Research Article
Print
Research Article
Myrothecium-like new species from turfgrasses and associated rhizosphere
expand article infoJunmin Liang, Guangshuo Li§, Shiyue Zhou|, Meiqi Zhao#, Lei Cai|
‡ Institute of Microbiology, Chinese Academy of Sciences, Beijing, China
§ Hebei University, Baoding, China
| University of Chinese Academy of Sciences, Beijing, China
¶ Agricultural University, Beijing, China
# Forwardgroup Turf Service & Research Center, Wanning, China
Open Access

Abstract

Myrothecium sensu lato includes a group of fungal saprophytes and weak pathogens with a worldwide distribution. Myrothecium s.l. includes 18 genera, such as Myrothecium, Septomyrothecium, Myxospora, all currently included in the family Stachybotryaceae. In this study, we identified 84 myrothecium-like strains isolated from turfgrasses and their rhizosphere. Five new species, i.e., Alfaria poae, Alf. humicola, Dimorphiseta acuta, D. obtusa, and Paramyrothecium sinense, are described based on their morphological and phylogenetic distinctions. Phylogenies were inferred based on the analyses of sequences from four DNA loci (ITS, cmdA, rpb2 and tub2). The generic concept of Dimorphiseta is broadened to include a third type of seta, i.e. thin-walled, straight with obtuse apices.

Keywords

Stachybotryaceae, soil fungi, turfgrass disease, multi-locus phylogeny, cup-shaped sporodochia

Introduction

Myrothecium was first introduced by Tode (1790) based on M. inundatum. The typical characters of these fungi are cup-shaped sporodochia covered by a mass of slimy, green to black conidia. The generic concept of Myrothecium has been emended several times (Link 1809; von Höhnel 1905; Pidoplichko and Kirilenko 1971). Decock et al. (2008) reported that the genus Myrothecium is not monophyletic based on internal transcribed spacer regions and the intervening 5.8S rDNA (ITS). Chen et al. (2015) re-evaluated the phylogeny of Myrothecium based on ITS and elongation factor 1-alpha (EF1-α) gene sequences, suggesting the polyphyly of Myrothecium within Stachybotryaceae. These studies did not make taxonomic conclusions accordingly. Lombard et al. (2016) constructed a backbone tree of Myrothecium s.l. based on a multi-locus phylogeny and resolved Myrothecium s.l. to 18 genera including 13 new genera introduced. Under the current concept of Myrothecium sensu stricto, only two species were included, M. inundatum and M. simplex (Lombard et al. 2016).

Most myrothecium-like species are saprobes in soils (Ellis and Ellis 1985). Many species were named referring to their substrates such as Alfaria terrestris, Albifimbria terrestris, Simorphiseta terrestris and Parvothecium terrestre. Some species were also reported as weak plant pathogens. For instance, Paramyrothecium roridum (syn. Myrothecium roridum) can infect coffee plants, causing bark canker (Tulloch 1972). Albifimbria verrucaria (syn. Myrothecium verrucaria) is pathogenic to mulberry causing leaf spot (Murakami et al. 2005). In addition, myrothecium-like species are also well-studied for their natural compounds, which are able to inhibit the activity of liver cancer and tumors (Pope 1944; Okunowo et al. 2010). Some myrothecium-like species can also produce a cocktail of secondary metabolites, which have strong antifungal and antibiotic activity (Kobayashi et al. 2004; Liu et al. 2006; Ruma et al. 2015). Hereto, more than 50 of these bioactive compounds have been reported from P. roridum and Alb. verrucaria (Wagenaar and Clardy 2001).

In a survey of turfgrass diseases from 2017, a number of myrothecium-like strains were collected from leaves and roots of turfgrasses and their rhizosphere. The aim of this study was to characterize these strains based on morphology and molecular phylogenetic analyses.

Materials and methods

Fungal isolates

From May 2017 to March 2018, turfgrass diseases were investigated on cold-season species in Beijing and on warm-season species in Hainan Province. Atotal of 130 samples were collected. Each sample was treated as an underground part of soil sample and a ground part of diseased grasses. Soil samples were isolated following the modified dilution plate method (Zhang et al. 2017). Five grams of each soil sample were suspended in 30 mL sterile water in a 50 mL bioclean centrifuge tube. The suspension was mixed thoroughly using Vortex-Genie 2 (Scientific Industries, New York) with maximum speed and then diluted to a series of concentration, i.e., 10-1, 10-2, 10-3 and 10-4. The 100 μL suspensions of each concentration were spread on to antibiotic potato dextrose agar (PDA, 4 g potato starch, 5 g dextrose and 15 g agar, 50 mg ampicillin and streptomycin sulfate in 1 L sterile water). The first few samples suggested that 10-2 was the best-diluted concentration for colony pickup. Diseased samples were isolated following a tissue isolation protocol (Chen et al. 2015). All plates were incubated at room temperature (23–25 °C) for 3–4 weeks, and from which all single colonies were picked up and transferred to clean PDA plates. Purified strains were stored at 4 °C for further studies. For phylogenetic analysis, associated sequences of 73 myrothecium-like strains and one outgroup strain were retrieved from GenBank (NCBI, https://www.ncbi.nlm.nih.gov/; Table 1).

Table 1.

Strains and NCBI GenBank accessions used in the phylogenetic analyses.

