured, glistening. Mycelium superficial or partly immersed, comprising septate, pale brown to brown, smooth-walledhyphae. Sexual morph:Undetermined. Asexual morph:Hyphomycetous. Conidiophores mononematous, macronematous, unbranched, erect, dark brown and gradually paler towards the apex, straight or flexuous, cylindrical, smooth, septate. Conidiogenous cells holoblastic, integrated, sympodially proliferating, terminal, pale brown to brown or subhyaline to hyaline. Conidia acrogenous, oblong or clavate to fusiform, with or without obtuse ends, hyaline to pale brown or brown, solitary,sometimes with secondary conidia, immature conidia aseptate, multi-septate at maturity,dry, smooth-walled. Conidial secession schizolytic. Notes: Minimelanolocus was introduced by Castañeda-Ruiz et al. (2001) and typified by M. navicularis (R.F.Castañeda) R.F.Castañeda based on segregation of some atypical species from the genus Pseudospiropes.The genus is characterized by conspicuous, mononematous, solitary or fasciculate, septate, erect, straight or flexuous, smooth or verrucose, cylindrical, sinuate or geniculate, brown to dark brown conidiophores, with amelanized base and hyaline to brown, oblong, cylindrical, clavatetofusiform, euseptate, acropleurogenous conidia (Castañeda- Ruiz et al.,2001; Hernández-Restrepo et al.,2013; Xia et al.,2014), while Pseudospiropes species have comparatively smaller,ellipsoidal and distoseptate conidia (Castañeda-Ruiz et al.,2001; Ma et al.,2011). Minimelanolocus is a morphologically well-studied genus, which has been described from awide range of hosts (Zhang et al.,2010; Ma et al.,2011a, b; Hernández-Restrepo et al.,2013; Xia et al.,2014). Presently,there are 24 species listed in Minimelanolocus (Index Fungorum, 2016) worldwide, and most are saprobes on rotten leaves or dead twigs, wood and bark. Liu et al. (2015) provided abackbone tree for Chaetothyriales and described four new species of Minimelanolocus with support from molecular data. Analysis of combined LSU and SSU was not adequate to distinguish species within Minimelanolocus. Minimelanolocus yunnanensis Qing Tian &K.D. Hyde, sp. nov. Index Fungorum number:IF552156; Facesoffungi number:FoF 02224, Figs 2, 3. Etymology:Inreference to the occurrence in Yunnan Province, China. Holotype:MFLU 16-1298. Saprobic on decaying wood in aquatic habitats. Sexual morph: Undetermined. Asexual morph: Colonies superficial, effuse, scattered, hairy,dark brown to black. Mycelium immersed, comprising septate, pale brown hyphae. Conidiophores 237–275 × 8-11.5 μm (x̅ =260 × 9.8 μm,n=10), mononematous, macronematous, unbranched, erect,straight or flexuous, cylindrical, smooth, dark brown, melanized at base, gradually paler and obtuse and rounded towards the apex, 7-9-septate. Conidiogenous cells holoblastic, integrated, sympodially proliferating, terminal and intercalary,pale brown or subhyaline. Conidia 20-26.5 × 5-7 μm (x̅ =23.2 × 6.2 μm,n=20), acrogenous, hyaline, cylindro-oblong, obtuse at the apex and base, solitary,1-septate when young, 3-septate at maturity,surrounded by anarrow gelatinous sheath, thick-walled, smooth-walled. Conidial secession schizolytic. Melanoctona tectonae gen. et sp. nov.and Minimelanolocus yunnanensis sp. nov.485

Fig. 2. Minimelanolocus yunnanensis (MFLU 16-1298, holotype) a. Conidiophore with conidia. b-d. Apex of conidiophores with conidia. e-h. Conidia. i. Germinating conidium. j. Surface view of culture on MEA. k. Reverse view of culture on MEA. Scale bars: a=20 μm, b–d, i=10 μm, e–h= 5μm. 486 Q. Tian et al.

