Mycological Progress (2020) 19:1031–1042 https://doi.org/10.1007/s11557-020-01609-0

ORIGINAL ARTICLE

Mycoenterolobium aquadictyosporium sp. nov. (, ) from a freshwater habitat in Thailand

Mark S. Calabon1,2 & Kevin D. Hyde1,3 & E. B. Gareth Jones4 & Mingkwan Doilom5,6 & Chun-Fang Liao5,6 & Saranyaphat Boonmee1,2

Received: 25 May 2020 /Revised: 25 July 2020 /Accepted: 28 July 2020 # German Mycological Society and Springer-Verlag GmbH Germany, part of Springer Nature 2020

Abstract A study of freshwater fungi in Thailand led to the discovery of Mycoenterolobium aquadictyosporium sp. nov. Evidence for the novelty and placement in Mycoenterolobium is based on comparison of morphological data. The new species differs from the type species, M. platysporum, in having shorter and wider conidia, and from M. flabelliforme in having much longer and wider conidia. The hyphomycetous Mycoenterolobium is similar to Cancellidium but differs in the arrangement of conidial rows of cells at the attachment point to the conidiophores. The conidia of the former are made up of rows of cells, radiating in a linear pattern from a single cell attached to the conidiophore, while in Cancellidium, adherent rows of septate branches radiate from the conidiophore. Cancellidium conidia also contain branched chains of blastic monilioid cells arising from the conidia, while these are lacking in Mycoenterolobium.AtmaturityinMycoenterolobium, the two conidial lobes unite and are closely appressed. Phylogenetic analyses based on a combined LSU, SSU, ITS, TEF1-α,andRPB2 loci sequence data support the placement of Mycoenterolobium aquadictyosporium close to the family within Pleosporomycetidae, Dothideomycetes. The novel species Mycoenterolobium aquadictyosporium is described and illustrated and is compared with other morphologically similar taxa.

Keywords 1 new taxon . Freshwater hyphomycete . Asian mycology . Multi-locus analysis . Thailand

Introduction Section Editor: Gerhard Rambold The incorporation of molecular data in phylogenetic and evo- * Saranyaphat Boonmee [email protected] lutionary studies has revolutionized the current knowledge of fungal systematics (Tanabe et al. 2005; Shenoy et al. 2006; 1 Center of Excellence in Fungal Research, Mae Fah Luang Taylor and Berbee 2006; Choi and Kim 2017; Liu et al. 2017; University, Chiang Rai 57100, Thailand Tedersoo et al. 2018; Naranjo-Ortiz and Gabaldón 2019; 2 School of Science, Mae Fah Luang University, Chiang Rai 57100, Samarakoon et al. 2019). Natural taxonomic placements of Thailand taxa and development of backbone trees based on molecular 3 Innovative Institute for Plant Health, Zhongkai University of phylogeny have been possible (Ebersberger et al. 2012;He Agriculture and Engineering, Guangzhou 510225, Guangdong et al. 2019; Luo et al. 2019; Hongsanan et al. 2020;Hydeetal. ’ Province, People s Republic of China 2020a; Wijayawardene et al. 2020). The knowledge of fungal 4 Department of Botany and Microbiology, College of Science, King and classification are however limited as only ap- Saud University, P. O Box 2455, Riyadh 11451, Kingdom of Saudi proximately 8% (120,000 described taxa) of the 2.2 to 3.8 Arabia million estimated global fungal species are described 5 CAS Key Laboratory for Plant Diversity and Biogeography of East (Hawksworth and Lücking 2017) and even fewer have been Asia, Kunming Institute of Botany, Chinese Academy of Science, Kunming 650201, Yunnan, People’s Republic of China sequenced. Several authors have suggested areas as to where to find the “missing fungi,” such as understudied countries 6 Honghe Innovation Center for Mountain Futures, Kunming Institute of Botany, Honghe County 654400, Yunnan, People’sRepublicof and hosts, particularly in tropical regions and biodiversity China hot spots, little explored habitats or niches, and cryptic species 1032 Mycol Progress (2020) 19:1031–1042 hidden under well-established names [see Hyde 2001;Jones samples were incubated for 5 days and observed using stereo- 2011; Hawksworth and Lücking 2017;Hydeetal.2018]. microscope for the presence of fruiting bodies. Macro- The taxonomy and classification of Dothideomycetes have morphological features were photographed using a Motic been comprehensively studied, and considerable changes have SMZ 168 Series dissection microscope for fungal structures occurred over the past decade (Schoch et al. 2009; Hyde et al. on the woody substrate, while microscopic characters were 2013; Hongsanan et al. 2020). Hyde et al. (2013) listed 105 documented using Nikon Eclipse 80i microscope. Single families under Dothideomycetes. Several published works spore isolation was used to obtain pure cultures and colonial have also focused on different ecological groups of characteristics described. Herbarium-type specimen was de- Dothideomycetes that include terrestrial (Ariyawansa et al. posited in Mae Fah Luang University (MFLU). Ex-type living 2013, 2014; Dai et al. 2014; Thambugala et al. 2014a, b; culture was deposited at Mae Fah Luang University Culture Jayasiri et al. 2018; Pem et al. 2019), rock-dwelling (Ruibal Collection (MFLUCC). The new species was registered in et al. 2009), freshwater (Shearer et al. 2009; Shearer et al. 2014; Faces of Fungi (http://www.facesoffungi.org; Jayasiri et al. Dong et al. 2020), and marine habitats (Suetrong et al. 2009; 2015) and MycoBank database (http://www.mycobank.org/). Jones et al. 2012). Divergence time estimates using molecular data and fossil records have helped to confirm the establish- ment of higher-level taxa, such as families, orders, and classes. DNA extraction, PCR amplification, and sequencing is the largest order of Dothideomycetes (Kirk et al. 2008; Zhang et al. 2009, 2012; Hyde et al. 2013; Fungal mycelia from pure cultures grown in malt extract agar Hongsanan et al. 2020) and comprises ascomycetes with (MEA) for 30 days were scraped off using sterilized scalpel bitunicate asci and ceolomycetous or hyphomycetous asexual and kept in a 1.5 ml microcentrifuge tube. Genomic DNA was morphs (Wijayawardene et al. 2016;Jayasirietal.2018;Pem extracted using the Biospin Genomic DNA Extraction et al. 2019;Lietal.2020). Twenty-eight families and 175 kit (BioFlux®, China) following the manufacturer’sprotocol. genera (with 12 genera listed as genera incertae sedis)were Polymerase chain reaction (PCR) was used to amplify five included in the Pleosporales by Lumbsch and Huhndorf genes: the large subunit (28SrDNA), small subunit (2010). Wijayawardene et al. (2018) updated the outline of (18SrDNA), internal transcribed spacers (ITS1-5.8S-ITS2), Lumbsch and Huhndorf and included 75 families and 52 gen- translation elongation factor 1-alpha (TEF1-α), and DNA- era in Pleosporales genera incertae sedis.IntheOutline of dependent RNA polymerase II second largest subunit Fungi and fungus-like taxa, Wijayawardene et al. (2020)in- (RPB2). The 28SrDNA was amplified using the primers cluded 87 families and 533 genera, with 49 genera listed as LROR and LR5 (Vilgalys and Hester 1990), while genera incertae sedis in Pleosporales. 18SrDNA was amplified using the primers NS1 and NS4 The asexual genus Mycoenterolobium was introduced by (White et al. 1990). For ITS, primers ITS5 and ITS4 were Goos (1970)withMycoenterolobium platysporum as the type used (White et al. 1990). The TEF1-α was amplified using species growing on decaying wood of Araucaria from primers EF1–983F and EF1–2218R (Rehner and Buckley Hawaii. Subsequently, Karandikar et al. (2015) introduced 2005), while RPB2 was amplified using primers fRPB2-5F/ Mycoenterolobium flabelliforme from dead bark of Tectona fRPB2-7cR (Liu et al. 1999). Polymerase chain reaction was grandis collected from Toranmal region of Maharashtra State, performed in a volume of 25 μl, which contained 12.5 μlof India. There is no sequence data for these two species; there- 2× Power Taq PCR MasterMix (Bioteke Co., China), 1 μlof fore, taxonomic placement of the genus was not possible until each primer (10 μM), 1 μl genomic DNA, and 9.5 μldeion- the current study. ized water. The PCR thermal cycle programs for LSU, SSU, In our survey of freshwater fungi in northern Thailand and TEF1-α amplification were as follows: initial denaturing (Hyde et al. 2016), we observed and isolated a saprobic hy- step of 94 °C for 3 min, followed by 40 cycles of denaturation phomycete from submerged wood. Phylogenetic analysis of at 94 °C for 45 s, annealing at 56 °C for 50 s, elongation at combined LSU, SSU, ITS, TEF1-α,andRPB2 shows that the 72 °C for 1 min, and final extension at 72 °C for 10 min. The genus Mycoenterolobium groups close to Testudinaceae. PCR thermal cycle program for the RPB2 was initial denatur- ingstepof95°Cfor5min,followedby40cyclesofdena- turation at 95 °C for 1 min, annealing at 52 °C for 2 min, Materials and methods elongation at 72 °C for 90 s, and final extension at 72 °C for 10 min. Agarose gel electrophoresis was done to confirm the Sample collection, morphological observation, and presence of amplicons at the expected molecular weight. PCR fungal isolation products were purified and sequenced with the primers men- tioned above at a commercial sequencing provider (BGI, Samples of submerged decayed wood were collected from a Shenzhen, China). A BLAST search of the newly generated freshwater stream in Phitsanulok Province, Thailand. The sequences was carried out to exclude contamination and to Mycol Progress (2020) 19:1031–1042 1033 search for related taxa in GenBank database (www.ncbi.nlm. Result nih.gov/blast/). Phylogenetic analysis

