Biodiversity Data Journal : Taxonomic Paper
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Taxonomic Paper
Hypomyces pseudolactifluorum sp. nov. (Hypocreales: Hypocreaceae) on Russula sp. from Yunnan, PR China
expand article infoFeng-ming Yu‡,§, Ruvishika S. Jayawardena§, Jianwei Liu, Kevin D. Hyde‡,§,|, Qi Zhao‡,
‡ Key Laboratory of Economic Plants and Biotechnology, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, China
§ Center of Excellence in Fungal Research, Mae Fah Luang University, Chiangrai, Thailand
| Institute of Plant Health, Zhongkai University of Agriculture and Engineering, Guangzhou, China
¶ Institute of Applied Fungi, Southwest Forestry University, Kunming, China
Open Access

Abstract

Background

Hypomyces is a large genus of fungicolous fungi, parasitising the fruiting bodies of Agaricales, Boletales, Helotiales, Pezizales and Polyporales. Hypomyces currently comprises of 147 species widely distributed in Australia, China, France, Germany, Italy, Japan, North America, Sri Lanka, Thailand and UK. Amongst them, 28 species have been recorded in China.

New information

Hypomyces pseudolactifluorum sp. nov., growing on the fruiting bodies of Russula sp. in subsect. Lactarioideae and collected from Yunnan, China, is described with illustrations and molecular phylogenetic data (combined ITS, LSU, TEF1-α and RPB2 sequence dataset). The new species is characterised by semi-immersed to immersed perithecia and fusiform, apiculate and verrucose ascospores. We also review the species diversity of the genus Hypomyces in China.

Keywords

Mycoparasite, species diversity, muti-gene phylogeny

Introduction

Fungicolous fungi are a large and diverse ecological group, currently containing more than 1500 taxa distributed in many lineages across the fungal kingdom (Põldmaa 2011, Sun et al. 2019a). Hypomyces (Fr.) Tul. & C. Tul. is an important genus of fungicolous fungi and placed in the family Hypocreaceae (Hypocreales, Sordariomycetes, Ascomycota) (Hyde et al. 2020). Hypomyces was originally introduced as a subgenus of Hypocrea Fr. (Fries 1825) and then Tulasne and Tulasne (1860) revised it to a genus and designated H. lactifluorum (Schwein.) Tul. & C. Tul. from the USA as its type. Hypomyces parasitises the fruiting bodies of Agaricales, Boletales, Helotiales, Pezizales and Polyporales (Rossman et al. 1999, Tamm and Põldmaa 2013, Sun et al. 2019a). Hypomyces is characterised by: superficial or immersed, spherical to ovate, pyriform, papillate and yellow, orange, tawny red or green perithecia in a subiculum; 8-spored, subcylindrical to cylindrical and with a thickened apical asci; and ellipsoid, lanceolate, fusiform to navicular, 0-1-septate or rarely 3-septate, hyaline, spinulose or verrucose and smooth-walled ascospores (Rossman et al. 1999, Zeng and Zhuang 2015). Its allied genera include Cladobotryum Nees, Mycogone Link, Sepedonium Link and Stephanoma Wallr (Wijayawardene et al. 2017) and its asexual morphs are Acremonium-, Dactylaria-, Papulaspora-, Trichothecium- or Verticillium-like (Jaklitsch et al. 2006, Hyde et al. 2020). Hypomyces currently comprises of 147 species in Species Fungorum (http://www.speciesfungorum.org/, accessed in April 2020) and is widely distributed in Australia, China, France, Germany, Italy, Japan, North America, Sri Lanka, Thailand and UK (Zhuang et al. 2012, Rossman et al. 2013, Zeng and Zhuang 2016, Zare and Gams 2016, Lechat et al. 2017, Wei and Kirschner 2017, Sun et al. 2019a, Sun et al. 2019b, Zeng and Zhuang 2019). Amongst them, 28 species have been reported in China (Table 1).

Table 1.

Species diversity of the genus Hypomyces in China (29 species in total).

Taxa names

Hosts

Distribution

References

Hypomyces amaniticola

Amanita sp.

China (Yunnan)

Zeng and Zhuang 2016

H. aurantius

Agaricus bisporus, Polyporales (Cymatoderma sp., Laetiporus sulphureus, Panellus sp., Polyporus picipes), Stereum sp.

China (Anhui, Fujian, Guangxi, Hainan, Hebei, Hunan, Jiangsu, Jiangxi, Shanghai, Sichuan, Zhejiang), New Zealand, USA

Chen and Fu 1989, Põldmaa 2011, Luo and Zhuang 2012

H. aureonitens

Phlebia tremellosa, Polyporus sp.

China (Fujian, Guangxi), Europe

Teng 1963, Sun et al. 2019a

H. chlorinigenus

Agaricaceae, Boletaceae

Belgium, China (Taiwan), Guyana; Indonesia, New Zealand, USA

Rogerson and Samuels 1989, Zeng and Zhuang 2016

H. chrysospermus

Boletus sp., Hemileccinum impolitum, Suillus americanus, Russula sp.