Fig. 3. Conidiophore and conidia of Minimelanolocus yunnanensis (Drawing form holotype). Scale bars =10μm.

Culturecharacteristics:Conidia germinating on MEA within 12 h. Colonies on MEA, fast growing, reaching up to 8mmdiam. in 7days at 25°C. Mycelium superficial, initially white, later becoming brown, hairy,with entire edge. Material examined:China, Yunnan Province, Dali, Wanhua stream, on decaying wood, 16 March 2014, Zonglong Luo, S-041 WHXM 24-1 (MFLU 16- 1298, holotype); ibid., (HKAS 84023, isotype); ex-type living culture MFLUCC 16-0764, KUMCC 16-0008. Notes:Among the accepted species of Minimelanolocus, Minimelanolocus yunnanensis is most similar to M. bambusae, M. leptotrichus, M. obscurus and M. olivaceus,which produce 3-septate, ellipsoidal or cylindrical, hyalineconidia (Cooke &Ellis1879; Sharma, 1980; Matsushima, 1983; Castañeda-Ruiz et al., 2003). However,the conidia of M. yunnanensis and M. leptotrichus are hyaline, while those of M. bambusae and M. obscurus are hyaline, then becoming pale brown when senescent and conidia of M. olivaceus are pale olivaceous. Minimelanolocus yunnanensis strongly resembles M. leptotrichus in conidial color, but differ in conidial shape. Conidia of M. yunnanensis are cylindro-oblong, with a strongly obtuse apex and base, while in M. leptotrichus conidiaare cylindrical, tapering at both ends. Moreover,the conidiogenous cells of M. yunnanensis are cylindrical with sympodially proliferating at the apex. In addition, the conidiaof M.yunnanensis have anarrow gelatinous sheathwhich is aunique character in Minimelanolocus. Melanoctona tectonae gen. et sp. nov.and Minimelanolocus yunnanensis sp. nov.487

Melanoctona Qing Tian, Doilom &K.D. Hyde, gen. nov. Index Fungorum number:IF552157; Facesoffungi number:FoF 02225. Etymology:Inreference to the characteristic of pigmentation and muriform conidia. Saprobic on decaying wood in terrestrial habitat. Colonies superficial, effuse, scattered, dark brown to black. Asexual morph:Hyphomycetous. Conidiophores mononematous, macronematous, unbranched, erect, straight or flexuous, smooth, dark brown. Conidiogenous cells holoblastic, integrated, sympodially proliferating, terminal and intercalary,pale brown or subhyaline. Conidia acrogenous, brown to black, ovoid, muriform, rounded at base and apex, smooth-walled. Sexual morph:Undetermined. Type species: Melanoctona tectonae Qing Tian, Doilom &K.D. Hyde, sp. nov. Notes:Anew genus Melanoctona is introduced here to accommodate an asexual species in Herpotrichiellaceae and is typified by M. tectonae which forms adistinct clade (Fig. 1). To date, the asexual morphs of Herpotrichiellaceae are predominantly black yeasts which are regarded as dematiaceous hyphomycetes in having holoblastic, conidiogenous cellswhich proliferate percurrently and aseptate or septate conidia, including Exophiala, Phialophora, Cladophialophora, Fonsecaea and Exophiala-Ramichloridium-Rhinocladiella species (Müller et al.,1987; Untereiner et al.,1995; Crous et al.,2007; Gueidan et al.,2014; Liu et al.,2015), and undetermined Pyricularia-like taxa (Klaubauf et al.,2014). Species of Exophiala are characterized by having brown aseptate to 1-septate conidia, forming successively from the apex of the sporogenous cell and aggregating in slime balls (Carmichael, 1966). Phialophora produces hyaline to pigmented, aseptate conidia,forming from flask-shaped to