Phylogenetic analysis The concatenated dataset (LSU, SSU, ITS, TEF1-α,and RPB2 loci) was composed of 59 taxa from Testudinaceae The gene sequences of closely related strains were assembled and related families in Pleosporales (Amniculicolaceae, based on the closest matches from the BLAST search results Anteagloniaceae, Aquasubmersaceae, Cryptocoryneaceae, and from recently published data in Wanasinghe et al. (2017) Hermatomycetaceae, Ligninsphaeriaceae, and Dayarathne et al. (2020). Sequences generated from the Lophiotremataceae, Pseudoastrosphaeriellaceae, three genes were analyzed along with other sequences re- Striatiguttulaceae, ), with Lophiostoma trieved from GenBank (Table 1). Five datasets, one for each arundinis CBS 621.86 and Lophiostoma crenatum CBS gene, were aligned with MAFFT v. 7 using the web server 629.86 as outgroup taxa (Table 1). (http://mafft.cbrc.jp/alignment/server; Katoh et al. 2019). The analyzed dataset, after trimming, comprised a total of Alignment was further refined manually, where necessary, 4214 characters including gaps (LSU = 830 bp; SSU = 1021; using BioEdit v.7.0.9.0 (Hall 1999). Aligned sequences were ITS = 474; TEF1-α = 866; RPB2 = 1007). The ML analysis automatically trimmed using TrimAl v. 1.3 on the web server for the combined dataset provided the best scoring tree (http://phylemon.bioinfo.cipf.es/utilities.html). The online (Fig. 1) with a final ML optimization likelihood value of − tool “ALTER” (Glez-Peña et al. 2010) was used to convert 32,311.80 (ln). The matrix had 1605 distinct alignment pat- the alignment file to phylip format. Phylogenetic analysis of terns and 24.63% proportion of gaps and completely undeter- both individual and combined gene data was performed using mined characters. The best ML phylogram resulted maximum likelihood (ML) and Bayesian inference (BI). with estimated base frequencies: A = 0.249042, C = Maximum parsimony (MP) analyses were performed using 0.247668, G = 0.270573, T = 0.232717, substitution rates: the heuristic search option with 1000 random taxa addition AC = 1.439606, AG = 3.875210, AT = 1.434240, CG = and tree bisection and reconnection (TBR) as the branch- 1.134209, CT = 8.381462, GT = 1.000000), and gamma dis- swapping algorithm in PAUP in CIPRES web portal tribution shape parameter: α = 0.195734. The Bayesian anal- (Swofford 2002;Milleretal.2010). All characters were un- ysis resulted in 4002 trees after 4,000,000 generations. The ordered and of equal weight and gaps were treated as missing first 1000 trees, representing the burn-in phase of the analyses, data.Maxtreeswereunlimited,branchesofzerolengthcol- were discarded, while the remaining 3002 trees were used for lapsed, and all multiple, equally parsimonious trees were calculating posterior probabilities in the majority rule consen- saved. Clade stability was assessed using a bootstrap (BS) sus tree. analysis with 1000 replicates, each with ten replicates of ran- The maximum parsimonious dataset consisted of 4527 dom stepwise addition of taxa (Hillis and Bull 1993). characters, of which 2732 were constant, 1193 parsimony- Descriptive tree statistics for parsimony (tree length [TL], informative (26.35%), and 289 parsimony-uninformative. consistency index [CI], retention index [RI], relative consis- The parsimony analysis of the data matrix resulted in 15 tency index [RC], and homoplasy index [HI]) were calculated equally most parsimonious trees with a length of 4356 steps for trees generated under different optimality criteria. (TL = 5935, CI = 0.396, RI = 0.641, RC = 0.254, HI = 0.604) Maximum likelihood analysis was performed using in the first tree. The tree topologies resulted from three analy- RAxML v. 8 software on the CIPRES web portal ses are similar. Support values for ML and MP above 50% and (Stamatakis 2006, 2014; Stamatakis et al. 2008)(http:// Bayesian posterior probabilities (PP) greater than 0.95 are www.phylo.org/portal2/; Miller et al. 2010). The GTR + given at the nodes. GAMMA model of nucleotide evolution was used. RAxML The ML phylogram generated based on sequence analysis rapid bootstrapping of 1000 replicates was performed. of the combined dataset indicates that Mycoenterolobium Bayesian inference analysis was performed using MrBayes aquadictyosporium formed a sister clade to v. 3.2.6 on XSEDE at the CIPRES web portal (Ronquist and nicotiae (CBS 101341) with 54% ML and 0.99 PP statistical Huelsenbeck 2003), using the parameter setting of 2 parallel support (Fig. 1). Muristudina chiangraiensis (MFLUCC 17– runs and 4 chains, run for 4,000,000 generations at which 2551), rosatii (CBS 331.78; CBS 690.82), point the standard deviation of split frequencies was below Verruculina enalia (BCC 18402; BCC 18401), 0.01. Trees were sampled every 1000 generations, and all bilgramii (CBS 333.78; AFTOL-ID 1598; CBS 101364), other parameters were left as default. Newly generated se- Trematosphaeria wegeliniana (CBS 123124), Angustospora quences have been deposited in GenBank (Table 1), and the nilensis (MFLU 15–1511), and Halotestudina muriformis finalized alignment and tree were deposited in TreeBASE, (MFLU 17–2634; MFLUCC 18–0392; MFLUCC 17–0395; submission ID: 26361 (http://www.treebase.org/). MFLU 18–0524) were grouped in Testudinaceae clade with 1034 Mycol Progress (2020) 19:1031–1042