China (Fujian, Jiling, Nanjing), Russia

Ma 2008, Luo and Zhuang 2012

H. completiopsis

Boletus sp.

China (Yunnan)

Zeng and Zhuang 2016

H. fistulina

Fistulina sp.

China (Guangxi)

Sun et al. 2019b

H. hubeiensis

Agaricus sp.

China (Hubei)

Zeng and Zhuang 2019

H. hyalinus

Agaricales (Amanita sp.), Polyporales

Canada, China (Jiangsu), Japan, USA

Teng 1934, Teng 1963, Rogerson and Samuels 1994

H. lateritius

Lactarius camphoratus, L. chelidonium, L. controversus, L. deliciosus, L. sanguifluus, L. thejogalus, L. trivialis, Lactarius sp.

Canada, China (Tibet), Europe, Japan, Mexico, New Zealand, USA

Rogerson and Samuels 1994, Luo and Zhuang 2012

H. luteovirens

Russula atropurpurea, R. rosea, R. sanguinaria, Russula sp.

Canada, China (Inner Mongolia), Europe, Japan, Russia, USA

Rogerson and Samuels 1994, Ma 2008

H. macrosporus

Russulaceae

China (Hubei), Mexico, USA

Rogerson and Samuels 1994, Luo and Zhuang 2012

H. microspermus

Boletaceae, Boletus sp., Imleria badia, Xanthoconium affine, Xerocomellus chrysenteron, Xerocomus sp.

Canada, China (Fujian, Guizhou, Hainan, Hubei, Jilin, Taiwan, Yunan), Indonesia, New Zealand, USA

Rogerson and Samuels 1989, Zeng and Zhuang 2016

H. mycophilus

Auricularia sp., Bulgari sp., Marasmius sp., Polyporus sp., Trametes versicolor

China (Guangdong), USA

Rogerson and Samuels 1993, Zeng et al. 2017

H. ochraceus

Decaying leaves, wood and fungi (e.g. Russula sp.)

China (Guangxi, Yunnan), Europe, USA

Teng 1963, Sun et al. 2019a

H. orthosporus

Polyporales

China (Tibet), Estonia, Finland, The Netherlands

Põldmaa 1996, Zeng and Zhuang 2019

H. papulasporae

Geoglossum difforme, G. fallax, G. glabrum, G. nigritum, G. simile, Glutinoglossum glutinosum, Trichoglossum hirsutum, T. walteri

China, USA, New Zealand

Rogerson and Samuels 1985, Sun et al. 2019a

H. polyporinus

Auricularia auricula-judae, Polyporales, Trametes versicolor, T. pubescens, Polyporus sp.

Canada, China (Guangxi), USA

Teng 1963, Rogerson and Samuels 1993

H. pseudolactifluorum sp. nov.

Russula sp.

China (Yunnan)

This study

H. rosellus

Agaricus bisporus, Armillaria sp., Hydnellum sp., Hyphoderma sp., Mycena sp., Polyporus sp., Russula sp., Trichaptum sp.

China (Gansu), Europe, Iran, Japan, Korea, USA

Tamm and Põldmaa 2013, Sun et al. 2019b

H. semicircularis

Ganoderma sichuanense, Microporus xanthopus

Cuba, China

Wei and Kirschner 2017, Sun et al. 2019a

H. sibirinae

Aphyllophorales, Boletus sp., Polyporales

China (Hunan), Indonesia, USA

Samuels et al. 1990, Zeng et al. 2017, Sun et al. 2019a

H. sinicus

Schizophyllum sp.

China (Anhui)

Zhuang et al. 2012

H. stephanomatis

Humaria hemisphaerica, Humaria sp.

Canada, China (Hubei), Germany, USA

Rogerson and Samuels 1985,Zeng and Zhuang 2016

H. subiculosus

Polyporaceae (Microporus affinis, Trametes versicolo)

China (Anhui, Beijing, Guangxi, Zhejiang), Cuba, Japan

Rogerson and Samuels 1993, Luo and Zhuang 2012

H. succineus

Pholiota sp.

China (Taiwan), USA

Rogerson and Samuels 1994, Zeng and Zhuang 2016

H. tegillum

Aphyllophorales, Polyporales

Brazil, China (Guangxi, Yunnan), Panama, USA

Rogerson and Samuels 1993, Luo and Zhuang 2012

H. triseptatus

Bark or associated with an ascomycete; Pyrenomycete

China (Hunan, Guangdong), Gabon

Rossman and Rogersson 1981, Zeng et al. 2017

H. yunnanensis

Boletus sp.

China (Yunnan)

Zeng and Zhuang 2016

Fungicolous fungi play important roles in the processes of the growths and degradations of their hosts. With the rapid development of mushroom industries, the fungicolous fungi on mushrooms have received more and more attention (Hyde et al. 2019). In this paper, we introduce a new member of fungicolous fungi, Hypomyces pseudolactifluorum sp. nov., on the fruiting bodies of Russula sp., collected from Yunnan Province, China. At the same time, we review the species diversity of the genus Hypomyces in China.