straight phialides (Harrington &Mcnew,2003). Cladophialophora has aseptate, hydrophobic conidia and is mostly isolated from human disease (Badali et al.,2011). Fonsecaea has melanized conidiophores with cylindrical denticles and aseptate, acrogenous conidia (de Hoog et al.,2000). Melanoctona however,has dark brown to black, muriform or multi-septate, acrogenous, conidia.In particular, Melanoctona tectonae was isolated from adead branch of Tectona grandis as a saprobe, while other asexual morphs in Herpotrichiellaceae have been isolated as human pathogens, such as human chromoblastomycosis (de Hoog et al.,2007). Therefore, anew genus is introduced in Herpotrichiellaceae to accommodate our fresh collection. Melanoctona tectonae Qing Tian, Doilom &K.D. Hyde, sp. nov. Index Fungorum number:IF552158, Facesoffungi number:FoF 02226, Fig. 4. Etymology:Inreference to the host genus. Holotype:MFLU 15-3528. Saprobic on decaying wood of Tectona grandis L.f. in terrestrial habitat. Colonies superficial, effuse, scattered, dark brown to black. Asexual morph: Hyphomycetous. Conidiophores 3-6 μm(x̅ =5.2 μm, n=7) width, mononematous, macronematous, unbranched, erect, straight or flexuous, smooth, dark brown. Conidiogenous cells holoblastic, integrated, sympodially proliferating, terminal and intercalary,pale brown or subhyaline. Conidia 28-39 × 43-49 μm(x̅ =34.5 × 47.3 μm, n=10), acrogenous, light brown to black, ovoid, muriform, rounded at base and apex, smooth-walled. Sexual morph:Undetermined. 488 Q. Tian et al.

Fig. 4. Melanoctonatectonae (MFLUCC 12-0389, holotype) a. Appearance of fungus on dead branch of Tectona grandis b, c. Conidia scattered on the surface of wood. d, e. Conidia. f-j. Conidiophores with conidia k. Germinating conidium. l. Surface view of culture on PDA. m. Reverse view of culture on PDA. Scale bars: b=1000 μm, c=200 μm, d-k =10μm.

Culturecharacteristics:Conidia germinating on PDA within 12 hand germ tubes produced from both ends. Colonies on PDA, fast growing, reaching up to 15 mm diam. in 14 days at 25°C. Mycelium superficial, circular,hairy,with entire edge, brown, dark brown at the periphery,completely dark brown from below. Material examined:THAILAND, Chiang Rai Province, Mae Chan District, on dead branch of Tectona grandis L.f. (Lamiaceae), 1July 2012, M. Doilom MKT062 (MFLU 15-3528, holotype); ibid., (HKAS 94893, isotype); ex-type living culture, MFLUCC 12-0389, KUMCC 16-0009.

dIScuSSIon

Members of Chaetothyriales have been found on various hosts and are an ecologically diverse group (Badali et al.,2008; Najafzadeh et al.,2011; Réblová et al., 2013; Feng et al.,2014b). Species in the family Herpotrichiellaceae mostly occur on decaying wood or are fungicolous on mushrooms (Barr,1987; Untereiner &Naveau, 1999; Untereiner,2000). Both sexual and asexual morphs are known in this family.The asexual morphs are often found as opportunistic pathogens of animals, including humans (Prenafeta-Boldú et al.,2006; Crous et al.,2007; Réblová et al.,2013; Vicente et al.,2014; Zeng et al.,2014). Minimelanolocus yunnanensis Melanoctona tectonae gen. et sp. nov.and Minimelanolocus yunnanensis sp. nov.489 was found on submerged wood in freshwater habitats. Species of Herpotrichiellaceae have small, superficial, setose ascomata, bitunicate asci and greenish-grey to brown, septate ascospores. The asexual morphs are predominantly black yeast or dematiaceous