Table 1 Taxa used in this study for the analysis of combined LSU, SSU, ITS rDNA, TEF1-α,andRPB2 sequence data and their GenBank accession numbers

Species Strain/voucher number LSU SSU ITS TEF1-α RPB2

Amniculicola immersa CBS 123083 FJ795498 GU456295 – GU456273 GU456358 Amniculicola parva CBS 123092 GU301797 GU296134 MH863272 GU349065 – Angustospora nilensis MFLU 15–1511 NG_057091 NG_061219 ––– Anteaglonium globosum ANM925.2 GQ221911 ––GQ221919 – Anteaglonium parvulum MFLUCC 14–0817 KU922913 KU922914 –– Anteaglonium thailandicum MFLUCC 14–0816 KU922909 KU922910 – KU922920 – Aquasubmersa japonica KT2813 LC061586 LC061581 LC061591 LC194383 LC194420 Aquasubmersa japonica KT2862 LC061587 LC061582 LC061592 LC194384 LC194421 Aquasubmersa japonica KT2863 LC061588 LC061583 LC061593 LC194385 LC194422 Aquasubmersa mircensis MFLUCC 11–0401 JX276955 JX276956 JX276954 Atrocalyx acutispora KT2436 LC194341 LC194299 LC194475 LC194386 LC194423 Atrocalyx lignicola CBS122364 LC194342 LC194300 LC194476 LC194387 LC194424 Cryptoclypeus oxysporus KT2772 LC194345 LC194303 LC194479 LC194390 LC194427 Cryptocoryneum brevicondensatum yone152 LC194349 LC194307 LC096155 LC096137 LC194431 Cryptocoryneum japonicum KT3300 LC194354 LC194312 LC096160 LC096142 LC194436 Cryptocoryneum pseudorilstonei CBS113641 LC194364 LC194322 LC096170 LC096152 LC194446 Halotestudina muriformis MFLUCC 18–0392 MN017894 MN017947 ––– Halotestudina muriformis MFLU 18–0524 MN017853 MN017919 ––MN077079 Halotestudina muriformis MFLU 17–2634 MN017852 MN017918 – MN077068 – Halotestudina muriformis MFLUCC 17–0395 MN017854 MN017920 ––– Hermatomyces tectonae MFLUCC 14–1141 KU764696 KU712466 KU144918 KU872758 – Hermatomyces thailandica MFLUCC 14–1143 KU764692 KU712468 KU144920 KU872754 KU712488 Lepidosphaeria nicotiae CBS 101341 DQ678067 ––DQ677910 DQ677963 Ligninsphaeria jonesii GZCC 15–0080 KU221038 –––– Ligninsphaeria jonesii MFLUCC 15–0641 NG_059642 –––– Longicorpus striataspora MFLUCC 17–2515 MK035990 MK035975 MK035967 MK034430 MK034438 Longicorpus striataspora MFLUCC 17–2516 MK035991 MK035976 MK035968 MK034431 MK034439 Longicorpus striataspora MFLUCC 18–0268 MK035989 MK035974 MK035966 MK034429 MK034437 Longicorpus striataspora MFLUCC 18–0267 MK035988 MK035973 MK035965 MK034428 MK034436 Lophiostoma arundinis CBS 621.86 DQ782384 DQ782383 AJ496633 DQ782387 DQ782386 Lophiostoma crenatum CBS 629.86 DQ678069 DQ678017 – DQ677912 DQ677965 Lophiotrema bambusae RP0030 KX672154 KX672159 KX672149 KX672162 KX672161 Lophiotrema eburnoides KT1424.1 LC001707 LC001706 LC001709 LC194403 LC194458 Lophiotrema fallopiae KT2748 LC149915 LC149911 LC149913 LC194404 LC194459 Massarina albocarnis CBS 119345 LC194379 LC194337 LC194503 LC194416 LC194471 Muristudina chiangraiensis MFLUCC 17–2551 MG602248 MG602249 MG602247 MG602251 MG602250 Mycoenterolobium aquadictyosporium MFLUCC 20–0118 MT447877 MT465324 MT465427 MT461428 MT477863 Neotestudina rosatii CBS 331.78 MH872904 – MH861143 –– Neotestudina rosatii CBS 690.82 DQ384107 DQ384069 ––– Polyplosphaeria fusca KT1616 AB524604 AB524463 AB524789 –– Pseudoastrosphaeriella bambusae MFLUCC 11–0205 KT955475 KT955455 – KT955437 KT955414 Pseudoastrosphaeriella longicolla MFLUCC 11–0171 KT955476 ––KT955438 KT955420 Pseudoastrosphaeriella thailandensis MFLUCC 14–0038 KT955479 KT955458 – KT955441 KT955409 Pseudocryptoclypeus yakushimensis KT2186 LC194380 LC194338 LC194504 LC194417 LC194472 Pseudolophiotrema elymicola KT1450 LC194381 LC194339 LC194505 LC194418 LC194473 Pseudotetraploa curviappendiculata HC 4930 AB524608 AB524467 AB524792 AB524823 – Quadricrura septentrionalis HC 4983 AB524615 AB524474 AB524799 AB524830 – Striatiguttula nypae MFLUCC 17–2517 MK035993 MK035978 MK035970 MK034433 MK034441 Mycol Progress (2020) 19:1031–1042 1035