Species Isolate no. a Host/Substrate Country NCBI accession numbers
cmdA ITS tub2 rpb2
Myrothecium simplex CBS 582.93T Decaying agaric Japan KU846439 NR145079 KU846537
CBS 100287 Russula nigricans Japan KU846440 KU846457 KU846538
M. inundatum CBS 275.48T = IMI 158855 Russula adusta England KU846435 KU846452 KU846533
CBS 116539 Agaric Canada KU846437 KU846454 KU846535
Albifimbria lateralis CBS117712T Unknown USA KU845865 KU845881 KU845957 KU845919
Alb. terrestris CBS 126186T Soil in mopane woodlands Namibia KU845867 KU845883 KU845959 KU845921
CBS 109378 = NRRL 31066 Dead hardwood USA KU845866 KU845882 KU845958 KU845920
CBS 127838 Soil Namibia KU845868 KU845884 KU845960 KU845922
LC12196 rhizosphere soils of Poa sp. China MK500260 MK478879 MK500277
Alb. verrucaria CBS 328.52T = NRRL 2003 = ATCC 9095 deteriorated baled cotton USA KU845875 KU845893 KU845969 KU845931
CBS 189.46 = IMI 140060 Solanum tubersum Cyprus KU845872 KU845889 KU845965 KU845927
LC12191 Rhizosphere soils of Poa sp. China MK500255 MK478874 MK500272 MK500264
LC12192 Rhizosphere soils of Poa sp. China MK500256 MK478875 MK500273 MK500265
LC12193 Rhizosphere soils of Poa sp. China MK500257 MK478876 MK500274 MK500266
LC12194 Rhizosphere soils of Poa sp. China MK500258 MK478877 MK500276 MK500267
LC12195 Rhizosphere soils of Poa sp. China MK500259 MK478878 MK500275 MK500268
Alb. viridis CBS 449.71T = BCC 37540 Unknown India KU845879 KU845898 KU845974 KU845936
CBS 127346 Soil USA KU845880 KU845899 KU845975 KU845937
Alfaria. ossiformis CBS 324.54T Prairie soil USA KU845977 KU845984 KU846015 KU846002
Alf. humicola sp. nov. CGMCC3.19213T = LC12143 Rhizosphere soils of Poa sp. Beijing, China MH885432 MH793291 MH793317 MH818829
LC12144 Rhizosphere soils of Poa sp. Beijing, China MH885434 MH793293 MH793318 MH818830
Alf. poae sp. nov. CGMCC3.19198T = LC12140 Leaves of Poa sp. Hainan, China MH885419 MH793278 MH793314 MH818826
LC12141 Rhizosphere soils of Poa sp. Hainan, China MH885420 MH793279 MH793315 MH818828
LC12142 Rhizosphere soils of Poa sp. Hainan, China MH885421 MH793280 MH793316 MH818827
Alf. putrefolia CBS 112037T Rotten leaf Brazil KU845985 KU846016 KU846003
CBS 112038 Rotten leaf Brazil KU845986 KU846017 KU846004
Alf. terrestris CBS 477.91T Soil Turkey KU845979 KU845988 KU846019 KU846006
Alf. thymi CBS 447.83T Thymus serpyllum The Netherlands KU845981 KU845990 KU846021
Capitofimbria compacta CBS 111739T Decaying leaf Brazil KU846261 KU846287 KU846404 KU846349
MUCL 50238 Bark Zimbabwe KU878556 KU878559 KU878558
Dimorphiseta terrestris CBS 127345T Soil collected in tallgrass prairie USA KU846284 KU846314 KU846431 KU846375
D. acuta sp. nov. CGMCC3.19208T = LC12122 Rhizosphere soils of Poa pratensis Beijing, China MH885429 MH793288 MH818815
LC12123 Leaves of Digitaria sanguinalis Beijing, China MH885417 MH793276 MH793300 MH818811
LC12124 Leaves of Poa pratensis Beijing, China MH885418 MH793277 MH793297 MH818812
D. acuta sp. nov. LC12125 Rhizosphere soils of Poa pratensis Beijing, China MH885427 MH793286 MH793298 MH818813
LC12126 Rhizosphere soils of Poa pratensis Beijing, China MH885428 MH793287 MH793299 MH818814
LC12127 Rhizosphere soils of Poa pratensis Beijing, China MH885430 MH793289 MH793301 MH818820
D. obtusa sp. nov. CGMCC3.19206T = LC12128 Poa pratensis Beijing, China MH885426 MH793285 MH793307 MH818816
LC12129 Rhizosphere soils of Agrostis stolonifera Beijing, China MH885415 MH793274 MH793303 MH818821
LC12130 Rhizosphere soils of Poa pratensis Beijing, China MH885431 MH793290 MH793308 MH818817
LC12131 rhizosphere soils of Poa sp. Beijing, China MH885416 MH793275 MH793304
LC12132 Rhizosphere soils of Festuca arundinacea Beijing, China MH885422 MH793281 MH793305 MH818818
LC12133 Rhizosphere soils of Poa pratensis Beijing, China MH885423 MH793282 MH793306 MH818819
LC12134 Roots of Poa pratensis Beijing, China MH885424 MH793283 MH793309
LC12135 Roots of Poa pratensis Beijing, China MH885425 MH793284 MH793302
Gregatothecium humicola CBS 205.96T Soil Papua New Guinea KU846285 KU846315 KU846432 KU846376
Peethambara sundara CBS 646.77T Dead twig India KU846471 KU846551 KU846509
CBS 521.96 = MUCL 39093 Dead twig Nepal KU846470 KU846550 KU846508
Inaequalispora prestonii CBS 175.73T Forest soil Malaysia KU846286 KU846316 KU846433 KU846377
MUCL 52636 rhizoplane and roots of plants Ecuador KY389317 KY366447 KY389355
Myxospora masonii CBS 174.73T Leaves of Glyceria sp. England KU846445 KU846462 KU846543 KU846500
My. graminicola CBS 116538T Decaying grass leaf USA KU846444 KU846461 KU846542 KU846499
My. aptrootii CBS 101263T Leaf litter China KU846441 KU846458 KU846539 KU846496
My. musae CBS 265.71T Musa sp. Madagascar KU846463 KU846544 KU846501
CPC 25150 Tarspot lesion South Africa KU846446 KU846464 KU846545 KU846502
My. crassiseta CBS 731.83T Dead twig Japan KU846442 KU846459 KU846540 KU846497
CBS 121141 = NRRL 45891 Pyrenomycete Hawaii KU846443 KU846460 KU846541 KU846498
Paramyrothecium humicola CBS 127295T Soil collected in tallgrass prairie USA KU846295 KU846412 KU846356
P. parvum CBS 257.35T Viola sp. United Kingdom KU846298 KU846415 KU846359
CBS 142.422= IMI 155923= MUCL 7582 Dune sand France KU846268 KU846297 KU846414 KU846358
P. foeniculicola CBS 331.51T Foeniculum vulgare leaf sheath The Netherlands KU846292 KU846409 KU846354
P. nigrum CBS 116537T Soil Spain KU846267 KU846296 KU846413 KU846357
LC12188 Rhizosphere soils of Poa sp. China MK500252 MK478871 MK500269 MK500261
P. cupuliforme CBS 127789T Surface soil in desert Namibia KU846264 KU846291 KU846408 KU846353
P. viridisporum CBS 873.85T Soil Turkey KU846278 KU846308 KU846425 KU846369
CBS 125835 Soil USA KU846280 KU846310 KU846427 KU846371
P. acadiense CBS 123.96T Tussilago farfara Canada KU846288 KU846405 KU846350
P. terrestris CBS 564.86T Soil Turkey KU846273 KU846303 KU846420 KU846364
CBS 566.86 Soil Turkey KU846275 KU846305 KU846422 KU846366
P. tellicola CBS 478.91T Soil Turkey KU846272 KU846302 KU846419 KU846363
P. foliicola CBS 113121T Decaying leaf Brazil KU846266 KU846294 KU846411
CBS 419.93 Air Cuba KU846265 KU846293 KU846410 KU846355
P. breviseta CBS 544.75T Unknown India KU846262 KU846289 KU846406 KU846351
P. roridum CBS 357.89T Gardenia sp. Italy KU846270 KU846300 KU846417 KU846361
CBS 212.95 Water The Netherlands KU846269 KU846299 KU846416 KU846360
CBS 372.50 = IMI 140050 Coffea sp. Colombia KU846271 KU846301 KU846418 KU846362
P. guiyangense GUCC 201608S01T Soil Guiyang, China KY196193 KY126418 KY196201
HGUP 2016-8001 Soil Guiyang, China KY196192 KY126417 KY196200
P. verruridum
HGUP 2016-8006T Soil Guizhou, China KY196197 KY126422 KY196205
P. sinense sp. nov. CGMCC3.19212T = LC12136 Rhizosphere soils of Poa sp. Beijing, China MH885437 MH793296 MH793313 MH818824
LC12137 Rhizosphere soils of Poa sp. Beijing, China MH885436 MH793295 MH793312 MH818822
LC12138 Rhizosphere soils of Poa sp. Beijing, China MH885433 MH793292 MH793310 MH818823
LC12139 Rhizosphere soils of Poa sp. Beijing, China MH885435 MH793294 MH793311 MH818825
Parvothecium terrestre CBS 198.89T Soil in virgin forest Brazil KU846449 KU846468 KU846548 KU846506
Neomyrothecium humicola CBS 310.96T Soil Papua New Guinea KU846448 KU846467 KU846505
Gregatothecium humicola CBS 205.96T Soi Papua New Guinea KU846285 KU846315 KU846432 KU846376
Xepicula crassiseta CBS 392.71T Soil Spain KU847222 KU847247 KU847337 KU847296
X. jollymannii CBS 276.48T= MUCL 11830 Nicotiana tabacum Malawi KU847223 KU847248 KU847338 KU847297
CBS 126168 Soil Namibia KU847224 KU847250 KU847340 KU847298
X. leucotricha CBS 131.64= IMI 103664= ATCC 16686 Soil India KU847225 KU847251 KU847341 KU847299
CBS 483.78 Soil Colombia KU847228 KU847254 KU847344 KU847302
Smaragdiniseta bisetosa CBS 459.82T Rotten bark India KU847206 KU847229 KU847319 KU847281
Striaticonidium brachysporum CBS 513.71 T = IMI 115293 Dune sand Iran KU847209 KU847232 KU847322 KU847284
S. brachysporum CBS 131.71= IMI 158441= ATCC 22270 Soil Ukrain KU847207 KU847230 KU847320 KU847282
LC12189 Rhizosphere soils of Poa sp. Beijing, China MK500253 MK478872 MK500270 MK500262
LC12190 Rhizosphere soils of Poa sp. Beijing, China MK500254 MK478873 MK500271 MK500263
S. synnematum CBS 479.85T Palm leaf Japan KU847218 KU847242 KU847332 KU847292
S. cinctum CBS 932.69T Soil The Netherlands KU847216 KU847239 KU847329 KU847290
CBS 277.48 = IMI 001526 Soil New Zealand KU847213 KU847236 KU847326 KU847288
S. humicola CBS 388.97 Soil Papua New Guinea KU847217 KU847241 KU847331 KU847291
Tangerinosporium thalictricola CBS 317.61T = IMI 034815 Thalictrum flavum UK KU847219 KU847243 KU847333
Xenomyrothecium tongaense CBS 598.80T Halimeda sp. Tonga KU847221 KU847246 KU847336 KU847295
Virgatospora echinofibrosa CBS 110115 Theobroma cacao Ecuador KU847220 KU847244 KU847334 KU847293
MUCL 39092 = ATCC 200437 Trewia nudiflora Nepal KU847245 KU847335 KU847294
Fusarium sambucinum CBS 146.95 Solanum tuberosum UK KM231391 KM231813 KM232078 KM232381