Materials and methods

Collections and Morphology

Hypomyces specimens, including their host mushrooms, were collected in an evergreen broad-leaved forest in Baihualing, Baoshan, Yunnan Province, China. The specimens, as well as collected host mushrooms, were placed on a piece of aluminium foil at first, then rolled the paper into a cylinder, twisted at the ends for sealing and lastly taken back to the laboratory for study (McKnight and McKnight 1997). Colour codes were recorded following those of Kornerup and Wanscher (1978). A Nikon Coolpix P510 camera was used to take photos in the wild. Dried specimens were observed and photographed using an Olympus SZ61 stereomicroscope and a Nikon ECLIPSE Ni compound microscope fitted with a Canon EOS 600D digital camera. Measurements were made using the Tarosoft® Image Frame Work programme v.0.9.7. The colour change of the perithecial wall was tested using 5% potassium hydroxide (KOH). Type specimens are deposited at the Herbarium of Mae Fah Luang University, Thailand (MFLU) and the Herbarium of Cryptogams Kunming Institute of Botany, Chinese Academy of Sciences, PR China (HKAS).

DNA extraction, PCR amplification and sequencing

The genomic DNA was extracted from the dried materials using the CTAB method (Doyle 1987). Tissues from the ascocarps of parasitic fungi and fruiting bodies of the host mushrooms were used to extract DNA, respectively. Primer pairs ITS1F/ITS4 (White et al. 1990), LR0R/LR5 (Rehner and Samuels 1994, Vilgalys and Hester 1990), TEF1-α 983f/TEF1-α 2218r (Carbone and Kohn 1999, Rehner and Buckley 2005) and RPB2-5f/RPB2-7cR (Liu et al. 1999) were used for amplification of the ITS, LSU, TEF1-α and RPB2 gene regions.

PCR was performed in a 25 μl reaction volume: 12.5 μl Taq PCR Master Mix (Abmgood, Richmond, BC, Canada), 1 μl forward primer, 1 μl reverse primer, 1 μl DNA template and 9.5 μl ddH2O. For ITS and LSU, PCR reaction conditions are: 8 min at 94ºC, followed by 30 s at 94ºC, 30 s at 52ºC and 1 min at 72ºC for 35 cycles and a final extension of 10 min at 72ºC. PCR reaction conditions of TEF1-α and RPB2 are: 8 min at 94ºC, followed by 1 min at 95ºC, 45 s at 59ºC for RPB2/55ºC for TEF1-α and 1 min at 72ºC for 35 cycles and a final extension of 10 min at 72ºC. The PCR products were detected using agarose gel electrophoresis and, in the gel documentation system, clear bands were observed. Sequencing was performed by Sangon Biotech (Shanghai) Co. Ltd., PR China; partial impure products were purified using the Cycle-pure-kit (Omega, America) and then cloned into pClone007 Simple vector (TSV-007S from Beijing TsingKe Biotech). Twenty clones of PCR products of each gene were sequenced using the universal primer pairs M13-47/M13-48.

Sequence alignment and phylogenetic analyses

The parasitic fungus: Hypomyces pseudolactifluorum sp. nov.

Molecular phylogenetic trees were constructed using our sequencing results of H. pseudolactifluorum sp. nov. and the voucher sequences of their allies obtained from NCBI GenBank (Table 2). Two species of Trichoderma, T. hamatum (DAOM 167057) and T. viride (CBS 119325) were used as outgroup taxa. All sequences were assembled and aligned using MAFFT v6.8 (Katoh et al. 2005) and manually edited via BioEdit version 7.0.9 (Hall 1999). Four sequence matrices of ITS, LSU, TEF1-α and RPB2 genes, respectively, were compiled. The optimal substitution model for each gene dataset was determined using jModelTest2 under the Akaike information criterion (AIC) (Darriba et al. 2012). The results indicated that the GTR+I+G model (-lnL = 8658.2624) is optimal for the ITS dataset, as well as the TIM1+I+G model (-lnL = 4392.5417) for LSU, the TrN+I+G model (-lnL = 5751.4959) for TEF1-α and the model SYM+I+G (-lnL = 6419.6669) for RPB2, respectively. Using the aligned sequence matrices, a combined gene sequence dataset (ITS, LSU, TEF1-α and RPB2, orderly) was assembled and aligned and was finally deposited in TreeBASE database (http://purl.org/phylo/treebase/phylows/study/TB2:S26593?x-access-code=152eadfc2292343af7627cfad5c2946c&format=html).

Table 2.

Voucher information and GenBank accession numbers for samples appearing in the Hypomyces phylogenetic tree. Our sequencing results are displayed in bold. (Label T indicate the sequences from ex-type strains.)

