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Myrothecium-like new species from turfgrasses and associated rhizosphere Myrothecium-like new species from turfgrasses and associated rhizosphere

Mycokeys

April 2019
51(2):29-53

DOI:10.3897/mycokeys.51.31957

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CC BY 4.0

Authors:

Junmin Liang

Chinese Academy of Sciences

Li Guangshuo

University of Copenhagen

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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.

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).

…

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

…

Dimorphiseta obtusa (from ex-type strain CGMCC3.19206) a–c colony on PDA, CMA, OAd conidioma on SNAe 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).

…

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

…

Alfaria poae (from ex-type strain CGMCC3.19198) a–c colony on PDA, CMA, OAd–e conidiomata on SNAf 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).

…

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Myrothecium-like new species from turfgrasses and associated rhizosphere 29

Myrothecium-like new species from turfgrasses and

associated rhizosphere

Junmin Liang1,*, Guangshuo Li1,2,*, Shiyue Zhou3, Meiqi Zhao4,5, Lei Cai1,3

1 State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, Beichen West Road,

Chaoyang District, Beijing 100101, China 2 College of Life Sciences, Hebei University, Baoding, Hebei Pro-

vince, 071002, China 3 College of Life Sciences, University of Chinese Academy of Sciences, Beijing 100049,

China 4 College of Plant Protection, China Agricultural University, Beijing 100193, China 5 Forwardgroup

Turf Service & Research Center, Wanning, Hainan Province, 571500, China

Corresponding author: Lei Cai (cail@im.ac.cn)

Academic editor: I. Schmitt|Received27 November 2018|Accepted 26 February 2019|Published 18 April2019

Citation: Liang J, Li G, Zhou S, Zhao M, Cai L (2019) Myrothecium-like new species from turfgrasses and associated

rhizosphere. MycoKeys 51: 29–53. https://doi.org/10.3897/mycokeys.51.31957

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 identied 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). e 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 rst introduced by Tode (1790) based on M. inundatum. e typical

characters of these fungi are cup-shaped sporodochia covered by a mass of slimy, green

to black conidia. e generic concept of Myrothecium has been emended several times

* ese authors contributed equally to this study.

Copyright Junmin Liang et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC

BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

MycoKeys 51: 29–53 (2019)

doi: 10.3897/mycokeys.51.31957

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RESEARCH ARTICLE

Junmin Liang et al. / MycoKeys 51: 29–53 (2019)

(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.

ese 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, Albimbria

terrestris, Simorphiseta terrestris and Parvothecium terrestre. Some species were also re-

ported as weak plant pathogens. For instance, Paramyrothecium roridum (syn. Myrothe-

cium roridum) can infect coee plants, causing bark canker (Tulloch 1972). Albimbria

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 antibi-

otic 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. e aim of

this study was to characterize these strains based on morphology and molecular phy-

logenetic 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 sam-

ples 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 modi-

ed 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. e suspension

was mixed thoroughly using Vortex-Genie 2 (Scientic 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. e 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

Myrothecium-like new species from turfgrasses and associated rhizosphere 31

and streptomycin sulfate in 1 L sterile water). e rst 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. Puried 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).

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 dierential 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 modied Cetyltrimethyl

Ammonium Bromide (CTAB) method (Rogers and Bendich 1994). Partial sequenc-

es of four genes, ITS, RNA polymerase II second largest subunit (rpb2), β-tubulin

(tub2) and calmodulin (cmdA) gene sequences were amplied 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. Amplication for each locus followed the PCR protocols as described in

Lombard et al. (2016). e PCR was performed in a 25 μL reaction volume including

2.5 μL 10 × PCR Buer (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 nal elongation at 72 °C for 5 min.

e puried PCR products were sequenced in both forward and reverse directions

on an ABI-3730 XL DNA Analyzer (Applied Biosystems, California, USA). e se-

Junmin Liang et al. / MycoKeys 51: 29–53 (2019)

Table 1. Strains and NCBI GenBank accessions used in the phylogenetic analyses.

Species Isolate no. aHost/Substrate Country NCBI accession numbers

cmdA ITS tub2 rpb2

Myrothecium

simplex

CBS 582.93TDecaying 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 –

Albimbria

lateralis CBS117712TUnknown 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 112037TRotten leaf Brazil –KU845985 KU846016 KU846003

CBS 112038 Rotten leaf Brazil –KU845986 KU846017 KU846004

Alf. terrestris CBS 477.91TSoil Turkey KU845979 KU845988 KU846019 KU846006

Alf. thymi CBS 447.83Tymus serpyllum e Netherlands KU845981 KU845990 KU846021 –

Capitombria

compacta

CBS 111739TDecaying leaf Brazil KU846261 KU846287 KU846404 KU846349

MUCL 50238 Bark Zimbabwe –KU878556 KU878559 KU878558

Dimorphiseta

terrestris CBS 127345TSoil 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

Myrothecium-like new species from turfgrasses and associated rhizosphere 33

Species Isolate no. aHost/Substrate Country NCBI accession numbers

cmdA ITS tub2 rpb2

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 646.77TDead twig India –KU846471 KU846551 KU846509

CBS 521.96 =

MUCL 39093 Dead twig Nepal –KU846470 KU846550 KU846508

Inaequalispora

prestonii

CBS 175.73TForest soil Malaysia KU846286 KU846316 KU846433 KU846377

MUCL 52636 rhizoplane and

roots of plants Ecuador –KY389317 KY366447 KY389355

Myxospora masonii CBS 174.73TLeaves of

Glyceriasp. England KU846445 KU846462 KU846543 KU846500

My. graminicola CBS 116538TDecaying grass

leaf USA KU846444 KU846461 KU846542 KU846499

My. aptrootii CBS 101263T Leaf litter China KU846441 KU846458 KU846539 KU846496

My. musae CBS 265.71TMusa 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 127295TSoil collected in

tallgrass prairie USA –KU846295 KU846412 KU846356

P. parvum

CBS 257.35TViola 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 e Netherlands –KU846292 KU846409 KU846354

P. nigrum

CBS 116537TSoil 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 873.85TSoil Turkey KU846278 KU846308 KU846425 KU846369

CBS 125835 Soil USA KU846280 KU846310 KU846427 KU846371

P. acadiense CBS 123.96TTussilago farfara Canada –KU846288 KU846405 KU846350

P. terrestris CBS 564.86TSoil Turkey KU846273 KU846303 KU846420 KU846364

CBS 566.86 Soil Turkey KU846275 KU846305 KU846422 KU846366

P. tellicola CBS 478.91TSoil Turkey KU846272 KU846302 KU846419 KU846363

P. foliicola CBS 113121T Decaying leaf Brazil KU846266 KU846294 KU846411 –

CBS 419.93 Air Cuba KU846265 KU846293 KU846410 KU846355

Junmin Liang et al. / MycoKeys 51: 29–53 (2019)