Table 1 (continued)

Species Strain/voucher number LSU SSU ITS TEF1-α RPB2

Striatiguttula nypae MFLUCC 17–2518 MK035994 MK035979 MK035971 MK034434 – Striatiguttula nypae MFLUCC 18–0265 MK035992 MK035977 MK035969 MK034432 MK034440 Striatiguttula phoenicis MFLUCC 18–0266 MK035995 MK035980 MK035972 MK034435 MK034442 Tetraploa sasicola KT 563 AB524631 AB524490 AB524807 AB524838 – Trematosphaeria wegeliniana CBS 123124 GU261722 GU261720 ––– Triplosphaeria maxima KT 870 AB524637 AB524496 AB524812 AB524843 – Ulospora bilgramii AFTOL-ID 1598 DQ678076 DQ678025 – DQ677921 DQ677974 Ulospora bilgramii CBS 101364 DQ678076 DQ678025 – DQ677921 DQ677974 Ulospora bilgramii CBS 333.78 MH872905 – MH861144 –– Verruculina enalia BCC 18401 GU479802 GU479770 – GU479863 GU479835 Verruculina enalia BCC 18402 GU479803 GU479771 – GU479864 GU479836

The newly generated sequences are indicated in bold

89% ML and 1.00 PP statistical support. The sequence data cultures, with dense mycelia. Sporulation in culture after placed Mycoenterolobium aquadictyosporium close to the 14 days of incubation. family Testudinaceae within Pleosporomycetidae, Material examined: Thailand, Phitsanulok Province, Wang Dothideomycetes. However, the family Testudinaceae is het- Thong District, Kaeng Sopha waterfall stream, on submerged erogeneous morphologically and phylogenetically weakly wood, 25 July 2019, S. Boonmee, PSL/WT-17 (MFLU 20– supported, and the new taxon is referred to Pleosporales 0443, holotype), ex-type living culture 20–0118. incertae sedis until further taxon sampling is undertaken. GenBank accession numbers: ITS = MT465427, LSU = MT447877, SSU = MT465324, TEF1-α = MT461428, Taxonomy RPB2 =MT477863 Notes: Mycoenterolobium,typifiedbyM. platysporum Mycoenterolobium aquadictyosporium M.S. Calabon, Goos, was isolated from decaying wood of Araucaria sp. Boonmee, E.B.G. Jones & K.D. Hyde, sp. nov. (Figs. 2 and 3) (Araucariaceae) in Hawaii (Goos 1970). Mycoenterolobium MycoBank number: MB835668; Faces of fungi number: is unique in the production of massive, flattened, fan-shaped FoF 08423 conidia that resemble Cancellidium species. However, they Etymology: referring to the aquatic habitat and shape of the differ in the arrangement of conidial rows of cells at the at- conidia tachment point to the conidiophores. The conidia of Saprobic on decaying wood submerged in freshwater hab- Mycoenterolobium are made up of rows of cells, radiating in itat. Sexual morph: undetermined. Asexual morph: hyphomy- a linear pattern from a basal cell attached to the conidiophore, cetous, on natural substrate, colonies effuse, black, shiny, uni- while Cancellidium is distinct in having parallel adherent rows formly spread, mycelium immersed. Conidiophores 7–22 × of septate branches radiating from the conidiophore (Goos 2–8 μm(̅x =11×5 μm, n = 7), micronematous, inconspicu- 1970; Seifert et al. 2011; Zhao et al. 2013). Cancellidium ous, short or absent. Conidiogenous cells monoblastic, inte- conidia also contain internal branched chains of blastic, grated, terminal, determinate, oval to irregularly shaped. cicatrized, and monilioid cells, developing from the base Conidia when immature 19–52 × 14–38 μm(̅x =30× (Pratibha et al. 2014), and these are lacking in 25 μm, n = 15); at mature 45–92 × 43–104 μm, and 10– Mycoenterolobium. Dictyosporous hyphomycetes wherein 30 μmthick(̅x =71×70×20μm, n = 30), acrogenous, soli- conidial morphology differs from Mycoenterolobium include tary, dictyosporous, strongly flattened, fan shaped at maturity, Aquadictyospora, Dictyopalmispora, Dictyosporium, composed of 15–25 rows of cells radiating from single cell at Dictyocheirospora, Digitodesmium, Jalapriya, the point of attachment, dark brown, smooth-walled. Pseudodictyosporium, and Vikalpa (Kirschner et al. 2013; Culture characteristics: conidia germinating on MEA Boonmee et al. 2016; Yang et al. 2018; Hyde et al. 2019; within 24 h and germ tubes produced from the basal part of Hongsanan et al. 2020). Aquadictyospora is characterized by the conidia. On MEA, colony circular with entire margin, broadly rounded conidia, composed of 4–6 compactly ar- reaching 10–15 mm in 25 days at 25 °C, dark brown to grayish ranged rows with a basal, subglobose, hyaline cell. brown from above, dark brown to black from below, surface Dictyopalmispora has complanate conidia, with unique hair- smooth, opaque, dry, raised to becoming umbonate in old like extensions produced from apical cells of all 4 rows. 1036 Mycol Progress (2020) 19:1031–1042 Mycol Progress (2020) 19:1031–1042 1037