Morphology and culture characteristics

Descriptions of macromorphological features are based on 7-d old materials incubated in the dark at room temperature (20–25 °C) and grown on potato dextrose agar (2% w/w; PDA), oatmeal agar (OA), cornmeal agar (CMA) and synthetic low-nutrient agar (SNA; Nirenberg 1981). Color description followed the color guide by Kornerup and Wanscher (1978). Digital images of colonies were made with a Nikon Eclipse 80i light microscope (Tokyo, Japan) with differential interference contrast (DIC) illumination and a LV2000 digital camera (Beijing, China). Slides mounted in clear lactic acid were also prepared to observe conidiogenesis, conidiophores and conidia.

DNA extraction and PCR amplification

Genomic DNA was extracted from 1–2 weeks’ old cultures grown on potato dextrose agar (2% w/w; PDA) incubated at room temperature using a modified Cetyltrimethyl Ammonium Bromide (CTAB) method (Rogers and Bendich 1994). Partial sequences of four genes, ITS, RNA polymerase II second largest subunit (rpb2), β-tubulin (tub2) and calmodulin (cmdA) gene sequences were amplified using the following pairs of primers, ITS1 and ITS4 (White et al. 1990) for ITS, RPB2-5F2 and RPB2-7cR (O’Donnell et al. 2007) for rpb2, Bt2a and Bt2b (Glass and Donaldson 1995) for tub2 and CAL-228F (Carbone and Kohn 1999) and CAL2Rd (Groenewald et al. 2013) for cmdA. Amplification for each locus followed the PCR protocols as described in Lombard et al. (2016). The PCR was performed in a 25 μL reaction volume including 2.5 μL 10 × PCR Buffer (Dingguo, Beijing, China), 2 mM MgCl2, 50 μM dNTPs, 0.1 μM of each primer, 0.5 U Taq DNA polymerase and 10 ng genomic DNA. PCR reactions were conducted in ProFlexTM PCR system (Applied Biosystems, California, USA) under the following reaction conditions: predenaturation at 94 °C for 5 min, followed by 35 cycles of denaturation at 94 °C for 30 s, annealing at 52 °C (for ITS) or 54 °C (for rpb2 and cmdA) or 56 °C (for tub2) for 40 s and elongation at 72 °C for 1 min, a final elongation at 72 °C for 5 min.

The purified PCR products were sequenced in both forward and reverse directions on an ABI-3730 XL DNA Analyzer (Applied Biosystems, California, USA). The sequences were checked and manually corrected where necessary. A consensus contig was assembled with BioEdit v. 7.0.9 (Hall 1999) and the reference sequences were downloaded from GenBank (Table 1). Sequences were aligned with MAFFT v. 7 (Kazutaka and Standley 2013) and manually trimmed to equal length by cutting the unaligned sequences at both ends.

Phylogenetic analyses

Phylogenetic analyses were based on Bayesian inference (BI) and Maximum Likelihood (ML). For BI analysis, the optimal evolutionary model was estimated in MrModeltest v. 2.3 (Nylander 2004) using the Akaike Information Criterion (AIC) for each locus. For the selected substitution models for each locus see Table 2. MrBayes v. 3.2.1 (Ronquist and Huelsenbeck 2003) was used to generate tree topology and a Markov Chain Monte Carlo (MCMC) algorithm of four chains was started with a random seed and a burn in of first 25% trees. The MCMC analysis lasted until the average standard deviation of split frequencies came below 0.01. The ML analysis was performed using RAxML servers (http://phylobench.vital-it.ch/raxml-bb/index.php), with a maximum likelihood bootstrap (LB) of 1,000 replicates, under the GTR-GAMMA model (Stamatakis 2006).