Species Isolate no. aHost/Substrate Country NCBI accession numbers

cmdA ITS tub2 rpb2

P. breviseta CBS 544.75TUnknown India KU846262 KU846289 KU846406 KU846351

P. roridum

CBS 357.89TGardenia sp. Italy KU846270 KU846300 KU846417 KU846361

CBS 212.95 Water e Netherlands KU846269 KU846299 KU846416 KU846360

CBS 372.50 = IMI

140050 Coea 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 2016-8006TSoil 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 198.89TSoil in virgin

forest Brazil KU846449 KU846468 KU846548 KU846506

Neomyrothecium

humicola CBS 310.96TSoil 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 459.82TRotten 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 932.69TSoil e 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 alictrum avum UK KU847219 KU847243 KU847333 –

Xenomyrothecium

tongaense CBS 598.80T Halimeda sp. Tonga KU847221 KU847246 KU847336 KU847295

Virgatospora

echinobrosa

CBS 110115 eobroma cacao Ecuador KU847220 KU847244 KU847334 KU847293

MUCL 39092 =

ATCC 200437 Trewia nudiora Nepal –KU847245 KU847335 KU847294

Fusarium

sambucinum CBS 146.95 Solanum

tuberosum UK KM231391 KM231813 KM232078 KM232381

† ATCC: American Type Culture Collection, Manassas, USA; BCC: BIOTEC Culture Collection, National Center for Genetic Engineer-

ing and Biotechnology (BIOTEC), Bangkok, ailand; CBS: CBS-KNAW Fungal Diversity Centre, Utrecht, e Netherlands; CGMCC:

China General Microbiological Culture Collection Center, Beijing, China; GUCC: Guizhou University Culture Collection, Guiyang,

China; HGUP: Herbarium of the Department of Plant Pathology, Guizhou University, China; IMI: International Mycological Institute,

England, UK; LC: Collection of Lei Cai, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China; MUCL: Mycothèque de

l’Université Catholique de Louvian, Belgium; NRRL: Northern Regional Research Laboratory, USA.

T Ex-type and ex-epitype cultures.

Myrothecium-like new species from turfgrasses and associated rhizosphere 35

quences 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 down-

loaded 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 Likeli-

hood (ML). For BI analysis, the optimal evolutionary model was estimated in Mr-

Modeltest 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 rst 25% trees. e MCMC analysis lasted until the

average standard deviation of split frequencies came below 0.01. e 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-GAM-

MA 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 myrotheci-

um-like strains were selected. Phylogenetic analyses of above 84 strains were per-

formed 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. e

characters of dierent alignments and statistics of phylogenetic analyses were shown

in Table 2. e four single locus trees of all strains showed essentially similar topol-

ogy (Supp. materials1–4), with only minor dierences aecting unsupported nodes

on the trees. e resulting multi-locus ML tree was presented in Fig. 1 together with

BI posterior probability values. Among 84 myrothecium-like strains, 14 strains were

identied as four known species, Albimbria verrucaria (10 strains), Alb. terrestris

(1 strain), Striaticonidium brachysporum (2 strains) and Paramyrothecium nigrum (1

strain). e rest of them were grouped into ve distinct clades with high supported

values. Based on the morphological and phylogenetic distinctions, ve novel species

(i.e. Alfaria humicola, Alf. poae, Dimorphiseta acuta, D. obtusa and Paramyrothecium

sinense) were described in this paper.

Junmin Liang et al. / MycoKeys 51: 29–53 (2019)

Figure 1. e 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). e type strains are labeled with “T”. Strains obtained from this

study are in red. e tree is rooted using Fusarium sambucinum (CBS 146.95).

Myrothecium-like new species from turfgrasses and associated rhizosphere 37

Table 2. Characteristics of the dierent 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

† ITS, the internal transcribed spacer regions and 5.8s rRNA gene; tub2, β-tubulin; cmdA, calmodulin; rpb2: RNA polymerase

II second largest subunit.

* B = Bayesian inference.

‡ G: Gamma distributed rate variation among sites. GTR: Generalised time-reverisble. I: Proportion of invariable sites. HKY:

Hasegawa-Kishino-Yano.

Figure 1. Continued.

Junmin Liang et al. / MycoKeys 51: 29–53 (2019)

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, exuous 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 MB 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 CG-

MCC3.19208 = 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 mass-

es, reverse on PDA bu. Conidiomata sporodochial, stromatic, supercial, 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, termi-

nating 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).

Distribution. China.

Etymology. Name refers to the setae with tapered and sharp apices.

Additional isolates examined. China, Beijing, from leaves of Digitaria sangui-

nalis, 21 Aug 2017, J.M. Liang, LC12123; China, Beijing, from leaves of Poa prat-

ensis, 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.

Myrothecium-like new species from turfgrasses and associated rhizosphere 39

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).

Notes. e multi-locus phylogenetic analyses indicated that D. acuta formed a

sister clade to D. terrestris, but diers 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, diers from D. acuta in producing 1-septate conidia (Alves et al.

2010), while M. xigazense diers in producing conidia that are truncate at both ends

(Wu et al. 2014).

Junmin Liang et al. / MycoKeys 51: 29–53 (2019)

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

MycoBank MB 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 con-

idiophores forming on the aerial mycelium, carrying slimy olivaceous green to black

conidial masses, reverse on PDA pale luteous to bu. Conidiomata sporodochial, stro-

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).

Myrothecium-like new species from turfgrasses and associated rhizosphere 41

matic, supercial, 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, un-

branched, hyaline, smooth, 80–250 μm long, 2–4 μm wide at the broadest, terminat-

ing 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).

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 MB 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 sporo-

dochial, stromatic, supercial, 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. Co-

nidiophores arising from the basal stroma, unbranched or branched, initially hya-

line and smooth, becoming pigmented and verrucose with age, 11–25 μm long.

Junmin Liang et al. / MycoKeys 51: 29–53 (2019)

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).

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). Al-

faria humicola lacks setae, distinguishing it from Alf. caricicola and Alf. thymi. Further-

more, 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 (Sac-

cardo, 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.

