IV ... MYCOTAXON 134(3)

MYCOTAXON

VOLUME ONE HUNDRED THIRTY-FOUR (3) — TABLE OF CONTENTS

134-3: TABLE OF CONTENTS, NOMENCLATURAL UPDATES, PEERS, EDITORIALS

Nomenclatural novelties & typifications 1.1... 0. eee eee vii

REVICW ETS GBa soe wb Seo cle ey KER ae wee es ahs y Enel ote eye a at aie WE A ix

SETAC ery asm, So An Tt OER Muley SMa Ey RRNA ame Sy A AP i BRA x

POTASH OR ATOURS, chet & LS AGRO Peal ohtmta De Shay, ester) Poul GMS Re as xi

ZO TD SUDMESSION PHO CCHUNE sos. 8y keels. ea pe DR ce ite Ge 9a ree xiii

RESEARCH ARTICLES

Amanita subjunquillea and its ectomycorrhizal association,

reported as new for Pakistan MuHAMMaAD IsHAQ, AROO] NASEER,

MUNAZZA KIRAN, MUHAMMAD FIAZ, ABDUL NASIR KHALID

New records of dictyostelid social amoebae from China

Pu Liu, SHUNHANG ZHANG, YUE ZOU, YU ZHANG, YONGJIA L1, Yu LI

Annulohypoxylon bawanglingense, A. diaoluoshanense,

A. guangxiense, and A. sichuanense spp. nov.

and new records from China WEI Li & Lin Guo

Pleurotheciopsis triseptata sp. nov. from China

CONG-CONG AI, JI-WEN XIA, XIU-GUO ZHANG, LI-GuUo Ma

Repetophragma verrucosum sp. nov. from China

CoONG-COoNG AI, JI-WEN XIA, XIU-GUO ZHANG, LI-Guo Ma

Pseudopyricularia cyperi, a new record for Iran

ADEL PORDEL, AMIRREZA AMIRMIJANI, MOHAMMAD JAVAN-NIKKHAH

Morganjonesia gen. nov. for two atypical Corynespora and

Teratosperma species Kal ZHANG, MIN QIAO, ZE-FEN

Yu, DE-WEI L1, RAFAEL E CASTANEDA-RUIZ

Botryosphaeria qinlingensis sp. nov. causing

oak frogeye leaf spot in China Linc-yu Liana, NING JIANG,

WEN-YAN CHEN, YING-MEI LIANG, CHENG-MING TIAN

Tretoheliocephala cylindrospora sp. nov., an asexual fungus

from Thailand CHARUWAN CHUASEEHARONNACHAI,

SAYANH SOMRITHIPOL, KANTHAWUT BOONMEE,

SALILAPORN NUANKAEW, NATTAWUT BOONYUEN

413

425

431

439

443

447

457

463

475

JULY-SEPTEMBER 2019... V

Novel cyphelloid fungi in Glabrocyphella, Heteroscypha,

and Rectipilus from Brazil LARISSA TRIERVEILER-PEREIRA,

JULIANO M. BALTAZAR, R. GREG THORN,

ADRIANA DE MELLO GUGLIOTTA 481

New species of Marthamyces and Ramomarthamyces gen. nov.

from New Zealand and the Cook Islands

PETER R. JOHNSTON & DUCKCHUL PARK 489

Septobasidium aquilariae sp. nov. from China

Ru1-Linc Ma, MENG GAo, XIANG-LIN ZHUANG, CHANG-LIN ZHAO 517

Limacella bangladeshana, first record of the genus in Thailand

JATURONG KuMLA, NAKARIN SUWANNARACH, SAISAMORN LUMYONG 529

New records of Acarospora and Psora from China

CHUN-XIAO WANG, CHUAN-FENG ZHENG, ZUN-TIAN ZHAO 535

Phylogenetic affinity and taxonomic reassessment of

Pseudoidium kalanchoes Monika GOTzZ, ELKE IpczaKx, Uwe BRauN 545

Beltraniomyces panthericolor and B. pulcher spp. nov.

from Brazil FLAVIA RODRIGUES BARBOSA, PATRICIA OLIVEIRA FIUZA,

GLEYSON CRISTIANO KORPAN BARBOSA, LOISE ARAUJO COSTA,

GABRIEL GINANE BARRETO, LUIS FERNANDO PASCHOLATI GUSMAO,

RAFAEL FELIPE CASTANEDA-RuIZ 555

Coenogonium hainanense sp. nov., and new records from China

X1A0-HAN Wu, WE-CHENG WANG,

MING-ZHU Dou, ZE-FENG JIA 561

Cystostereum sirmaurense sp. nov. from India

RAMANDEEP Kaur, AVNEET P. SINGH, G.S. DHINGRA 577

MycoBIoTA (FUNGA) NEW TO THE MYCOTAXON WEBSITE

Diversity of macrofungi in Yushan, Jiangsu, China (summary)

BING Xu, Haoyu Lu, DONGLEI ZHAO, WEI WANG, HonG Jit 581

BOOK REVIEWS AND NOTICES ELSE C. VELLINGA 583

vi... MYCOTAXON 134(3)

PUBLICATION DATE FOR VOLUME ONE HUNDRED THIRTY-FOUR (2)

MYCOTAXON for APRIL-JUNE 2019 (I-XIv + 223-412)

was issued on July 3, 2019

JULY-SEPTEMBER 2019... VII

NOMENCLATURAL NOVELTIES AND TYPIFICATIONS

PROPOSED IN MYCOTAXON 134(3)

Annulohypoxylon bawanglingense Wei Li bis & L. Guo

[FN 570628], p. 431

Annulohypoxylon diaoluoshanense Wei Li bis & L. Guo

[FN 570630], p. 432

Annulohypoxylon guangxiense Wei Li bis & L. Guo

[FN 570633], p. 434

Annulohypoxylon sichuanense Wei Li bis & L. Guo

[FN 570632], p. 434

Beltraniomyces panthericolor L.A. Costa, G.G. Barreto, ER. Barbosa, Fiuza &

R.E. Castafieda [MB 830313], p. 556

Beltraniomyces pulcher ER. Barbosa, Fiuza, G.C.K. Barbosa, R.F. Castafieda &

Gusmao [MB 830314], p. 558

Botryosphaeria qinlingensis C.M. Tian & L.Y. Liang

[MB 828967], p. 469

Coenogonium hainanense X.H. Wu & Z.F. Jia

[FN 570605], p. 563

Cystostereum sirmaurense R. Kaur, Avn. P. Singh & Dhingra

[MB 829964], p. 578

Marthamyces culmigenus (Ellis & Everh.) P.R. Johnst.

[IF 556322], p. 496

Glabrocyphella cyathiformis Trierv.-Per. & Thorn

[MB 828713], p. 482

Heteroscypha malmei (W.B. Cooke) Trierv.-Per., Baltazar & Thorn

[MB 828714], p. 484

Marthamyces harakeke P.R. Johnst.

[IF 556323], p. 499

Marthamyces maccormackii P.R. Johnst.

[IF 556324], p. 503

Marthamyces metrosideri P.R. Johnst.

[IF 556325], p. 505

Marthamyces renga P.R. Johnst.

[IF556326], p. 508

Morganjonesia R.F. Castaneda, K. Zhang & D.W. Li

[MB 830288], p. 458

vill ... MYCOTAXON 134(3)

Morganjonesia calophylli (Hol.-Jech. & R.F. Castaneda.) R.F. Castaneda,

K. Zhang & D.W. Li [MB 830290], p. 461

Morganjonesia litchii (Matsush.) K. Zhang & R.F. Castaneda

[MB 830291, p. 461

Oidium kalanchoes Listner ex U. Braun 1987 (neotypified)

[MBT 385509], p. 549

Pleurotheciopsis triseptata L.G. Ma, X.G. Zhang

[MB 831794], p. 440

Repetophragma verrucosum L.G. Ma & X.G. Zhang

[MB 831795], p. 444

Ramomarthamyces P.R. Johnst.

[IF 556327], p. 510

Ramomarthamyces barbatus (P.R. Johnst.) P.R. Johnst.

[IF 556329], p. 513

Ramomarthamyces dracophylli (P.R. Johnst.) P.R. Johnst.

[IF 556330], p. 513

Ramomarthamyces gilvus (Rodway) P.R. Johnst.

[IF 556331], p. 513

Ramomarthamyces tuku P.R. Johnst.

[IF 556333], p. 511

Septobasidium aquilariae C.L. Zhao

[MB 832332], p. 522

Tretoheliocephala cylindrospora Chuaseehar., Somrith. & Boonyuen

[MB 828099], p. 476

JULY-SEPTEMBER 2019...

REVIEWERS — VOLUME ONE HUNDRED THIRTY-FOUR (3)

The Editors express their appreciation to the following individuals who

have, prior to acceptance for publication, reviewed one or more of the

papers prepared for this issue.

Artur Alves Josiane Santana Monteiro

Akira Nakagiri

Lorelei L. Norvell

Flavia Rodrigues Barbosa

Rafael FE. Castaneda-Ruiz

Roger T.A. Cook

Lin Gao

Sergio P. Gorjén

Shouyu Guo

Danny Haelewaters

Nils Hallenberg

Chenglin Hou

Sana Jabeen

Kerry Knudsen

John C. Landolt

Marc-Henri Lebrun

De-Wei Li

Quan Lu

Jian Ma

Eric H.C. McKenzie

Shaun R. Pennycook

Liliane Petrini

Huzefa A. Raja

Mario Rajchenberg

Qiang Ren

B.M. Sharma

Chi Song

Jie Song

Steven L. Stephenson

Marcelo A. Sulzbacher

Susumu Takamatsu

Xinli Wei

Ze-Fen Yu

Xiu-Guo Zhang

Andrea Paola Zuluaga Cruz

x ... MYCOTAXON 134(3)

ERRATA FROM PREVIOUS VOLUMES

VOLUME 104

pp. 97-101 FOR: Skeletocutis luteolus READ: Skeletocutis luteola

The 2008 paper entitled “Skeletocutis luteolus sp. nov. from southern and eastern China”

by Cui & Dai incorrectly presents the masculine form of the epithet. The species

name should be corrected throughout (and elsewhere) to Skeletocutis luteola,

the feminine form, to agree with the feminine genus.

ERRATA IN CURRENT ISSUE (134-3)

Cited below are mistakes present in files submitted for final review but not

detected by the authors until after the paper was in press.

p. 535, line 24 FOR: porpidia-type READ: porpidia-type

JULY-SEPTEMBER 2019... XI

FROM THE EDITOR-IN-CHIEF

MyYCOTAXON WELCOMES TWO FINAL EDITORIAL ADVISORS—Our heretofore

understaffed advisory board successfully completed its first task for 2019 by

nominating and electing two new members to bring itself up to full advisory

strength. We are extremely pleased to welcome Elaine Malosso and Xinli Wei,

who join Chair BRANDON MATHENY, KAREN HANSEN, ELSE VELLINGA, and

TODD OSMUNDSON on the journal's masthead (p. ii).

Profa. ELAINE MALOosso, currently with UFPE (Universidade Federal de

Pernambuco) in Recife, Brazil, received her Ph.D. in Molecular Microbial

Ecology from the University of Newcastle upon Tyne (England) in 2003. Fluent

in English, Portuguese, and Spanish, Elaine has both co-authored and reviewed

numerous papers on hyphomycetes for MycoTaxon and is also interested in

fungal diversity, decomposition, molecular phylogenetic analyses, soils, fresh

water, and arbuscular mycorrhizal fungi.

Prof. X1nt1 WEI, who served as a postdoctoral fellow at Sunchon National

University, South Korea, during 2006-2007 and Visiting Scholar at Chicago's

Field Museum in Illinois (USA) during 2016-2017, began her study of lichens

in 2000. As professor at the State Key Laboratory of Mycology & Lichenology

at the Chinese Academy of Sciences in Beijing, she also served as Chief Editor

of the LICHEN FLORA OF CHINA (English version). Xinli is especially interested

in the taxonomy and biogeography of Hypogymnia, and has also contributed to

numerous papers for MyCcoTAXON.

MycoTaxon 134(3) contains 20 papers by 78 authors (representing 12

countries) and examined by 34 pre-submission expert reviewers as well as by

the NOMENCLATURE EDITOR and EDITOR-IN-CHIEE.

The 2019 July-September Mycotaxon proposes two new genera

(Morganjonesia from CHINA and Ramomarthamyces from NEW ZEALAND & THE

Cook IsLaANDs) plus 19 newly described species representing Annulohypoxylon,

Botryosphaeria, Coenogonium, Pleurotheciopsis, Repetophragma, and

Septobasidium from Cuina; Beltraniomyces & Glabrocyphella from BRAZIL;

Cystostereum from INbD1A; Marthamyces & Ramomarthamyces from NEw

ZEALAND & the Cook IsLANDs; and Tretoheliocephala from THAILAND. We also

publish seven new combinations in Heteroscypha, Marthamyces, Morganjonesia,

and Ramomarthamyces and present a newly established synonymy for Erysiphe

sedi (accompanied by neotypification of Oidium kalanchoes).

xi ... MYCOTAXON 134(3)

New species range extensions are reported for [ascomycetes]

Pseudopyricularia cyperi in Iran; [basidiomycetes] Amanita subjunquillea and

its ectomycorrhizae in PAKISTAN, Limacella for the first time in THAILAND, and

the cyphelloid fungus Rectipilus natalensis in BRAzIL; and in CuINna [lichens]

Acarospora & Psora and Coenogonium (accompanied by a key to all species in

the country); and [myxomycetes] dictyostelid social amoebae.

This issue is rounded out by the announcement of a new mycobiota covering

macrofungal diversity in Yushan and Else Vellinga’s reviews of three new

substantial field identification guides.

Warm regards,

Lorelei L. Norvell (Editor-in-Chief)

28 September 2019

JULY-SEPTEMBER 2019... XIII

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MY COTAXON

July-September 2019—Volume 134, pp. 413-423

https://doi.org/10.5248/134.413

Amanita subjunquillea and its ectomycorrhizal association,

reported as new for Pakistan

MUHAMMAD IsHAQ’, AROOJ NASEER?, MUNAZZA KIRAN’,

MUHAMMAD FIAz’, ABDUL NASIR KHALID?

' Department of Botany, Hazara University, Mansehra, Pakistan

? Department of Botany, University of the Punjab,

Quaid-e-Azam Campus-54590, Lahore, Pakistan

“CORRESPONDENCE TO: ishaqm783@gmail.com

AsBstRAcT—Amanita subjunquillea and its ectomycorrhizal association are reported for

the first time from moist temperate Himalayan forests of Pakistan. The sample was studied

based on morphological characters and nucleotide sequence analyses of the ITS region

generated from basidiomata and ectomycorrhizal roots of Quercus floribunda. Our collection

differs from the type in its dark orange pileus disc and pale yellow margins. Remaining

morphological and molecular data are consistent with previously reported specimens. This

represents the first report of A. subjunquillea from Pakistan.

Keyworps—Amanitaceae, Basidiomycota, Phalloideae, ribosomal DNA, taxonomy

Introduction

Amanita Pers. (Amanitaceae, Agaricales, Basidiomycota) is a large genus

comprising about 1000 described species that are important economically and

ecologically (Bas 2000, Tulloss 2005, Yang 2005, Kirk & al. 2008, Li & al. 2015,

Zhang & al. 2015, Cui & al. 2018, Yang & al. 2018). Amanita contains both edible

and poisonous species, and some species form ectomycorrhizal associations

with vascular plants, which play a key role in maintaining healthy ecosystems

(Zhang & al. 2004, 2010; Thongbai & al. 2016; Cui & al. 2018; Yang & al. 2018).

Poisonous Amanita species are classified mostly in A. sect. Phalloideae (Fr.)

Konrad & Maublanc, which is characterized by the absence of striation and

414... Ishag & al.

lack of appendiculate pileus and the possession of attenuate lamellulae, an

annulus, a stipe with a bulbous base and limbate shaped volva, and amyloid

basidiospores (Corner & Bas 1962, Bas 1969, Yang 2005, Cai & al. 2016,

Cui & al. 2018). Amanita subjunquillea, also known as the East Asian death cap,

is a poisonous mushroom broadly distributed in diverse habitats in China, India,

Japan, and South Korea (Bhatt & al. 2003, 2007; Yang 2005; Zhang & al. 2010;

Cui & al. 2018). Symptoms of A. subjunquillea poisonings are gastrointestinal

in nature and similar to A. phalloides, and ingestion can result in death (Rho &

al. 2000, Kim & al. 2008).

A total of 24 species of Amanita have been reported from Pakistan, but it

is estimated that the true number of Amanita from Pakistan could be above

50 (Kiran & al. 2018, Jabeen & al. 2019, Saba & al. 2019). Many collections

of Amanita have been identified locally based on morphological features;

however, such identifications are not reliable because some poisonous and

edible mushrooms are morphologically similar. The Korea Forest Service

reported the occurrence of twenty-three fatal mushroom poisonings in

Korea during 2004-13 (Cho & al. 2015; www.forest.go.kr/). Such cases also

occur every year in rural areas of Pakistan, especially in hilly areas of Swat,

Khyber Pakhtunkhwa. The aim of this study was to confirm our collection

of A. subjunquillea from Buner District as a new record for Pakistan using

morphological, molecular and phylogenetic data.