R Fig. 1 Phylogenetic tree based on RAxML analyses of combined LSU, apex, apical cells with or without appendages. Dictyosporium α SSU, ITS, TEF1- ,andRPB2 sequence data. Bootstrap support values differs in having cheiroid, digitate, and complanate conidia for ML and MP higher than 50% and PP greater than 0.95 are indicated above the nodes as MP/ML/PP. The new isolate is represented in blue. without separating arms, while Dictyocheirospora is charac- The tree is rooted to Lophiostoma arundinis (CBS 621.86) and terized by non-complanate conidia with arms arising from the Lophiostoma crenatum (CBS 629.86) (). Bar = 0.05 basal cell and closely gathered at the apex and compact. estimated number of nucleotide substitutions per site per branch Pseudodictyosporium has cheiroid conidia with 2–3rowsof cells, while Vikalpa has conidia with 3 rows of cells each in a Digitodesmium has cheiroid, digitate conidia with apical ge- different plane. latinous caps, while Jalapriya has cheiroid, euseptate conidia Three species are included in Mycoenterolobium: with 5–7 rows of cells, rows converging or not converging at M. aquadictyosporium, M. flabelliforme K.G. Karand., P.N.

Fig. 2 Mycoenterolobium aquadictyosporium (MFLUCC 20–0118, holotype). a Host. b, c Appearance of colonies in natural substrate. d Conidia. e, f Developing conidia attached to conidiogenous cells. g, h Immature conidia. and i–l Mature conidia. Scale bars b =1mm;c, d =200μm; e–l =10μm 1038 Mycol Progress (2020) 19:1031–1042 Mycol Progress (2020) 19:1031–1042 1039

R Fig. 3 Mycoenterolobium aquadictyosporium (MFLUCC 20–0118, freshwater species, M. platysporum and M. flabelliforme were holotype). a Germinated conidium. b Colony on MEA from above and collected from terrestrial trees of Araucaria sp. and Tectona below. c–f Development of conidia in culture. g–k developing conidia attach to conidiogenous cells. l–o conidia. Scale bars a, k–o =50μm; c, grandis (Goos 1970; Karandikar et al. 2015), while d =1mm;e, f =100μm; g–j =15μm M. platysporum has also been collected from submerged man- grove wood of Rhizophora stylosa (Nakagiri 1993). Singh & S.K., Singh and M. platysporum Goos (Goos 1970; Karandikar et al. 2015). Mycoenterolobium aquadictyosporium can be distinguished from Discussion M. platysporum in having shorter but wider conidia (45– 92 μm long, 43–104 μm wide vs. 110–130 μm long, 75– Phylogenetic results showed a close relationship between the 80 μm wide). The conidia of Mycoenterolobium new taxon and the family Testudinaceae, typified by aquadictyosporium are longer than in M. flabelliforme (45– Testudina terrestris Bizz. and established by von Arx (1971) 92 μmlong,43–104 μm wide vs. 23.5–37.5 μm long, 24.5– to accommodate Argynna, Lepidosphaeria, Neotestudina, 45.5 μm wide). Mycoenterolobium aquadictyosporium is a Pseudophaeotrichum,andTestudina. Argynna was