hyphomycetes, with holoblastic conidia, proliferating percurrently conidiogenous loci and unicellular or septate conidia (Müller et al.,1987). Capronia, the largest genus, has black yeast-like asexual morphs belonging to Exophiala, Ramichloridium and Rhinocladiella (Untereiner et al.,1995). Based on recent studies (Crous et al.,2007; Badali et al.,2008; Feng et al., 2014b; Vicente et al.,2014; Zeng et al.,2014; Liu et al.,2015), we include the majority of species of Herpotrichiellaceae with available sequence data in the phylogenetic analyses. Melanoctona tectonae is placed in Herpotrichiellaceae and distinguished from other genera in Herpotrichiellaceae based on the phylogenetic analyses (Fig. 1) and morphological characters. Minimelanolocus yunnanensis is clearly distinguished from other species in Minimelanolocus,and phylogenetically clusters in the genus Minimelanolocus (Fig. 1). Acknowledgments. The Mushroom Research Foundation, Chiang Rai, Thailand is thanked for supporting this research. Kevin D. Hyde thanks the Chinese Academy of Sciences, project number 2013T2S0030, for the award of Visiting Professorship for Senior International Scientists at Kunming Institute of Botany.Qing Tian and Putarak Chomnunti thank the grant of the Thailand Research Foundation (TRG5780008) for supporting studies on sooty moulds and the National Research Council of Thailand (grant for Dothideomycetes No: 58201020010) for supporting studies on Dothideomycetes. Dr.Shaun Pennycook is gratefully thanked for checking and revising the Latin name of new taxa.

reFerenceS

BADALI H., GUEIDAN C., NAJAFZADEH M.J., BONIFAZ A., GERRITS VANDEN ENDE A.H.G. &DEHOOG G.S., 2008 — Biodiversity of the genus Cladophialophora. Studies in Mycology 61: 175-191. BADALI H., PRENAFETA-BOLDU F.X., GUARRO J., KLAASSEN C.H., MEIS J.F.&DE HOOG G.S., 2011 — Cladophialophora psammophila,anovel species of Chaetothyriales with apotential use in the bioremediation of volatile aromatic hydrocarbons. Fungal Biology 115(10): 1019-1029. BARR M.E., 1987 — Prodromus to class Loculoascomycetes. University of Massachusetts, Amherst. BIGELOW H.E. &BARR M.E., 1963 — Contribution to the fungus flora of northeastern North America. III. Rhodora 65(764): 289-309. CARMICHAEL J.W., 1966 — Sabouraudia 5: 122. CASTAÑEDA-RUIZ R.F., GUARRO J., VELÁSQUEZ-NOA S. &GENÉ J., 2003 — Anew species of Minimelanolocus and some hyphomycete records from rain forests in Brazil. Mycotaxon 85: 231-239. CASATÑEDA-RUÍZR.F.C., HEREDIA G., REYES M., ARIAS R.M. &DECOCK C., 2001 — A revision of the genus Pseudospiropes and some new taxa. Cryptogamie Mycologie 22: 3-18. CHOMNUNTI P.,KOT.W.K., CHUKEATIROTE E., CAI L., JONES E.B.G., KODSUEB R., CHEN H., HASSAN B.A. &HYDE K.D., 2012 — Phylogeny of Chaetothyriaceae in northern Thailand including three new species. Mycologia 104: 382-395. CHOMNUNTI P.,HONGSANAN S., AGUIRRE-HUDSON B., TIAN Q., PERŠOH D., DHAMI M.K., ALIAS A.S., XU J.C., LIU X.Z., STADLER M. &HYDE K.D., 2014 — The sooty moulds. Fungal Diversity 66: 1-36. COOKE M.C. &ELLIS J.B., 1879 — New Jersey fungi. Grevillea 8(45): 11-16. CROUS P.W.,SCHUBERTK., BRAUN U., DE HOOG G.S., HOCKING A.D., SHIN H.D. & GROENEWALD J.Z., 2007 — Opportunistic, human-pathogenic species in the Herpotrichiellaceae are phenotypically similar to saprobic or phytopathogenic species in the Venturiaceae. Studies in Mycology 58: 185-217. 490 Q. Tian et al.