Materials & methods

Sampling site

Collections were made at Bottle Green Mountains, Gokand Valley, Buner

District, Khyber Pakhtunkhwa, Pakistan, at 3183 m a.s.l. The climate of Gokand is

subtropical at lower elevations and temperate above. This area is covered by mixed

forests of conifers (Pinus roxburghii Sarg., P. wallichiana A. B. Jacks.) and broadleaf

trees (Populus ciliata Wall. ex Royle, Q. floribunda Lindl. ex A. Camus [= “Q. dilatata”

Royle], Q. oblongata D. Don [= Q. incana Roxb.], Zanthoxylum armatum DC., and

Olea europaea subsp. cuspidata (Wall. & G. Don) Cif. [= O. ferruginea Royle]). Blue

pine (PB wallichiana) and chir pine (P. roxburghii) forests predominate (Khan & al.

2003).

The area was sampled during rainy seasons in July and August in 2017. Specimens

were tagged and photographed on site. Morphological characters were recorded

from fresh fruiting bodies in field, with colors coded according to Munsell Soil Color

Charts (1975). Specimens were then dried using a fan heater and placed in small zip-

lock bags for long-term preservation.

Amanita subjunquillea and its ectomycorrhizae in Pakistan ... 415

Sampling, Isolation, and morphological characterization of ectomycorrhizal roots

Soil cores (15 cm*) were dug from moist temperate oak forests close enough to

Quercus floribunda tree trunks to ensure that they contained Quercus roots. Twenty

to twenty-five soil cores were taken from every sampling site and stored in polythene

bags to prevent evaporation. In the laboratory, each soil core was drenched in water

up to 1-2 hours and then held over a 2 mm sieve beneath running water to detach

roots from soil particles. Ectomycorrhizal roots were sorted into morphotypes using

a Meiji Techno EMZ-5TR stereomicroscope. The morphotypes were then kept in

distilled water in McCartney bottles, with replicates reserved in 2% CTAB buffer

at 8°C for molecular analysis. The material was deposited in Lahore Herbarium,

Department of Botany, University of the Punjab, Quaid-e-Azam Campus, Lahore,

Pakistan (LAH).

Morphological analyses of basidiomata

For microscopic studies, tissues of pileus, gills, annulus, stipe, and volva were

mounted in 5% KOH, stained with 1% aqueous Congo red (w/v), and observed under

a Labomed light microscope. Basidiospore dimensions include length and width

ranges (with parenthetical extreme values), range of Q coefficients (L/W), mean

length and width (L,, x W.,), and mean Q value (Q_). The collection was vouchered

and deposited in Lahore Herbarium, Department of Botany, University of the Punjab,

Quaid-e-Azam Campus, Lahore, Pakistan (LAH).

DNA extraction, amplification, and sequencing

DNA was extracted using a modified CTAB method (Bruns 1995). The nuclear

ribosomal internal transcribed spacer (ITS) locus was amplified using the fungal-

specific ITSIF primer (5-cCTTGGTCATTTAGAGGAAGTAA-3; Gardes & Bruns 1993)

and the eukaryotic ITS4 primer (5-TccTccGCTTATTGATATGC-3; White & al. 1990).

The amplified products were sent to Macrogen Inc. in Korea for purification and

sequencing.

For identification of ectomycorrhiza, DNA was extracted using the Qiagen DNeasy

Plant Mini Kit, and the ITS was amplified using the ITS1F/ITS4 primers. Amplicons

obtained from ectomycorrhizal roots were purified with Exonuclease I and Shrimp

Alkaline Phosphatase enzymes (Werle & al. 1994) and sequenced at the University of

Florida Interdisciplinary Center for Biotechnology Research (www.biotech.ufl.edu/).

Sequences were BLAST searched at NCBI (blast.ncbi.nlm.nih.gov/Blast.cgi), and

closely related Amanita sequences were retrieved from GenBank. All the sequences

were assembled in BioEdit (Hall 1999) and trimmed to facilitate alignment. A

maximum likelihood tree was inferred for the ITS alignment using RAxML-HPC2

v 8.1.11 (Stamatakis 2014) with a GTRCAT nucleotide substitution model. Rapid

bootstrapping was performed with 1000 bootstrap iterations. All phylogenetic

analyses were performed on the CIPRES Science Gateway v3.1 (Miller & al. 2010).

The phylogeny from ML analysis was displayed with FigTree v1.4.2 (tree.bio.ed.ac.uk/

software/figtree/) and then edited in Adobe Illustrator. All newly generated sequences

were submitted to GenBank.

416... Ishaq & al.

Fic. 1. Amanita subjunquillea (LAH35854).

Basidiomata. Scale bars: A = 0.79 cm; B = 1.17 cm; C = 1.02 cm.

Amanita subjunquillea and its ectomycorrhizae in Pakistan ... 417

Taxonomy

Amanita subjunquillea S. Imai, Bot. Mag., Tokyo 47: 424 (1933) Fics 1, 2

Piteus 60-70 mm diam, plano-convex with a slightly depressed disc

and slightly uplifted margin, dark orange (10YR9/2) at disc, becoming

light brown (5Y 9/4) toward margin; context white (5Y9/2) when cut.

LAMELLAE free, close, entire, margins whitish (10Y9/2) to pale yellow

(10Y 9/4). LAMELLULAE truncated, abundant, irregularly distributed. STrPE

120-130 x 5-15 mm, cylindrical, slightly tapering upwards, whitish (5Y9/2)

at the base and pale yellowish (5Y9/4) toward the apex, covered by pale

yellowish (5Y9/4) fibrillose squamules. ANNULUS superior, membranous,

persistent, pendant, whitish (SY 9/2) to pale yellow (5Y 9/4). Votva limbate,

membranous, white.

BASIDIOSPORES (3.8-)4.3—5.7(-6.1) x (3.8-)4.1-5.2(-5.6) um, Q = 1-1.2(-1.3),

Lx W,,=5.1 x 4.6 um, Q_ = 1.1, globose to subglobose, hyaline in 5% KOH,

amyloid in Melzer’s reagent, thin-walled, smooth, monoguttulate; apiculus

0.6-1 um long. BAsrp1a 20-27 x 8-9 um, 2-4-spored, thin-walled, hyaline,

narrowly clavate to clavate; sterigmata 1-2 um long. PILEIPELLIS hyphae 4-6

um diam, branched, filamentous, septate, ellipsoid to subcylindrical, hyaline.

Volva cells globose, ovoid to broadly lageniform 7-35 um diam, 19-68 um

long; filamentous hyphae 3.1-4.5 um diam. STIPITIPELLIS hyphae 12-16 um

diam, septate, unbranched, thin-walled, hyaline in 5% KOH. PARTIAL VEIL

ELEMENTS 34-55 x 10-21 um, ellipsoid to clavate intermixed with short

septate filaments 1-2 um diam. CLAMP CONNECTIONS absent throughout.

MATERIAL EXAMINED: PAKISTAN. KHYBER PAKHTUNKHWA PROVINCE, Buner

District, Gokand Valley, Bottle Green Mountains, 3183 m a.s.l., under Quercus

floribunda [= “Q. dilatata” Royle], solitary, 13 August 2017, Muhammad Ishaq IB124

(LAH-35854; GenBank MH620472).

Hasitat: Solitary on soil under Quercus floribunda.

Ectomycorrhizal morphology FIG. 3

MycorRRHIZAL SYSTEM monopodial pinnate, 20-45 mm, 0.7-1 mm thick,

profusely branched, surface smooth, light brown (5YR8/4), older parts dark

reddish brown (5YR3/4). Unramified ends slightly straight to (infrequently)

bent, cylindrical. Mantle brown (5YR8/5), not translucent. OUTER MANTLE

HYPHAE 2.7-3.4 um diam, thin-walled, septate, branched, cells similar in size

to inner mantle hyphae, sub-cylindrical to ellipsoid, end cells obtuse, no clamp

connections. INNER MANTLE HYPHAE 2.4-3.5 um diam, thin-walled, septate,

418... Ishaq & al.

E — | —

Fic. 2. Amanita subjunquillea (LAH35854). A. Basidia; B. Basidiospore; C. Partial veil elements;

D. Volva; E. Pileipellis; F. Stipitipellis. Scale bars: A = 7.5 um; B = 3 um; C, D = 7 um; E = 4.5 um;

F=1 um.

Amanita subjunquillea and its ectomycorrhizae in Pakistan ... 419

Fic. 3. Amanita subjunquillea/Quercus floribunda mycorrhizae (LAH-EM101, LAH-EM102).

A., B. Ectomycorrhizal roots; C. Inner mantle layer cells; D. Outer mantle layer cells. Scale bars:

A=1.91 mm; B= 0.54 mm; C = 4.5 um; D = 10.2um.

unbranched, narrowly clavate to clavate, 6-11 um long, end cells obtuse,

cystidia infrequent. EMANATING HYPHAE numerous, white. RHIZOMORPHS

white (2.5R9/2), thick, frequent, abundant.

MATERIAL EXAMINED: PAKISTAN. KHYBER PAKHTUNKHWA PROVINCE, Shangla/

Swat boundary, Toa, 2100 m a.s.l, associated with roots of Quercus floribunda, 15

420... Ishaq & al.

July 2015, Arooj Naseer & Abdul Nasir Khalid ANT210 (LAH-EM101; GenBank

MH998629); ANT211 (LAH-EM102; GenBank MH998628); ANT212 (LAH-EM103;

GenBank, MH998627).

Phylogenetic results

FIG. 4

The aligned ITS dataset, including sequences retrieved from GenBank,

comprised 30 nucleotide sequences. Of the 665 characters in the final dataset,

275 were conserved, 363 were variable and parsimony-uninformative, and

228 were parsimony-informative. Our ML tree contains three sections:

Amanita sect. Validae, A. sect. Amanita, and A. sect. Phalloideae. The

AY436455 Amanita flavipes HKAS36582

AY436457 Amanita fritillaria HKAS38331

ay AB015696 Amanita flavipes LEM960088a

AY436456 Amanita flavoconia HKAS34047

Sect.

AY436453 Amanita excelsa HKAS31510 Validae

400} AB015679 Amanita citrina LEM960298

AY 436449 Amanita af citrina HKAS34170

4 AY436471 Amanita porphyria HKAS31531

ABO15677 Amanita porphyria LEM960303

40q AB015701 Amanita pantherina LEM970680

AY436469 Amanita parvipantherina HKAS38334

AY436466 Amanita cf pantherina HKAS26746

AY436459 Amanita griseopantherina HKAS32523

AY436476 Amanita subfrostiana HKAS34551

KR996715 Amanita subjunquillea MHHNU7751 C

FJ176730 Amanita subjunquillea MHHNU7041 C

FJ176731 Amanita subjunquillea MHHNU7049 C

KJ466424 Amanita subjunquillea HKAS74993 C

JX998033 Amanita subjunquillea HKAS75772 C Sect.

MH998627 Amanita subjunquillea LAHEM103

MH998628 Amanita subjunquillea LAHEM102

MH998629 Amanita subjunquillea LAHEM101

FJ176729 Amanita subjunquillea MHHNU6827 C

KM052527 Amanita subjunquillea ASIS22769 K

FJ176733 Amanita subjunquillea HKAS52315 C

KT894848 Amanita subjunquillea ASIS26536 K

FJ176732 Amanita subjunquillea HKAS50910 C

AY436480 Limacella glioderma_HKAS31576

0.2

Sect.

Amanita

MH620472 Amanita subjunquillea LAH35854 Phalloideae

Fic. 4. Molecular phylogenetic reconstruction of Amanita spp. based on ITS sequences. The best

scoring ML tree (-InL = 1591.4482) is shown. Only bootstrap values >50 are shown. Amanita

subjunquillea sequences from China are annotated as [C] and those from Korea as [K].

Amanita subjunquillea and its ectomycorrhizae in Pakistan ... 421

sequences from our Pakistani specimen and ectomycorrhizal roots,

grouped within A. sect. Phalloideae with previously generated sequences of

A. subjunquillea from China and South Korea.

Comparison of the ITS sequences revealed 19 polymorphic positions

among different collections of A. subjunquillea from Pakistan, China, and

Korea. The Pakistani sequence contains eleven polymorphic positions (at

positions 2, 7, 32, 35, 107, 204, 504, 507, 553, 648, and 667), with most

nucleotides at these allelic positions resembling Chinese samples. However,

one Chinese sample is closer to the Korean specimen KT894848. Maximum

nucleotide differences were observed in Korean sample KM052527, with

seven positions differing from all other collections, including Korean sample

KT894848. It is possible that the KM052527 ITS sequence needed additional

purification, but more sequences are required to verify these seven nucleotide

differences.

Discussion

Amanita subjunquillea is a deadly poisonous mushroom distinguished by

a brownish yellow to mustard yellow pileus, non-appendiculate margins, and

a subglobose to subclavate to turnip-shaped basal bulb (Yang 1997, 2005;

Cai & al. 2016; Cui & al. 2018).

Macroscopic and microscopic details of the examined specimens match

the protologue description of A. subjunquillea (Imai 1933), except for pileus

coloration patterns. The pileus color of the Pakistani specimen is dark orange

at the disc and pale yellow toward the margin, while the type specimen is

described with the pileus darker at the center and brownish yellow, dirty

citrine yellow, or mustard yellow toward the margin. This is the only

macroscopic difference separating our collection from the type; the nearly

cylindrical stipe that tapers slightly upward and is covered with yellowish

fibrillose scales matches the type description. The spore and basidial shapes

are similar among all collections, but the basidia and spore sizes are smaller

in our specimen compared with Indian and East Asian specimens. Clamp

connections are absent in all specimens.

Our phylogenetic analysis clustered ITS sequences from our Pakistani

collection and ectomycorrhizal roots in A. sect. Phalloideae with ten other

sequences belonging to A. subjunquillea. We found some variation among

the Chinese, Korean, and Pakistani nucleotides; nucleotide differences

within same species from different localities of China were also observed by

Zhang & al. (2010). The type collection was sampled under warm temperate

422 ... Ishaq & al.

conditions in Tokyo whereas our sampling site lies in a moist temperate

region. These environmental differences can affect gene function so that

same species collected from different geographic regions may show minor

variation in both morphological and molecular characters.

Amanita subjunquillea is endemic to East Asia (Zhang & al. 2010). This

is the first published report of its ectomycorrhizal association with Quercus

floribunda based on morphological, anatomical, and phylogenetic data.

Amanita subjunquillea has previously been reported from Japan, Korea,

India, and China, to which we now add Pakistan.

Acknowledgments

The authors are grateful to Prof. Zhu Liang Yang (Kunming Institute of Botany,

Chinese Academy of Sciences, China) and Dr. David Hibbett (Clark University,

Worcester, Massachusetts, USA) for their valuable comments and suggestions. We

are also grateful to Dr. Danny Haelewaters (Faculty of Science, University of South

Bohemia, Ceske Budejovice, Czech Republic) and Dr. Sana Jabeen (Division of

Science and Technology, University of Education, Lahore, Pakistan) for their efforts

to improve this manuscript.

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https://doi.org/10.3852/16-008

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higher-rank taxonomy and the species in China. Fungal Diversity 91(1): 5-230.

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Cho HJ, Park MS, Lee H, Oh SY, Jang Y, Fong JJ, Lim YW. 2015. Four new species of Amanita in Inje

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MY COTAXON

July-September 2019—Volume 134, pp. 425-429

https://doi.org/10.5248/134.425

New records of dictyostelid social amoebae from China

Pu Liu, SHUNHANG ZHANG, YUE ZOU, YU ZHANG, YONGJIA Li, Yu Lr

Engineering Research Center of Chinese Ministry of Education for Edible and Medicinal Fungi,

Jilin Agricultural University, Changchun, 130118, P. R. China

* CORRESPONDENCE TO: fungi966@126.com

ABSTRACT—Two species of dictyostelids (Dictyostelium aureocephalum and Heterostelium

filamentosum) new to China were isolated from samples of forest soils; and a second Chinese

record of Coremiostelium polycephalum (previously recorded from Taiwan) was also isolated.

Descriptions and illustrations based on these isolates are provided.

Key worps—Acytosteliaceae, Dictyosteliaceae, Dictyostelida, Mycetozoa, taxonomy

Introduction

Dictyostelid cellular slime molds (dictyostelids), with both animal-like

(protozoan) and fungus-like characteristics, are the second largest group

of slime molds. These organisms play a role in maintaining the balance that

exists between bacteria and other organisms in soils (Raper 1984). The original

system that placed dictyostelids in genera based on morphology was revised

by Sheikh & al. (2018), who proposed a new classification based on unique 18S

rRNA sequence signatures.

In sampling from Chinese soils, two dictyostelid species, Dictyostelium

aureocephalum and Heterostelium filamentosum, were recorded as new from

China, and Coremiostelium polycephalum, previously isolated from Taiwan, was

recorded as new from Fujian Province. Descriptions and illustrations based on

these newly recovered isolates are provided.