Taxa names

Specimen/Strain

number

GenBank accession numbers

References

ITS

LSU

TEF1-α

RPB2

Cladobotryum amazonense

CBS 470.80

MH861285

MH873051

/

/

Vu et al. 2019

C. apiculatum

CBS 174.56 T

NR_159770

MH869109

/

/

Vu et al. 2019

C. asterophorum

CBS 676.77 T

FN859395

MH872869

FN868712

FN868649

Põldmaa 2011

C. croceum

CBS 231.95

MH862511

MH874154

/

/

Vu et al. 2019

C. cubitense

CBS 416.85

FN859396

/

FN868713

FN868650

Põldmaa 2011

G.A. m643.w

FN859397

/

FN868714

FN868651

Põldmaa 2011

C. indoafrum

TFC 201295

FN859403

FN859403

FN868721

FN868657

Põldmaa 2011

C. multiseptatum

CBS 472.71 T

FN859405

MH871991

FN868723

FN868659

Põldmaa 2011

C. obconicum

CBS 528.81

MH861373

MH873126

/

/

Vu et al. 2019

C. paravirescens

TFC 97-23 T

FN859406

FN859406

FN868724

FN868660

Põldmaa 2011

C. penicillatum

CBS 407.80 T

FN859407

MH873046

FN868725

FN868661

Põldmaa 2011

C. protrusum

CBS 118999

FN859408

FN859408

FN868726

FN868662

Põldmaa 2011

C. purpureum

CBS 154.78 T

FN859415

/

FN868733

FN868669

Põldmaa 2011

C. rubrobrunnescens

CBS 176.92 T

FN859416

MH874016

FN868734

FN868670

Põldmaa 2011

Cladobotryum sp.

FSU 5046

FN859421

/

FN868739

FN868675

Põldmaa 2011

C. stereicola

CBS 457.71 T

MH860217

MH871984

/

/

Vu et al. 2019

C. tchimbelense

TFC 201146 T

FN859419

FN859419

FN868737

FN868673

Põldmaa 2011

C. tenue

CBS 152.92 T

FN859420

FN859420

FN868738

FN868674

Põldmaa 2011

Hypomyces aconidialis

TFC 201334 T

FN859457

FN859457

FN868775

FN868711

Põldmaa 2011

TFC 201215

FN859456

FN859456

FN868774

FN868710

Põldmaa 2011

H. albidus

CBS 460.71

MH860220

MH871987

/

/

Vu et al. 2019

H. armeniacus

TFC 02-86/2 T

FN859424

FN859424

FN868742

FN868678

Põldmaa 2011

H. aurantius

TFC 95-171

FN859425

FN859425

FN868743

FN868679

Põldmaa 2011

H. australasiaticus

TFC 03-8 T

FN859428

FN859428

FN868746

FN868681

Põldmaa 2011

TFC 99-95

FN859427

/

FN868745

FN868680

Põldmaa 2011

H. australis

TFC 2007-18

AM779860

AM779860

FN868747

/

Põldmaa 2011

H. boletiphagus

MFLU 17-1391

MH459152

MH459168

/

MH464785

Sun et al. 2019b

H. boletus

MFLU 17-1392

MH459153

MH459170

/

MH464787

Sun et al. 2019b

H. chlorinigenus

KSH511

KT946843

/

KU041505

KU041493

Otto et al. 2016

KSH512 T

KT946844

/

KU041506

KU041494

Otto et al. 2016

H. completus

KSH411 (S172)