Results

In this study, 603 fungal strains were isolated. Based on colony morphologies and preliminary sequence comparison of ITS via BLASTn in GenBank, 84 myrothecium-like strains were selected. Phylogenetic analyses of above 84 strains were performed on single locus and concatenated datasets (ITS, cmdA, tub2 and rpb2), with 70 strains in Myrothecium s.l. as reference and Fusarium sambucinum (CBS 146.95) as outgroup. After alignment, the concatenated datasets of four loci contained 569 characters (with gaps) for ITS, 318 for tub2, 732 for cmdA and 724 for rpb2. The characters of different alignments and statistics of phylogenetic analyses were shown in Table 2. The four single locus trees of all strains showed essentially similar topology (Supp. materials 1–4), with only minor differences affecting unsupported nodes on the trees. The resulting multi-locus ML tree was presented in Fig. 1 together with BI posterior probability values. Among 84 myrothecium-like strains, 14 strains were identified as four known species, Albifimbria verrucaria (10 strains), Alb. terrestris (1 strain), Striaticonidium brachysporum (2 strains) and Paramyrothecium nigrum (1 strain). The rest of them were grouped into five distinct clades with high supported values. Based on the morphological and phylogenetic distinctions, five novel species (i.e. Alfaria humicola, Alf. poae, Dimorphiseta acuta, D. obtusa and Paramyrothecium sinense) were described in this paper.

Figure 1. 

The ML consensus tree inferred from a four-locus concatenated alignment (ITS, cmdA, rpb2 and tub2). Bootstrap values (1,000 replicates) over 70% for ML and posterior probability (PP) over 0.95 are added to the left of a node (ML/PP). The type strains are labeled with “T”. Strains obtained from this study are in red. The tree is rooted using Fusarium sambucinum (CBS 146.95).

Table 2.

Characteristics of the different datasets and statistics of phylogenetic analyses used in this study.

Locus† Number of sites* Evolutionary model‡ Number of tree sampled in B Maximum-likelihood statistics
Total Conserved Phylogenetically informative B unique patterns Best tree optimised likelihood Tree length
ITS 569 334 193 247 GTR+I+G 7501 -32666.73 5.36
tub2 318 168 140 159 HKY+I+G
cmdA 732 258 381 490 HKY+I+G
rpb2 724 360 367 367 GTR+I+G

Taxonomy

Dimorphiseta L. Lombard & Crous., Persoonia. 36: 188. 2016. emend. J.M.Liang & L.Cai.

Dimorphiseta terrestris L. Lombard & Crous. Persoonia. 36: 188. 2016. (Type species)

Note

Dimorphiseta was a monotypic genus, introduced based on D. terrestris, which showed both type I (thin-walled, flexuous to circinate, narrowing to a sharp apex) and type II (thick-walled, straight to slightly curved, narrowing to a sharp apex) setae. Our study demonstrated that there is a third type of setae (type III: thin-walled, straight, terminating in an obtuse apex) in the genus.

Dimorphiseta acuta J.M. Liang, G.S. Li & L. Cai, sp. nov.

MycoBank No: 829693
Fig. 2

Type

China, Beijing, isolated from rhizosphere soils of Poa pratensis, 26 Aug 2017, J.M. Liang, holotype HMAS 247957, dried culture on PDA, ex-holotype culture CGMCC3.19208 = LC12122.

Description

Colonies on PDA, CMA and OA approx. 7–8 cm diam. after 7 d at room temperature (approx. 25 °C), mycelium white and abundant, with conidiophores forming on the aerial mycelium, carrying slimy olivaceous green to black conidial masses, reverse on PDA buff. Conidiomata sporodochial, stromatic, superficial, cupulate to discoid, scattered, rarely gregarious, irregular in outline, 50–300 μm diam., 60–150 μm deep, consisting of bundles of parallel, longitudinal, closely compacted hyphae, terminating in whorls of 3–5 conidiogenous cells, covered by an olivaceous green to black slimy mass of conidia without marginal hyphae. Stroma poorly developed, hyaline, of a textura angularis. Setae arising from the conidial mass, thick-walled, subhyaline, smooth, 5–15-septate, tapering to sharp apices, 120–370 μm long, 10–13 μm wide at the broadest part, 2–4 μm wide at the apex. Conidiophores macronematous, irregularly, unbranched, smooth to lightly verrucose, arising from the basal stroma. Conidiogenous cells phialidic, subcylindrical, hyaline, smooth, 10–20 μm long, 2–3 μm wide. Conidia aseptate, smooth, hyaline, ellipsoidal, rounded at the base, pointed at the apex with a funnel-shaped appendage, 7–12 × 2–3 μm (av. 10 ± 0.7 × 3 ± 1.3 μm, n = 50).

Figure 2. 

Dimorphiseta acuta (from ex-type strain CGMCC3.19208) a–c colony on PDA, CMA, OA d conidiomata on SNA e conidiophores f conidiogenous cells g setae h–k conidia. Scale bars: 5 μm (e, f, h): 50 μm (g); 2 μm (i, j, k).

Distribution

China.

Etymology

Name refers to the setae with tapered and sharp apices.

Additional isolates examined

China, Beijing, from leaves of Digitaria sanguinalis, 21 Aug 2017, J.M. Liang, LC12123; China, Beijing, from leaves of Poa pratensis, 21 Aug 2017, J.M. Liang, LC12124; China, Beijing, from rhizosphere soils of P. pratensis, 21 Aug 2017, J.M. Liang & G.S. Li, LC12125, 21 Jul 2017, J.M. Liang, LC12126, 25 Jul 2017, J.M. Liang, LC12127.

Notes

The multi-locus phylogenetic analyses indicated that D. acuta formed a sister clade to D. terrestris, but differs from the latter in the type and size of setae. Dimorphiseta terrestris produces both types of setae, the thin-walled and circinate type (Type I) and the thick-walled sharp-edged type (Type II), whereas D. acuta only produces the type I setae. In addition, the setae of D. acuta are much longer and wider than that in D. terrestris (120–370 μm × 10–13 μm vs. 70–95 × 3–4 μm) (Lombard et al. 2016). Morphologically, D. acuta should also be compared with M. miconiae and M. xigazense, which also produce sharp-edged setae. Myrothecium miconiae, however, differs from D. acuta in producing 1-septate conidia (Alves et al. 2010), while M. xigazense differs in producing conidia that are truncate at both ends (Wu et al. 2014).

Dimorphiseta obtusa J.M. Liang, G.S. Li & L. Cai, sp. nov.

MycoBank No: 829694
Fig. 3

Type

China, Beijing, isolated from rhizosphere soils of P. pratensis, 23 Jun 2017, J.M. Liang, holotype HMAS 247954, ex-holotype culture CGMCC3.19206 = LC12128.