Materials & methods

Samples used for dictyostelid isolations were collected from Shandong Province,

Fujian Province, and Yunnan Province, China, in 2012 and 2016. Each sample

426 ... Liu & al.

consisted of 30-50 g of soil that was placed in a sterile whirl-pack plastic bag. Each

sample bag was numbered and the sample itself preserved at 4 °C in the herbarium

of the Mycological Institute of Jilin Agricultural University, Changchun, China

(HMJAU). Isolation methods followed Cavender & Raper (1965), with some

minor modifications outlined by Liu & al. (2019). Isolates were identified using the

descriptions in Raper (1984) and the new classification system proposed by Sheikh

& al. (2018). The characteristic stages in the life cycle, including cell aggregation and

formation of pseudoplasmodia and sorocarps, were observed under a Zeiss Axio

Zoom V16 dissecting microscope with a 1.5x objective and a 10x ocular. Slides with

sorocarps were prepared with water as the mounting medium. Spores, sorophores,

and sorocarps were observed on the slides and measured using a Zeiss Axio

Imager A2 light microscope with 10x ocular and 10, 40, and 100x (oil) objectives.

Photographs were taken with Zeiss Axiocam 506 color microscope camera. Vouchers

were conserved in the Herbarium of the Mycological Institute of Jilin Agricultural

University, Changchun, China (HMJAU).

Taxonomy

Dictyostelium aureocephalum H. Hagiw.,

Bull. Nat. Sci. Mus., Tokyo, B 17(3): 103. 1991. PLATE la-g

When cultured at 23 °C on non-nutrient agar with Escherichia coli,

sorocarps gregarious, solitary, prostrate, phototropic, normally 0.7-2.6 mm.

Sorophores colorless, sinuose, with hook-like structures if prostrate, tapering

from conical bases to clavate tips, basal disks small. Sori yellow, globose,

commonly 70-210 um in diam. Spores hyaline, elliptical, 5.7-8.0 x 2.9-3.9

um, without polar granules. Aggregations with radiate streams.

SPECIMEN EXAMINED: CHINA, YUNNAN PROVINCE, Lijiang, Yulong Mountains, from

soil (S5204) collected 22 Oct. 2016 in a mixed forest, isolated 2016, P. Liu & al. (HMJAU

MR228).

CoMMENTS—Dictyostelium aureocephalum was first isolated from a sample

collected from the humus layer of a scrub and meadow in the alpine region of

Nepal (Hagiwara 1991). This species is characterized by its yellow sori. It has

PG- spores and somewhat smaller sorocarps, features that distinguish it from

other dictyostelids with yellowish sori.

Heterostelium filamentosum (F. Traub & al.) S. Baldauf, S. Sheikh & Thulin,

Protist 169(1): 11. 2018. PLATE 1h-l

When cultured at 23 °C on non-nutrient agar with E. coli, sorocarps white,

erect to semi-erect, with whorls of both primary and secondary branches, the

terminal segments always elongate. Sorophores colorless, tapering from bases

to tips. Terminal sori always small, white, globose, commonly 30-80 um diam.

Dictyostelium & Heterostelium spp. new for China ... 427

PLaTE 1. Dictyostelium aureocephalum: a—c. Sorocarps; d. Sorophore tip; e. Sorophore base;

f. Aggregation; g. Spores. Heterostelium filamentosum: h. Sorocarps; i. Sorophore branches;

j. Spores; k. Aggregation; 1. Sorophore base. Coremiostelium polycephalum: m. Sorocarps;

n. Sorophore base; 0. Sorophore; p. Spores. Scale bars: a, b = 2 mm; c, f, h, k, m= 1 mm; d = 100 um;

e= 50 um; g,i,n, o= 20 um; j, |, p= 10/um.

Lateral sori white, globose, commonly 45-160 um diam. Spores elliptical to oval,

mostly 7.5-11 x 3.5-5 um, with unconsolidated polar granules. Aggregations

radiate.

SPECIMEN EXAMINED: CHINA, SHANDONG PROVINCE, Jinan, Jinxiang Mountain,

from soil (S2832) collected 9 Aug. 2012 in a broadleaf forest, isolated 2013, P. Liu & al.

(HMJAU MR182).

ComMENTS—Heterostelium filamentosum was originally isolated and reported

as widespread in Switzerland (Traub & al. 1981). It has also been found in

428 ... Liu & al.

other countries such as the United States, Japan (Raper 1984; Hagiwara 1989,

2008), and China (this study). This species is characterized by its large spores,

secondary branches, and elongation of branches.

Coremiostelium polycephalum (Raper) S. Baldauf, S. Sheikh, Thulin & Spiegel,

Protist 169(1): 24. 2018. PLATE 1m-p

When cultured at 23 °C on non-nutrient agar with E. coli, sorocarps

white, erect, solitary, commonly less than 1 mm tall, typically clustered to

form coremiform sorocarps, normally consisting 2-10 branches per cluster.

Sorophores tapering from bases to tips. Sori white or hyaline, globose, 45-85 um

diam. Spores elliptical, usually 6.0-7.5 x 3.0-3.5 um, with unconsolidated

granules. Aggregations radiate.

SPECIMEN EXAMINED: CHINA, FUJIAN PROVINCE, Fuzhou, from soil ($2052) collected

24 Mar. 2012 in a coniferous forest, isolated 2012, P. Liu & al. (HMJAU MRI177).

ComMENtTS—The type of Coremiostelium polycephalum was isolated from

the soil of a mixed forest in South Carolina in the United States (Raper 1984;

Hagiwara 1989). It is a worldwide species which has been found in Africa,

America, Asia, and Europe. However, this is only the second report from

China. Both of these two localities (Fujian Province and Taiwan) are located in

subtropical China, so this species seems to prefer warmer habitats. This species

has small clustered sorocarps developed in a coremium-like manner, a feature

which allows it to be distinguished from other dictyostelids.

Acknowledgments

We wish to express our appreciations to two peer reviewers Prof. Steven L.

Stephenson (University of Arkansas, USA) and Prof. John C. Landolt (Shepherd

University, WV USA), for their valuable comments relating to this manuscript.

This study was supported by the National Natural Science Foundation of China

(No. 31870015, 31300016), the Science and Technology Research Programs of

the Education Department of Jilin Province in the Thirteenth Five-Year Plan

(No. JJKH20180671KJ), Science and Technology Development Program of Jilin

Province (No. 20180101273JC), and 111 Project (No. D17014).

Literature cited

Cavender JC, Raper KB. 1965. The Acrasieae in nature. I. Isolation. American Journal of Botany 52:

294-296. https://doi.org/10.1002/j.1537-2197.1965.tb06788.x

Liu P, Zou Y, Li Shu, Stephenson SL, Li Y. 2019. Two new species of dictyostelid cellular slime molds

in high-elevation habitats on the Qinghai-Tibet Plateau, China. Scientific Reports 9:5 [13 p.].

https://doi.org/10.1038/s41598-018-37896-7

Hagiwara H. 1989. The taxonomic study of Japanese dictyostelid cellular slime molds. National

Science Museum, Tokyo.

Dictyostelium & Heterostelium spp. new for China ... 429

Hagiwara H. 1991. Dictyostelium aureocephalum, a new dictyostelid cellular slime mold from

Nepal. Bulletin of the National Science Museum, B 17(3): 103-107.

Hagiwara H. 2008. Taxonomic studies on dictyostelids. 3. Sexuality of the Polysphondylium

candidum Complex. Bulletin of the National Science Museum, B 34(1): 17-21.

Raper KB. 1984. The dictyostelids. Princeton University Press, Princeton, New Jersey.

Sheikh S, Thulin M, Cavender JC, Escalante R, Kawakami S, Lado C, Landolt JC, Nanjundiah V,

Queller DC, Strassmann JE, Spiegel FW, Stephenson SL, Vadell EW, Baldauf SL. 2018. A new

classification of the dictyostelids. Protist 169: 1-28. https://doi.org/10.1016/j.protis.2017.11.001

Traub F, Hohl HR, Cavender JC. 1981. Cellular slime molds of Switzerland. I. Description of

new species. American Journal of Botany 68: 162-171.

https://doi.org/10.1002/j.1537-2197.1981.tb12375.x

MY COTAXON

July-September 2019—Volume 134, pp. 431-438

https://doi.org/10.5248/134.431

Annulohypoxylon bawanglingense, A. diaoluoshanense,

A. guangxiense, and A. sichuanense spp. nov.

and new records from China

WEI LI & LIN GUO"

State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences,

Beijing 100101, China

* CORRESPONDENCE TO: guol@im.ac.cn

AxBsTRACT—Four new species—Annulohypoxylon bawanglingense, A. diaoluoshanense,

A. guangxiense, and A. sichuanense—are described, and A. maeteangense and A. substygium

are recorded as new to China.

KEY worRDS—Ascomycota, pyrenomycetous fungi, taxonomy, Hypoxylaceae, Xylariales

Introduction

We describe four new species of Annulohypoxylon and two new

Annulohypoxylon records from China. These specimens are conserved in the

Herbarium Mycologicum Academiae Sinicae, Beijing, China (HMAS).

Materials & methods

The specimens were examined using a Motic SMZ-168 stereomicroscope.

Microscopic features of asci and ascospores were described from slide preparations

mounted in water and Melzer’s reagent, and measurements were made from 20

ascospores using a Zeiss Axioskop 2 plus compound microscope. Stromata were

photographed using a Sony NEX-7 digital camera.

Taxonomy

Annulohypoxylon bawanglingense Wei Li bis & L. Guo, sp. nov. Figs 1-3

EN 570628

Differs from Annulohypoxylon moriforme by its larger ascospores, and from A.

microdisicum by its larger disc on the ostiole.

432 ... Li& Guo

Type: China, Hainan, Changjiang, Bawangling, alt. 490 m, on the wood of the

Adenanthera microsperma Teijsm. & Binn. (Leguminosae), 17.11.2017, L. Guo 12112

(Holotype, HMAS 248019).

ErymMo ocy: The epithet refers to the type locality, Bawangling.

Stromata pulvinate to effused-pulvinate, with conspicuous perithecial

mounds, up to 4 exposed, 0.2-17 x 0.15-3.5 cm, 0.9-1.5 mm thick; surface

brownish black; blackish brown granules immediately beneath surface, shiny,

with KOH-extractable greenish olivaceous pigment; the tissue below the

perithecial layer inconspicuous, black. Perithecia subspherical to obovate,

0.35-0.5 mm diam. Ostioles papillate, encircled with a truncatum-type disc,

0.35-0.5 mm diameter. Asci 119-170 tm long overall, the spore-bearing

parts 80-100 x 4-5 um above 40-70 um long stipes; apical ring bluing in

Melzer’s iodine reagent, discoid, 1-1.5 um high, 1.5-2 um broad. Ascospores

brown, unicellular, ellipsoid-inequilateral, with narrowly rounded ends,

8-11 x 3.5-4 um, with straight germ slit spore-length; perispore dehiscent in

10% KOH, smooth, epispore smooth.

CoMMENTs: Annulohypoxylon bawanglingense is similar to A. moriforme

(Henn.) Y.M. Ju & al., which differs in its smaller (6-9 x 2.5-4 um) spores

(Ju & Rogers 1996). The new species resembles A. microdiscum (Y.M. Ju &

J.D. Rogers) Sir & Kuhnert, which differs in its smaller (0.1-0.17 mm diam)

discs surrounding the ostioles (Kuhnert & al. 2017).

Annulohypoxylon diaoluoshanense Wei Li bis & L. Guo, sp. nov. Fics 4-6

EN 570630

Differs from Annulohypoxylon macrodiscum by its thicker stromata.

Type: China, Hainan, Diaoluoshan, on wood, 26.1X.1958, RY. Zheng & al. 227

(Holotype, HMAS 29644).

ErymMo.oey: The epithet refers to the type locality, Diaoluoshan.

Stromata hemispherical, with inconspicuous to conspicuous _perithecial

mounds, 1.2-4 x 1-3.3 cm, 4-10 mm thick; surface blackish brown, blackish

brown granules immediately beneath surface, with KOH-extractable greenish

olivaceous pigment; the tissue below the perithecial layer blackish brown,

conspicuous, 3-7 mm thick. Perithecia spherical or obovate, 0.8-1.2 mm diam.

Fics 1-9. Annulohypoxylon bawanglingense (HMAS 248019, holotype). 1. Stromata on wood;

2, 3. Ascospores. Annulohypoxylon diaoluoshanense (HMAS 29644, holotype). 4. Stromata on

wood; 5, 6. Ascospores. Annulohypoxylon guangxiense (HMAS 33768, holotype). 7. Stromata on

wood; 8, 9. Ascospores.

Annulohypoxylon spp. nov. & new records (China) ... 433

434 ... Li& Guo

Ostioles conical-papillate, encircled with a truncatum-type disc, 0.5-0.8 mm

diam. Asci not seen. Ascospores dark brown, unicellular, ellipsoid-inequilateral,

with broadly rounded ends, 8-11.5 x 4-5.5 um, with straight germ slit spore-

length; perispore dehiscent in 10% KOH, smooth, epispore smooth.

Comments: Annulohypoxylon diaoluoshanense is similar to A. macrodiscum

Jad. Pereira & al., which differs in its thinner (0.8-1 mm thick) stroma and

thinner tissue (0.3-0.5 mm thick) below the perithecial layer (Pereira & al.

2010).

Annulohypoxylon guangxiense Wei Li bis & L. Guo, sp. nov. Fics 7-9

FN 570633

Differs from Annulohypoxylon archeri by its tissues without apparent KOH-extractable

pigments and its perispore indehiscent in 10% KOH.

Type: China, Guangxi, Donglan, on wood, 20.1.1958, LW. Xu 822 (Holotype, HMAS

33768).

ErymMo.ocy: The epithet refers to the type locality, Guangxi Province.

Stromata pulvinate, with inconspicuous perithecial mounds; 1-4 x 0.6-1.8 cm,

0.5-1 mm thick; surface blackish brown, blackish brown granules immediately

beneath surface and lacking KOH-extractable pigments; the tissue below

the perithecial layer inconspicuous. Perithecia spherical, 0.4-0.5 mm diam.

Ostioles papillate, encircled with a truncatum-type disc, 0.1-0.2 mm diam.

Asci not seen. Ascospores brown, unicellular, ellipsoid-inequilateral, with

narrowly rounded ends, 8-12.5 x 3-5 um, with straight germ slit spore-length;

perispore indehiscent in 10% KOH, smooth, epispore smooth.

CoMMENTs: Annulohypoxylon guangxiense is similar to A. archeri (Berk.) Y.M.

Ju & al, which differs in its tissues with KOH-extractable greenish olivaceous

pigments and perispore dehiscent in 10% KOH (Ju & Rogers 1996).

Annulohypoxylon sichuanense Wei Li bis & L. Guo, sp. nov. FIGs 10-12

EN 570632

Differs from Annulohypoxylon multiforme by its tissues with KOH-extractable pigments

brown or reddish brown and its ostioles occasionally encircled with an inconspicuous

truncatum-type disc.

Fics 10-18. Annulohypoxylon sichuanense (HMAS 281263, holotype). 10. Stromata on wood;

11, 12. Ascospores. Annulohypoxylon maeteangense (HMAS 247821). 13. Stromata on wood;

14, 15. Ascospores. Annulohypoxylon substygium (HMAS 247819). 16. Stromata on wood;

17, 18. Ascospores.

Annulohypoxylon spp. nov. & new records (China) ... 435

436 ... Li& Guo

Type: China, Sichuan, Xiangtang, Hazhai, on decayed wood, 11.VHI.2013, W. Li & G.

Huang 2278 (Holotype, HMAS 281263).

EryMo.ocy: The epithet refers to type locality, Sichuan Province.

Stromata peltate, with constricted base, usually confluent, with inconspicuous

perithecial mounds, 1.8-2.3 x 1-1.7 cm, 1.5-5 mm thick; surface dark brown

to blackish brown; blackish brown granules immediately beneath surface,

with KOH-extractable brown or reddish brown pigment; the tissue below the

perithecial layer conspicuous, blackish brown, 0.7-3 mm thick. Perithecia

subspherical to obovate, 0.6-0.8 mm high, 0.3-0.7 mm broad. Ostioles

conical-papillate, often without encircling disc or occasionally encircled with

an inconspicuous truncatum-type disc, 0.1-0.2 mm diam. Asci 136-210

um long overall, the spore-bearing parts 64-121 x 5-6 um above 65-140

um long stipes. Ascus apical ring bluing in Melzer’s iodine reagent, discoid,

1-1.5 um high, 2-2.5 um diam. Ascospores brown, unicellular, ellipsoid-

inequilateral, with narrowly rounded ends, 8-12 x 3.5-5 um, with straight

germ slit, shorter than spore-length; perispore dehiscent in 10% KOH,

smooth, epispore smooth.

CoMMENTSs: Annulohypoxylon sichuanense is similar to A. multiforme (Fr.)

Y.M. Ju & al., which differs in its tissues with KOH-extractable greenish

olivaceous pigments and ostioles without an encircling disc.

Annulohypoxylon maeteangense J. Fourn. & M. Stadler,

Fungal Diversity 40: 32, 2010. Figs 13-15

Stromata irregularly pulvinate, with conspicuous perithecial mounds,

0.2-4 x 0.1-2.3 cm, 0.6-0.75 mm thick, often rosellinoid at the margins,

surface dark brown to black with an olivaceous tones; olivaceous to brown

granules immediately beneath surface, with KOH-extractable dull green

pigment; the tissue below the perithecial layer inconspicuous, black.