Fig. 4 Fan-shaped conidia of Mycoenterolobium and Cancellidium. a, b rows of septate branches radiating from the point of attachment to the Conidia of Mycoenterolobium aquadictyosporium (MFLUCC 20–0118) conidiophore. e Strings of monilioid cells of Cancellidium sp. (MFLU showing rows of cells radiating in a linear pattern from a basal cell. c, d 20–0429). Scale bars a–e =20μm Conidia of Cancellidium sp. (MFLU 20–0429) with parallel adherent 1040 Mycol Progress (2020) 19:1031–1042 transferred to Argynnaceae by Shearer and Crane (1980), and 2020), and a new order has been introduced to accommodate Pseudophaeotrichum was synonymized as Neotestudina the genus (Hyde et al. 2020b).Thescanningelectronmicro- (Dayarathne et al. 2020; Hongsanan et al. 2020). Currently, graph study of Nakagiri (1993) on conidium development of Testudinaceae includes nine genera, i.e., Angustospora, Mycoenterolobium platysporum contributed to the current Halotestudina, Lepidosphaeria, Lojkania, Muritestudina, knowledge on the septation of the lobes in the genus and its Neotestudina, Testudina, Ulospora,andVerruculina muriform morphology. Mycoenterolobium platysporum has a (Suetrong et al. 2009; Dayarathne et al. 2020; stipitate cup morphology at an early stage of conidial devel- Wijayawardene et al. 2020). Members in Testudinaceae are opment then the opposing sides of the cup elongates until it mainly characterized by globose to subglobose ascomata, a reaches both apical edges attaining a bivalve-like morphology. peridium comprising cells of textura angularis, clavate to cy- At maturity in Mycoenterolobium, the two conidial lobes unite lindrical asci with short pedicels and ellipsoidal, verrucose to and are closely appressed (Seifert et al. 2011). verruculose ascospores with mostly one longitudinal septum. Halotestudina, Lepidosphaeria,andVerruculina were isolat- Acknowledgments Mark S. Calabon is grateful to the Mushroom ed from marine habitats, while Angustospora species are Research Foundation and the Department of Science and Technology, Science Education Institute (Philippines). MS Calabon is thankful to freshwater taxa (Hawksworth 1979; Hyde et al. 2013;Li Dr. Eleni Gentekaki for her comments and suggestions in the phyloge- et al. 2016; Dayarathne et al. 2020). Doilom et al. (2018) netic tree. The authors are grateful to DF Bao for the Cancellidium provided descriptions, illustrations, and notes for the type photo. The two anonymous reviewers are also acknowledged for their species of Lepidosphaeria, Neotestudina,andUlospora useful comments. from their type specimens. Wanasinghe et al. (2017)and Funding Saranyaphat Boonmee would like to thank the Thailand Dayarathne et al. (2020) provided a synopsis and key to the Research Fund (Grant No. TRG6180001) and Plant Genetic genera in Testudinaceae. Conservation Project under the Royal Initiation of Her Royal Highness In the phylogenetic analysis of Wanasinghe et al. (2017), Princess Maha Chakri Sirindhorn-Mae Fah Luang University. Kevin D. “ the placement of several genera within Testudinaceae was not Hyde thanks the Thailand Research Fund grant entitled Impact of cli- mate change on fungal diversity and biogeography in the Greater Mekong congruent and varied between the estimated ML and MP trees. Subregion” (Grant No. RDG6130001). E. B. Gareth Jones is supported For instance, Muritestudina chiangraiensis grouped with under the Distinguished Scientist Fellowship Program (DSFP), King Neotestudina rosatii in ML analysis but clustered with Saud University, Kingdom of Saudi Arabia. Mingkwan Doilom thanks Verruculina enalia in MP analysis. The inconsistencies in the 5th batch of Postdoctoral Orientation Training Personnel in Yunnan Province (Grant No. Y934283261) and the 64th batch of China Post- the estimated trees may be due to weak phylogenetic signal doctoral Science Foundation (Grant No. Y913082271). of the analyzed multi-loci, unequal evolutionary rates and dif- ferent genealogical histories (Wanasinghe et al. 2017). Two fungal species from marine habitats, Massarina ricifera and References Quintaria lignatilis, were clustered with other genera in Testudinaceae but with weak bootstrap support (Schoch Ariyawansa HA, Kang JC, Alias SA et al (2013) Towards a natural et al. 2009; Suetrong et al. 2009). Resolving the taxonomic classification of Dothideomycetes: the genera Dermatodothella, placement of both marine species needs additional taxon sam- Dothideopsella, Grandigallia, Hysteropeltella and Gloeodiscus pling (Hyde et al. 2013). We therefore refer (Dothideomycetes incertae sedis). Phytotaxa 147:35–47. https:// doi.org/10.11646/phytotaxa.147.2.1 Mycoenterolobium aquadictyosporium to the Pleosporales Ariyawansa HA, Thambugala KM, Kang JC et al (2014) Towards a incertae sedis until further taxon sampling is undertaken. natural classification of Dothideomycetes 2: the genera Mycoenterolobium aquadictyosporium is similar to Cucurbidothis, Heterosphaeriopsis, Hyalosphaera, Navicella and Cancellidium pinicola in having dictyoseptate, muriform, Pleiostomellina (Dothideomycetes incertae sedis). Phytotaxa 176: – strongly flattened, fan-shaped conidia that are wider at the 7 17. https://doi.org/10.11646/phytotaxa.176.1.4 Boonmee S, D’Souza MJ, Luo Z et al (2016) Dictyosporiaceae fam. nov. apex and narrower at the base (Yeung et al. 2006;Hyde Fungal Divers 80:457–482. https://doi.org/10.1007/s13225-016- et al. 2020b). Mycoenterolobium aquadictyosporium differs 0363-z from the latter in the arrangement of conidial rows of cells at Choi JJ, Kim SH (2017) A genome tree of life for the fungi kingdom. the attachment point to the conidiophores. The conidia of Proc Natl Acad Sci U S A 114:9391–9396. https://doi.org/10.1073/ Mycoenterolobium are made up of rows of cells, radiating in pnas.1711939114 Dai DQ, Bahkali AH, Jayarama Bhat D et al (2014) Towards a natural a linear pattern from a basal cell attached to the conidiophore, classification of dothideomycetes 3: the genera Muellerites, while Cancellidium is distinct in having parallel adherent rows Trematosphaeriopsis, Vizellopsis and Yoshinagella of septate branches radiating from the conidiophore (Seifert (Dothideomycetes incertae sedis). Phytotaxa 176:18–27. https:// et al. 2011;Zhaoetal.2013)(Fig.4). Furthermore, doi.org/10.11646/phytotaxa.176.1.5 Cancellidium is not closely related to the Dothideomycetes Dayarathne M, Jones EBG, Maharachchikumbura S et al (2020) Morpho- molecular characterization of microfungi associated with marine as it belongs to Sordariomycetidae genera incertae sedis based habitats. Mycosphere 11:1–188. https://doi.org/10.5943/ (Sordariomycetes) (Hyde et al. 2020a; Wijayawardene et al. mycosphere/11/1/1 Mycol Progress (2020) 19:1031–1042 1041