Description

Colonies on PDA, OA and CMA approx. 5–6 cm diam. after 7 d at room temperature (approx. 25 °C), mycelium white and abundant, with conidiophores forming on the aerial mycelium, carrying slimy olivaceous green to black conidial masses, reverse on PDA pale luteous to buff. Conidiomata sporodochial, stromatic, superficial, scattered, rarely gregarious, oval to elongate or irregular in outline, 60–280 µm diam., 40–120 µm deep, with a setose fringe surrounding green to black slimy mass of conidia. Stroma poorly developed, hyaline, smooth to verrucose, of textura angularis. Setae arising from the basal stroma, thin-walled, 3–6-septate, unbranched, hyaline, smooth, 80–250 µm long, 2–4 µm wide at the broadest, terminating in a blunt apex. Conidiophores macronematous, irregularly, unbranched, smooth to lightly verrucose, arising from the basal stroma, up to 18 μm long. Conidiogenous cells phialidic, hyaline, smooth to verrucose, cylindrical, 7–19 × 2–3 μm, becoming narrowed at the tip with collarette. Conidia aseptate, ellipsoidal or cylindrical, hyaline, smooth, rounded both ends, with a funnel-shaped apical appendage, 9–11 × 2–4 μm (av. 10 ± 0.5 × 3 ± 0.3 μm, n = 50).

Figure 3. 

Dimorphiseta obtusa (from ex-type strain CGMCC3.19206) a–c colony on PDA, CMA, OA d conidioma on SNA e setae f conidiophores g conidiogenous cells h–k conidia. Scale bars: 50 μm (e); 10 μm (f, g); 5 μm (h); 2 μm (i, j, k).

Distribution

China.

Etymology

Named refers the setae with obtuse apices.

Additional isolates examined

China, Beijing, from rhizosphere soils of Agrostis stolonifera, 24 Jul 2017, J.M. Liang, LC12129; China, Beijing, from rhizosphere soils of P. pratensis, 25 Aug 2017, J.M. Liang & G.S. Li, LC12130, 19 Jul 2017, J.M. Liang, LC12133; China, Beijing, from rhizosphere soils of Poa sp., 19 Jul 2017, J.M. Liang, LC12131; China, Beijing, from rhizosphere soils of Festuca arundinacea, 19 Jul 2017, J.M. Liang, LC12132; China, Beijing, from leaves of P. pratensis, 23 Jun 2017, J.M. Liang, LC12134, LC12135.

Notes

Dimorphiseta obtusa formed a highly supported cluster with D. terrestris and D. acuta, but can be distinguished from the latter two by having setae with erect and obtuse apices. In addition, D. obtusa is also morphologically similar to two old un-sequenced Myrothecium taxa, i.e. M. biforme and M. dimorphum, but both of these two taxa have two types of conidia. Myrothecium biforme produces short cylindrical and ellipsoidal to navicular conidia (Jiang et al. 2014) and M. dimorphum has ovate and ellipsoidal conidia (Watanabe et al. 2003).

Alfaria humicola J.M. Liang, G.S. Li & L. Cai, sp. nov.

MycoBank No: 829696
Fig. 4

Type

China, Beijing, Olympic Park, from rhizosphere soil of Poa sp., 13 Dec 2017, S.Y. Zhou, holotype HMAS 247955, ex-holotype culture CGMCC3.19213 = LC12143.

Description

Colonies on PDA, CMA and OA approx. 7–8 cm diam. after 7 d at 25 °C. Hyphae hyaline, smooth, branched, 1–2 μm wide. Conidiomata sporodochial, stromatic, superficial, cupulate to discoid, scattered to gregarious, oval to elongate or irregular in outline, 50–200 μm diam., 70–150 μm deep, without setose hyphae, covered by a green to black agglutinated slimy mass of conidia. Stroma well-developed, hyaline, of textura globulose or textura angularis. Setae absent. Conidiophores arising from the basal stroma, unbranched or branched, initially hyaline and smooth, becoming pigmented and verrucose with age, 11–25 µm long. Conidiogenous cells phialidic, cylindrical to allantoid, initially hyaline and smooth becoming pigmented and verrucose with age, 14–33 × 2–3 µm. Conidia aseptate, smooth, hyaline, elongated ellipsoidal to limoniform, straight, 7–9(–10) × 2–3 µm (av. 8 ± 0.6 × 3 ± 0.2 µm, n = 50).

Figure 4. 

Alfaria humicola (from ex-type CGMCC3.19213) a–c colony on PDA, CMA, OA d conidiomata on SNA e sporodochial conidioma, arrows showing branched conidiosphores and conidiogenous cells f conidia. Scale bars: 10 µm (e); 5 µm (f).

Distribution

China.

Etymology

Name refers the substrate, soil, from which this fungus was isolated.

Additional isolate examined

China, Beijing, Olympic Park, from rhizosphere soil of Poa sp., 13 Dec 2017, S.Y. Zhou, LC12144.

Notes

Alfaria humicola represents another distinct lineage in Alfaria (Fig. 1). Alfaria humicola lacks setae, distinguishing it from Alf. caricicola and Alf. thymi. Furthermore, the conidiogenous cells of Alf. humicola (14–33 × 2–3 µm) are much longer than that of Alf. arenosa (5–10 × 1–2 µm), Alf. ossiformis (5–10 × 2–3 µm) and Alf. terrestris (5–11 × 1–3 µm). Compared with those old Myrothecium taxa lacking sequences, Alf. humicola is morphologically similar to M. atrocarreum (Berkeley & Broome, 1877), M. conicum (Fuckel, 1870), M. ellipsosporum (Fuckel, 1866), M. fragosianum (Saccardo, 1917), M. leucomelas (Höhnel, 1925) and M. oryza (Saccardo, 1917), but Alf. humicola produces limoniform conidia which makes it distinguishable. In addition, the conidiogenous cells of Alf. humicola show conspicuous collarettes which were not described in previous old taxa.

Alfaria poae J.M. Liang, G.S. Li & L. Cai, sp. nov.

MycoBank No: 829697
Fig. 5

Type

China, Hainan Province, Haikou, isolated from leaves of Imperata cylindrica, 10 Mar 2018, J.M. Liang and L. Cai, holotype HMAS 247953, ex-holotype culture CGMCC3.19198 = LC12140.

Description

Colonies on PDA, CMA and OA with white aerial mycelium, approx. 6–7 cm diam. after 7 d at 25 °C, giving rise to dark green or blank sporodochia scattered or gregarious on the surface, covered by olivaceous green pillars of conidia, reverse on PDA sienna. Hyphae hyaline, smooth, branched, 1–2 μm wide. Conidiomata synnematous, solitary, 60–250 μm high, 30–80 μm wide at the base, 60–150 μm at the apex, with setose hyphae surrounding a green agglutinated mass of conidia. Stroma well developed, hyaline, of textura angularis. Setae absent. Conidiophores arising from the basal stroma, branched, initially hyaline and becoming pigmented and verrucose with age covered by an olivaceous green mucoid layer, up to 30 µm long. Conidiogenous cell phialidic, clavate to cylindrical, hyaline, smooth, 5–10 × 1–2 µm, becoming pigmented and verrucose with age, with conspicuous collarettes and periclinal thickenings. Conidia aseptate, smooth, hyaline, ellipsoidal to fusiform, 6–8 × 2–3 µm (av. 7 ± 0.4× 2 ± 0.2 µm, n = 50).