Figure 4. Alfaria humicola (from ex-type CGMCC3.19213) a–c colony on PDA, CMA, OA d conidi-

omata on SNA e sporodochial conidioma, arrows showing branched conidiosphores and conidiogenous

cells f conidia. Scale bars: 10 μm (e); 5 μm (f).

Myrothecium-like new species from turfgrasses and associated rhizosphere 43

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

MycoBank MB 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, ap-

prox. 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,

Figure 5. Alfaria poae (from ex-type strain CGMCC3.19198) a–c colony on PDA, CMA, OA d–eco-

nidiomata on SNA f synnematous conidioma g conidiogenous cells, the arrow showing conspicuous col-

larette h aged conidiophores i conidia. Scale bars: 50 μm (f); 5 μm (g); 10 μm (h, i ).

Junmin Liang et al. / MycoKeys 51: 29–53 (2019)

reverse on PDA sienna. Hyphae hyaline, smooth, branched, 1–2 μm wide. Conidioma-

ta 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. Conidiog-

enous cell phialidic, clavate to cylindrical, hyaline, smooth, 5–10 × 1–2 μm, becom-

ing 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).

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 pro-

duces ellipsoidal to fusiform conidia, which are dierent from the ossiform conidia

produced by Alf. ossiformis (Lombard et al. 2016). e conidia of Alf. terrestris have

basal hilum which was not observed in Alf. poae. In addition, Alf. poae shares mor-

phological characters with several un-sequenced Myrothecium taxa, such as M. atro-

carneum (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 epitypied by fresh collections. Although M. leucomelas

(host: Sumbaviae rotttleroidis; location: Bulacan, Luzon) had a detailed description,

it cannot be epitypied by Alf. Poae, because Alf. poae was collected from a distinct

location and plant host. Taking the above special characters into account, we con-

sidered introducing a new species, Alfaria poae.

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

MycoBank MB 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, supercial, 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

Myrothecium-like new species from turfgrasses and associated rhizosphere 45

of conidia. Stroma poorly developed, hyaline, of textura angularis. Setae arising from

stroma, thin-walled, hyaline, 1–3-septate, straight to exuous, 45–90 μm long, 1–3

μm wide, tapering to an acutely rounded apex. Conidiophores arising from the ba-

sal stroma, consisting of a stipe and a penicillately branched conidiogenous appara-

tus; stipes unbranched, hyaline, septate, smooth, 20–30 × 2–3 μm; primary branches

aseptate, unbranched, smooth, 13–40 × 2–3 μm; secondary branches aseptate, un-

branched, 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 thicken-

ings. Conidia aseptate, hyaline, smooth, cylindrical, 6–7 × 2–3 μm (av. 7 ± 0.3 × 2 ±

0.2 μm, n = 40), rounded at both ends.

Distribution. China.

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).

Junmin Liang et al. / MycoKeys 51: 29–53 (2019)

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 origi-

nal description of Myrotheciella catenuligera suggested that it lacks seta (Spegazzini

1911), thus is clearly dierent from the morphological circumscription of P. roridum.

erefore, 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. e 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. dimor-

phum show seta with obtuse apices, but both taxa produce two types of conidia (Jiang

et al. 2014; Watanabe et al. 2003).

Discussion

e ITS has been shown to be insucient to delineate the myrothecium-like spe-

cies. 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 ve novel species in

Stachybotryaceae.

By comparing the topologies of the four single-locus trees, incomplete lineage sort-

ing 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 Albimbria spe-

cies (Supp. materials 2, 3). ree Dimorphiseta species are similar in the conidial shape

and size (7–19 μm long), which are distinct from the shorter conidia in Albimbria

(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 Albimbria

species (Lombard et al. 2016). Furthermore, the rpb2 and 28S ribosomal DNA com-

bined dataset, which was suggested to delimit generic boundaries of myrothecium-like

Myrothecium-like new species from turfgrasses and associated rhizosphere 47

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 Dimor-

phiseta, Capitombria, Gregatothecium and Neomyrothecium. In this study, we reported two

new species in Dimorphiseta (D. acuta and D. obtusa). With this addition, the generic con-

cept of Dimorphiseta is slightly expanded for including a third type of setae. Hereto, Di-

morphiseta 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 co-

nidial size should be dened 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. thailan-

dicum (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). ese above species were identied, either based on morphology

only or with a single molecular locus (ITS), and should be better conrmed 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., Capitombria, Melanconis, Striaticonidium, Xepicula

(Lombard et al. 2016), Digitiseta (Gordillo and Decock 2018). Only a limited num-

ber 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 de-

scriptions are suciently clear to recognize a species. ey should be epitypied 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

is study was nancially supported by National Natural Science Foundation of Chi-

na (NSFC 31600405).

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Supplementary material 1

Figure S1. e ML consensus tree inferred based on ITS partial sequence with

bootstrap values for ML (> 70%) and posterior probability (PP) (PP > 0.95) la-

beled to the left of a node (ML/PP)

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

Data type: phylogenetic data

Explanation note: e type strains were labeled with “T”. Strains obtained from this

study are in red.

(http://opendatacommons.org/licenses/odbl/1.0/). e 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.

Link: https://doi.org/10.3897/mycokeys.51.31957.suppl1

Myrothecium-like new species from turfgrasses and associated rhizosphere 51

Supplementary material 2

Figure S2. e ML consensus tree inferred based on tub2 partial sequence with

bootstrap values for ML (> 70%) and posterior probability (PP) (PP > 0.95) la-

beled to the left of a node (ML/PP)

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

Data type: phylogenetic data

Explanation note: e type strains were labeled with “T”. Strains obtained from this

study are in red.

(http://opendatacommons.org/licenses/odbl/1.0/). e 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.

Link: https://doi.org/10.3897/mycokeys.51.31957.suppl2

Supplementary material 3

Figure S3. e ML consensus tree inferred based on cmdA partial sequence with

bootstrap values for ML (> 70%) and posterior probability (PP) (PP > 0.95) la-

beled to the left of a node (ML/PP)

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

Data type: phylogenetic data

Explanation note: e type strains were labeled with “T”. Strains obtained from this

study are in red.

(http://opendatacommons.org/licenses/odbl/1.0/). e 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.

Link: https://doi.org/10.3897/mycokeys.51.31957.suppl3

Junmin Liang et al. / MycoKeys 51: 29–53 (2019)

Supplementary material 4

Figure S4. e ML consensus tree inferred based on rpb2 partial sequence with

bootstrap values for ML (> 70%) and posterior probability (PP) (PP > 0.95) la-

beled to the left of a node (ML/PP)

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

Data type: phylogenetic data

Explanation note: e type strains were labeled with “T”. Strains obtained from this

study are in red.