CROUS P.W.,WINGFIELD M.J., LE ROUX J.J., RICHARDSOND.M., STRASBERG D., SHIVAS R.G., ALVARADO P.,EDWARDS J., MORENO G., SHARMA R., SONAWANE M.S., TANY.P., ALTéSA., BARASUBIYE T.,BARNES C.W., BLANCHETTE R.A., BOERTMANN D., BOGO A., CARLAVILLA J.R., CHEEWANGKOON R., DANIEL R., DE BEER Z.W., DE JESÚSYáÑEZ-MORALES M., DUONG T.A., FERNáNDEZ-VICENTE J., GEERING A.D.W., GUEST D.I., HELD B.W., HEYKOOP M., HUBKA V. ,ISMAIL A.M., KAJALE S.C., KHEMMUK W.,KOLAŘÍKM., KURLI R., LEBEUF R., LÉVESQUE C.A., LOMBARD L., MAGISTAD., MANJÓNJ.L., MARINCOWITZ S., MOHEDANO J.M., NOVáKOVá A., OBERLIES N.H., OTTOE.C., PAGUIGAN N.D., PASCOE I.G., PÉREZ-BUTRÓNJ.L., PERRONE G., RAHI P., RAJA H.A., RINTOUL T.,SANHUEZA R.M.V., SCARLETT K., SHOUCHEY.S., SHUTTLEWORTH L.A., TAYLOR P.W.J., THORN R.G., VAWDREY L.L., SOLANO- VIDAL R., VOITK A., WONG P.T.W.,WOOD A.R., ZAMORA J.C. &GROENEWALD J.Z., 2015 — Fungal Planet description sheets: 371-399. Persoonia-Molecular Phylogeny and Evolution of Fungi 35: 264-327. DE HOOG G.S., GUARRO J., GENÉ J. &FIGUERAS M.J., 2000 — Atlas of Clinical Fungi, 2nd ed. Centraalbureau voor Schimmelcultures, Utrecht, Universitat Rovira iVirgili, Reus. DE HOOG G.S., VICENTE V. A., NAJAFZADEH M.J., HARRAK M.J., BADALI H. & SEYEDMOUSAVI S., 2011 — Waterborne Exophiala species causing disease in cold- blooded animals. Persoonia-Molecular Phylogeny and Evolution of Fungi:27: 46-72. FENG P.Y. ,DEHOOG G.S., NAJAFZADEH M.J., GERRITS VANDEN ENDE A.H.G., STIELOW J.B., BADALI H., BOEGER W.A. &VICENTE V.A., 2014(a) — Cladophialophoraabundans,a novel species of Chaetothyriales isolated from the natural environment. Mycological Progress 13: 381-391. FENG P.Y. ,LUQ.Y., NAJAFZADEH M.J., GERRITS VANDEN ENDE A.H.G., SUN J.F., LI R.Y., XI L.Y., VICENTE V. A., LAI W.,LUC.&DEHOOG G.S., 2014(b) — Cyphellophora and its relatives in Phialophora:Biodiversity and possible role in human infection. Fungal Diversity 65(1): 17-45. GEISER D.M., GUEIDAN C., MIADLIKOWSKAJ., LUTZONI F.,KAUFF F.,HOFSTETTER V. , FRAKER E., SCHOCH C., TIBELL L., UNTEREINER W.A. &APTROOT A., 2006 — Eurotiomycetes: Eurotiomycetidae and Chaetothyriomycetidae. Mycologia 98: 1054-1065. GUEIDAN C., APTROOT A., DA SILVA CÁCERES M.E., BADALI H. &STENROOS S., 2014 — A reappraisal of orders and families within the subclass Chaetothyriomycetidae (Eurotiomycetes, Ascomycota). Mycological Progress 13: 1027-1039. HALL T.A., 1999 — BioEdit: auser-friendly biological sequence alignmenteditor and analysis program for Windows 95/98/NT. Nucleic Acids Symposium Series:95-98. HARRINGTON T.C. &MCNEW D.L., 2003 — Phylogenetic analysis places the Phialophora-like anamorph genus Cladophora in the Helotiales. Mycotaxon 87: 141-151. HERNÁNDEZ-RESTREPO M., CASTAǸEDA-RUIZ R.F., GENÉ J., GUARRO J., MINTER D.W.