Figure 5. 

Alfaria poae (from ex-type strain CGMCC3.19198) a–c colony on PDA, CMA, OA d–e conidiomata on SNA f synnematous conidioma g conidiogenous cells, the arrow showing conspicuous collarette h aged conidiophores i conidia. Scale bars: 50 μm (f); 5 μm (g); 10 μm (h, i).

Distribution

China.

Etymology

Name refers the host, Poa sp., from which this fungus was isolated.

Additional isolate examined

China, Hainan, from leaves of Imperata cylindrica, 10 Mar 2018, J.M. Liang & Lei Cai, LC12141, LC12142.

Notes

Alfaria poae formed a well-supported clade in Alfaria (Fig. 1). Similar to Alf. ossiformis and Alf. terrestris, Alf. poae does not produce setae surrounding the sporodochia, distinguishing it from Alf. caricicola and Alf. thymi. Alfaria poae produces ellipsoidal to fusiform conidia, which are different from the ossiform conidia produced by Alf. ossiformis (Lombard et al. 2016). The conidia of Alf. terrestris have basal hilum which was not observed in Alf. poae. In addition, Alf. poae shares morphological characters with several un-sequenced Myrothecium taxa, such as M. atrocarneum (Berkeley & Broom, 1877), M. conicum (Fuckel, 1870), M. ellipsosporum (Fuckel, 1866) and M. leucomelas (Höhnel, 1925). Because the descriptions of M. atrocarneum, M. conicum and M. ellipsosporum were not elaborate enough, these old species are not distinct from Alf. poae yet. Future comparisons should be made when these old species are epitypified by fresh collections. Although M. leucomelas (host: Sumbaviae rotttleroidis; location: Bulacan, Luzon) had a detailed description, it cannot be epitypified by Alf. Poae, because Alf. poae was collected from a distinct location and plant host. Taking the above special characters into account, we considered introducing a new species, Alfaria poae.

Paramyrothecium sinense J.M. Liang, G.S. Li & L. Cai, sp. nov.

MycoBank No: 829698
Fig. 6

Type

China, Beijing, Olympic Park, from rhizosphere soil of Poa sp., 13 Dec 2017, S.Y. Zhou, holotype HMAS 247956, ex-holotype culture CGMCC3.19212 = LC12136.

Description

Colonies on PDA, CMA and OA approx. 5–6 cm diam. after 7 d at 25 °C. Hyphae white, hyaline, smooth, branched, 1–2 μm wide, reverse on PDA pale luteous. Conidiomata sporodochial, stromatic, cupulate, superficial, scattered or gregarious, oval or irregular in outline, 80–600 μm diam., 50–150 μm deep, with a white setose fringe surrounding an olivaceous green to black agglutinated slimy mass of conidia. Stroma poorly developed, hyaline, of textura angularis. Setae arising from stroma, thin-walled, hyaline, 1–3-septate, straight to flexuous, 45–90 μm long, 1–3 μm wide, tapering to an acutely rounded apex. Conidiophores arising from the basal stroma, consisting of a stipe and a penicillately branched conidiogenous apparatus; stipes unbranched, hyaline, septate, smooth, 20–30 × 2–3 μm; primary branches aseptate, unbranched, smooth, 13–40 × 2–3 μm; secondary branches aseptate, unbranched, smooth, 8–15 × 2–3 μm; terminating in a whorl of 3–6 conidiogenous cells; conidiogenous cell phialidic, cylindrical to subcylindrical, hyaline, smooth, straight to slightly curved, 7–16 × 1–3 μm, with conspicuous collarettes and periclinal thickenings. Conidia aseptate, hyaline, smooth, cylindrical, 6–7 × 2–3 μm (av. 7 ± 0.3 × 2 ± 0.2 μm, n = 40), rounded at both ends.

Figure 6. 

Paramyrothecium sinense (from ex-type CGMCC3.19212) a–c colony on PDA, CMA, OA d conidiomata on SNA e sporodochial conidioma f setae g conidia h conidiogenous cells. Scale bars: 20 μm (e, f) ; 10 μm (g); 5 μm (h).

Distribution

China.

Etymology

Named after the country of collection, China.

Additional isolate examined

China, Beijing, Olympic Park, from rhizosphere soils of Poa sp., 13 Dec 2017, S.Y. Zhou, LC12137, LC12138, LC12139.

Notes

Lombard et al. (2016) introduced a new genus, Paramyrothecium, based on an epitype of Myrothecium roridum Tode, 1790. Gams (2016) pointed out that Myrotheciella catenuligera, the type species of Myrotheciella was listed as a synonym of P. roridum by Lombard et al. (2016), thus Paramyrothecium is illegitimate and Myrotheciella should be the correct name for Paramyrothecium. However, the original description of Myrotheciella catenuligera suggested that it lacks seta (Spegazzini 1911), thus is clearly different from the morphological circumscription of P. roridum. Therefore, we do not agree with the treatment of Lombard et al. (2016) of listing Myrotheciella catenuligera as a synonym of P. roridum.

Paramyrothecium sinense formed a highly supported distinct clade closely related to P. humicola. The setae of this species are terminated with obtuse apices, dissimilar to the acute apices in P. humicola. In addition, the conidiophore stipes (20–30 μm long) and primary branches (13–40 μm long) of P. sinense are much longer than those of P. humicola (stipe, 12–22 μm long; primary branches, 7–17 μm long) (Lombard et al. 2016). Among old un-sequenced taxa in Myrothecium, only M. biforme and M. dimorphum show seta with obtuse apices, but both taxa produce two types of conidia (Jiang et al. 2014; Watanabe et al. 2003).

Discussion

The ITS has been shown to be insufficient to delineate the myrothecium-like species. With the additions of partial sequences of rpb2, cmdA and tub2, phylogenetic relationships within Stachybotryaceae could be better resolved (Lombard et al. 2016). In this study, we isolated fungi from rhizosphere soils, leaves and roots of several turfgrasses, and our phylogenetic analyses based on concatenated four loci together with the morphological characters supported the recognition of five novel species in Stachybotryaceae.

By comparing the topologies of the four single-locus trees, incomplete lineage sorting was discovered in Dimorphiseta. Based on the single-locus trees of ITS and rpb2, D. acuta, D. obtusa and D. terrestris grouped together (Supp. materials 1, 4). Whereas in the single-locus phylogenetic analyses based on tub2 and cmdA, D. obtusa grouped distantly from D. acuta and D. terrestris, but close to Myxospora and Albifimbria species (Supp. materials 2, 3). Three Dimorphiseta species are similar in the conidial shape and size (7–19 μm long), which are distinct from the shorter conidia in Albifimbria (4–8 μm long) and Myxospora (4–6 μm long) species (Tulloch 1972; Lombard et al. 2016). Conidia with a funnel-shaped apical appendage are a distinct feature of three Dimorphiseta species, but they are absent in all Myxospora species and most Albifimbria species (Lombard et al. 2016). Furthermore, the rpb2 and 28S ribosomal DNA combined dataset, which was suggested to delimit generic boundaries of myrothecium-like species (Lombard et al. 2016) revealed that the three Dimorphiseta species clustered together (Supp. material 6: Table S1, Supp. material 5).