(http://opendatacommons.org/licenses/odbl/1.0/). e 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.

Link: https://doi.org/10.3897/mycokeys.51.31957.suppl4

Supplementary material 5

Figure S5. e ML consensus tree inferred based on LSU and rpb2 partial se-

quences with bootstrap values for ML (> 70%) and posterior probability (PP) (PP

> 0.95) labeled to the left of a node (ML/PP)

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

Data type: phylogenetic data

Explanation note: e type strains were labeled with “T”. Strains obtained from this

study are in red.

(http://opendatacommons.org/licenses/odbl/1.0/). e 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.

Link: https://doi.org/10.3897/mycokeys.51.31957.suppl5

Myrothecium-like new species from turfgrasses and associated rhizosphere 53

Supplementary material 6

Table S1. NCBI GenBank accessions of 28S ribosomal DNA large-subunit se-

quences (LSU) used in the phylogenetic analyses

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

Data type: phylogenetic data

(http://opendatacommons.org/licenses/odbl/1.0/). e 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.

Link: https://doi.org/10.3897/mycokeys.51.31957.suppl6

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Original Article THE INTERNATIONAL JOURNAL ON NEOTROPICAL BIOLOGY THE INTERNATIONAL JOURNAL ON GLOBAL BIODIVERSITY AND ENVIRONMENT

Article

Full-text available

Apr 2022
Braz J Biol

This study aimed to identify by molecular analysis, morphology, chemistry and antioxidant extracts of filamentous fungi isolated from the digestive tract of Phylloicus sp, an aquatic insect that lives on leaf packages in tropical streams and participates together with fungi of the decomposition of plant substrates in aquatic habitats. Insect larvae of Phylloicus sp. were collected in streams in the state of Tocantins, Brazil. Fungi were isolated from the digestive tract of larvae after disinfection and dissection, then described and purified for identification purposes and testing for antioxidant activity. Molecular identity was performed of ITS1 and ITS4, TUB e TEF sequencing. Fungal extracts were produced in 70% ethanol solution and later lyophilized. For analysis of chemical groups of extracts, thin layer chromatography (TLC) was performed in two mobile phases and different developers. Morphology was performed by optical microscopy stained with Toluidine Blue and measurement performed using the ImageJ program. Antioxidant activity performed in TLC and by quantitative method for DPPH and hydrogen peroxide (H2O2) radicals. Four fungi were identified: Endomelanconiopsis endophytica, Myxospora musae, Neopestalotiopsis cubana and Fusarium pseudocircinatum. The TLC showed several spots with acetone/chloroform mobile phase and UV 254 nm developers and I2 vapor. Fungal extracts demonstrate antioxidant action to reduce the DPPH free radical and especially for H2O2 above 50%, E. endophytica 91.6%, M. musae 87.8%, N. cubana 89.5% and 92.3% for F. pseudocircinatum. This study demonstrated that the molecular technique by PCR was satisfactory for identifying fungi, and extracts with numerous chemical groups and potent reducing agents. Thus future work, should be carried out evaluating these four species for industrial use.

Plant pathogenic Paramyrothecium species: distribution, biology, epidemiology and disease management

Article

Jun 2024

Paramyrothecium comprises saprobic and plant pathogenic members. Eight plant-pathogenic Paramyrothecium species have been recorded in Asia, America, and some parts of Africa and Europe. Among the commonly reported species are P. roridum and P. foliicola. Several Paramyrothecium species are associated with coffee leaf spots, muskmelon crown rot, and eggplant crater rot. Paramyrothecium is commonly found in soil, decaying plant material, and diseased fruits, stems, and leaves of several plant species. The life cycle of Paramyrothecium species includes an asexual stage throughout disease development, with no sexual morphs reported. Environmental factors, such as temperature and humidity, influence the distribution and prevalence of Paramyrothecium. Paramyrothecium-associated diseases occur through various mechanisms, including wind and rain dispersal of conidia, contaminated soil, and plant debris. Paramyrothecium disease development can be exacerbated when the soil is wet and plant tissues are damaged, which served as pathogen entry. Adequate water management, soil sanitation, and proper handling of crops are important to minimize losses in commercial crop production. Several biological control agents and pesticides have also been reported to control the pathogen and the associated disease.

Paramyrothecium eichhorniae sp. nov., Causing Leaf Blight Disease of Water Hyacinth from Thailand

Article

Full-text available

Feb 2022

Paramyrothecium eichhorniae sp. nov. was observed and collected from Chiang Mai and Phetchaburi Provinces, Thailand. This new species is introduced based on morphological and molecular evidence. This fungus is characterized by its production of sporodochium conidiomata with a white setose fringe surrounding an olivaceous green to dark green slimy mass of conidia, penicillately branched conidiophores, and aseptate and cylindrical to ellipsoid conidia. Phylogenetic analyses of combined LSU rDNA, ITS rDNA, tef1, rpb2, tub2 and cmdA sequence data using maximum parsimony, maximum likelihood and Bayesian approaches placed the fungus in a strongly supported clade with other Paramyrothecium species in Stachybotryaceae (Hypocreales, Sordariomycetes). The descriptions of the species are accompanied by illustrations of morphological features, and a discussion of the related taxa is presented.

Biodeterioration and cellulolytic activity by fungi isolated from a nineteenth-century painting at the National Theatre of Costa Rica

Article

Nov 2021

Living organisms can induce deterioration of cultural heritage. Conservation strategies aimed at avoiding damage and aiding restoration, require a comprehensive knowledge of structure, chemical composition, and identity of microorganisms that colonize artworks. The National Theatre of Costa Rica (NTCR), a building with historic architecture, houses several oil paintings from the nineteenth century, some with visible signs of biodeterioration. One of them is a large format painting on canvas called La Danza (size 9.83 × 5.13 m) from 1896 by Italian artist Vespasiano Bignami, located on the ceiling of the theatre's foyer. In the present study, we undertook a physicochemical and microbiological study of La Danza to identify the fungal species that inhabit the artwork and are responsible for the damage observed. Scanning electron microscope (SEM) images and attenuated total reflectance - Fourier transform infrared (ATR-FTIR) spectroscopic data indicated that the canvas material is made of hemp, the binder contains linseed oil and lead white, and a material in the inner face of the canvas is mainly composed of beeswax. Fungi were isolated onto potato dextrose agar (PDA) and carboxymethyl cellulose (CMC) agar, and then identified with molecular (BTUB, nrDNA ITS, and TEF1 regions) and morphological methods. Four isolates belonging to the genera Myxospora, Pestalotiopsis, Ustilago, and aff. Penicillium, were obtained. Qualitative tests showed cellulolytic activity in all isolated specimens, confirming their possible role in biodeterioration of the canvas. Phylogenetic and morphological data revealed a new species of Myxospora we name here as Myxospora theatro sp. nov., in reference to NTCR. The findings broaden the knowledge of fungi capable of inhabiting and damaging cultural heritage. They also provide valuable information to develop strategies for conservation and restoration of oil paintings on canvas.