KT946842

/

KU041504

KU041492

Otto et al. 2016

KSH410 (S171) T

KT946841

/

KU041503

KU041491

Otto et al. 2016

H. corticiicola

CBS 137.71 T

MH860037

MH871817

/

/

Vu et al. 2019

H. dactylarioides

CBS 141.78 T

FN859429

MH872879

FN868748

FN868683

Põldmaa 2011

H. fistulina

HMAS 279800 T

MH459154

MH459171

MH464781

/

Sun et al. 2019b

H. gabonensis

TFC 201156 T

FN859430

FN859430

FN868749

FN868684

Põldmaa 2011

H. heterosporus

CBS 719.88 T

FN859398

MH873844

FN868716

FN868653

Põldmaa 2011

H. khaoyaiensis

G.J.S. 01-304 T

FN859431

AJ583483

FN868750

FN868685

Põldmaa 2011

H. lactifluorum

TAAM 170476 T

FN859432

EU710768

FN868751

EU710773

Põldmaa 2011

H. laeticolor

JCM 10758 T

LC228655

LC228712

/

/

Sun et al. 2019b

H. luteovirens

CBS 128483

MH864958

MH876402

/

/

Vu et al. 2019

H. mycophilus

CBS 175.56

MH857567

MH869110

/

/

Vu et al. 2019

H. odoratus

G.A. m329

FN859434

FN859434

FN868753

FN868688

Põldmaa 2011

TFC 200887

FN859439

/

FN868757

FN868693

Põldmaa 2011

H. orthosporus

10736

MK478468

MN044763

MK484609

/

Zeng and Zhuang 2019

H. penicillatus

NBRC 100524

LC146740

LC146740

/

/

Sun et al. 2019b

H. pseudolactifluorum sp.nov.

MFLU 20-0265 T

MT260402

MT260399

MT259361

MT259359

This study

MFLU 20-0266

MT260403

MT260400

MT259362

MT259360

This study

H. polyporinus

ATCC 76479

AF543771

AF543793

AF543784

/

Currie et al. 2003

H. pseudocorticiicola

JCM 12654 T

LC228663

LC228721

/

/

Sun et al. 2019b

H. rosellus

TFC 201071

FN859443

FN859443

FN868762

FN868697

Põldmaa 2011

H. samuelsii

CBS 536.88

FN859444

/

FN868763

FN868698

Põldmaa 2011

TFC 2007-23

FN859451

FN859451

FN868769

FN868705

Põldmaa 2011

H. semicircularis

CBS 705.88 T

FN859417

MH873843

FN868735

FN868671

Põldmaa 2011

H. semitranslucens

CBS 458.71

MH860218

MH871985

/

/

Vu et al. 2019

CBS 821.70

MH859960

MH871759

/

/

Vu et al. 2019

H. sibirinae

CBS 744.88

MH862151

AJ459304

/

/

Vu et al. 2019

H. sinicus

HMAS 251317 T

NR_156252

MN044986

MK484610

/

Zhuang et al. 2012

H. stephanomatis

G.J.S. 88-50

/

AF160243

AF534632

AF545566

Põldmaa et al. 2000

H. subiculosus

TFC 97.166

FN859452

/

FN868770

FN868706

Põldmaa 2011

H. tubariicola

CBS 115.79 T

KU382164

MH872953

/

/

Vu et al. 2019

CBS 225.84

KU382162

KU382220

/

/

Zare and Gams 2016

H. virescens

G.A. i1906 T

FN859454

/

FN868772

FN868708

Põldmaa 2011

G.A. i1899

FN859453

/

FN868771

FN868707

Põldmaa 2011

Sepedonium ampullosporum

CBS 392.52 T

MH857094

MH868629

/

/

Vu et al. 2019

S. chalcipori

CBS 278.92

MH862358

MH874023

/

/

Vu et al. 2019

CBS 148.92 T

MH862347

MH874014

/

/

Vu et al. 2019

S. tulasneanum

CBS 940.69

MH859489

MH871270

/

/

Vu et al. 2019

Trichoderma hamatum

DAOM 167057 T

EU280124

HM466686

AF534620

AF545548

Hoyos-Carvajal et al. 2009

T. viride

CBS 119325 T

DQ677655

/

DQ672615

EU711362

Jaklitsch et al. 2006

Maximum Likelihood (ML) analysis was performed using IQ-Tree (Nguyen et al. 2014, Chernomor et al. 2016) with the computing models listed above and a bootstrap test of 1000 replicates. Bayesian Inference (BI) analysis was carried out using MrBayes v3.2.6 (Ronquist et al. 2012). The TIM1 and TrN substitution models were replaced by the GTR model (Huelsenbeck and Rannala 2004). Four simultaneous Markov Chain Monte Carlo (MCMC) chains were run for random trees of 10,000,000 generations and were sampled by every 100 generations. The computing was stopped when the standard deviation of the split frequencies fell below 0.01 and ESS values > 200. Subsequently, phylogenetic trees were summarised and posterior probabilities (PP) were performed using MCMC by discarding the first 25% generations as “burn-in” (Huelsenbeck and Ronquist 2001). Gaps were treated as missing data. Phylogenetic trees were viewed in FigTree v.1.4.2 (http://tree.bio.ed.ac.uk/software/figtree).

The host mushroom: Russula sp.

Voucher sequences (ITS gene) for phylogenetic analyses of the host mushroom and its allies were obtained from our sequencing results and GenBank databases (Li et al. 2020) (Table 3). Five species of Russula subg. Compactae, R. acrifolia, R. adusta, R. eccentrica, R. nigricans and R. subnigricans were selected as the outgroup taxa. Sequence alignment and phylogenetic analyses followed those of the parasitic fungus above. ML analysis was performed using IQ-Tree with TVM+I+G model (-lnL = 5298.7964) (Nguyen et al. 2014, Chernomor et al. 2016). The ITS sequence matrix of the host mushroom and its allies were deposited in the TreeBASE database (http://purl.org/phylo/treebase/phylows/study/TB2:S26693?x-access-code=2e445b17aebe1f93266051a8920ae62f&format=html).

Table 3.

Voucher information and GenBank accession numbers for samples appearing in the Russula phylogenetic tree. Our sequencing results are displayed in bold.