Perithecia spherical, 0.3-0.4 mm diam. Ostioles conical-papillate, black

and shiny, encircled by a slightly concave truncatum-type disc, 0.15-0.25

mm diam. Asci 100-118 um long overall, the spore-bearing parts 60-83 x

4-5 um above 32-40 um long stipes. Ascus apical ring bluing in Melzer’s

iodine reagent, discoid, 0.8-1 um high, 1.5-2 um broad. Ascospores brown,

unicellular, ellipsoid-inequilateral, with narrowly to broadly rounded ends,

6.5-8.5 x 3-4 um, with straight germ slit almost spore-length; perispore

dehiscent in 10% KOH, smooth, epispore smooth.

Annulohypoxylon spp. nov. & new records (China) ... 437

SPECIMENS EXAMINED: CHINA, HaINAN, Qionghai, Niululing, alt. 116 m, on wood,

4.X11.2009, Y.F Zhu & L. Guo 11 (HMAS 247821); Wanning, Shimeiwan, alt. 3 m, on

wood, 6.XII.2009, Y.F. Zhu & L. Guo 53 (HMAS 255362).

COMMENTs: Our specimens are morphologically similar to the holotype, but

our asci have longer spore-bearing parts; in the holotype from Thailand they

measure only 45-62 um (Fournier & al. 2010).

Annulohypoxylon substygium Sir & Kuhnert, Fungal Diversity. 85:20, 2017.

Figs 16-18

Stromata pulvinate to effused-pulvinate, with inconspicuous perithecial

mounds up to % exposed, 1-9 x 0.5-3 cm, 0.6-1.1 mm thick; surface black

with reddish brown tones to black; black granules immediately beneath

surface, with KOH-extractable light brownish black pigment; the tissue below

the perithecial layer inconspicuous, black. Perithecia spherical to obovate,

0.4-0.7 mm high, 0.3-0.5 mm broad. Ostioles papillate, encircled by a bovei-

type disc, 0.25-0.3 mm diam. Asci 96-110 um total length, the spore-bearing

parts 66-68 um long, 4-4.5 um broad, the stipes 30-46 um long. Ascus

apical ring weak bluing in Melzer’s iodine reagent, discoid, 0.2-0.5 um high,

1-1.5 um broad. Ascospores light brown to brown, unicellular, ellipsoid

nearly equilateral, with broadly rounded ends, 6.5-8 x 3-3.5 um, with

straight germ slit running almost the entire spore length; perispore dehiscent

in 10% KOH, smooth, epispore smooth.

SPECIMENS EXAMINED: CHINA, JIANGSU, Jurong, Baohuashan, on decayed wood,

15.IX.1935, H.N. Shen 101 (HMAS 9684); HAINAN, Ledong, Jianfengling, alt. 850

m, 29.VII.2007, S.H. He 1173 (HMAS 255340); Changjiang, Bawangling, alt. 600 m,

10.V.2009, W. Li & X.Y. Liu 2234 (HMAS 247819); Bawangling, alt. 500 m, 17.11.2017, L.

Guo 12136 (HMAS 255358).

CoMMENTs: In our specimens the tissue below the perithecial layer was

inconspicuous, while it is conspicuous (0.5-0.6 mm thick) in the holotype

(Kuhnert & al. 2017).

Discussion

Including the six species reported in this paper, we now recognize 26

Annulohypoxylon species in China (Teng 1963; Tai 1979; Bi & al. 1990; Ju &

Rogers 1996, 1999; Ju & al. 2004; Ma 2011; Kuhnert & al. 2017).

Acknowledgments

The authors would like to express their deep thanks to Prof. Xiuguo Zhang (Shandong

Agricultural University, China) and to Dr. L.E. Petrini (Breganzona, Switzerland)

438 ... Li& Guo

for serving as pre-submission reviewers, to Dr. Shaun Pennycook (Auckland, New

Zealand) for nomenclatural review, and for the support by the Ministry of Science and

Technology of the People’s Republic of China (No. 2013FY110400).

Literature cited

Bi ZS, Zheng GY, Li TH, Wang YZ. 1990. Macrofungus Flora of the Mountainous District of

North Guangdong. Guangdong Science and Technology Press, Guangzhou. 450 p.

Fournier J, Stadler M, Hyde KD, Duong ML. 2010. The new genus Rostrohypoxylon and

two new Annulohypoxylon species from Northern Thailand. Fungal Diversity 40: 23-36.

https://doi.org/10.1007/s13225-010-0026-4

Ju YM, Rogers JD. 1996. A revision of the genus Hypoxylon. Mycological Memoirs 20. 365 p.

Ju YM, Rogers JD. 1999. The Xylariaceae of Taiwan (excluding Anthostomella). Mycotaxon 73:

343-440.

Ju YM, Rogers JD, Hsieh HM. 2004. New Hypoxylon species and notes on some names

associated with or related to Hypoxylon. Mycologia 96(1): 154-161.

Kuhnert E, Sir EB, Lambert C, Hyde KD, Hladki AI, Romero AI, Rohde M, Stadler M. 2017.

Phylogenetic and chemotaxonomic resolution of the genus Annulohypoxylon (Xylariaceae)

including four new species. Fungal Diversity 85: 1-43.

https://doi.org/10.1007/s13225-016-0377-6

Ma HX. 2011. Taxonomy and molecular phylogeny of several genera of Xylariaceae from

China. Jilin Agricultural University, Changchun. 176 p.

Pereira J, Rogers JD, Bezerra JL. 2010. New Annulohypoxylon species from Brazil. Mycologia

102(1): 248-252. https://doi.org/10.3852/09-116

Tai FL. 1979. Sylloge Fungorum Sinicorum. Science Press, Beijing. 1527 p.

Teng SC. 1963. Fungi of China. Science Press, Beijing. 808 p.

MY COTAXON

July-September 2019—Volume 134, pp. 439-442

https://doi.org/10.5248/134.439

Pleurotheciopsis triseptata sp. nov. from China

ConG-Conc At’, JI-WEN XIA’, XIU-GUO ZHANG’, LI-Guo Ma”

"Shandong Provincial Key Laboratory for Biology of Vegetable Diseases and Insect Pests,

College of Plant Protection, Shandong Agricultural University,

Taian, Shandong, 271018, China

? Shandong Key Laboratory of Plant Virology, Institute of Plant Protection,

Shandong Academy of Agricultural Sciences, Jinan, Shandong, 250100, China

“CORRESPONDENCE TO: maliguo809@163.com

ABSTRACT—A new anamorphic species, Pleurotheciopsis triseptata, is described and

illustrated from specimens collected on dead stems in China. This fungus is characterized

by polyblastic and sympodial conidiogenous cells producing catenate, fusiform, 3-euseptate,

versicolored conidia.

Key worps—microfungi, hyphomycete, taxonomy

Introduction

Pleurotheciopsis B. Sutton was erected by Sutton (1973) for P pusilla (the

type species) and P bramleyi. The genus is characterized by distinct and

brown conidiophores with polyblastic, integrated, sympodial, and denticulate

conidiogenous cells that produce euseptate and hyaline to brown conidia that

form acropetal unbranched chains (Sutton 1973, Ellis 1976, Castaneda & al.

2001). Assigning species in Pleurotheciopsis is primarily based on conidial

morphological characters, including shape, septation, size, and pigmentation.

Five additional species have been described (Castaneda 1985, Cazau & al. 1993,

Castaneda & Iturriaga 1999, Castaneda & al. 2001, Rambelli & al. 2008), but

P. setiformis (with synchronous and ampulliform conidiogenous cells and setae

around the conidiophores) was subsequently transferred to Ampullicephala by

R.F. Castaneda & al. (2009). Castaneda & al. (2001) provided a key to five of the

accepted Pleurotheciopsis species.

440 ... Ai & al.

During a survey on wood-inhabiting microfungi in China, a new

species that belongs to the genus Pleurotheciopsis was found. Specimens are

deposited in the Herbarium of Department of Plant Pathology, Shandong

Agricultural University, Taian, Shandong, China (HSAUP) and the

Mycological Herbarium, Institute of Microbiology, Chinese Academy of

Sciences, Beijing, China (HMAS).

Taxonomy

Pleurotheciopsis triseptata L.G. Ma & X.G. Zhang, sp. nov. FIG. 1

MycoBAnk MB 831794

Differs from Pleurotheciopsis sylvestris by its 3-euseptate and smaller conidia, and from

P. bramleyi and P. websteri by its 3-euseptate and versicolored conidia that are bigger

than P bramleyi and shorter than P. websteri.

Type: China, Sichuan Province, Longchi National Forest Park, on dead stems of an

unidentified broadleaf tree, 22 Apr. 2012, L.G. Ma (Holotype, HSAUP H2239; isotype,

HMAS 146166).

EryMmo ocy: triseptata, referring to the number of conidial septa.

COLONIES on the natural substratum effuse, brown to dark brown, hairy.

Mycelium partly superficial, partly immersed, composed of branched,

septate, pale brown, smooth hyphae, 2.0-3.5 um diam. CONIDIOPHORES

macronematous, mononematous, single, unbranched, erect, straight to

flexuous, cylindrical, smooth, thick-walled, brown to dark brown, pale

brown towards the apex, 4—9-septate, 105-240 um long, 6.5-13.5 um diam

at the base, 4-6.5 um wide at the tapered apex. CONIDIOGENOUS CELLS

integrated, terminal, polyblastic, sympodial, brown, cylindrical, smooth,

thick-walled, with 1-4 short, unthickened denticles, 29.5-60 x 4.5-6.5

um. Conidial secession schizolytic. Conrp1A in distinct, unbranched,

acropetal chains, fusiform, sometimes curved, thick-walled, middle cells

brown and end cells pale brown to subhyaline, tapered towards the ends,

3-euseptate, intercalary conidia 24-33.5 x 5.5-7.5 um, truncate at the

ends, apical conidia 22.5-33 x 6-7.5 um, obtuse at the apex and truncate

at the base.

CoMMENTS—Pleurotheciopsis triseptata is most similar to P_ sylvestris

R.F. Castaneda & Iturr. in conidial shape and pigmentation, but P sylvestris

has conidia that are 4—7-septate and larger (29-49 x 7—8.5 um; Castafieda

& Iturriaga 1999). Pleurotheciopsis bramleyi B. Sutton and P. websteri Cazau

& al. also resemble our new species, but P bramleyi has conidia that are

hyaline, 2—-3-septate, and smaller (16.5-25 x 4.5-6.5 um; Sutton 1973),

Pleurotheciopsis triseptata sp. nov. (China) ... 441

20um 20um

20m 20um

Fic. 1. Pleurotheciopsis triseptata (holotype, HSAUP H2239).

A, B. Conidiophores, conidiogenous cells, and conidia;

C. Conidiophore with conidiogenous cell; D. Conidia.

while conidia in P websteri are 5-septate, hyaline, longer, and narrower

(32-38 x 5-6 um; Cazau & al. 1993).

Acknowledgments

The authors express gratitude to Dr. Jian Ma and Dr. Ze-Fen Yu for serving as

pre-submission reviewers and for their valuable comments and suggestions. This

project was supported by Young Talents Training Program of Shandong Academy

of Agricultural Sciences (CXGC2018E04), National Key R&D Program of China

(2017YFD0201700), National Natural Science Foundation of China (31400019,

31230001, 31093440), and the Ministry of Science and Technology of the People’s

Republic of China (2006FY 120100).

442 ... Ai & al.

Literature cited

Castafieda RE. 1985. Deuteromycotina de Cuba. Hyphomycetes III. Instituto de Investigaciones

Fundamentales en Agricultura Tropical “Alejandro de Humboldt’, Havana, Cuba.

Castaneda RF, Iturriaga T. 1999. A new species of Pleurotheciopsis from a rainforest in Venezuela.

Mycotaxon 70: 63-68.

Castaneda RE, Calduch M, Garcia D, Izquierdo Z. 2001. A new species of Pleurotheciopsis from leaf

litter. Mycotaxon 77: 1-5.

Castafieda RF, Gusmao LFP, Moraes Junior VO de, Ledo-Ferreira SM, Saikawa M, Minter DW,

Stadler M. 2009. Two setose anamorphic fungi: Ampullicephala gen. nov. and Venustosynnema

grandiae sp. nov. Mycotaxon 109: 275-288. https://doi.org/10.5248/109.275

Cazau MC, Arambarri AM, Cabello MN. 1993. New hyphomycetes from Santiago River. VI.

(Buenos Aires Province, Argentina). Mycotaxon 46: 235-240.

Ellis MB. 1976. More dematiaceous hyphomycetes. Commonwealth Mycological Institute, Kew,

Surrey, England.

Rambelli A, Venturella G, Ciccarone C. 2008. Dematiaceous hyphomycetes from Pantelleria

mediterranean maquis litter. Flora Mediterranea 18: 441-467.

Sutton BC. 1973. Some hyphomycetes with holoblastic sympodial conidiogenous cells. Transactions

of British Mycological Society 61(3): 417-429. https://doi.org/10.1016/S0007-1536(73)80112-3

MY COTAXON

July-September 2019—Volume 134, pp. 443-446

https://doi.org/10.5248/134.443

Repetophragma verrucosum sp. nov. from China

ConG-ConG At’, JI-WEN X14’, XIU-GUO ZHANG’, L1-Guo Ma”

' Shandong Provincial Key Laboratory for Biology of Vegetable Diseases and Insect Pests,

College of Plant Protection, Shandong Agricultural University,

Taian, Shandong, 271018, China

? Shandong Key Laboratory of Plant Virology, Institute of Plant Protection,

Shandong Academy of Agricultural Sciences, Jinan, Shandong, 250100, China

“CORRESPONDENCE TO: maliguo809@163.com

ABSTRACT—A new anamorphic species, Repetophragma verrucosum, is described and

illustrated from specimens collected on decaying branches in China. This fungus is

distinguished by monoblastic and annellidic conidiogenous cells that produce fusiform to

subobclavate, 3-euseptate, verrucose, and yellowish brown conidia.

KEY worps—hyphomycete, taxonomy, microfungi

Introduction

The vegetation in China is composed of a great variety of evergreen

and deciduous broadleaf forests where many different kinds of wood-

inhabiting fungi have been recently discovered (e.g., Ma & al. 2016a,b,

2018). During our ongoing study of anamorphic fungi from this region,

we collected a species with morphological characters of Repetophragma

Subram. (Subramanian 1992) from decaying branches. Our specimen, which

differed morphologically from previously described Repetophragma species,

is proposed here as a new species.

The specimens are deposited in the Herbarium of Department of Plant

Pathology, Shandong Agricultural University, Taian, Shandong, China

(HSAUP) and the Mycological Herbarium, Institute of Microbiology,

Chinese Academy of Sciences, Beijing, China (HMAS).

444 ... Ai & al.

Taxonomy

Repetophragma verrucosum L.G. Ma & X.G. Zhang, sp. nov. FIG. 1

MB 831795

Differs from Repetophragma dennisii by its narrower, longer, verrucose conidia; and

from R. laxisporum by its 3-euseptate, wider, verrucose conidia.

Type: China, Guangdong Province, Liuxihe National Forest Park, on dead branches

of an unidentified broadleaf tree, 12 Oct. 2010, L.G. Ma (Holotype, HSAUP H7082;

isotype, HMAS 146165).

ETYMOLOGY: verrucosum, referring to the verrucose conidial wall.

CoLonlies on the natural substratum effuse, brown to dark brown, hairy.

Mycelium partly superficial, partly immersed, composed of septate, pale

brown, smooth hyphae, 2.5-3.5 um diam. CONIDIOPHORES macronematous,

mononematous, single, unbranched, erect, straight to slightly flexuous,

cylindrical, smooth, thick-walled, dark brown to black, brown towards

the apex, multiseptate, 190-335 x 6.0-8.5 tum. CONIDIOGENOUS CELLS

holoblastic, monoblastic, integrated, terminal, indeterminate, brown,

cylindrical, smooth, thick-walled, with numerous annellidic percurrent

extensions towards the apex. Conidial secession schizolytic. CoNrpIA

acrogenous, solitary, fusiform to subobclavate, thick-walled, yellowish

brown, slightly paler at the end cells, straight or slightly curved, verrucose,

3-euseptate, 23-37.5 x 7.5-10.5 um, 2.5-4.5 um diam. at the truncate base,

rounded or slightly acute at the apex.

CoMMENTS—Subramanian (1992) segregated Repetophragma with

R. biseptatum (M.B. Ellis) Subram. as its type species from Sporidesmium

Link. The genus is diagnosed by macronematous and single conidiophores

possessing integrated and terminal conidiogenous cells with annellidic

percurrent extensions and solitary and euseptate conidia (Wu & Zhuang

2005, Castaneda & al. 2011). Castafieda & al. (2011) reviewed the genus,

proposed a new species and twelve new combinations, and provided a

comparative table and a key to 32 accepted Repetophragma species. Among

three additional species since added are R. hainanense Jian Ma & X.G.

Zhang and R. elegans J.W. Xia & X.G. Zhang, described from China (Ma &

al. 2014, Wang & al. 2017).