Effects of Biogas Slurry Combined With Chemical Fertilizer on Soil Bacterial and Fungal Community Composition in a Paddy Field

Article

Full-text available

Aug 2021

The application of biogas slurry and chemical fertilizer in paddy fields can be a practical method to reduce the environmental risk and utilize the nutrients of biogas slurry. The responses of bacterial and fungal communities to the application of biogas slurry and chemical fertilizer are important reflections of the quality of the ecological environment. In this study, based on a 3-year field experiment with different ratios of biogas slurry and chemical fertilizer (applying the same pure nitrogen amount), the Illumina MiSeq platform was used to investigate the bacterial and fungal community diversity and composition in paddy soil. Our results revealed that compared with the observations under regular chemical fertilization, on the basis of stable paddy yield, the application of biogas slurry combined with chemical fertilizer significantly enhanced the soil nutrient availability and bacterial community diversity and reduced the fungal community diversity. Dissolved organic carbon (DOC), DOC/SOC (soil organic carbon), available nitrogen (AN) and available phosphorus (AP) were positively correlated with the bacterial community diversity, but no soil property was significantly associated with the fungal community. The bacterial community was primarily driven by the application of biogas slurry combined with chemical fertilizer (40.78%), while the fungal community was almost equally affected by the addition of pure biogas slurry, chemical fertilizer and biogas slurry combined with chemical fertilizer (25.65–28.72%). Biogas slurry combined with chemical fertilizer significantly enriched Proteobacteria , Acidobacteria , Planctomycetes , Rokubacteria , and Ascomycota and depleted Chloroflexi , Bacteroidetes , Crenarchaeota , Basidiomycota , and Glomeromycota . The observation of the alteration of some bacteria- and fungus-specific taxa provides insights for the proper application of biogas slurry combined with chemical fertilizer, which has the potential to promote crop growth and inhibit pathogens.

Leptosphaerulina species isolated from golf turfgrass in China, with description of L. macrospora, sp. nov

Article

Full-text available

Jul 2021

Leptosphaerulina leaf blight occurs on most turfgrasses. Hitherto, Leptosphaerulina species associated with this disease include L. americana, L. argentinensis, L. australis, and L. trifolii. However, following Koch’s postulates, L. australis was confirmed as saprobes but not pathogens, and the other three species have not been tested. The pathogenicity of Leptosphaerulina spp. is still questionable. In this study, we isolated 19 Leptosphaerulina strains from diseased golf turfgrasses in China, and they were identified as L. gaeumannii, L. saccharicola, and a new species, L. macrospora, through multilocus (ITS, 28S, rpb2, and tub2) phylogenetic analyses and morphological observations. Pathogenicity test revealed that the three Leptosphaerulina species identified in this study cannot infect live/healthy turfgrass tissues of Poa pratensis and Agrostis stolonifera and only produced pseudothecia on the dead leaves of stressed seedlings. Considering the results of pathogenicity tests in this and previous studies, we speculate that most Leptosphaerulina species isolated from diseased turfgrass are not pathogens but saprobes. Applying proper management practices to prevent severe turfgrass stress is a key measure to reduce or eliminate the effects of Leptosphaerulina on golf turfgrass.

Taxonomic and phylogenetic contributions to Celtis formosana, Ficus ampelas, F. septica, Macaranga tanarius and Morus australis leaf litter inhabiting microfungi

Article

Full-text available

Jun 2021

This article provides descriptions and illustrations of microfungi associated with the leaf litter of Celtis formosana, Ficus ampelas, F. septica, Macaranga tanarius and Morus australis collected from Taiwan. These host species are native to the island and Celtis formosana is an endemic tree species. The study revealed 95 species, consisting of two new families (Cylindrohyalosporaceae and Oblongohyalosporaceae), three new genera (Cylindrohyalospora, Neodictyosporium and Oblongohyalospora), 41 new species and 54 new host records. The newly described species are Acrocalymma ampeli (Acrocalymmaceae), Arthrinium mori (Apiosporaceae), Arxiella celtidis (Muyocopronaceae), Bertiella fici (Melanommataceae), Cercophora fici (Lasiosphaeriaceae), Colletotrichum celtidis, C. fici, C. fici-septicae (Glomerellaceae), Conidiocarpus fici-septicae (Capnodiaceae), Coniella fici (Schizoparmaceae), Cylindrohyalospora fici (Cylindrohyalosporaceae), Diaporthe celtidis, D. fici-septicae (Diaporthaceae), Diaporthosporella macarangae (Diaporthosporellaceae), Diplodia fici-septicae (Botryosphaeriaceae), Discosia celtidis, D. fici (Sporocadaceae), Leptodiscella sexualis (Muyocopronaceae), Leptospora macarangae (Phaeosphaeriaceae), Memnoniella alishanensis, M. celtidis, M. mori (Stachybotryaceae), Micropeltis fici, M. ficina (Micropeltidaceae), Microthyrium fici-septicae (Microthyriaceae), Muyocopron celtidis, M. ficinum, Mycoleptodiscus alishanensis (Muyocopronaceae), Neoanthostomella fici (Xylariales genera incertae sedis), Neodictyosporium macarangae (Sordariales genera incertae sedis), Neofusicoccum moracearum (Botryosphaeriaceae), Neophyllachora fici (Phyllachoraceae), Nigrospora macarangae (Apiosporaceae), Oblongohyalospora macarangae (Oblongohyalosporaceae), Ophioceras ficinum (Ophioceraceae), Parawiesneriomyces chiayiensis (Wiesneriomycetaceae), Periconia alishanica, P. celtidis (Periconiaceae), Pseudocercospora fici-septicae (Mycosphaerellaceae), Pseudoneottiospora cannabacearum (Chaetosphaeriaceae) and Pseudopithomyces mori (Didymosphaeriaceae). The new host records are Alternaria burnsii, A. pseudoeichhorniae (Pleosporaceae), Arthrinium hydei, A. malaysianum, A. paraphaeospermum, A. rasikravindrae, A. sacchari (Apiosporaceae), Bartalinia robillardoides (Sporocadaceae), Beltrania rhombica (Beltraniaceae), Cladosporium tenuissimum (Cladosporiaceae), Coniella quercicola (Schizoparmaceae), Dematiocladium celtidicola (Nectriaceae), Diaporthe limonicola, D. millettiae, D. pseudophoenicicola (Diaporthaceae), Dictyocheirospora garethjonesii (Dictyosporiaceae), Dimorphiseta acuta (Stachybotryaceae), Dinemasporium parastrigosum (Chaetosphaeriaceae), Discosia querci (Sporocadaceae), Fitzroyomyces cyperacearum (Stictidaceae), Gilmaniella bambusae (Ascomycota genera incertae sedis), Hermatomyces biconisporus (Hermatomycetaceae), Lasiodiplodia thailandica, L. theobromae (Botryosphaeriaceae), Memnoniella echinata (Stachybotryaceae), Muyocopron dipterocarpi, M. lithocarpi (Muyocopronaceae), Neopestalotiopsis asiatica, N. phangngaensis (Sporocadaceae), Ophioceras chiangdaoense (Ophioceraceae), Periconia byssoides (Periconiaceae), Pestalotiopsis dracaenea, P. formosana, P. neolitseae, P. papuana, P. parva, P. portugallica, P. trachycarpicola (Sporocadaceae), Phragmocapnias betle (Capnodiaceae), Phyllosticta capitalensis (Phyllostictaceae), Pseudopestalotiopsis camelliae-sinensis (Sporocadaceae), Pseudopithomyces chartarum, P. sacchari (Didymosphaeriaceae), Pseudorobillarda phragmitis (Pseudorobillardaceae), Robillarda roystoneae (Sporocadaceae), Sirastachys castanedae, S. pandanicola (Stachybotryaceae), Spegazzinia musae (Didymosphaeriaceae), Stachybotrys aloeticola, S. microspora (Stachybotryaceae), Strigula multiformis (Strigulaceae), Torula fici (Torulaceae), Wiesneriomyces laurinus (Wiesneriomycetaceae) and Yunnanomyces pandanicola (Sympoventuriaceae). The taxonomic placement of most taxa discussed in this study is based on morphological observation of specimens, coupled with multi-locus phylogenetic analyses of sequence data. In addition, this study provides a host-fungus database for future studies and increases knowledge of fungal diversity, as well as new fungal discoveries from the island.