& STADLER M., 2013 — Microfungi from Portugal: Minimelanolocus manifestus sp. nov.and Vermiculariopsiella pediculata comb. nov. Mycotaxon 122: 135-143. HILLIS D.M. &BULL J.J., 1993 — An empirical test of bootstrapping as amethod for assessing confidence in phylogenetic analysis. Systematic Biology 42: 182-192. Index Fungorum (2016) www.indexfungorum.org/names/names.asp. ISOLA D., ZUCCONI L., ONOFRI S., CANEVAG., DE HOOG G.S. &SELBMANN L., 2016 — Extremotolerant rock inhabiting black fungi from Italian monumental sites. Fungal Diversity 76: 75-96. JAYASIRI S.C., HYDE K.D., ARIYAWANSA H.A., BHATD.J., BUYCK B., CAI L., DAI Y.C., ABD- ELSALAM K.A., ERTZ D., GIBERTONI T.B., HIDAYAT I., JEEWON R., JONES E.B.G., KARUNARATHNA S.C., KIRK P.,LIW.J., LIU J.K., LUANGSA-ARD J.J., MAHARACHCHIKUMBURA S.S.N., MANAMGODA D.S., MCKENZIE E.H.C., MONCALVO J.M., GHOBAD-NEJHAD M., NILSSON H., PANG K.L., PEREIRA O.L., RASPé O., ROLLINS A.W., ROMERO A.I., SALAZAR J.A.E., STEPHENSON S., SUETRONG S., TAYLOR J.E., TSUI C.K.M., VIZZINI A., ABDEL-WAHAB M.A., WEN T.C., BOONMEE S., DAI D.Q., DARANAGAMA D.A., DISSANAYAKE A.J., EKANAYAKA A.H., HONGSANAN S., JAYAWARDENA R.S., PERERA R.H., PHOOKAMSAK R., DE SILVA N.I., THAMBUGALAK.M., TIAN Q., WANASINGHE D.N., WIJAYAWARDENE N.N., ZHAO R.L., KANG J.C. &PROMPUTTHA I., 2015 — The Faces of Fungi database-A unique perspective: Fungal names linked with morphology, phylogeny and human impacts. Fungal Diversity 74: 3-18. Melanoctona tectonae gen. et sp. nov.and Minimelanolocus yunnanensis sp. nov.491

KATOHK.&STANDLEY D.M., 2013 — MAFFT multiple sequence alignment software version 7: improvements in performance and usability. Molecular Biology and Evolution 30:772-780 (outlines version 7). KIRK P.M., CANNON P.F.,MINTER D.W.&STAPLERS J.A., 2008 — Dictionary of the fungi, 10th edn. CABI Bioscience, UK. KLAUBAUF S., THARREAU D., FOURNIER E., GROENEWALD J.Z., CROUS P.W.,DEVRIES R.P. &LEBRUN M.H., 2014 — Resolving the polyphyletic nature of Pyricularia (Pyriculariaceae). Studies in Mycology 79: 85-120. LIU J.K., PHOOKAMSAK R., JONES E.B.G., ZHANG Y.,KO-KO T.W.,HUH.L., BOONMEE S., DOILOM M., CHUKEATIROTE E., BAHKALI A.H., WANG Y. &HYDE K.D., 2011 — Astrosphaeriella is polyphyletic, with species in Fissuroma gen. nov., and Neoastrosphaeriella gen. nov. Fungal Diversity 51: 135-154. LIU X.Y., UDAYANGA D., LUO Z.L., CHEN L.J., ZHOU D.Q., SU H.Y.&HYDE K.D., 2015 — Backbone tree for Chaetothyriales with four new species of Minimelanolocus from aquatic habitats, Fungal Biology 119(11): 1046-1062. MA J., MA L.G., ZHANG Y.D., CASTAǸEDA-RUÍZR.F.