In the multi-locus sequence analysis of Myrothecium s.l. by Lombard et al. (2016), thirteen new genera were introduced including several monotypic genera, such as Dimorphiseta, Capitofimbria, Gregatothecium and Neomyrothecium. In this study, we reported two new species in Dimorphiseta (D. acuta and D. obtusa). With this addition, the generic concept of Dimorphiseta is slightly expanded for including a third type of setae. Hereto, Dimorphiseta is the genus with the most variable types of seta among Myrothecium s.l., which might be useful in the generic delimitation in Myrothecium s.l. (Lombard et al. 2016).

Lombard et al. (2016) narrowed the concept of Myrothecium s.s. to only include species with sporodochia or mononematous conidiophores producing conidia shorter than 5 μm in green slimy masses without mucoid appendages. Whether or not a conidial size should be defined in the generic concept remained debatable. Because many Myrothecium published recently produced much longer conidia, e.g. M. chiangmaiense (4–7 μm) (Dai et al. 2017), M. uttaraditense (10–15 μm) (Dai et al. 2017), M. thailandicum (6.5–10 μm) (Dai et al. 2017), M. septentrionale (8.5–12 μm) (Tibpromma et al. 2017), M. variabile (12.5–16.5 μm) (Wu et al. 2014) and M. xigazense (2.5–15 μm) (Wu et al. 2014). These above species were identified, either based on morphology only or with a single molecular locus (ITS), and should be better confirmed for their generic placement when more data are available. Currently, there are 90 records of Myrothecium in Index Fungorum (Jan 10, 2019), and 25 names have been successively transferred to other genera, i.e., Capitofimbria, Melanconis, Striaticonidium, Xepicula (Lombard et al. 2016), Digitiseta (Gordillo and Decock 2018). Only a limited number of the remaining species in Myrothecium have available molecular data (Dai et al. 2017; Tibpromma et al. 2017), as most of these taxa have no living cultures. We agree with Gams (2016) that these unvisited taxa are still important when the original descriptions are sufficiently clear to recognize a species. They should be epitypified in future studies when fresh collections with living cultures are available, and before that, descriptions of new taxa in this group should be made carefully with the inclusion of these un-sequenced taxa in morphological comparisons.

Acknowledgements

This study was financially supported by National Natural Science Foundation of China (NSFC 31600405).

References

  • Alves JL, Barreto RW, Pereira OL, Soares DJ (2010) Additions to the mycobiota of the invasive weed Miconia calvescens (Melastomataceae). Mycologia, 102(1): 69–82. https://doi.org/10.3852/09-070
  • Carbone I, Kohn LM (1999) A method for designing primer sets for speciation studies in filamentous ascomycetes. Mycologia 91: 553–556. https://doi.org/10.2307/3761358
  • Decock C, Huret S, Bivort C (2008) Anamorphic fungi from French Guyana. Septomyrothecium sp. nov. and S. setiramosum comb. nov. (anamorphic Hypocreales, Ascomycota). Cryptogamie Mycologie 29: 321–331. https://doi.org/10.1093/ml/gcm091
  • Ellis MB, Ellis JP (1985) Microfungi on Land Plants-An Identification Handbook. Bulletin of the Torrey Botanical Club 113: 61. https://doi.org/10.2307/2996241
  • Fuckel L (1866) Fungi Rhenani exsiccati Cent. 12–17 (2), no 1450–1632. Hedwigia. 5: 23–30.
  • Fuckel L (1870) Symbolae mycologicae. Beiträge zur Kenntniss der Rheinischen Pilze. Jahrbücher des Nassauischen Vereins für Naturkunde. 23–24: 1–459.
  • Glass NL, Donaldson G (1995) Development of primer sets designed for use with PCR to amplify conserved genes from filamentous ascomycetes. Applied and Environmental Microbiology 61: 1323–1330. https://doi.org/10.0000/PMID7747954
  • Gams W (2016) Are old taxa without living authenticated cultures losing their status? IMA Fungus. 7(2): 72–73.
  • Gordillo A, Decock C (2018) Myrothecium-like (Ascomycota, Hypocreales) species from tropical areas: Digitiseta gen. nov. and additions to Inaequalispora and Parvothecium. Mycological Progress 17: 179–190. https://doi.org/10.1007/s11557-017-1302-4
  • Groenewald JZ, Nakashima C, Nishikawa J, Shin HD, Park JH, Jama AN, Groenewald M, Braun U, Crous PW (2013) Species concepts in Cercospora: spotting the weeds among the roses. Studies in Mycology 75: 115–170. https://doi.org/10.3114/sim0012
  • Höhnel (1925) Mitt. Bot. Inst. Techn. Hochsch. Wien 2(3): 96.
  • Jiang YL, Wang HF, Pan HQ, Zhang TY (2014) Myrothecium (Hyphomycetes): three new species, one new variety and a key to species and varieties of the genus known from soils in China. Mycosystema, 33(1): 7–14.
  • Kazutaka K, Standley DM (2013) MAFFT Multiple Sequence Alignment Software Version 7: Improvements in Performance and Usability. Molecular Biology and Evolution 30: 772–780. https://doi.org/10.1093/molbev/mst010
  • Kobayashi M, Sato I, Abe F, Nitta K, Hashimoto M, Fujie A, Hino M (2004) FR227244, a novel antifungal antibiotic from Myrothecium cinctum No. 002 I. Taxonomy, fermentation, isolation and physio-chemical properties. Journal of Antibiotics 57: 780–787. https://doi.org/10.7164/antibiotics.57.788
  • Kornerup A, Wanscher JH (1978) Methuen Handbook of Colour. Methuen.
  • Link HF (1809) Observationes in ordines plantarum naturales. Dissertatio I.3: 3–42.
  • Liu JY, Huang LL, Ye YH, Zou WX, Guo ZJ, Tan RX (2006) Antifungal and new metabolites of Myrothecium sp. Z16, a fungus associated with white croaker Argyromosum argentatus. Journal of Applied Microbiology 100: 195–202. https://doi.org/10.1111/j.1365-2672.2005.02760.x
  • Lombard L, Houbraken J, Decock C, Samson R.A, Meijer M, Réblová M, Groenewald JZ, Crous PW (2016) Generic hyper-diversity in Stachybotriaceae. Persoonia 36: 156–246. https://doi.org/10.3767/003158516X691582
  • Murakami R, Kobayashi T, Takahashi K (2005) Myrothecium leaf spot of mulberry caused by Myrothecium verrucaria. Journal of General Plant Pathology 71: 153–155. https://doi.org/10.1007/s10327-004-0178-8
  • Nirenberg HI (1981) A simplified method for identifying Fusarium spp. occurring on wheat. Canadian Journal of Botany 59: 1599–1609. https://doi.org/10.1139/b81-217
  • Nylander JAA (2004) MrModelTest (program distributed by the author). Evolutionary Biology Centre, Uppsala University.
  • O’Donnell K, Sarver BA, Brandt M, Chang DC, Noble-Wang J, Park BJ, Sutton DA, Benjamin L, Lindsley M, Padhye A, Geiser DM, Ward TJ (2007) Phylogenetic diversity and microsphere array-based genotyping of human pathogenic Fusaria, including isolates from the multistate contact lens-associated U.S. keratitis outbreaks of 2005 and 2006. Journal of Clinical Microbiology 45: 2235–2248. https://doi.org/10.1128/JCM.00533-07
  • Okunowo WO, Gbenle GO, Osuntoki AA, Adekunle AA, Ojokuku SA (2010) Production of cellulolytic and xylanolytic enzymes by a phytopathogenic Myrothecium roridum and some avirulent fungal isolates from water hyacinth. African Journal of Biotechnology 9: 1074–1078. https://doi.org/10.5897/AJB09.1598
  • Pidoplichko NM, Kirilenko TS (1971) On the taxonomy of the genus Myrothecium. In: Pidoplichko NM (Ed.) Metabolites of soil micromycetes. Dumka, Naukova, Kiev, Ukrain, 157–171.
  • Rogers SO, Bendich AJ (1994) Extraction of total cellular DNA from plants, algae and fungi. In: Gelvin SB, Schilperoort RA (Eds) Plant Molecular Biology Manual. Springer, Dordrecht, 183–190. https://doi.org/10.1007/978-94-011-0511-8_12
  • Ruma K, Sunil K, Kini KR, Prakash HS (2015) Genetic diversity and antimicrobial activity of endophytic Myrothecium spp. isolated from Calophyllum apelatum and Garcinia morella. Molecular Biology Reports 42: 1533–1543. https://doi.org/10.1007/s11033-015-3884-8
  • Saccardo PA (1917) Notae mycologicae series XXIII. Fungi Philippinenses. Atti della Accademia Scientifica Veneto-Trentino-Istriana. 10: 57–94.
  • Spegazzini C (1911) Mycetes Argentinenses (Series V). Anales del Museo Nacional de Historia Natural Buenos Aires. ser. 3, 13: 329–467.
  • Tibpromma S, Hyde KD, Jeewon R, Maharachchikumbura SSN, Liu JK, Bhat DJ et al. (2017) Fungal diversity notes 491–602: taxonomic and phylogenetic contributions to fungal taxa. Fungal Diversity 83: 1–261 https://doi.org/10.1007/s13225-017-0378-0
  • Tulloch M (1972) The genus Myrothecium Tode ex Fr. Mycological Papers 130: 1–42.
  • Von Höhnel FV (1905) Über Myrothecium und Formverwandte Gattungen. Annales Mycologici 3: 559–560.
  • Watanabe T, Watanabe Y, Nakamura K (2003) Myrothecium dimorphum sp. nov. a soil fungus from beach sand in the Bonin (Ogasawara) Islands, Japan. Mycoscience, 44(4): 283–286. https://doi.org/10.1007/s10267-003-0112-5
  • White TJ, Burns T, Lee S, Taylor F, White TJ, Lee S-H, Taylor L, Shawe-Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis MA, Gelfand DH, Sninsky JJ et al. (Eds) PCR protocols: a guide to methods and applications: 282–287. https://doi.org/10.1016/B978-0-12-372180-8.50042-1
  • Wu YM, Jiang YL, Ma YN, Zhang TY (2014) Two new species of Myrothecium from the Qinghai-Tibet Plateau Area, China. Mycotaxon 129: 403–406. https://doi.org/10.5248/122.171