Xenomyrothecium tongaense PTS8: a rare endophyte of Polianthes tuberosa with salient antagonism against multidrug-resistant pathogens

Article

Full-text available

Feb 2024

Introduction Endophytes refer to microorganisms residing within the endosphere of plants, particularly perennials, without inflicting noticeable injury or inducing obvious morphological variations to their host plant or host organism. Endophytic fungi, although often overlooked microorganisms, have garnered interest due to their significant biological diversity and ability to produce novel pharmacological substances. Methods In this study, fourteen endophytic fungi retrieved were from the stem of the perennial plant Polianthes tuberosa of the Asparagaceae family. These fungal crude metabolites were tested for antagonistic susceptibility to Multi-Drug Resistant (MDR) pathogens using agar well diffusion, Minimum Inhibitory Concentration (MIC), and Minimum Bactericidal Concentration (MBC) assays. The chequerboard test was used to assess the synergistic impact of active extract. Results and discussion In early antibacterial screening using the Agar plug diffusion test, three of fourteen endophytes demonstrated antagonism against Methicillin-resistant Staphylococcus aureus (MRSA) and Vancomycin-resistant Enterococcus (VRE). Three isolates were grown in liquid medium and their secondary metabolites were recovered using various organic solvents. Eight extracts from three endophytic fungi displayed antagonism against one or more human pathogens with diameters ranging from 11 to 24 mm. The highest antagonistic effect was obtained in ethyl acetate extract for PTS8 isolate against two MRSA (ATCC 43300, 700699) with 20 ± 0.27 and 22 ± 0.47 mm zones of inhibition, respectively, among different solvent extracts. The extract had MICs of 3.12 ± 0.05 and 1.56 ± 0.05 μg/mL, and MBCs of 50 ± 0.01 and 12.5 ± 0.04 μg/mL, respectively. Antagonism against VRE was 18 ± 0.23 mm Zone of Inhibition (ZOI) with MIC and MBC of 6.25 ± 0.25 and 25 ± 0.01 μg/mL. When ethyl acetate extract was coupled with antibiotics, the chequerboard assay demonstrated a synergistic impact against MDR bacteria. In an antioxidant test, it had an inhibitory impact of 87 ± 0.5% and 88.5 ± 0.5% in 2,2-Diphenyl-1-Picrylhydrazyl and reducing power assay, respectively, at 150 μg/mL concentration. PTS8 was identified as a Xenomyrothecium tongaense strain by 18S rRNA internal transcribed spacer (ITS) sequencing. To our insight, it is the foremost study to demonstrate the presence of an X. tongaense endophyte in the stem of P. tuberosa and the first report to study the antibacterial efficacy of X. tongaense which might serve as a powerful antibacterial source against antibiotic-resistant human infections.

Paramyrothecium amorphophalli sp. nov., a causal agent of leaf blight on elephant foot yam in northern Thailand

Article

Full-text available

Feb 2023

Paramyrothecium was introduced in 2016 containing 20 species and many species have been reported as plant pathogens, causing leaf blight and leaf spots on different plant species. This study revealed a new species Paramyrothecium amorphophalli from symptomatic leaves of elephant foot yam (Amorphophallus sp.), collected from Chiang Rai, Thailand based on distinct morphological characteristics and phylogenetic analyses. The isolate formed a distinct lineage from Paramyrothecium eichhorniae and P. vignicola based on maximum likelihood, maximum parsimony and Bayesian posterior probability analyses of ITS, cmdA, rpb2 and tub2 sequence data. Furthermore, pathogenicity assays confirmed that P. amorphophalli is pathogenic to the original host, causing leaf blight.