&ZHANG X.G., 2011(a) — Pseudospiropes linderae sp. nov.and notes on Minimelanolocus (both anamorphic Strossmayeria)new to China. Nova Hedwigia 93: 465-473. MA J., ZHANG Y. D., MA L.G. &ZHANG X.G., 2011(b) — Twonew Minimelanolocus species from southern China. Mycotaxon 117: 131-135. MATSUSHIMA T.,1983 —“Matsushima mycological memoirs. No. 3”, Matsushima Fungus Collection, Kobe:90. MÜLLER E. &VON ARX J. A., 1962 — Die Galtungen der didymosporen Pyrenomyceten. Beitrage zur Kryptogamenflora der Schweiz 11(2): 1-922. MÜLLER E., PETRINI O., FISHER P.J., SAMUELS G.J. &ROSSMAN A.Y., 1987 — Taxonomy and anamorphs of the Herpotrichiellaceae with notes on generic synonymy. Transactions of the British Mycological Society 88: 63-74. MUNK A., 1953 — The system of pyrenomycetes. Dansk Botanisk Arkiv 15: 1-163. MUNK A., 1957 — Danish pyrenomycetes. Dansk Botanisk Arkiv 17: 1-491. NAJAFZADEH M.J., VICENTE V.A., SUN J., MEIS J.F.&DEHOOG G.S., 2011 — Fonsecaea multimorphosa sp. nov,anew species of Chaetothyriales isolated from afeline cerebral abscess. Fungal Biology 115: 1066-1076. NYLANDER J.A.A., 2004 — MrModeltest 2.0. Program distributed by the author.Evolutionary Biology Centre, Uppsala University. PAGE R.D., 1996 — TREEVIEW:anapplication to display phylogenetic trees on personal computers. Computer Applications in the Biosciences12: 357-358. PATTENGALE N.D., ALIPOUR M., BININDA-EMONDS O.R.P., MORET B.M.E. &STAMATAKIS A., 2009 — How many bootstrap replicates are necessary? LNCS 5541: 184-200. PRENAFETA-BOLDÚ F.X., SUMMERBELL R. &DEHOOG G.S., 2006 — Fungi growing on aromatic hydrocarbons: biotechnology’sunexpected encounter with biohazard? FEMS Microbiol Reviews 30: 109-130. RANNALA B. &YANGZ., 1996 — Probability distribution of molecular evolutionary trees: anew method of phylogenetic inference. Journal of Molecular Evolution 43: 304-311. RÉBLOVÁ M., UNTEREINER W.A. &RÉBLOVÁ K., 2013 — Novel evolutionary lineages revealed in the Chaetothyriales (Fungi) based on multigene phylogenetic analyses and comparison of ITS secondary structure. PLoS ONE 8(5): e63547. REHNER S.A. &SAMUELS G.J., 1994 — Taxonomy and phylogeny of Gliocladium analysed from nuclear large subunit ribosomal DNA sequences. Mycological Research 98: 625-634. RONQUIST F. &HUELSENBECK J.P., 2003 — MrBayes 3: Bayesian phylogenetic inference under mixed models. Bioinformatics 19: 1572-1574. SAMUELS G.J. &MÜLLER E., 1978 — Life-history studies of Brazilian ascomycetes. 3. Melanomma radicans sp. nov.and its Apiosphaeria anamorph, Trematosphaeria perrumpens sp. nov.and Berlesiella fungicola sp. nov.and its Ramichloridium anamorph. Sydowia 31: 142-156. SHARMA N.D., 1980 — Some additions to fungi of India – VIII. Journal of the Indian Botanical Society 57 102-105. SILVESTRO D. &MICHALAK I., 2011 — raxmlGUI: agraphical front-end for RAxML. Organisms Diversity &Evolution 12: 335-337. STAMATAKIS A., HOOVER P. &ROUGEMONT J., 2008 — Arapid bootstrap algorithm for the RAxML WebServers. Systematic Biology 57: 758-771. SWOFFORD D.L., 2002 — PAUP 4.0b10: phylogenetic analysis using parsimony.Sinauer Associates, Sunderland. 492 Q. Tian et al.