Taxa names

Specimen/Strain number

GenBank accession

References

Russula acrifolia

TUB UE12.09.2003-3

DQ421998

Eberhardt 2002

R. adusta

PC 547RUS27

AY061652

Miller and Buyck 2002

R. aff. chloroides

FH 12273

KT934015

Looney et al. 2016

R. brevipes

SMI329

FJ845429

Kranabetter et al. 2009

R. brevipes

JS160927-01

MG407682

GenBank

R. brevipes

TENN 070667

KY848511

Looney et al. 2018

R. brevipes var. acrior

JMP 0058

EU819422

Palmer et al. 2008

R. byssina

HGAS-MF 009907

MN648951

Li et al. 2020

R. byssina

HGAS-MF009921

MN648949

Li et al. 2020

R. byssina

HGAS-MF 009913

MN648950

Li et al. 2020

R. cascadensis

UBC F30189

KX812838

Bazzicalupo 2018

R. cascadensis

UBC F19691

HM240541

Buyck et al. 2017

R. cf. angustispora

PC BB2004-252

EU598152

GenBank

R. cf. brevipes

F 28785

MH718203

GenBank

R. cf. brevipes

F CDW47

GQ166868

GenBank

R. cf. brevipes

GO 2009-276

KC152212

GenBank

R. cf. delica

UBC F30260

KX812852

Bazzicalupo 2018

R. chloroides

PC 205RUS24

AY061663

Miller and Buyck 2002

R. chloroides

UBC F20353

KC581331

GenBank

R. chloroides

RUS-12091401

KF432954

Wisitrassameewong et al. 2014

R. cremicolor

HGAS-MF 009901

MN648955

Li et al. 2020

R. cremicolor

HGAS-MF 009908

MN648952

Li et al. 2020

R. cremicolor

HGAS-MF 009912

MN648953

Li et al. 2020

R. cremicolor

HGAS-MF 009919

MN648954

Li et al. 2020

R. delica

hue22 (TUB)

AF418605

Eberhardt 2002

R. delica

FH 12-272

KF432955

Wisitrassameewong et al. 2014

R. delica

HA 2015-004

KX263000

Aghajani et al. 2017

R. delica

PC 496RUS26

AY061671

Miller and Buyck 2002

R. delica

TUB hue22

AF418605

Eberhardt 2002

R. delica

UBC F30263

KX812842

Bazzicalupo 2018

R. delica

RMUKK 37

KX267630

GenBank

R. delica

KA 12-1327

KR673555

Kim et al. 2015

R. delica

HMJAU 32182

KX094989

Liu et al. 2017

R. eccentrica

HCCN 23685

KC699778

Park et al. 2014

R. japonica

MHHNU 31049

MK167414

Chen and Zhang 2019

R. japonica

HGAS-MF 009923

MN648957

Li et al. 2020

R. japonica

HGAS-MF 009915

MN648956

Li et al. 2020

R. leucocarpa

HGAS-MF 009910

MN648948

Li et al. 2020

R. leucocarpa

HGAS-MF 009916

MN648947

Li et al. 2020

R. littoralis

PC 1222IS87

AY061702

Miller and Buyck 2002

R. marangania

MEL 2293694

EU019930

Lebel and Tonkin 2007

R. nigricans

TUB fo46761

AF418607

Eberhardt 2002

R. pallidospora

PC 2-1221IS85

AY061701

Miller and Buyck 2002

R. pumicoidea

MEL T-14771

EU019931

Lebel and Tonkin 2007

R. sinuata

MEL H4755

EU019943

Lebel and Tonkin 2007

R. subnigricans

MHHNU ZP6932

EF534351

Yin et al. 2008

R. vesicatoria

PC 0124666

KY800359

Buyck et al. 2017

Russula sp.

MFLU 20-0265 (host)

MT755627

In this study

Taxon treatment

Hypomyces pseudolactifluorum F. M. Yu, Q. Zhao & K. D. Hyde, sp. nov.

Materials    Download as CSV 
Holotype:
  1. scientificName:
    Hypomyces pseudolactifluorum
    ; country:
    China
    ; stateProvince:
    Yunnan
    ; locality:
    Baoshan, Longyang, Baihualing
    ; verbatimElevation:
    2094m
    ; locationRemarks:
    label transliteration: "Yunnan, Baoshan, Longyang, Baihualing, on Russula sp., 20 July 2018, Jian-Wei Liu; [云南保山百花岭 2094 m, 2018.07.20, 刘建伟]
    ; verbatimCoordinates:
    25°17.931’N, 98°47.0718’E
    ; decimalLatitude:
    25.2989
    ; decimalLongitude:
    98.7845
    ; georeferenceProtocol:
    label
    ; lifeStage:
    Telemorph
    ; catalogNumber:
    MFLU 20-0265
    ; recordedBy:
    Jian-Wei Liu
    ; identifiedBy:
    Feng-Ming Yu
    ; dateIdentified:
    2019
Paratype:
  1. scientificName:
    Hypomyces pseudolactifluorum
    ; country:
    China
    ; stateProvince:
    Yunnan
    ; locality:
    Baoshan, Longyang, Baihualing
    ; verbatimElevation:
    2094m
    ; locationRemarks:
    label transliteration: "Yunnan, Baoshan, Longyang, Baihualing, on Russula sp., 20 July 2018, Jian-Wei Liu; [云南保山百花岭 2094 m, 2018.07.20, 刘建伟]
    ; verbatimCoordinates:
    25°17.931’N, 98°47.0718’E
    ; decimalLatitude:
    25.2989
    ; decimalLongitude:
    98.7845
    ; georeferenceProtocol:
    label
    ; lifeStage:
    Telemorph
    ; catalogNumber:
    MFLU 20-0266
    ; recordedBy:
    Jian-Wei Liu
    ; identifiedBy:
    Feng-Ming Yu
    ; dateIdentified:
    2019
Isotype:
  1. scientificName:
    Hypomyces pseudolactifluorum
    ; country:
    China
    ; stateProvince:
    Yunnan
    ; locality:
    Baoshan, Longyang, Baihualing
    ; verbatimElevation:
    2094m
    ; locationRemarks:
    label transliteration: "Yunnan, Baoshan, Longyang, Baihualing, on Russula sp., 20 July 2018, Jian-Wei Liu; [云南保山百花岭 2094 m, 2018.07.20, 刘建伟]
    ; verbatimCoordinates:
    25°17.931’N, 98°47.0718’E
    ; decimalLatitude:
    25.2989
    ; decimalLongitude:
    98.7845
    ; georeferenceProtocol:
    label
    ; lifeStage:
    Telemorph
    ; catalogNumber:
    HKAS 107300
    ; recordedBy:
    Jian-Wei Liu
    ; identifiedBy:
    Feng-Ming Yu
    ; dateIdentified:
    2019