Identification and Pathogenicity of Paramyrothecium Species Associated with Leaf Spot Disease in Northern Thailand

Article

Full-text available

May 2022

Species of Paramyrothecium that are reported as plant pathogens and cause leaf spot or leaf blight have been reported on many commercial crops worldwide. In 2019, during a survey of fungi causing leaf spots on plants in Chiang Mai and Mae Hong Son provinces, northern Thailand, 16 isolates from 14 host species across nine plant families were collected. A new species Paramyrothecium vignicola sp. nov. was identified based on morphology and concatenated (ITS, cmdA, rpb2, and tub2) phylogeny. Further, P. breviseta and P. foliicola represented novel geographic records to Thailand, while P. eichhorniae represented a novel host record (Psophocarpus sp., Centrosema sp., Aristolochia sp.). These species were confirmed to be the causal agents of the leaf spot disease through pathogenicity assay. Furthermore, cross pathogenicity tests on Coffea arabica L., Commelina benghalensis L., Glycine max (L.) Merr., and Dieffenbachia seguine (Jacq.) Schott revealed multiple host ranges for these pathogens. Further research is required into the host–pathogen relationship of Paramyrothecium species that cause leaf spot and their management. Biotic and abiotic stresses caused by climate change may affect plant health and disease susceptibility. Hence, proper identification and monitoring of fungal communities in the environment are important to understand emerging diseases and for implementation of disease management strategies.

Fungal systematics and evolution: FUSE 3

Article

Full-text available

Dec 2017
SYDOWIA

The present study introduces seven new species, one new combination, one new variety and several interesting taxonomical notes and/or geographical records. Most of the new taxa are Ascomycetes, but the study also includes a new variety of a Basidiomycete. Novel species include Gyromitra khanspurensis (Discinaceae, Pezizales, Pezizomycetes) from Pakistan growing near Cedrus deoadara and Paramyrothecium guiyangense and Paramyrothecium verruridum (Stachybotriaceae, Hypocreales, Sordariomycetes) both isolated from soil in China. New species from South Africa are Sclerostagonospora elegiae on culm litter of Elegia equisetacea, Sclerostagonospora fusiformis on culm litter of Thamnochortus spicigerus, Sclerostagonospora pinguis on culm litter of Cannomois virgata and Sclerostagonospora sulcata on culm litter of Ischyrolepis subverticellata (Phaeosphaeriaceae, Pleosporales, Dothideomycetes). Hapalocystis berkeleyi var. kickxii with its basionym Hypoxylon kickxii is shown to be a taxon on species level and thus recombined as Hapalocystis kickxii (Sydowiellaceae, Diaporthales, Sordariomycetes), and it is lecto- and epitypified. The new variety Pluteus romellii var. luteoalbus (Pluteaceae, Agaricales, Agaricomycetes) growing on a mossy fallen stem of a deciduous tree is described from Czech Republic. Cortinarius scaurocaninus (Cortinariaceae, Agaricales, Agaricomycetes) is new for Austria, Humicola grisea (Chaetomiaceae, Sordariales, Sordariomycetes) is an interesting new record for Chile. Two taxa are reported as new for Turkey: the lichenicolous fungus Opegrapha parasitica (Opegraphaceae, Arthoniales, Arthoniomycetes) growing partly immersed in the thallus of Aspicilia and the lichen Rinodina zwackhiana (Physciaceae, Teloschistales, Lecanoromycetes) from calcareous rock. Finally, Xerula strigosa (Physalacriaceae, Agaricales, Agaricomycetes), described from China, is confirmed to be present also in Pakistan.

Culturable mycobiota from Karst caves in China, with descriptions of 20 new species

Article

Full-text available

Dec 2017

Karst caves are distinctly characterised by darkness, low to moderate temperatures, high humidity, and scarcity of organic matter. During the years of 2014-2015, we explored the mycobiota in two unnamed Karst caves in Guizhou province, China, and obtained 563 fungal strains via the dilution plate method. Preliminary ITS analyses of these strains suggested that they belonged to 246 species in 116 genera, while 23.5 % were not identified to species level. Among these species, 85.8 % (211 species) belonged to Ascomycota; 7.3 % (18 species) belonged to Basidiomycota; 6.9 % (17 species) belonged to Mucoromycotina. The majority of these species have been previously known from other environments, mostly from plants or animals as pathogens, endophytes or via a mycorrhizal association. We also found that 59 % of these species were discovered for the first time from Karst caves, including 20 new species that are described in this paper. The phylogenetic tree based on LSU sequences revealed 20 new species were distributed in six different orders. In addition, ITS or multi-locus sequences were employed to infer the phylogenetic relationships of new taxa with closely related allies. We conclude that Karst caves encompass a high fungal diversity, including a number of previously unknown species. Novel species described include: Amphichorda guana, Auxarthronopsis guizhouensis, Biscogniauxia petrensis, Cladorrhinum globisporum, Collariella quadrum, Gymnoascus exasperatus, Humicola limonisporum, Metapochonia variabilis, Microascus anfractus, Microascus globulosus, Microdochium chrysanthemoides, Paracremonium variiforme, Pectinotrichum chinense, Phaeosphaeria fusispora, Ramophialophora globispora, Ramophialophora petraea, Scopulariopsis crassa, Simplicillium calcicola, Volutella aeria, and Wardomycopsis longicatenata. © 2017 Naturalis Biodiversity Center & Westerdijk Fungal Biodiversity Institute.

Fungal diversity notes 491–602: taxonomic and phylogenetic contributions to fungal taxa