THOMPSON J.D., GIBSON T.J., PLEWNIAK F.,JEANMOUGIN F. &HIGGINS D.G., 1997 — The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Research 25(24): 4876. TIAN Q., LIU J.K., HYDE K.D., WANASINGHE D.N., BOONMEE S., JAYASIRI S.C., LUO Z.L., TAYLOR J.E., PHILLIPS A.J.L., BHATD.J., LI W.J., ARIYAWANSA H., THAMBUGALA K.M., JONES E.B.G., CHOMNUNTI P.,BAHKALI A.H., XU J.C. &CAMPORESI E., 2015 — Phylogenetic relationships and morphological reappraisal of Melanommataceae (Pleosporales). Fungal Diversity 74: 267-324. UNTEREINER W.A., STRAUS N.A. &MALLOCH D., 1995 — Amolecular-morphotaxonomic approach to the systematics of the Herpotrichiellaceae and allied black yeasts. Mycological Research 99(8): 897-913. UNTEREINER W.A., 2000 — Capronia and its anamorphs: exploring the value of morphological and molecular characters in the systematics of the Herpotrichellaceae. Studies in Mycology 45: 141-149. UNTEREINER W.A. &NAVEAU F.A., 1999 — Molecular systematics of the Herpotrichiellaceae with an assessment of the phylogenetic positions of Exophiala dermatitidis and Phialophora americana. Mycologia 91: 67-83. VICENTE V. A., NAJAFZADEH M.J., SUN J.F., GOMES R.R., ROBL D., MARQUES S.G., AZEVEDO C.M.P.S. &DEHOOG G.S., 2014 — Environmental siblings of black agents of human chromoblastomycosis. Fungal Diversity 65: 47-63. VILGALYS R. &HESTER M., 1990 — Rapid genetic identification and mapping of enzymatically amplified ribosomal DNA from several Cryptococcus species. Journal of Bacteriology 172: 4238-4246. VON ARX J.A. &MÜLLER E., 1975 — Are-evaluation of the bitunicate ascomycetes with keys to families and genera. Studies in Mycology 9: 1-159. WHITE T.J., BRUNS T.,LEE S. &TAYLOR J., 1990 — Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ (eds), PCR Pro- tocols: aguide to method and applications. Academic Press, New York, pp. 315-322. XIA J.W., MA L.G., CASATÑEDA-RUÍZR.F.C. &ZHANG X.G., 2014 — Minimelanolocus bicolorata sp. nov., Paradendryphiopsis elegans sp. nov.and Corynesporella bannaense sp. nov.from southern China. Mycoscience 55: 299-307. ZENG J.S., FENG P.Y. ,GERRITS VANDEN ENDE A.H.G., XI L.Y., HARRAK M.J. &DE HOOG G.S., 2014 — Multilocus analysis of the Exophiala jeanselmei clade containing black yeasts involved in opportunistic disease in humans. Fungal Diversity 65: 3-16. ZHANG Y.D., MA J., MA L.G. &ZHANG X.G., 2010 — Anew species of Minimelanolocus from Fujian, China. Mycotaxon 114: 373-376. ZHAXYBAYEVAO.&GOGARTEN J.P., 2002 — Bootstrap, Bayesian probability and maximumlikelihoodmapping: exploring newtools for comparative genome analyses. BMC Genomics 3(1): 4.