Doilom M, Hyde KD, Phookamsak R et al (2018) Mycosphere notes Jones EBG, Hyde KD, Pang KL, Suetrong S (2012) Phylogeny of the 225–274: types and other specimens of some genera of Dothideomycetes and other classes of marine . In: Jones Ascomycota. Mycosphere 9:647–754. https://doi.org/10.5943/ EBG, Pang KL (eds) Marine fungi: and fungal-like organisms. mycosphere/9/4/3 Walter de Gruyter, Berlin, pp 17–34 Dong W, Wang B, Hyde KD et al (2020) Freshwater Dothideomycetes. Karandikar KG, Singh PN, Singh SK (2015) Mycoenterolobium Fungal Divers (in prep.) flabelliforme: a new anamorphic fungus from India. Plant Pathol Ebersberger I, De Matos SR, Kupczok A et al (2012) A consistent phy- Quar 5:49–51. https://doi.org/10.5943/ppq/5/2/3 logenetic backbone for the fungi. Mol Biol Evol 29:1319–1334. Katoh K, Rozewicki J, Yamada KD (2019) MAFFT online service: mul- https://doi.org/10.1093/molbev/msr285 tiple sequence alignment, interactive sequence choice and visualiza- Glez-Peña D, Gómez-Blanco D, Reboiro-Jato M et al (2010) ALTER: tion. Brief Bioinform 20:1160–1166. https://doi.org/10.1093/bib/ program-oriented conversion of DNA and protein alignments. bbx108 Nucleic Acids Res 38(Web Server issue):W14–W18. https://doi. Kirk PM, Cannon PF, Minter DW, Stalpers JA (2008) Ainsworth & org/10.1093/nar/gkq321 Bisby’s dictionary of the fungi, 10th edn. CABI Europe, UK Goos RD (1970) A new genus of the hyphomycetes from Hawaii. Kirschner R, Pang KL, Jones EBG (2013) Two cheirosporous hyphomy- Mycologia 62:171–175. https://doi.org/10.2307/3757719 cetes reassessed based on morphological and molecular examina- Hall TA (1999) BioEdit: a user-friendly biological sequence alignment tion. Mycol Prog 12:29–36 editor and analysis program for Windows 95/98/NT. 41(1999):95– Li GJ, Hyde KD, Zhao RL et al (2016) Fungal diversity notes 253–366: Nucl Acids Symp Ser 98. https://doi.org/10.14601/Phytopathol_ taxonomic and phylogenetic contributions to fungal taxa. Fungal Mediterr-14998u1.29 Divers 78:1–237. https://doi.org/10.1007/s13225-016-0366-9 Hawksworth DL (1979) Ascospore sculpturing and generic concepts in Li WJ, McKenzie EHC, Liu JK et al (2020) Taxonomy and phylogeny of the Testudinaceae (syn. ). Can J Bot 57:91–99. https:// hyaline-spored coelomycetes. Fungal Divers 100:279–801. https:// doi.org/10.1139/b79-017 doi.org/10.1007/s13225-020-00440-y Hawksworth DL, Lücking R (2017) Fungal diversity revisited: 2.2 to 3.8 Liu YJ, Whelen S, Hall BD (1999) Phylogenetic relationships among million species. Microbiol Spectr 5:FUNK-0052-2016. https://doi. ascomycetes: evidence from an RNA polymerse II subunit. Mol org/10.1128/microbiolspec.funk-0052-2016 Biol Evol 16:1799–1808. https://doi.org/10.1093/oxfordjournals. He MQ, Zhao RL, Hyde KD et al (2019) Notes, outline and divergence molbev.a026092 times of Basidiomycota. Fungal Divers 99:105–367. https://doi.org/ Liu JK, Hyde KD, Jeewon R et al (2017) Ranking higher taxa using 10.1007/s13225-019-00435-4 divergence times: a case study in Dothideomycetes. Fungal Divers Hillis DM, Bull JJ (1993) An empirical test of bootstrapping as a method 84:75–99. https://doi.org/10.1007/s13225-017-0385-1 for assessing confidence in phylogenetic analysis. Syst Biol 42:182– Lumbsch HT, Huhndorf SM (2010) Myconet volume 14. Part one. 192. https://doi.org/10.1093/sysbio/42.2.182 Outline of Ascomycota—2009. Part two. Notes on Ascomycete Hongsanan S, Hyde KD, Phookamsak R et al. (2020) Refined families of Systematics. Nos. 4751–5113. Fieldiana Life Earth Sci 1:1–64. Dothideomycetes. Mycosphere (in prep.) https://doi.org/10.3158/1557.1 Hyde KD (2001) Where are the missing fungi? Does Hong Kong have Luo ZL, Hyde KD, Liu JK et al (2019) Freshwater Sordariomycetes. any answers? Mycol Res 105:1514–1518 Fungal Divers 99:451–660. https://doi.org/10.1007/s13225-019- Hyde KD, Jones EBG, Liu JK et al (2013) Families of Dothideomycetes. 00438-1 Fungal Divers 63:1–313. https://doi.org/10.1007/s13225-013-0263- Miller MA, Pfeiffer W, Schwartz T (2010) Creating the CIPRES Science 4 Gateway for inference of large phylogenetic trees. In: 2010 Gateway Hyde KD, Fryar S, Tian Q et al (2016) Lignicolous freshwater fungi computing environments workshop, GCE 2010 along a north-south latitudinal gradient in the Asian/Australian re- Nakagiri A (1993) Intertidal mangrove fungi from Iriomote Island. IFO gion; can we predict the impact of global warming on biodiversity Res Commun 16:24–62 and function? Fungal Ecol 19:190–200. https://doi.org/10.1016/j. Naranjo-Ortiz MA, Gabaldón T (2019) Fungal evolution: diversity, tax- funeco.2015.07.002 onomy and phylogeny of the Fungi. Biol Rev 94:2101–2137. Hyde KD, Norphanphoun C, Chen J et al (2018) Thailand’samazing https://doi.org/10.1111/brv.12550 diversity: up to 96% of fungi in northern Thailand may be novel. Pem D, Jeewon R, Bhat DJ et al (2019) Mycosphere notes 275-324: a Fungal Divers 93:215–239. https://doi.org/10.1007/s13225-018- morpho-taxonomic revision and typification of obscure 0415-7 Dothideomycetes genera (incertae sedis). Mycosphere 10:1115– Hyde KD, Tennakoon DS, Jeewon R et al (2019) Fungal diversity notes 1246. https://doi.org/10.5943/mycosphere/10/1/22 1036–1150: taxonomic and phylogenetic contributions on genera Pratibha J, Prabhugaonkar A, Hyde KD, Bhat DJ (2014) Phylogenetic and species of fungal taxa. Fungal Divers 96:1–242. https://doi. placement of Bahusandhika, Cancellidium and Pseudoepicoccum org/10.1007/s13225-019-00429-2 (asexual Ascomycota). Phytotaxa 176:68–80. https://doi.org/10. Hyde KD, Norphanphoun C, Maharachchikumbura SSN et al (2020a) 11646/phytotaxa.176.1.9 Refined families of Sordariomycetes. Mycosphere 11:305–1059. Rehner SA, Buckley E (2005) A Beauveria phylogeny inferred from https://doi.org/10.5943/mycosphere/11/1/7 nuclear ITS and EF1-α sequences: evidence for cryptic diversifica- Hyde KD, Bao DF, Hogsanan S et al (2020b) Evolution of freshwater tion and links to Cordyceps teleomorphs. Mycologia 97:84–98. Diaporthomycetidae (Sordariomycetes); implications for freshwater https://doi.org/10.3852/mycologia.97.1.84 fungi species numbers. Fungal Divers (in press) Ronquist F, Huelsenbeck JP (2003) MrBayes 3: Bayesian phylogenetic Jayasiri SC, Hyde KD, Ariyawansa HA et al (2015) The Faces of Fungi inference under mixed models. Bioinformatics 19:1572–1574. database: fungal names linked with morphology, phylogeny and https://doi.org/10.1093/bioinformatics/btg180 human impacts. Fungal Divers 74:3–18. https://doi.org/10.1007/ Ruibal C, Gueidan C, Selbmann L et al (2009) Phylogeny of rock- s13225-015-0351-8 inhabiting fungi related to Dothideomycetes. Stud Mycol 64:123– Jayasiri SC, Hyde KD, Jones EBG et al (2018) Taxonomic novelties of 133. https://doi.org/10.3114/sim.2009.64.06 hysteriform Dothideomycetes. Mycosphere 9:803–837. https://doi. Samarakoon MC, Hyde KD, Hongsanan S et al (2019) Divergence time org/10.5943/mycosphere/9/4/8 calibrations for ancient lineages of Ascomycota classification based Jones EBG (2011) Are there more marine fungi to be described? Bot Mar on a modern review of estimations. Fungal Divers 96:285–346. 54:343–354 https://doi.org/10.1007/s13225-019-00423-8 1042 Mycol Progress (2020) 19:1031–1042