Article

Full-text available

Mar 2017

This is a continuity of a series of taxonomic and phylogenetic papers on the fungi where materials were collected from many countries, examined and described. In addition to extensive morphological descriptions and appropriate asexual and sexual connections, DNA sequence data are also analysed from concatenated datasets to infer phylogenetic relationships and substantiate systematic positions of taxa within appropriate ranks. Wherever new species or combinations are proposed, we apply an integrative approach using morphological and molecular data as well as ecological features wherever applicable. Notes on 112 fungal taxa are compiled in this paper including Biatriosporaceae and Roussoellaceae, Didysimulans gen. nov., 81 new species, 18 new host records and new country records, five reference specimens, two new combinations, and three sexual and asexual morph reports. The new species are Amanita cornelii, A. emodotrygon, Angustimassarina alni, A. arezzoensis, A. italica, A. lonicerae, A. premilcurensis, Ascochyta italica, A. rosae, Austroboletus appendiculatus, Barriopsis thailandica, Berkleasmium ariense, Calophoma petasitis, Camarosporium laburnicola, C. moricola, C. grisea, C. ossea, C. paraincrustata, Colletotrichum sambucicola, Coprinopsis cerkezii, Cytospora gelida, Dacrymyces chiangraiensis, Didysimulans italica, D. mezzanensis, Entodesmium italica, Entoloma magnum, Evlachovaea indica, Exophiala italica, Favolus gracilisporus, Femsjonia monospora, Fomitopsis flabellata, F. roseoalba, Gongronella brasiliensis, Helvella crispoides, Hermatomyces chiangmaiensis, H. chromolaenae, Hysterium centramurum, Inflatispora caryotae, Inocybe brunneosquamulosa, I. luteobrunnea, I. rubrobrunnea, Keissleriella cirsii, Lepiota cylindrocystidia, L. flavocarpa, L. maerimensis, Lophiotrema guttulata, Marasmius luculentus, Morenoina calamicola, Moelleriella thanathonensis, Mucor stercorarius, Myrmecridium fluviae, Myrothecium septentrionale, Neosetophoma garethjonesii, Nigrograna cangshanensis, Nodulosphaeria guttulatum, N. multiseptata, N. sambuci, Panus subfasciatus, Paraleptosphaeria padi, Paraphaeosphaeria viciae, Parathyridaria robiniae, Penicillium punicae, Phaeosphaeria calamicola, Phaeosphaeriopsis yuccae, Pleurophoma italica, Polyporus brevibasidiosus, P. koreanus, P. orientivarius, P. parvovarius, P. subdictyopus, P. ulleungus, Pseudoasteromassaria spadicea, Rosellinia mearnsii, Rubroboletus demonensis, Russula yanheensis, Sigarispora muriformis, Sillia italica, Stagonosporopsis ailanthicola, Strobilomyces longistipitatus, Subplenodomus galicola and Wolfiporia pseudococos. The new combinations are Melanomma populina and Rubroboletus eastwoodiae. The reference specimens are Cookeina tricholoma, Gnomoniopsis sanguisorbae, Helvella costifera, Polythrincium trifolii and Russula virescens. The new host records and country records are Ascochyta medicaginicola, Boletellus emodensis, Cyptotrama asprata, Cytospora ceratosperma, Favolaschia auriscalpium, F. manipularis, Hysterobrevium mori, Lentinus sajor-caju, L. squarrosulus, L. velutinus, Leucocoprinus cretaceus, Lophiotrema vagabundum, Nothophoma quercina, Platystomum rosae, Pseudodidymosphaeria phlei, Tremella fuciformis, Truncatella spartii and Vaginatispora appendiculata and three sexual and asexual morphs are Aposphaeria corallinolutea, Dothiora buxi and Hypocrella calendulina.

Bambusicolous fungi

Article

Full-text available

May 2016

Fourty-three species of microfungi from bamboo are treated, including one new family, Occultibambusaceae, three new genera, Neoanthostomella, Occultibambusa and Seriascoma, 27 new species, one renamed species and 15 re-described or re-illustrated species, and four designated reference specimens are treated in this paper, the majority of which are saprobic on dead culms. To determine species identification, separate phylogenetical analyses for each group are carried out, based on molecular data from this study and sequences downloaded from GenBank. Morphologically similar species and phylogenetically close taxa are compared and discussed. In addition a list of bambusicolous fungi published since Hyde and colleagues in 2002 is provided.

Generic hyper-diversity in Stachybotriaceae

Article

Full-text available

Apr 2016

The family Stachybotriaceae was recently introduced to include the genera Myrothecium, Peethambara and Stachybotrys. Members of this family include important plant and human pathogens, as well as several species used in industrial and commercial applications as biodegraders and biocontrol agents. However, the generic boundaries in Stachybotriaceae are still poorly defined, as type material and sequence data are not readily available for taxonomic studies. To address this issue, we performed multi-locus phylogenetic analyses using partial gene sequences of the 28S large subunit (LSU), the internal transcribed spacer regions and intervening 5.8S nrRNA (ITS), the RNA polymerase II second largest subunit (rpb2), calmodulin (cmdA), translation elongation factor 1-alpha (tef1) and β-tubulin (tub2) for all available type and authentic strains. Supported by morphological characters these data resolved 33 genera in the Stachybotriaceae. These included the nine already established genera Albosynnema, Alfaria, Didymostilbe, Myrothecium, Parasarcopodium, Peethambara, Septomyrothecium, Stachybotrys and Xepicula. At the same time the generic names Melanopsamma, Memnoniella and Virgatospora were resurrected. Phylogenetic inference further showed that both the genera Myrothecium and Stachybotrys are polyphyletic resulting in the introduction of 13 new genera with myrothecium-like morphology and eight new genera with stachybotrys-like morphology. © 2015-2016 Naturalis Biodiversity Center & Centraalbureau voor Schimmelcultures.

Anamorphic fungi from French Guyana. Septomyrothecium maraitiense sp. nov. and S. setiramosum comb. nov. (anamorphic Hypocreales, Ascomycota)

Article

Full-text available

Dec 2008

Septomyrothecium maraitiense sp. nov., isolated from a decaying leaf in French Guyana, South America, is described and illustrated. The species is compared to Septomyrothecium uniseptatum and Myrothecium setiramosum, and the latter is transferred into Septomyrothecium.

MrModeltest V2. Program Distributed by the Author

Article

Full-text available

Jan 2004

Johan Nylander

Myrothecium-like (Ascomycota, Hypocreales) species from tropical areas: Digitiseta gen. nov. and additions to Inaequalispora and Parvothecium

Article

May 2017

Inaequalispora and Parvothecium are two myrothecium-like, closely related genera of Hypocreales. They are also morphologically similar, sharing sporodochial conidiomata, penicillate conidiophores, fusiform to ellipsoidal conidia accumulating in a green slimy drop, and hypha-like setoid extensions emerging through the conidial mass. During a revision of myrothecium-like isolates originating from rainforest areas of South America (Ecuador, Brazil) and Southeast Asia (Singapore), multilocus phylogenetic inferences (based on DNA sequence data of ITS, partial nuc 28S, and partial tef1a, rpb2 and tub2) and morphological studies concordantly revealed the occurrence of two undescribed species of Inaequalispora (I. longiseta sp. nov. and I. cylindrospora sp. nov.) and one undescribed species of Parvothecium (P. amazonensesp. nov.). Myrothecium setiramosum, M. dimorphum, and two undescribed taxa form the base of a new lineage, sister to the current Parvothecium lineage. This lineage is recognized as Digitiseta gen. nov., typified by D. setiramosa comb. nov. Digitiseta dimorpha comb. nov. is also proposed, and the new species D. parvodigitata sp. nov. and D. multidigitata sp. nov. are described.

The genus Myrothecium Tode ex FR

Article

Jan 1972

M. Tulloch

Mycetes argentinenses (Series IV)

Article

C. Spegazzini

Myrothecium-like new species from turfgrasses and associated rhizosphere Myrothecium-like new species from turfgrasses and associated rhizosphere

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