Repetophragma verrucosum resembles R. dennisii (M.B. Ellis) Subram. and

R. laxisporum (R.F. Castafieda) R.E. Castafeda in conidial shape. However,

the conidia of R. dennisii are shorter and wider (23-32 x 11-13 um), smooth,

and brown with paler apical cells (Subramanian 1992); and R. laxisporum

Repetophragma verrucosum sp. nov. (China) ... 445

20um

Fic. 1. Repetophragma verrucosum (holotype, HSAUP H7082).

A. Conidiophores, conidiogenous cells, and conidia;

B. Conidiophores with conidiogenous cells; C. Conidia.

differs from our species by producing 6-7-septate, slightly narrower (7-9

um), smooth, brown conidia with subhyaline end cells (Castafeda & al. 2011).

446 ... Ai & al.

Acknowledgments

The authors express gratitude to Dr. Jian Ma and Dr. Ze-Fen Yu for serving as

pre-submission reviewers and for their valuable comments and suggestions. This

project was supported by Young Talents Training Program of Shandong Academy

of Agricultural Sciences (CXGC2018E04), National Key R&D Program of China

(2016YFD0300700), National Natural Science Foundation of China (31400019,

31230001, 31093440), the Ministry of Science and Technology of the People’s

Republic of China (2006FY120100).

Literature cited

Castafieda RE, Heredia G, Arias RM, McKenzie EHC, Hyde KD, Stadler M, Saikawa M, Gené J,

Guarro J, Iturriaga T, Minter DW, Crous PW. 2011. A new species and re-disposed taxa in

Repetophragma. Mycosphere 2: 273-289.

Ma J, Zhang XG, Castafieda RF. 2014. New species of Acrodictys and Repetophragma from dead

branches in China. Mycotaxon 127: 129-134. https://doi.org/10.5248/127.129

Ma J, Zhang K, Zhang XG, Castafieda RF. 2016a. Three new species of Spadicoides from Lushan

Mountain, China. Mycological Progress 15: 43. https://doi.org/10.1007/s11557-016-1185-9

Ma YR, Xia JW, Gao JM, Li XY, Castafeda RF, Zhang XG, Li Z. 2016b. Atrokylindriopsis, a new

genus of hyphomycetes from Hainan, China, with relationship to Chaetothyriales. Mycological

Progress 14: 77. https://doi.org/10.1007/s11557-015-1071-x

Ma LG, Li HH, Xia JW, Castafeda RE, Zhang XG. 2018. A new anamorphic fungus of Nakataea

from China. Nova Hedwigia 107: 189-193. https://doi.org/10.1127/nova_hedwigia/2017/0463

Subramanian CV. 1992. A reassessment of Sporidesmium (hyphomycetes) and some related taxa.

Proceedings of the Indian National Science Academy, Part B, Biological Sciences 58: 179-189.

Wang XM, Chen SS, Liu XM, Zhao ZJ, Li HH, Zhang XG, Xia JW. 2017. Repetophragma elegans sp.

nov. from Hainan Province, China. Mycotaxon 132: 881-884. https://doi.org/10.5248/132.881

Wu WP, Zhuang WY. 2005. Sporidesmium, Endophragmiella and related genera from China. Fungal

Diversity Research Series 15. 351 p.

MY COTAXON

July-September 2019—Volume 134, pp. 447-455

https://doi.org/10.5248/134.447

Pseudopyricularia cyperi, a new record for Iran

ADEL PORDEL', AMIRREZA AMIRMIJANI,

MOHAMMAD JAVAN-NIKKHAH™

' Department of Plant Protection, College of Agriculture and Natural Science,

University of Tehran, Karaj 31587-77871, Iran

? Department of Plant Protection, Faculty of Agriculture, University of Jiroft,

Jiroft, Iran

* CORRESPONDENCE TO: jnikkhah@ut.ac.ir

AssTrRACcT—During a survey of sedge plants in northern Iran, three specimens of

Pseudopyricularia cyperi were isolated from Cyperus sp. Their taxonomical identity

was established by their spore and conidiophore morphology and ITS rDNA sequence

analysis. The specimens are described and illustrated. Pseudopyricularia cyperi is a new

record for Iran.

Key worps—Magnaporthales, phylogeny, Pyriculariaceae, taxonomy

Introduction

Pyriculariaceae was established as a new family of the Magnaporthales

by Klaubauf & al. (2014). The family contains ten genera: Bambusicularia,

Barretomyces, Deightoniella, Macgarvieomyces, Neocordana, Neopyricularia,

Proxipyricularia, Pseudopyricularia, Pyricularia, and Xenopyricularia

(Klaubauf & al. 2014, Hernandez-Restrepo & al. 2015). Some species

originally described in Pyricularia have been transferred to these new genera;

e.g., Macgarvieomyces borealis, Proxipyricularia zingiberis, Neopyricularia

commelinicola, Pseudopyricularia higginsii, and Xenopyricularia zizaniicola.

Pyricularia higginsii Luttr. was transferred to Dactylaria by Ellis (1976), but

Bussaban & al. (2003) maintained the species in Pyricularia based on rDNA

ITS sequence analysis. On the basis of analyses of five gene regions, Klaubauf

448 ... Pordel, Amirmijani, Javan-Nikkhah

& al. (2014) placed some isolates previously identified as Pyricularia higginsii

in a new genus Pseudopyricularia and separated them into Pseudopyricularia

higginsii and two new species, P. cyperi and P. kyllingae.

Additional species since described or combined in Pseudopyricularia

include P bothriochloae, P hagahagae, P. hyrcaniana, P. iraniana, and

P. persiana (Crous & al. 2015, 2018; Pordel & al. 2017; Marin-Felix & al.

2017; Jayawardena & al. 2019). Pseudopyricularia species are primarily

distinguished from Pyricularia sensu stricto by having short, determinate,

brown conidiophores with an apical rachis with flat-tipped denticles; they are

similar to each other in conidial size and can be resolved only by phylogenetic

analysis (Klaubauf & al. 2014; Pordel & al. 2015, 2017). Pordel & al. (2017)

reassessed the morphological criteria, and re-described Pseudopyricularia

conidia as obclavate, fusiform, cylindrical, and 1-2-septate.

Here we describe and illustrate Pseudopyricularia cyperi from Iranian

specimens and compare them with other species in Pseudopyricularia.

Materials & methods

Sampling and fungal isolates

Plant material was obtained from Mazandaran Province in Iran during the

summer and fall of 2012 and 2015. Leaves of Cyperus sp. with leaf spot symptoms

were collected and stored in a refrigerator at 4 °C until used. To induce sporulation

of Pseudopyricularia, leaf pieces were surface sterilized for 2 min in 1% sodium

hypochlorite, dried on filter paper, and then incubated on wet filter paper at

25 °C. Conidia produced on these surface-sterilized leaf pieces were transferred to

water agar (WA). Single hyphal tips emerging from germinating conidia were then

transferred to potato dextrose agar (PDA) medium (Pordel & al. 2015).

Living cultures have been deposited in the herbarium in the Department of Plant

Protection, Faculty of Agricultural Sciences & Engineering, University of Tehran,

Karaj, Iran (UTFC).

Morphological characterization

Cultures were grown on oatmeal agar (OA) and PDA to determine overall colony

morphology and characteristics. Micromorphological characters were determined

from colonies grown on synthetic nutrient-poor agar (SNA; Nirenberg 1976), and

WA supplemented with pieces of rice leaves. Plates were incubated at 23-25 °C

under a regime of 12 h dark/12 h near-ultraviolet light and examined for sporulation

after 1-3 wks. Measurements and microphotographs were taken from slide mounts

in lactophenol and lactophenol cotton blue. Measurements were taken from 70-100

conidiophores and 200 conidia. Photographs were taken using a Sony digital camera

mounted on an Olympus BH2 microscope.

Pseudopyricularia cyperi new for Iran ... 449

Pyricularia oryzae BF0028

Pyricularia oryzae CRO0029

Pyricularia oryzae JP0028

Pyricularia oryzae CBS 365.52

Pyricularia oryzae CBS 375.54

Pyricularia sp. CBS 133598

Pyricularia zingibericola RNOO01

Pyricularia penniseticola BF0017

Xenopyricularia zizaniicola CBS 133593

100 ' Xenopyricularia zizaniicola CBS 132356

Bambusicularia brunnea CBS 133599

Proxipyricularia Zingiberis CBS 132355

Pyricularia grisea BRO029

Pyricularia grisea JP0034

"| Pyricularia grisea CROO24

Pyricularia ctenantheicola GROO01

99 ' Pyricularia ctenantheicola GRO002

Neopyricularia commelinicola CBS 128303

99 ' Neopyricularia commelinicola CBS 128306

Macgarvieomyces borealis CBS 461.65

Macgarvieomyces juncicola CBS 610.82

Pseudopyricularia iraniana UTFC-P012

Pseudopyricularia iraniana_ \RAN 2761C

Pseudopyricularia iraniana \RAN 2762C

Pseudopyricularia kyllingae CBS 133597 '

Pseudopyricularia cyperi PHO0053 Ny LQ

Pseudopyricularia cyperi UTFC-PO17 | ii

Pseudopyricularia cyperi UTFC-PO8

Pseudopyricularia cyperi UTFC-PO16

Pseudopyricularia cyperi CBS 133595

Pseudopyricularia cyperi CBS 665.79

Pseudopyricularia persiana UTFC-PO22 \\

Pseudopyricularia persiana UTFC-PO21 [| 4

Pseudopyricularia persiana UTFC-PO20

Pseudopyricularia higginsii CBS 121934

Pseudopyricularia hagahagae CPC 25635

Pseudopyricularia bothriochloae CPC 21650

Pseudopyricularia hyrcaniana \RAN 2759C

Pseudopyricularia hyrcaniana \RAN 2760C

Pseudopyricularia hyrcaniana \RAN 2758C

Barretomyces calatheae CBS 129274

InowAdopnesd

elle

0.01

Fic. 1. Maximum likelihood tree inferred from ITS sequence of Pyriculariaceae species.

Branch values of ML >80% are noted above internodes. Strains of Pseudopyricularia cyperi

from Iran are in bold.

450 ... Pordel, Amirmijani, Javan-Nikkhah

Phylogenetic analysis

The three isolates of Pseudopyricularia cyperi collected in Iran were used for

phylogenetic analysis. Total genomic DNA was extracted using the protocol of

Zhong & Steffenson (2001) and the ITS region was amplified according to Klaubauf

& al. (2014). Additional ITS sequences were downloaded from GenBank. A total of

41 nucleotide sequences (including GenBank sequences originating from Klaubauf

& al. 2014) were analyzed. Barretomyces calatheae was used as outgroup (Klaubauf

& al. 2014).

The sequences were edited by CLC Genomic Workbench 10.1 and aligned using

MUSCLE (Edgar 2004). Phylogenetic analyses were performed with MEGA v. 6

(Tamura & al. 2013). A phylogenetic tree was constructed using the maximum

likelihood method based on the Kimura two-parameter model (Kimura 1980). The

percentage of replicate trees in which the associated taxa clustered together in the

bootstrap test (1000 replicates) are shown next to the branches. The tree is drawn to

scale, with branch lengths in the same units as those of the evolutionary distances

used to infer the phylogenetic tree. The evolutionary distances were computed using

the maximum composite likelihood method and are in the units of the number

of base substitutions per site. All positions with less than 95% site coverage were

eliminated, permitting fewer than 5% alignment gaps, missing data, and ambiguous

bases at any position. The final ITS dataset comprised 296 positions.

Results

Phylogenetic analysis

Our phylogenetic analyses grouped Pseudopyricularia species together in

one clade within Pyriculariaceae with a high (bootstrap >98%) statistical

support (Fic. 1). These species have variously shaped 1-2-septate conidia.

All are pathogenic to plants but have different hosts and have likely evolved

separately. The phylogeny (Fic. 1) demonstrates a 100% MLBP support for

a close relationship between known P. cyperi isolates and the strains isolated

from Cyperus sp. in Iran, supporting classification of the Iranian samples

as Pseudopyricularia cyperi. In particular, isolate UTFC-P016 has the same

ITS sequences as P. cyperi CBS 133595 (ex-holotype; from Japan) and isolate

UTFC-P017 has the same ITS sequences as P. cyperi PHO053 (ex-paratype;

from Philippines).

Taxonomy

Pseudopyricularia cyperi Klaubauf, M.-H. Lebrun & Crous,

Stud. Mycol. 79: 110. 2014. Fics 2, 3

Colonies grown on PDA after 1 wk at 23-25 °C white, reaching 40 mm

diam. Conidiophores solitary, erect, straight or curved to geniculate, branched,

Pseudopyricularia cyperi new for Iran... 451

Fic. 2. Pseudopyricularia cyperi (UTFC-PO8). A-C. Solitary, erect, branched, and unbranched

conidiophores; D-F. Conidia. Scale bars = 10 um.

medium brown, smooth, 40-100 x 3-4 um. Conidiogenous cells integrated,

terminal and intercalary, pale brown, smooth, forming a rachis with several

protruding, flat-tipped denticles. Conidia solitary, obclavate, medium brown,

smooth to finely roughened, 2-septate, 22-29 x 5-6 um; hilum truncate,

slightly protruding, unthickened, not darkened.

SPECIMENS EXAMINED—On infected leaves of Cyperus sp. IRAN, MAZANDARAN

PROVINCE, Amol, 25 July 2012, Adel Pordel cy2k (UTFC-PO8; GenBank KP 144446),

Adel Pordel cy4-1 (UTFC-PO16; GenBank MF768983), Adel Pordel cy2k1 (UTFC-O17

GenBank MF768984).

CoMMENTS— The morphology of the Iranian specimens cited above agree

with the protologue description of Pseudopyricularia cyperi (Klaubauf &

al. 2014). This species is morphologically similar to P higginsii (Luttr.)

452 ... Pordel, Amirmijani, Javan-Nikkhah

Fic. 3. Pseudopyricularia cyperi (UTFC-PO8). A. Conidia; B. Conidiophores

Pseudopyricularia cyperi new for Iran ... 453

Klaubauf & al., which differs by its longer and slightly wider conidia

(17.5-36.5 x 5.3-6.5 um, Luttrell 1955; 20-36 x 5-6 um, Ellis 1976). This

represents the first report of Pseudopyricularia cyperi in Iran.

Discussion

The order Magnaporthales has been reclassified into three families,

Magnaporthaceae, Pyriculariaceae, and Ophioceraceae (Klaubauf & al. 2014,

Luo & al. 2015, Pordel & al. 2017). The Pyriculariaceae is characterized

by Pyricularia-like asexual states and pathogenicity to Poaceae or other

monocotylenous plants. Phylogenetic analysis based on five gene regions

allowed definition of several new genera and species in Pyriculariaceae

(Klaubauf & al. 2014).

Previously, several species of Pseudopyricularia were unrecognized

and placed in the P higginsii complex. Based on phylogenetic analysis,

new Pseudopyricularia species were identified in this clade (Klaubauf &

al. 2014, Pordel & al. 2017). Pseudopyricularia and Macgarvieomyces are

morphologically and phylogenetically similar (Klaubauf & al. 2014, Pordel

& al. 2017). Macgarvieomyces is distinguished from Pseudopyricularia

and Pyricularia sensu stricto by its production of chlamydospores, mostly

unbranched conidiophores, and narrowly obclavate, granular, guttulate

and 1-septate conidia (Pordel & al. 2017). Pseudopyricularia bothriochloae

(Crous & Cheew.) Y. Marin & Crous is closely related to P hyrcaniana

Pordel & Jav.-Nikkh (both with l-septate conidia), while P hagahagae

Crous & M.J. Wingf. (with 2-septate conidia) is clearly separated from

them phylogenetically (Fic. 1). The phylogeny also supports five species

(P. higginsii, P iraniana Pordel & Jav.-Nikkh, P cyperi, P persiana

G. Ghorbani & al., and P. kyllingae Klaubauf & al.) as separate from species

having 1-septate conidia. Pseudopyricularia cyperi strains from Iran, which

lie in P cyperi clade, are morphologically similar to all previous records

of this species from Cyperus sp. in other countries (Klaubauf & al. 2014;

Borromeo & al. 1993, as “P. higginsii”).

Acknowledgments

We gratefully acknowledge the High Council for Research of the University of

Tehran for financially supporting and providing materials under grant number

7110022/6/29. We thank Marc Henri Lebrun (UMR BIOGER INRA AgroParisTech,

Thiverval-Grignon, France), Eric H.C. McKenzie (Manaaki Whenua Landcare

Research, Auckland, New Zealand), and Andrea Paola Zuluaga Cruz (Centro De

Investigacion Tibaitata, Mosquera, Colombia) for their expert reviews.

454 ... Pordel, Amirmijani, Javan-Nikkhah

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Pyricularia infecting rice and weed hosts. Phytopathology 83: 393-399.

https://doi.org/10.1094/Phyto-83-393

Bussaban B, Lumyong S, Lumyong P, Hyde KD, McKenzie EHC. 2003. Three new species

of Pyricularia are isolated as zingiberaceous endophytes from Thailand. Mycologia 95:

519-524. https://doi.org/10.2307/3761895

Crous PW, Wingfield MJ, Roux JJ Le, Richardson DM, Strasberg D, Shivas RG & al. 2015. Fungal

Planet description sheets: 371-399. Persoonia 35: 264-327.

https://doi.org/10.3767/003158515X690269

Crous PW, Luangsa-ard JJ, Wingfield MJ, Carnegie AJ, Hernandez-Restropo M & al. 2018.