Schoch CL, Crous PW, Groenewald JZ et al (2009) A class-wide phylo- Thambugala KM, Singtripop C, Chunfang Y et al (2014b) Towards a genetic assessment of Dothideomycetes. Stud Mycol 64:1–15-S10. natural classification of Dothideomycetes 7: the genera Allosoma, https://doi.org/10.3114/sim.2009.64.01 Austropleospora, Dangeardiella, Griggsia and Karschia Seifert K, Morgan-Jones G, Gams W, Kendrick B (2011) The genera of (Dothideomycetes incertae sedis). Phytotaxa 181:34–46. https:// hyphomycetes. CBS-KNAW Fungal Biodiversity Centre, doi.org/10.11646/phytotaxa.181.1.2 Netherlands Vilgalys R, Hester M (1990) Rapid genetic identification and mapping of Shearer CA, Crane JL (1980) Taxonomy of two cleistothecial ascomy- enzymatically amplified ribosomal DNA from several Cryptococcus cetes with papilionaceous ascospores. Trans Br Mycol Soc 75:193– species. J Bacteriol 172:4238–4246. https://doi.org/10.1128/jb.172. 200. https://doi.org/10.1016/s0007-1536(80)80079-9 8.4238-4246.1990 Shearer CA, Raja HA, Miller AN et al (2009) The molecular phylogeny von Arx J (1971) Testudinaceae, a new family of ascomycetes. Persoonia of freshwater Dothideomycetes. Stud Mycol 64:145–153. https:// 6:365–369 doi.org/10.3114/sim.2009.64.08 Wanasinghe D, Jeewon R, Tibpromma S et al (2017) Saprobic Shearer CA, Pang KL, Suetrong S et al (2014) Phylogeny of the Dothideomycetes in Thailand: Muritestudina gen. et sp. nov. Dothideomycetes and other classes of freshwater fissitunicate (Testudinaceae) a new terrestrial pleosporalean ascomycete, with Ascomycota. In: Jones EBG, Hyde KD, Pang KL (eds) Freshwater hyaline and muriform ascospores. Stud Fungi 2:219–234. https:// fungi and Fungal-like organisms. Walter de Gruyter, Berlin, pp 25– doi.org/10.5943/sif/2/1/26 46 White TJ, Bruns T, Lee S, Taylo JW (1990) Amplification and direct Shenoy BD, Jeewon R, Wu WP et al (2006) Ribosomal and RPB2 DNA sequencing of fungal ribosomal RNA genes for phylogenetics. In: sequence analyses suggest that Sporidesmium and morphologically Innis MA, Gelfand DH, Sninsky JJ, White TJ (eds) PCR protocols: a similar genera are polyphyletic. Mycol Res 110:916–928. https:// guide to methods and applications. Academic Press, New York, pp doi.org/10.1016/j.mycres.2006.06.004 315–322 Stamatakis A (2006) RAxML-VI-HPC: maximum likelihood-based phy- Wijayawardene NN, Hyde KD, Wanasinghe DN et al (2016) Taxonomy logenetic analyses with thousands of taxa and mixed models. and phylogeny of dematiaceous coelomycetes. Fungal Divers 77:1– – Bioinformatics 22:2688 2690. https://doi.org/10.1093/ 316. https://doi.org/10.1007/s13225-016-0360-2 bioinformatics/btl446 Wijayawardene NN, Hyde KD, Lumbsch HT et al (2018) Outline of Stamatakis A (2014) RAxML version 8: a tool for phylogenetic analysis Ascomycota: 2017. Fungal Divers 88:167–263. https://doi.org/10. – and post-analysis of large phylogenies. Bioinformatics 30:1312 1007/s13225-018-0394-8 1313. https://doi.org/10.1093/bioinformatics/btu033 Wijayawardene NN, Hyde KD, Al-ani LKT et al (2020) Outline of fungi Stamatakis A, Hoover P, Rougemont J (2008) A rapid bootstrap algo- – – and fungi-like taxa. Mycosphere 11:1 367. https://doi.org/10.5943/ rithm for the RAxML web servers. Syst Biol 57:758 771. https:// mycosphere/11/1/8 doi.org/10.1080/10635150802429642 Yang J, Liu JK, Hyde KD et al (2018) New species in Dictyosporium, Suetrong S, Schoch CL, Spatafora JW et al (2009) Molecular systematics new combinations in Dictyocheirospora and an updated backbone of the marine Dothideomycetes. Stud Mycol 64:155–173S6. https:// tree for Dictyosporiaceae. MycoKeys 36:83–105. https://doi.org/10. doi.org/10.3114/sim.2009.64.09 3897/mycokeys.36.27051 Swofford DL (2002) PAUP: phylogenetic analysis using parsimony, ver- Yeung QSY, Jeewon R, Hyde KD (2006) Cancellidium pinicola sp. nov. sion 4.0 b10. Sinauer Associates, Sunderland from Pinus massoniana and its phylogeny. Cryptogam Mycol 27: Tanabe Y, Watanabe MM, Sugiyama J (2005) Evolutionary relationships 295–304 among basal fungi (Chytridiomycota and Zygomycota): insights from molecular phylogenetics. J Gen Appl Microbiol 51:267–276 Zhang Y, Schoch CL, Fournier J et al (2009) Multi-locus phylogeny of Pleosporales: a taxonomic, ecological and evolutionary re-evalua- Taylor JW, Berbee ML (2006) Dating divergences in the fungal tree of – life: review and new analyses. Mycologia 98:838–849 tion. Stud Mycol 64:85 102. https://doi.org/10.3114/sim.2009.64. Tedersoo L, Sánchez-Ramírez S, Kõljalg U et al (2018) High-level clas- 04 sification of the fungi and a tool for evolutionary ecological analy- Zhang Y, Crous PW, Schoch CL, Hyde KD (2012) Pleosporales. Fungal – ses. Fungal Divers 90:135–159. https://doi.org/10.1007/s13225- Divers 53:1 221. https://doi.org/10.1007/s13225-011-0117-x 018-0401-0 Zhao G, Yu P, Liu X (2013) Cancellidium and Canalisporium – Thambugala KM, Ariyawansa HA, Liu ZY et al (2014a) Towards a (hyphomycetes) from China. Nov Hedwigia 96:221 236. https:// natural classification of dothideomycetes 6: the genera Dolabra, doi.org/10.1127/0029-5035/2012/0048 Placostromella, Pleosphaerellula, Polysporidiella and Pseudotrichia (Dothideomycetes incertae sedis). Phytotaxa 176: Publisher’snoteSpringer Nature remains neutral with regard to jurisdic- 55–67. https://doi.org/10.11646/phytotaxa.176.1.8 tional claims in published maps and institutional affiliations.