Description

Index Fungorum number: IF557817

Sexual morph. Subiculum light yellow (4A4–5) when fresh and pale orange, light orange to brownish-orange (5A3–4, 5C4, 6C6) after being dried, usually covering the pileus, stipe and deformed gills of the host mushroom. Perithecia aggregated, semi-immersed to immersed in subiculum, except for their erumpent papilla, yellowish-brown to dark brown (5E6, 6E6, 6F6–8), pyriform to subglobose, 262–484 × 136–284 μm; perithecial wall 12–25 μm thick, single-layer, cells 9–22 × 4–8 μm. Papilla prominent, 129–177 μm high, at base 135–284 μm wide. Asci 8-spored, cylindrical, 147–222 × 4–9 μm; apex thickened, 4.9–6.0 wide and 2.5-3.0 μm high. Ascospores uniseriate and with ends overlapping, fusiform, 30–38 × 6–8 μm, single-septate, septum median and with dense verrucae and prominently apiculate, apiculi 4.5–8.0 μm long, straight or curved. Asexual morph: unknown. (Fig. 1)

Figure 1.  

Hypomyces pseudolactifluorum sp. nov.. a: The host mushroom (Russula sp.); b-e: Perithecia embedded in subiculum effused over the substratum; d-e: Median sections of an ascoma; f: Section of peridium; g-l: Asci with ascospores; m-t: Ascospores. Scale bars: a = 5 cm; b = 1 mm; c = 200 μm; d, e = 100 μm; f, g= 50 μm; h - l = 20 μm; m - t = 10 μm.

Diagnosis

The new species is similar to Hypomyces lactifluorum on Russula and Lactarius spp. from North America (Rogerson and Samuels 1994), but has smaller perithecia and shorter asci. The main differences of the two species are compared in Table 4.

Table 4.

Main differences between Hypomyces lactifluorum and H. pseudolactifluorum sp. nov..

H. lactifluorum

(Rogerson and Samuels 1994)

H. pseudolactifluorum

Subiculum

Pale yellowish-orange to bright orange (young), in age becoming deep red, reddish-purple to very dark purple (old), occasionally fading to pink, turning purple in 3% KOH.

Light yellow (4A4–5) when fresh, and pale orange to light orange to brownish-orange (5A3–4, 5C4, 6C6) after being dried, KOH (-).

Perithecia

Ovate to obpyriform, deep orange to reddish-purple, 400–600 × 200–450 μm

Pyriform to subglobose, yellowish-brown to dark brown (5E6, 6E6, 6F6–8), 262–484 × 136–284 μm

Embedded type

Immersed except for papilla

Semi-immersed to immersed except for papilla

Papilla

Averaging 120 μm high, 120 μm wide

129–177 μm high and 135 –284 μm wide at base

Asci

Long cylindrical, 200–260 × 5–10 μm

Cylindrical, 147–222 × 4–8.5 μm

Ascospores

Fusiform, 1-septate, 35–40 × 4.5–7 μm

Fusiform, 1-septate, 30–38 × 5.5–8 μm

Apiculi

4.5–7.5 μm long

4–6 μm long

Hosts

Russula and Lactarius spp.

Russula sp.

Distribution

North America

P.R. China (Yunnan)

Etymology

Referring to the most closely-related species, Hypomyces lactifluorum.

Distribution

PR CHINA (Yunnan).

Host

On the fruiting bodies of Russula sp. that grew on the humus layer in an evergreen broad-leaved forest of a rainforest. The host mushrooms: basidiocarps medium-sized and infundibuliform, pilei 63−77 mm in diameter. As serious degradation has occurred, the colour and other characters of the host mushrooms cannot be determined. Molecular phylogenetic evidence indicates it is a Russula species.

Notes

Only sexual morph had been discovered on the hosts (Russula sp.) of the new species.

Analysis

Phylogenetic analyses

Parasitic fungus: Hypomyces pseudolactifluorum sp. nov

The combined ITS+LSU+TEF1-α+RPB2 sequence dataset (excluding the outgroup taxa) contains 3,262 characters (709 for ITS, 893 for LSU, 921 for TEF1-α and 739 for RPB2) from 56 Hypomyces species and two Trichoderma species. Amongst them, 2,246 characters are constant, 209 variable characters are parsimony-uninformative and 807 characters are parsimony-informative. The ML and BI analyses resulted in trees with similar topology and support values and the ML tree is shown in Fig. 2.