Fungal Planet description sheets: 785-867. Persoonia 41: 238-417.

https://doi.org/10.3767/persoonia.2018.41.12

Edgar RC. 2004. MUSCLE: a multiple sequence alignment method with reduced time and space

complexity. BMC Bioinformatics 5: 113.

Ellis MB. 1976. More dematiaceous hyphomycetes. Commonwealth Mycological Institute, Kew,

Surrey, England. 507 p.

Hernandez-Restrepo M, Groenewald JZ, Crous PW. 2015. Neocordana gen. nov., the causal

organism of cordana leaf spot on banana. Phytotaxa 205(4): 229-238.

https://doi.org/10.11646/phytotaxa.205.4.2

Jayawardena RS, Hyde KD, Jeewon R, Ghobad-Nejhad M, Wanasinghe DN & al. 2019. One stop

shop II: taxonomic update with molecular phylogeny for important phytopathogenic genera:

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Kimura M. 1980. A simple method for estimating evolutionary rates of base substitutions

through comparative studies of nucleotide sequences. Journal of Molecular Evolution

16: 111-120. https://doi.org/10.1007/bf01731581

Klaubauf S, Tharreau D, Fournier E, Groenewald JZ, Crous PW, de Vries RP, Lebrun MH. 2014.

Resolving the polyphyletic nature of Pyricularia (Pyriculariaceae). Studies in Mycology

79: 85-120. https://doi.org/10.1016/j.simyco.2014.09.004

Luo J, Walsh E, Zhang N. 2015. Toward monophyletic generic concepts in Magnaporthales: species

with Harpophora asexual states. Mycologia 107: 641-646. https://doi.org/10.3852/14-302.

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46(6): 810-814. https://doi.org/10.1080/00275514.1954.12024417

Marin-Felix Y, Groenewald JZ, Cai L, Chen Q, Marincowitz S, Barnes I & al. 2017. Genera of

phytopathogenic fungi: GOPHY 1. Studies in Mycology 86: 99-216.

https://doi.org/10.1016/j.simyco.2017.04.002

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Pordel A, Javan-Nikkhah M, Khodaparast SA. 2015. A reappraisal of the Pyriculariaceae in Iran.

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https://doi.org/10.1007/s11557-017-1307-z

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MYCOTAXON

July-September 2019—Volume 134, pp. 457-461

https://doi.org/10.5248/134.457

Morganjonesia gen. nov. for two atypical

Corynespora and Teratosperma species

Kal ZHANG”, MIN QIAO’, ZE-FEN YU’,

DE-WEI LI*4, RAFAEL F. CASTANEDA-RUIZ>

' Department of Landscaping, Shandong Yingcai University, Jinan, Shandong 250104, China

? Laboratory for Conservation and Utilization of Bio-resources,

Key Laboratory for Microbial Resources of the Ministry of Education,

Yunnan University, Kunming, Yunnan, 650091, China

> The Connecticut Agricultural Experiment Station,

Valley Laboratory, 153 Cook Hill Road, Windsor, CT 06095, USA

*Co-Innovation Center for Sustainable Forestry in Southern China,

Nanjing Forestry University, Nanjing, Jiangsu 210037, China

° Instituto de Investigaciones Fundamentales en Agricultura Tropical Alejandro de Humboldt

(INIFAT), Calle 1 Esq. 2, Santiago de Las Vegas, C. Habana, Cuba, C.P. 17200

* CORRESPONDENCE TO: kaise0907@126.com

ABSTRACT—Morganjonesia is proposed as a new genus for two species, Corynespora

calophylli and Teratosperma litchii, which have monotretic conidial ontogeny on the terminal

conidiogenous cells and solitary, euseptate, verruculose, brown conidia with a single, subulate

or acute, hyaline conidial appendage at the apex (sometimes with another downwardly-direct

conidial appendage). Illustrations of both species are provided.

KEY worDs—asexual Ascomycota, hyphomycetes, systematics

Introduction

Corynespora Giissow, typified by C. mazei Giissow [= C. cassiicola (Berk. &

M.A. Curtis) C.T. Wei], is distinguished by macronematous, mononematous,

and generally unbranched, erect conidiophores and monotretic, terminal,

integrated, determinate or with several doliiform, enteroblastic percurrent

extension conidiogenous cells. The conidiogenous loci are apical, slightly

458 ... Zhang & al.

depressed, somewhat melanized, and thickened around the pore to produce

solitary or blastocatenate, multidistoseptate, obclavate, cylindrical, obovate,

oval, smooth-walled or verruculose, pale pigmented conidia usually with

a slightly melanized basal scar (Morgan-Jones 1988, Seifert & al. 2011).

Corynespora calophylli clearly deviates from the generic concept of Corynespora,

because it is characterized by monotretic conidial ontogeny and solitary,

2-euseptate, obclavate to obpyriform, brown conidia with an apical, acute,

hyaline conidial appendage, slightly papillate at the truncate base (Holubova-

Jechova & Castafieda-Ruiz 1986).

Teratosperma Syd. & P. Syd., typified by T. singulare Syd & P. Syd., is

characterized by macronematous, mononematous, generally unbranched,

erect conidiophores and monoblastic, terminal, integrated, indeterminate

conidiogenous cells with several annellidic enteroblastic percurrent extensions.

The conidia are staurospore, tetraradiate, rostrate, brown to olivaceous brown,

septate at the main axis, with 1-3-basal arms, and pale pigmented (Ellis 1971,

Seifert & al. 2011). Teratosperma litchii clearly deviates from the generic

concept of Teratosperma, because it is distinguished by a monotretic conidial

ontogeny and solitary, 3(-4)-euseptate, obclavate, brown conidia with 1-2

apical, subulate, hyaline appendage that are slightly papillate at the truncate

base (Matsushima 1980).

Here we propose a new genus, Morganjonesia, to accommodate these two

atypical species.

Taxonomy

Morganjonesia R.E. Castaneda, K. Zhang & D.W. Li, gen. nov.

MB 830288

Differs from Corynespora species by its euseptate conidia with an acute or subulate,

hyaline apical or downwardly direct conidial appendage; and differs from Teratosperma

species by its monotretic conidial ontogeny.

TYPE SPECIES: Corynespora calophylli Hol.-Jech. & R.E. Castafieda [= Morganjonesia

calophylli (Hol.-Jech. & R.F. Castafieda) R.E Castafieda & al.]

Erymo ocy: Latin, Morganjonesia, in honor of Dr. Gareth Morgan-Jones in recognition

of his contributions to the study of hyphomycetes.

CONIDIOPHORES macronematous, mononematous, unbranched, erect, straight,

cylindrical, septate, smooth, brown to dark brown. CONIDIOGENOUS CELLS

monotretic, integrated, terminal, cylindrical, determinate or sometimes with

one or several oblong enteroblastic percurrent extensions. Conidial secession

schizolytic. Conrp1A solitary, acrogenous, obclavate to obpyriform, euseptate,

459

Morganjonesia gen. & spp. nov. ...

Fic. 1. Morganjonesia calophylli (holotype, INIFAT C84/150).

A. Conidia with single apical conidial appendages; B. Conidiogenous cells with tretic loci.

460 ... Zhang & al.

thd

asks

EI ss

ta . a

Soa

Stree ies

ek «

235,

Fic. 2. Morganjonesia litchii (holotype, MFC 9048).

A. Conidia with two conidial appendages; B. Conidiogenous cells with tretic loci.

constricted at the septa, slightly papillate at the truncate base, brown or dark

brown, smooth-walled or verruculose, with 1-2 acute or subulate, hyaline,

smooth conidial appendages at the apex.

Morganjonesia gen. & spp. nov. ... 461

Morganjonesia calophylli (Hol.-Jech. & R.F. Castafieda.) R.E. Castafieda,

K. Zhang & D.W. Li, comb. nov. FIG. 1

MB 830290

= Corynespora calophylli Hol.-Jech. & R.F. Castafieda, Ceska Mykol. 40(2): 83 (1986).

Morganjonesia litchii (Matsush.) K. Zhang & R.F. Castafieda, comb. nov. Fi. 2

MB 830291

= Teratosperma litchii Matsush., Matsush. Mycol. Mem. 1: 73 (1980).

NoTE: Solicorynespora R.F. Castaneda & W.B. Kendr., typified by S. zapatensis

R.E Castaneda & W.B. Kendr., is similar to Morganjonesia in the monotretic

conidial ontogeny and the solitary, euseptate conidia (Hernandez-Restrepo &

al. 2014, Ma & al. 2016), but its protologue (Castaheda-Ruiz & Kendrick 1990)

did not describe a hyaline, acute or subulate conidial appendage.

Acknowledgments

This work was financed by the National Natural Science Foundation Program of

PR China (31870016). The authors express their sincere gratitude to Dr. Josiane S.

Monteiro and Dr. Flavia Rodrigues Barbosa for their critical review of the manuscript.

We acknowledge the facilities provided by Dr. P.M. Kirk and Dr. K. Bensch through the

Index Fungorum and MycoBank websites, respectively. Dr. Lorelei Norvell’s editorial

review and Dr. Shaun Pennycook’s nomenclature review are greatly appreciated.

Literature cited

Castafeda-Ruiz RF, Kendrick B. 1990. Conidial fungi from Cuba: II. University of Waterloo

Biology Series 33. 62 p.

Ellis MB. 1971. Dematiaceous hyphomycetes. Kew, Commonwealth Mycological Institute.

Hernandez-Restrepo M, Castafieda-Ruiz RF, Gené J, Silvera-Simén C, Cano J, Guarro J.

2013. Two new species of Solicorynespora from Spain. Mycological Progress 13: 157-164.

https://doi.org/10.1007/s11557-013-0903-9

Holubova-Jechova V, Castafeda Ruiz RE. 1986. Studies on Hyphomycetes from Cuba III. New

and interesting dematiaceous taxa from leaf litter. Ceska Mykologie 40: 74-85.

Ma J, Xia JW, Zhang XG, Luo YQ, Castafieda-Ruiz RF. 2016. Solicorynespora species associated

with dead branches of subtropical forests in southern China. Cryptogamie, Mycologie

37: 37-44. https://doi.org/10.7872/crym/v37.iss1.2016.37

Matsushima T. 1980. Saprophytic microfungi from Taiwan, part 1. Hyphomycetes. Matsushima

Mycological Memoirs 1. 82 p.

Morgan-Jones G. 1988. Notes on hyphomycetes. LX. Corynespora matuszakii, an undescribed

species with narrow, cylindrical, catenate conidia and highly reduced conidial cell lumina.

Mycotaxon 33: 483-487.

Seifert K, Morgan-Jones G, Gams W, Kendrick B. 2011. The genera of hyphomycetes. CBS

Biodiversity Series 9. 997 p.

MY COTAXON

July-September 2019—Volume 134, pp. 463-473

https://doi.org/10.5248/134.463

Botryosphaeria ginlingensis sp. nov.

causing oak frogeye leaf spot in China

LING-YU LIANG’, NING JIANG’, WEN-YAN CHEN’,

YING-MEI LIANG?, CHENG-MING TIAN"

"The Key Laboratory for Silviculture and Conservation of Ministry of Education &

? Museum of Beijing Forestry University,

'? Beijing Forestry University, Qinghua Eastern Road 35, Haidian District, Beijing, China

* CORRESPONDENCE TO: chengmt@bjfu.edu.cn

ABSTRACT — During our investigation on plant diseases in Qinling Mountain in China, an

oak disease with frogeye leaf spot symptoms was discovered. ‘The associated pathogen was

characterised by morphological and molecular approaches and is described here as a new

species, Botryosphaeria qinlingensis.

Key worps — Botryosphaeriaceae, Botryosphaeriales, taxonomy

Introduction

Botryosphaeria was firstly proposed by Cesati & De Notaris (1863) without

designating a type species. Subsequently, B. berengeriana (Hohnel 1909) and

B. quercuum (‘Theissen & Sydow 1915) were designated as the type, but neither

species was included in the original description of the genus. Barr (1972)

proposed B. dothidea (Moug. ex Fr.) Ces. & De Not. as the type species, because

it was one of the original species and an earlier synonym of B. berengeriana.

Botryosphaeria dothidea is now universally accepted as the type of the genus.

Slippers & al. (2004) provided a revised description of B. dothidea based on

the type specimen and fresh collections and designated a neotype and epitype.

Botryosphaeria was restricted to B. dothidea and B. corticis (Crous &al. 2006),

but subsequently, B. agaves, B. auasmontanum, B. fabicerciana, B. fusispora,

B. kuwatsukai, B. minutispermatia, B. pseudoramosa, B. gingyuanensis,

464 ... Liang & al.

B. quercus, B. ramosa, B. rosaceae, B. scharifii, B. sinensis, and B. wangensis

have been accepted in this genus (Abdollahzadeh & al. 2013; Ariyawansa & al.

2016; Chen & al. 2011; Li & al. 2018; Liu & al. 2012; Pavlic & al. 2008; Slippers

& al. 2014; Wijayawardene & al. 2016; Xu & al. 2015; Zhou & al. 2016, 2017).

Quercus is an important tree genus that contains many commercial taxa

widespread from temperate to subtropical broad-leaved forests in China,

such as Q. acutissima, Q. aliena, Q. aliena var. acutiserrata, Q. variabilis, and

Q. wutaishanica. A disease with frogeye leaf spot symptoms was discovered

on the leaf surface of Q. aliena var. acutiserrata and Q. acutissima (PLATE 1).

Similar symptoms have been recorded on Q. dentata, Q. acutissima, Q. aliena,

Q. aliena var. acutiserrata, and Q. variabilis, and the agents causing those

symptoms were identified as Macrophoma species (Tian & al. 1991). Here we

present an accurate identification based on combined morphology and DNA

sequence data, and describe the pathogen as a new species, Botryosphaeria

ginlingensis.

Materials & methods

Samples and isolates

In this study, diseased oak leaves were collected from Quercus aliena var.

acutiserrata and Q. acutissima in Shaanxi Province in China. Single conidial isolates

were obtained from conidiomata by removing a mucoid conidial mass from pycnidial

ostioles following Jiang & al. (2018). Specimens of the new species were deposited in

the Museum of Beijing Forestry University, Beijing, China (BJFC). Axenic cultures

were maintained in the China Forestry Culture Collection Center, Beijing, China

(CFCE).

Morphological studies

Species identification was based on conidial morphology and fruiting bodies were

observed according to Slippers & al. (2013). Fifty spores were selected randomly for

measurement using a Leica compound microscope (LM, DM 2500). Morphological or

cultural characteristics were observed on PDA after incubation at 25 °C and recorded

at 5 and 10 days, including colony color and texture.

DNA extraction, PCR amplification, and sequencing

Genomic DNA was extracted from fungal mycelium growing on MEA using a

modified CTAB method (Doyle & Doyle 1990). The internal transcribed spacer of

rDNA (ITS) was amplified using primers ITS4 and ITS5 (White & al. 1990). The

translation elongation factor-1 alpha (TEF1-a) was amplified using primers EF1-688F

and EF1-1251R (Alves & al. 2008). The B-tubulin gene (TUB) was amplified using

primers Bt2a and Bt2b (Glass & Donaldson 1995). These regions were amplified and

sequenced following Zhang & al. (2009). The PCR products were estimated visually by

Botryosphaeria qinlingensis sp. nov. (China) ... 465

PLATE 1. Botryosphaeria ginlingensis:

Frogeye disease symptoms on leaves of Quercus aliena var. acutiserrata.

electrophoresis in 2% agarose gel set to 60 V for 90 min. DNA was sequenced using

an ABI PRISM® 3730XL DNA Analyzer with BigDye® Terminater Kit v.3.1 at the

Shanghai Invitrogen Biological Technology Company Limited.

DNA sequence analysis

The new sequences generated in this study and the reference sequences of

Botryosphaeria isolates included in the phylogenetic analyses are summarized in

TABLE 1. Neofusicoccum luteum (CBS 110299) was used as outgroup (Li & al. 2018).

These sequences were aligned with MAFFT v. 7 (Katoh & Standley 2013) and manually

adjusted. Phylogenetic analyses were generated from the combined ITS-TEFla-TuB

sequences using PAUP v. 4.0b10 (Swofford 2003) for maximum parsimony (MP),

PhyML v. 3.0 (Guindon & al. 2010) for maximum likelihood (ML), and MrBayes

v.3.1.2 for Bayesian Inference (BI) (Ronquist & Huelsenbeck 2003) following Zhou &

al. (2017) and Li & al. (2018).

Results

Molecular phylogeny

The combined ITS, TEFl-a, and Tus alignment comprised 32 sequences

(including one outgroup) and 1344 characters including alignment gaps, of

which 1127 were parsimony informative, 95 were variable and parsimony-

466 ... Liang & al.