Supplementary materials

Supplementary material 1 

Figure S1. The ML consensus tree inferred based on ITS partial sequence with bootstrap values for ML (> 70%) and posterior probability (PP) (PP > 0.95) labeled to the left of a node (ML/PP)

Junmin Liang, Guangshuo Li, Shiyue Zhou, Meiqi Zhao, Lei Cai

Data type: phylogenetic data

Explanation note: The type strains were labeled with “T”. Strains obtained from this study are in red.

This dataset is made available under the Open Database License (http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License (ODbL) is a license agreement intended to allow users to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited.
Download file (314.12 kb)
Supplementary material 2 

Figure S2. The ML consensus tree inferred based on tub2 partial sequence with bootstrap values for ML (> 70%) and posterior probability (PP) (PP > 0.95) labeled to the left of a node (ML/PP)

Junmin Liang, Guangshuo Li, Shiyue Zhou, Meiqi Zhao, Lei Cai

Data type: phylogenetic data

Explanation note: The type strains were labeled with “T”. Strains obtained from this study are in red.

This dataset is made available under the Open Database License (http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License (ODbL) is a license agreement intended to allow users to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited.
Download file (311.92 kb)
Supplementary material 3 

Figure S3. The ML consensus tree inferred based on cmdA partial sequence with bootstrap values for ML (> 70%) and posterior probability (PP) (PP > 0.95) labeled to the left of a node (ML/PP)

Junmin Liang, Guangshuo Li, Shiyue Zhou, Meiqi Zhao, Lei Cai

Data type: phylogenetic data

Explanation note: The type strains were labeled with “T”. Strains obtained from this study are in red.

This dataset is made available under the Open Database License (http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License (ODbL) is a license agreement intended to allow users to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited.
Download file (321.59 kb)
Supplementary material 4 

Figure S4. The ML consensus tree inferred based on rpb2 partial sequence with bootstrap values for ML (> 70%) and posterior probability (PP) (PP > 0.95) labeled to the left of a node (ML/PP)

Junmin Liang, Guangshuo Li, Shiyue Zhou, Meiqi Zhao, Lei Cai

Data type: phylogenetic data

Explanation note: The type strains were labeled with “T”. Strains obtained from this study are in red.

This dataset is made available under the Open Database License (http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License (ODbL) is a license agreement intended to allow users to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited.
Download file (328.77 kb)
Supplementary material 5 

Figure S5. The ML consensus tree inferred based on LSU and rpb2 partial sequences with bootstrap values for ML (> 70%) and posterior probability (PP) (PP > 0.95) labeled to the left of a node (ML/PP)

Junmin Liang, Guangshuo Li, Shiyue Zhou, Meiqi Zhao, Lei Cai

Data type: phylogenetic data

Explanation note: The type strains were labeled with “T”. Strains obtained from this study are in red.

This dataset is made available under the Open Database License (http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License (ODbL) is a license agreement intended to allow users to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited.
Download file (861.06 kb)
Supplementary material 6 

Table S1. NCBI GenBank accessions of 28S ribosomal DNA large-subunit sequences (LSU) used in the phylogenetic analyses

Junmin Liang, Guangshuo Li, Shiyue Zhou, Meiqi Zhao, Lei Cai

Data type: phylogenetic data

This dataset is made available under the Open Database License (http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License (ODbL) is a license agreement intended to allow users to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited.
Download file (21.72 kb)