Figure 2.  

ML tree of Hypomyces pseudolactifluorum sp. nov. and its allies generated from a combined ITS, LSU,TEF1-α and RPB2 gene sequence dataset. Supporting values of MLBP (left, greater than 75%) and BIBP (right, greater than 0.9) are shown at the nodes, respectively. The new species is marked in red.

In the phylogenetic tree, the parasitic fungi MFLU 20-0265 and MFLU 20-0266 are clustered together and formed a distinct lineage with the same branch length and strong supportive values (MLBP = 100%, BIPP = 1), which support them to be conspecific. The parasitic fungi are closely related H. lactifluorum and they form a sister clade also with strong supportive values (MLBP = 100%, BIPP = 1). Comparing the gene sequences of the two species, there are 25 bp (4.3%) differences across 582 bp in ITS, 28 bp (3.2%) differences across 870 bp in LSU, 24 bp (2.6%) differences across 921 bp in TEF1-α and 24 bp (3.2%) differences across 739 bp in RPB2 (Suppl. material 1). Following the recommendations from Jeewon and Hyde (2016), we assign the parasitic fungi as H. pseudolactifluorum sp. nov.

The host mushroom: Russula sp.

According to the ITS phylogenetic tree of the host mushroom and its allies, the host mushroom (MFLU 20-0265) is clustered together with Russula leucocarpa (HGAS-MF 009910 and HGAS-MF 009916) (MLBP = 100%) in subsect. Lactarioideae. However, their ITS sequences have 24 bp (3.5%) differences across 694 bp, which indicated they may be two distinct species. Due to lack of sufficient morphological evidence, the host mushroom was temporarily identified as Russula sp. (Fig. 3).

Figure 3.  

ML tree of Russula sp. (in red) and it allies inferred from the ITS sequence dataset. Five species of Russula subg. Compactae were used as the outgroup taxa. Supporting values of MLBP (greater than 75%) are shown at the nodes.

Discussion

Zeng and Zhuang (2016) described H. amaniticola on Amanita sp. and H. completiopsis and H. yunnanensis on Boletus sp., also from China. Though with similar colour and shapes of perithecia, the host of H. pseudolactifluorum sp. nov. is decidedly different from those of these three species. Furthermore, H. pseudolactifluorum sp. nov. (KOH-) has smaller perithecia and larger ascospores than those of H. completiopsis (KOH+) and H. pseudolactifluorum sp. nov. has larger perithecia, asci and ascospores than those of H. amaniticola (KOH+) and H. yunnanensis (KOH-). Unfortunately, these three species all lack molecular data.

With the rapid development of mushroom industries, fungal pathogens on mushrooms have received more and more attention (Hyde et al. 2019). The fungicolous fungi Hypomyces is an important group of mushroom pathogens. Many Hypomyces species, for example, H. aurantius, H. perniciosus, H. rosellus, H. odoratus etc., have all been recorded as the causes of Cobweb or Web bubble disease which seriously influence mushroom industries (Fletcher and Gaze 2007, Carrasco et al. 2017, Zhang et al. 2017, Zhang et al. 2017). Russula is the largest subgenus in agaric with approximately 800 species (Li et al. 2020) and many Russula species are important edible mushrooms. Since growing on Russula sp., H. pseudolactifluorum sp. nov., as well as H. lactifluorum from North America (Rogerson and Samuels 1994), could be one of the potential pathogens of some Russula species in Asia.

Acknowledgements

The research is supported by the Second Tibetan Plateau Scientific Expedition and Research (STEP) Program (Grant No. 2019QZKK0503); the Open Research Project of “Cross-Cooperative Team” of the Germplasm Bank of Wild Species, Kunming Institute of Botany, Chinese Academy of Sciences (Grant No. 292019312511043); Science and Technology Service Network Initiative of the Chinese Academy of Sciences (KFJ-STS-QYZD-171); the Biodiversity Survey and Assessment Project of the Ministry of Ecology and Environment, PR China (2019HJ2096001006) and Impact of climate change on fungal diversity and biogeography in the Greater Mekong Subregion (Grant No. RDG6130001).

References

Supplementary material

Suppl. material 1: Hypomyces pseudolactifluorum sp. nov. (Hypocreales: Hypocreaceae) on Russula sp. from Yunnan, PR China 
Authors:  FENG-MING YU, RUVISHIKA S. JAYAWARDENA, JIAN-WEI LIU, KEVIN D. HYDE, QI ZHAO
Data type:  word
Brief description: 

Sequence differences of ITS, LSU, TEF1-α and RPB2 genes between H. lactifluorum (TAAM 170476) and H. pseudolactifluorum sp. nov.. The locus’ numbers refer to the nucleotide positions of the gene sequences of H. lactifluorum from GenBank. Gap is replaced by ‘-’.