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uninformative, and 122 were constant. The MP analysis produced 50 equally

most parsimonious trees, with the first tree (TL = 261, CI = 0.893, RI = 0.905,

RC = 0.808) shown in PLaTE 2. The ML and BI phylogenetic trees were

topologically similar to the MP tree. The new species appeared in a single clade

with high bootstrap support (PLATE 2).

95/100/1

98/100/1

r =

92/95/0.99

CBS 122069 B. ramosa |

69/94/0.97) CGMCC3.18742 B. gingyuanensis

| CGMCC3.18743 B. qingyuanensis

100/100/1

87/100/0.99

70/-/-

-/59/-

sent at CBS 135219 B. kuwatsukai_

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72/62/- LY

64/99/0.95 — MFLUCC 10-0098 B. fusispora |

73/88/0.99

(CBS 990.70 Dichomera saubinetii

100/100/1 — .

194/99/1| GZCC 16-0013 B. minutispermatia

|GZCC 16-0014 B. minutispermatia

100/100/1

61/77/0.97 |) CEMCC3.18744 B. oe

oy _ CGMCC3.18745 B. wangensis

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| CGMCC3.18007 B. rosaceae

92/100/1| | CGMCC3.18008 B. rosaceae

|p CBS 133992 B. agaves”

97/99/1 || MFLUCC 10-0051 B. agaves

74/88/1

20.0

PLATE 2. Phylogram of Botryosphaeria based on combined ITS, tefl, and tub genes. Bootstrap

support values >50% and posterior probability values 20.95 are presented as MP/ML/PP, above

branches. Scale bar = 20 nucleotide substitutions.

Botryosphaeria qinlingensis sp. nov. (China) ... 469

PLATE 3. Botryosphaeria qinlingensis from Quercus aliena var. acutiserrata (BJFC-S1576, holotype).

A-D. Conidiomata; E. Transverse sections through conidiomata; F. Longitudinal section through

a conidioma; G. Conidiogenous cells and developing conidia; H. Conidia. Scale bars: A-C = 1 cm;

D, E= 2 mm; F-H = 10 um.

Taxonomy

Botryosphaeria qinlingensis C.M. Tian & L.Y. Liang, sp. nov. PLATE 3

MB 828967

Differs from Botryosphaeria wangensis and B. minutispermatia by its wider conidia.

Type: China, Shaanxi Prov., Ningshan County, Qinling Mountain, Huoditang forest

park, 33°26'07”N 108°26’48’E, 1570 ma.s.l, on leaves of Quercus aliena var. acutiserrata,

3 July 2018, coll. N. Jiang & C.M. Tian (Holotype, BJFC-S1576; ex-type culture CFCC

52984; GenBank MK434301, MK425020, MK425022).

Erymo ocy: ginlingensis, named after the Qinling Mountain.

470 ... Liang & al.

Sexual morph: Undetermined. Asexual morph: Conidiomata formed on

oak leaf spot, stromatic, brown to black, 120-250 um diam, forming a

botryose aggregate of up to 15 (sometimes solitary) globose with a central

ostiole, semi-immersed to superficial. Paraphyses, hyaline, aseptate, up to

35 um long, 3-5 um diam. at the base tapering to acutely rounded apices,

1-2 um diam. at the tip. Conidiophores absent. Conidiogenous cells

holoblastic, discrete, hyaline, cylindrical to lageniform, phialidic with

periclinal thickening, 4.5-10.5 x 1.5-3 um. Conidia hyaline, thin-walled,

smooth with granular contents, unicellular, aseptate, ellipsoid to fusoid, base

subtruncate to bluntly rounded, (14.3—)19.2-24.8(-30.0) x (6.1-)7.1- 8.4(-

9.5) um, L/W = (2.0-)2.4—3.3(-3.7) (n = 100). Colonies initially white with

fluffy, aerial mycelium, becoming black on the surface after 5 days; reverse

side of the colonies dark-brown.

ADDITIONAL SPECIMEN EXAMINED: CHINA, SHAANXI PRov., Ningshan County,

Qinling Mountain, Shibazhang village, 33°26’10”N 108°26’42”E, 1700 m a.s.l, on leaves

of Quercus acutissima, 7 July 2018, coll. N. Jiang & C.M. Tian (BJFC-S1577; living

culture CFCC 52985; GenBank MK434302, MK425021, MK425023).

Host/DistRIBUTION: on leaves of Quercus aliena var. acutiserrata and Q. acutissima in

China.

Note: The two isolates of B. ginlingensis clustered in a well-supported clade

(MP/ML/BI = 100/100/1) in PLaTE 2 and appeared closely related to B.

wangensis G.Q. Li & S.F. Chen and B. minutispermatia Ariyaw. & al. However,

B. ginlingensis can be morphologically distinguished from B. wangensis and

B. minutispermatia, which differ by their narrower conidia (B. wangensis,

5.5-6.5 um, Li & al. 2018; B. minutispermatia, 3-4 um, Ariyawansa & al.

2016).

Discussion

In this study, disease samples with typical frogeye leaf spot symptom were

observed and collected from Quercus aliena var. acutiserrata and Quercus

acutissima in Qinling Mountain in China. Based on DNA sequence data of

combined ITS, TEF1l-a, and Tus loci, Botryosphaeria qinlingensis is proposed

here as the pathogen causing oak frogeye leaf spot disease.

Although the sexual state of Botryosphaeria qinlingensis was absent, we can

still distinguish this pathogen from other Botryosphaeria species by conidial

dimension (TABLE 2), supplemented by host information. Another species,

Botryosphaeria quercus, was recorded on oak leaves collected from Italy and

described with only Dichomera-like conidia, but we can separate B. quercus and

B. qinlingensis in the phylogenetic tree (PLATE 2).

TABLE 2. Conidial size of accepted Botryosphaeria species.

SPECies

B. agaves

B. auasmontanum

B. corticis

B. dothidea

B. fabicerciana

B. fusispora

B. kuwatsukai

B. minutispermatia

B. pseudoramosa

B. gingyuanensis

B. qinlingensis

B. quercus

B. ramosa

B. rosaceae

B. scharifii

B. sinensis

B. wangensis

Acknowledgments

Botryosphaeria qinlingensis sp. nov. (China) ... 471

CONIDIAL SIZE (um)

(8.1-)8.8-11.3(-13) x

(2.5-)2.9-3.9(-5)

(20.5-)23.5-32.5(-34.5) x

(5.0-)5.5-7(-7.5)

(20-)23-27(-30) x

4—5(-6)

(16.5-)19.5-24.5(-26) x

(4.5—)5-6.5(-7.5)

16-22 x

4-5.5

(18.5-)20-24.5(—26) x

5-7(-8)

8-14 x

3-4

(8—)10-13(-16) x

(4-)4.5-5(-6)

(15-)19.5-24.5(-28.5) x

(5-)6-6.5(-7.5)

(14.3-)19.2-24.8(-30) x

(6.1-)7.1-8.4(-9.5)

10-24 x

6-10

(11-)12-15(-16) x

(4.7-)5-6(-7)

20-31 x

6-8

(11.5-)13-17(-19) x

4-6.5

(15-)19-29 x

5-7

(20.5-)22—26(-29) x

(4.5—)5.5-6.5(-7.5)

L/W

4.5

4.9

3.8

3.6

3.7.

2.5

3.5

2.4-3.3

2.3

3.9

27;

4.1

3.9

REFERENCE

Liu & al. 2012

Slippers & al. 2014

Phillips & al. 2006

Slippers & al. 2004

Chen & al. 2011

Liu & al. 2012

Xu & al. 2015

Ariyawansa & al. 2016

Li & al. 2018

This study

Wijayawardene & al. 2016

Pavlic & al. 2008

Zhou & al. 2017

Abdollahzadeh & al. 2013

Zhou & al. 2016

Li & al. 2018

This study was financed by National Natural Science Foundation of China (Project

31670647). We thank Drs Artur Alves (University of Aveiro, Portugal) and Quan Lu

(Chinese Academy of Forestry, Beijing) for expert presubmission review. We are also

grateful for the assistance of Chungen Piao and Minwei Guo (China Forestry Culture

Collection Center (CFCC) in Beijing).

472 ... Liang & al.

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MYCOTAXON

July-September 2019—Volume 134, pp. 475-480

https://doi.org/10.5248/134.475

Tretoheliocephala cylindrospora sp. nov.,

an asexual fungus from Thailand

CHARUWAN CHUASEEHARONNACHAI ”?', SAYANH SOMRITHIPOL ”,

KANTHAWUT BOONMEE 2, SALILAPORN NUANKAEW ”?,

NATTAWUT BOONYUEN ”?

"National Biobank of Thailand &

? National Center for Genetic Engineering and Biotechnology (BIOTEC):

'&2 National Science and Technology Development Agency (NSTDA),

113 Thailand Science Park, Thanon Phahonyothin, Tambon Khlong Nueng,

Amphoe Khlong Luang, Pathum Thani 12120, Thailand

° Master of Science Program in Biotechnology, College of Agricultural Innovation

and Biotechnology and Food, Faculty of Biotechnology, Rangsit University,

Pathum Thani 12000, Thailand

* CORRESPONDENCE TO: charuwan.chu@biotec.or.th

ABsTRACT—Tretoheliocephala cylindrospora is described and illustrated as a new species

collected from a decaying twig of an unidentified plant in Phu Laenkha National Park,

Chaiyaphum Province, northeastern Thailand. The new fungus, which is compared with

T. compacta, the type species of the previously monotypic genus, is characterized by

unbranched conidiophores that produce solitary, oblong or cylindrical, unicellular conidia.

Key worps—microfungi, morphology, plant debris, taxonomy

Introduction

Recent work on fungal biodiversity in Thailand has prompted an increase

in the mycobiota of the country with several new species (e.g., Boonyuen & al.

2016; Chuaseeharonnachai & al. 2013, 2014, 2016, 2017; Sri-indrasutdhi & al.

2010, 2015; Yamaguchi & al. 2012). During continuing surveys of microfungi

associated with plant materials in Thailand, a Periconia-like fungus was

collected on an unidentified decaying twig. The conidiogenesis and conidial

476 ... Chuaseeharonnachai & al.

morphology placed this fungus in the genus Tretoheliocephala Gusmao & al.,

but it demonstrated clear differences from the single Tretoheliocephala species

previously described (Gusmao et & al. 2017) and is therefore proposed as new

to science.

Materials & methods

Samples of decaying leaves and twigs were collected in October 2013 from Phu

Laenkha viewpoint (16°01’N 101°53’E) in Phu Laenkha National Park, Chaiyaphum

Province, Thailand. Samples were transported to the laboratory and washed with tap

water to remove soil particles and adhering debris. The samples were soaked with sterile

distilled water and incubated in moist chambers at room temperature (~20-25°C) for

at least 2 weeks. The fungus was isolated under an Olympus SZ61 stereomicroscope

using the single spore technique outlined by Goh (1999). Spores were transferred to

different media—cornmeal agar (CMA), malt extract agar (MEA), potato carrot agar

(PCA), potato dextrose agar (PDA), and water agar (WA) with added 50 ug/mL of two

antibiotics (penicillin and streptomycin)—as recommended by Choi & al. (1999) and

Kirk & al. (2008), but isolation was unsuccessful.

Fresh material was mounted in water and examined and measured using an Olympus

CX31 compound microscope equipped with a drawing tube. Microphotographs were

obtained with an Olympus DP70 equipped with Nomarski differential interference

optics. Permanent slides are deposited at the BIOTEC Bangkok Herbarium, Thailand

(BBH).

Taxonomy

Tretoheliocephala cylindrospora Chuaseehar., Somrith. & Boonyuen, sp. nov.

MB 828099 PLATES 1, 2

Differs from T. compacta (type of the monotypic genus) by its solitary, oblong or

cylindrical, unicellular conidia on unbranched conidiophores.

Type: Thailand, Chaiyaphum Province, Phu Laenkha viewpoint in Phu Laenkha

National Park, 16°01’N 101°53’E, on dead twig of an unidentified plant, 31 Oct. 2013, C.

Chuaseeharonnachai (Holotype, BBH 38817).

ErymMo.oey: The specific epithet refers to the cylindrical shape of the conidia.

Saprobic on dead twig. Asexual morph: CoLontzs on natural substratum

effuse, brown or black, hairy. MyceLtum mostly immersed in the substratum,

hyphae branched, septate, smooth-walled, pale brown to brown, 1.25-2.5 um

diam. CONIDIOPHORES macronematous, mononematous, unbranched,

capitate at the apex (rarely spathulate), cylindrical below, erect, sometimes

regenerating at the terminal end with one enteroblastic percurrent extension,

6-10-septate, smooth-walled, brown to dark brown toward the base, 100-227

um long, 3.75-5 um diam. at the apex, 7.5-10 diam. at the base (x = 180.16 x

Tretoheliocephala cylindrospora sp. nov. (Thailand) ... 477

& D

PLaTE 1. Tretoheliocephala cylindrospora (holotype, BBH 38817). A-E. Conidiophores with

conidiogenous cells and conidia (the arrow in B indicates regeneration point of the conidiophores);

F, G. Conidiogenous cells and conidia. H. Conidia (arrows indicate pore-like detachment scars at

the basal end). Scale bars: A, B = 15 um; C-E= 10 um; F-H = 5 um.

4.87 x 8.75 um; n = 25). CONIDIOGENOUS CELLS monotretic, discrete, globose,

determinate, terminal, brown to dark brown, smooth-walled, aggregating

within a compact system of globose to subglobose cells (2.5-5 um diam)

to form a capitulum with a diameter of 12.5-32.5 um. Conidial secession

schizolytic. Conip14 solitary, oblong or cylindrical with rounded apices,

unicellular, dry, brown, smooth-walled with a minute pore-like detachment

scar at the basal end, 7.5 x 3 um (n = 50).

Sexual Morph: Unknown.

ComMMENT— Tretoheliocephala was erected by Gusmao & al. (2017), typified by

its only species, T: compacta. The genus was characterized by macronematous,

erect dark brown conidiophores with a compact cluster of globose, monotretic

conidiogenous cells that produce solitary or blastocatenate, ovoid, oblong,

dolabriform, sub-obpyriform, or obcuneiform, 0-2-septate, brown conidia.

Our new species has features that fit well within the generic description of

Tretoheliocephala, forming stalked conidiophores with a more or less spherical

compact terminal cluster of globose and monotretic conidiogenous cells

producing oblong or cylindrical conidia.

Morphologically, T. cylindrospora is distinguished from T: compacta, which

has larger, unbranched conidiophores (320-700 x 15-19 um) or occasionally

478 ... Chuaseeharonnachai & al.

PLATE 2. Tretoheliocephala cylindrospora (holotype, BBH 38817).

Conidiophores, conidiogenous cells, and conidia. Scale bar = 20 um.

branched conidiophores (200-300 x 13-15 um), larger conidiogenous cells

(8-12 um diam), and larger morphologically variable, 0-2-septate (mostly

1-septate) conidia (12-21 x 6-7 tum) ranging from ovoid, oblong, dolabriform,

subobpyriform to obcuneiform, formed solitarily or blastocatenately (Gusmao

&al. 2017).

Tretoheliocephala cylindrospora sp. nov. (Thailand) ... 479

We regard T! cylindrospora as a rare species, in view of the fact that it has

been found only once since 2010 during our study of microfungi of Thailand.

Despite several attempts to isolate this fungus onto synthetic media, the new

fungus failed to grow on CMA, MEA, PCA, PDA and WA, suggesting that the

nutritional requirements in these media may not be enough for germination

and viability of the fungus. Thus, we were unable to undertake sequencing of

this fungus. Further collections are required to enable culture and sequencing

of Tretoheliocephala species to determine their taxonomic placement in the

Ascomycota.

Acknowledgments

This work was financially supported by National Center for Genetic Engineering

and Biotechnology (BIOTEC), National Science and Technology Development

Agency (NSTDA) with the grant number P-14—51395. Professor E.B. Gareth Jones is

thanked for reviewing, correcting, and improving the English of the draft manuscript.

The authors are deeply grateful to Prof. Dr. Akira Nakagiri (Fungus/Mushroom

Resource and Research Center, Tottori University, Japan) and Prof. Dr. R.E Castafeda-

Ruiz (INIFAT Alejandro de Humboldt, Habana, Cuba) for serving as reviewers and

providing valuable suggestions. Dr. Lorelei L. Norvell’s editorial review and Dr. Shaun

Pennycook’s nomenclature review are greatly appreciated.

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garethii sp. nov. (Fuscosporellales) from a rivulet in a community-based northern forest, in

Thailand. Mycosphere 7: 1265-1272. https://doi.org/10.5943/mycosphere/7/9/2

Choi YW, Hyde KD, Ho WH. 1999. Single spore isolation of fungi. Fungal Diversity 3: 29-38.

Chuaseeharonnachai C, Yamaguchi K, Sri-indrasutdhi V, Somrithipol $, Okane I, Nakagiri

A, Boonyuen N. 2013. Diversity of aero-aquatic hyphomycetes from six streams in Doi

Inthanon and Khao Yai tropical forests, Thailand. Cryptogamie, Mycologie 34: 183-197.

https://doi.org/10.7872/crym.v34.iss2.2013.183

Chuaseeharonnachai C, Somrithipol $, Boonyuen N. 2014. A new species of Fusticeps from

Thailand. Mycosphere 5: 313-317. https://doi.org/10.5943/mycosphere/5/2/5

Chuaseeharonnachai C, Somrithipol S, Boonyuen N. 2016. Periconia notabilis sp. nov.

and a new record and notes on the genus in Thailand. Mycotaxon 131: 491-502.

https://doi.org/10.5248/131.491

Chuaseeharonnachai C, Somrithipol S, Suetrong S, Klaysuban A, Pornputtapong N, Jones EBG,

Boonyuen N. 2017. Conioscypha nakagirii sp. nov., from naturally submerged wood in

Thailand based on nuclear ribosomal and protein-coding genes. Mycoscience 58: 424-431.

https://doi.org/10.1016/j.myc.2017.06.003

Goh TK. 1999. Single-spore isolation using a hand-made glass needle. Fungal Diversity 2: 47-63.

Gusmao LFP, Monteiro JS, Castafteda-Ruiz RF. 2017. Tretoheliocephala compacta gen. & sp. nov.

from the Brazilian semi-arid region. Mycotaxon 132: 453-458. https://doi.org/10.5248/132.453

Kirk PM, Cannon PF, Minter DW, Stalpers JA. 2008. Ainsworth & Bisby’s dictionary of the fungi.

10th edn. CAB International, Wallingford Oxon, UK.

480 ... Chuaseeharonnachai & al.

Sri-indrasutdhi V, Boonyuen N, Suetrong S, Chuaseeharonnachai C, Sivichai S, Jones EBG. 2010.

Wood-inhabiting freshwater fungi from Thailand: Ascothailandia grenadoidia gen. et sp. nov.,

Canalisporium grenadoidia sp. nov. with a key to Canalisporium species (Sordariomycetes

Ascomycota). Mycoscience 51: 411-420. https://doi.org/10.1007/S10267-010-0055-6

Sri-indrasutdhi V, Tsui CKM, Chuaseeharonnachai C, Yamaguchi K, Suetrong S, Okane I, Nakagiri

A, Boonyuen N. 2015. Helicocentralis hyalina gen. et sp. nov., an aero-aquatic helicosporous

fungus (Leotiomycetes, Ascomycota) in ‘Thailand. Mycological Progress 14:81 [12 p.].

https://doi.org/10.1007/s11557-015-1103-6

Yamaguchi K, Tsurumi Y, Suzuki R, Chuaseeharonnachai C, Sri-indrasutdhi V, Boonyuen N,

Okane I, Suzuki K, Nakagiri A. 2012. Trichoderma matsushimae and T: aeroaquaticum:

two aero-aquatic species with Pseudaegerita-like propagules. Mycologia 104: 1109-1120.

https://doi.org/10.3852/11-253

MY COTAXON

July-September 2019—Volume 134, pp. 481-488

https://doi.org/10.5248/134.481

Novel cyphelloid fungi in Glabrocyphella,

Heteroscypha, and Rectipilus from Brazil

LARISSA TRIERVEILER-PEREIRA’ , JULIANO M. BALTAZAR?’,

R. GREG THORN?, ADRIANA DE MELLO GUGLIOTTA’

'Nucleo de Pesquisa em Micologia, Instituto de Botanica (IBt),

Av. Miguel Stéfano, 3687, Agua Funda, CEP 04301-902, Sao Paulo, SP, Brazil

?Centro de Ciéncias da Natureza, Campus Lagoa do Sino, Universidade Federal de Sado Carlos,

Rodovia Lauri Sim6es de Barros (SP-189) Km 12, Buri, Sado Paulo, Brazil

° Department of Biology, University of Western Ontario, London, ON N6A 5B7, Canada

* CORRESPONDENCE TO: Lt_pereira@yahoo.com.br

ABSTRACT—New records of cyphelloid fungi from Brazil are presented, based on specimens

collected and identified by J. Rick and conserved at Herbarium PACA (Sao Leopoldo, Rio

Grande do Sul). Glabrocyphella cyathiformis sp. nov. is characterized by small (0.1-0.5 mm

diam.), pale yellow, turbinate basidiomes and ellipsoid to fusoid basidiospores (5-6 x 2-2.5

um). Flagelloscypha malmei is recombined as Heteroscypha malmei comb. nov., and Rectipilus

natalensis is reported for the first time from the Americas. Descriptions, photographs, and

illustrations are presented for the three species.

Key worps—Agaricomycetes, Auriculariales, Marasmiaceae, mycobiota, taxonomy

Introduction

The cyphelloid fungi are an artificial assemblage characterized by

basidiomes morphologically similar to the original concept of the genus

Cyphella Fr. The type species [currently accepted as Aleurodiscus digitalis (Alb.

& Schwein.) Donk] was originally described as a Peziza; indeed, cyphelloid

fungi can be characterized as discomycetes with basidia, according to Donk

(1966). One difference that can be noted in the field is that discomycetes have

their hymenium directed upward, while those of cyphelloid fungi are directed

downward, due to differences in the release of spores from asci and basidia.

482 ... Trierveiler-Pereira & al.

Cyphelloid fungi have cupulate, discoid, tubular to flattened basidiomes

and a smooth to more or less wrinkled hymenophore. Basidiomes are usually

less than 1 mm long and grow on litter (grass, leaves, twigs, and other small

substrates) and rotten wood. They are externally covered with specialized

hyphae, which serve as important characters for circumscribing genera (Donk

1959, Cooke 1961, Agerer 1983, Bodensteiner & al. 2004). According to

Agerer (1986), the outer hyphae form the most striking features of cyphelloid

basidiomes.

Initially, these fungi were grouped in three genera—Cyphella, Solenia, and

Porotheleum (Cooke 1961)—but currently more than 40 diverse genera are

recognized for the group (Bodensteiner 2007). Several new cyphelloid species

have been based on Brazilian collections (Spegazzini 1889; Hennings 1897,

1908; Rick 1906, 1931, 1959, 1960; Cooke 1961; Singer 1989; Gorjon & Jesus

2014) and a few reports have been published in the past decades (Putzke 2002;

Meijer 2006; Sulzbacher & al. 2008, 2009).

During a taxonomical review of cyphelloid type specimens described by

J. Rick from Southern Brazil (Trierveiler-Pereira & al. unpubl. data),

additional collections of cyphelloid fungi kept at Herbarium PACA (Sao

Leopoldo, Rio Grande do Sul, Brazil) were also examined. Here we present

novelties resulting from this research on cyphelloid fungi.

Materials & methods

Dried basidiomes were measured, photographed, and morphologically analyzed.

Colors of macrostructures were coded according to Kornerup & Wanscher (1978). For

microscopical analyses, basidiomes were hand-sectioned with the aid of a razor blade

under a stereoscopic microscope (Agerer 2002). Sections were mounted in distilled

water and 3% KOH or 3% KOH with 1% aqueous phloxine to observe and measure

the microstructures. The coloring of pigmented elements (e.g., external hyphae and

spores) was evaluated in distilled water (Bodensteiner 2007). Amyloid and dextrinoid

reactions were tested with Melzer’s iodine reagent. Dimensions recorded correspond

to the hyphal diameter without crystal covering, spore length without apiculus and

basidia without sterigmata.

Taxonomy

Glabrocyphella cyathiformis Trierv.-Per. & Thorn, sp. nov. FIG. 1

MB 828713

Differs from Glabrocyphella palmarum by its smaller, turbinate basidiomes and

its smaller, subfusiform basidiospores; and from G. rubescens by its grayish yellow

basidiomes and its narrower, subfusiform basidiospores.

Glabrocyphella cyathiformis sp. nov. (Brazil) ... 483

Fic 1. Glabrocyphella cyathiformis (PACA 20938, holotype). A. Basidiomes on rotten wood; B, C.

Basidiomes in detail; D. Basidiospores. Scale bars: A = 1 mm; B = 0.5 mm; C = 0.3 mm; D = 5 um.

Type: Brazil, Rio Grande do Sul, Salvador do Sul, 19.[X.1944, J. Rick (Holotype, PACA

20938, as Solenia sp.)

ErymMo.oey: Latin, cyathiformis: cup-shaped—slightly wider at the top than at the

bottom, referring to the shape of the basidiomes.

BASIDIOMEs Clustered in small groups that are in contact, forming a reticulate

pattern (rarely scattered), short-cylindrical (when young) to turbinate, 0.1-0.5

mm diam. x 0.3-0.5 mm high; cup margins curving inwards; externally grayish

yellow (3B4) to light yellowish (4A4), velvety; hymenium smooth, pastel yellow

(1A4). Subiculum absent. Growing on a small piece of rotten wood.

EXTERNAL HYPHAE adpressed to the surface, some projecting ca 40-100 um

from the basidiome, pale yellow, thick-walled, tortuous, 1-4 um diam., fibulate,

non-dextrinoid and non-amyloid, lacking encrustations or distinctive features,

apices round. SUBHYMENIAL LAYER 10-25 um thick; hyphae agglutinated,

hyaline, ca 2 um diam., fibulate. HyMENIAL LAYER 10-15 um; basidia not

seen; basidioles tightly packed together, clavate to short-clavate, hyaline,

9-15 x 4-5 um, with clamp connections at the base; sterile elements not

seen. BAsip1ospores subfusiform, hyaline, smooth, thin-walled, (4.8-)5-6 x

2-2.5 um, with a distinct apiculus, non-dextrinoid, non-amyloid.

ECOLOGY & DISTRIBUTION: growing on rotten wood; known only from the

type locality.

ADDITIONAL SPECIMENS EXAMINED:

Glabrocyphella rubescens: BRAZIL, R10 GRANDE DO SUL: Sao Leopoldo, 1932, J. Rick

(PACA 15276, isotype, labeled ‘Solenia rubescens Rick’).

Glabrocyphella palmarum: BRAZIL, R10 GRANDE DO SUL: Sao Leopoldo, 1931, J. Rick

(PACA 12863, as Cyphella palmarum).

484 ... Trierveiler-Pereira & al.

REMARKS—Glabrocyphella W.B. Cooke included 12 species when introduced

by Cooke (1961), and a 13th species, G. stercoraria Barrasa & al., has since been

proposed.

Glabrocyphella cyathiformis is characterized by turbinate, yellowish

basidiomes velvety on the exterior with external hyphae lacking encrustations

or any other distinctive feature and without a subiculum and ellipsoid to fusoid

hyaline basidiospores [(4.8-)5-6 x 2-2.5 um].

Glabrocyphella cyathiformis is morphologically close to G. palmarum (Berk.

& M.A. Curtis) W.B. Cooke, which differs by its ovate to tear-shaped, larger (6-7

x 3-4 um) basidiospores (Cooke 1961) and larger (<2 mm high) basidiomes

(Burt 1914, as Cyphella palmarum). From our examination, Rick's PACA 12863

collection of G. palmarum has basidiospores (7.5-)8-9 x 2-3 um and cupulate-

stipitate basidiomes.

Glabrocyphella rubescens Rick ex W.B. Cooke was described based on

specimens collected in Southern Brazil. This species is distinguished from

G. cyathiformis by its cream-colored basidiomes that turn flesh-color upon

bruising and its ovoid basidiospores (4.5 x 3.5 um, Cooke 1961; 5-6 x 3-4 um,

from our examination of the isotype).

Heteroscypha malmei (WB. Cooke) Trierv.-Per., Baltazar & Thorn, comb. nov.

MB 828714 Fic. 2

= Flagelloscypha malmei W.B. Cooke, Beih. Sydowia 4: 62, 1961.

BASIDIOMES scattered or mainly in clusters of 2-30 basidiomes, 0.2-0.9 mm

diam., cupuliform, external surface yellowish white to pale yellow (1A3, 1A2),

tomentose; cup margins hairy, curving inwards; external hairs ca 0.1 mm long;

hymenial surface pastel yellow (1A4), smooth. Subiculum absent. Growing on

dead palm tree (peduncular bract).

EXTERNAL HYPHAE (hairs) 100-140 x 2-4 um, hyaline to pale yellow, more

pigmented towards the base, thick-walled with narrow lumen, non-dextrinoid

and non-amyloid, encrusted with acicular <4 um long crystals; apical end

flagelliform, smooth, 20-40 um long. SUBHYMENIAL LAYER ca 15 um thick;

hyphae agglutinated, hyaline, ca 2 um diam. HYMENIAL LAYER ca 25 um; basidia

short clavate, hyaline, 15-22 x 4-6 um, with clamp connection at the base,

4-sterigmate, longitudinally septate at the apex; sterile elements (leptocystidia)

observed in the hymenial layer, especially near the margins, tortuous, hyaline,

smooth, ca 40 x 4 um. Basip1osporEs fusoid to asymmetrical-ellipsoid,

hyaline, smooth, thin-walled, (7.5-)8-10 x (3.5-)4.0-4.5 um, Q = 2.2 (n = 13),

with a distinct apiculus, some germinating with hyphae, non-dextrinoid, non-

amyloid; some adhering in groups of 3-4.

Glabrocyphella cyathiformis sp. nov. (Brazil) ... 485

Fic 2. Heteroscypha malmei (PACA 12865). A. Basidiomes on rotten wood; B. Basidiospores;

C. Basidioles and basidia; D. External hyphae; E. Hymenial sterile elements. Scale bars: A = 1 mm;

B-E = 10 um.

SPECIMEN EXAMINED: BRAZIL. R10 GRANDE DO SUL: Salvador do Sul, 1943, J. Rick

(PACA 12865, as Cyphella palmarum).

ECOLOGY & DISTRIBUTION: growing on rotten wood; known only from

Southern Brazil, State of Rio Grande do Sul, from the municipalities of Santo

Angelo (holotype of Flagelloscypha malmei; Cooke 1961) and Sao Salvador

(this study).

REMARKS—In the examined material, the hymenial layer of most of the

basidiomes has been eaten by insects and only the external layer and hairs

remain, therefore, few basidia and basidiospores were observed.

Heteroscypha Oberw. & Agerer was proposed as a monotypic genus, typified

by Cyphella applanata [=H. applanata (P.H.B. Talbot) Oberw. & Agerer]; detailed

illustrations and description of the type specimen were presented, showing the

apically septate basidia (Agerer & Oberwinkler 1979). Heteroscypha malmei

was previously considered a member of Flagelloscypha Donk, since the external

hairs have acicular crystal encrustations, with the exception of the flagelliform

apical end (Cooke 1961).

Our Brazilian specimen of H. malmei has slightly longer basidiospores with

a Q ratio higher compared with those previously described for F malmei (8-9 x

4-5 um, Cooke 1961; 7.5-8.5 x 4-5 um, Q = 1.7, Agerer & Oberwinkler 1979).

Heteroscypha applanata differs from H. malmei by its white, discoid-petaliform

basidiomes, its thin-walled external hairs, its fully septate basidia, and

A486 ... Trierveiler-Pereira & al.

its longer basidiospores (9-12.6 x 3.2-5.4 um, Cooke 1961; 8-10 x 4-5 um,

Q = 2.1, Agerer & Oberwinkler 1979).

Rick’s PACA 12865 collection of Heteroscypha malmei was labelled Cyphella

palmarum; however Glabrocyphella palmarum differs from H. malmei by its

holobasidia, its larger (<3 mm diam.) hairless cups, and its smaller basidiospores

(Cooke 1961).

As indicated by Agerer & Oberwinkler (1979), additional fresh materials

of these species suitable for cultural, ultrastructural, and molecular studies are

highly desirable.

Rectipilus natalensis (W.B. Cooke) Agerer, Persoonia 7(3): 425, 1973. FIG. 3

BASIDIOMES agglutinated, tubuliform, long, 0.5-1.0 mm high; 0.1-0.3 mm

diam.; externally light orange (5A5) to grayish yellow (4B4) covered by

scattered whitish hyphae, pale yellow near the margins (2A3); margins hairy;

hymenium light orange (5A4, 5A5), smooth. Subiculum absent. Growing on

small pieces of wood.

EXTERNAL HYPHAE compact, closely adpressed to the surface of the

basidiome, pale yellow, thick-walled, not encrusted, 2-3 um diam., clamp

connections present, hyphal tips coiled and twisted near the margins, non-

dextrinoid and non-amyloid. SUBHYMENIAL LAYER 10-20 um thick; hyphae

agglutinated, pale yellow, ca 2 um diam. HYMENIAL LAYER 15-20 um; basidioles

yellowish, basidia clavate, 10 x 4 um, 4-sterigmata, with clamp connection at

the base; sterile elements not seen. Bastp1ospores globose to subglobose,

hyaline, smooth, 3-4(-4.5) x 2.2-3 um, non-dextrinoid, non-amyloid.

SPECIMEN EXAMINED: BRAZIL. R10 GRANDE DO SUL: Salvador do Sul, 9.IX.1944,

J. Rick (PACA 22683, as Solenia sp.).

ECOLOGY & DISTRIBUTION: growing on rotten wood; known from Southern

Africa (Talbot 1956, as Solenia natalensis) and Southern Brazil (this study).

REMARKS-Rectipilus Agerer contains eleven species (Lucas & Dentinger 2015).

Gorjon & Jesus (2014) provided an identification key to nine of these species.

Rectipilus natalensis is characterized by cylindrical, tubuliform basidiomes

with external hairs from the upper portions with spirally coiled apices, the lack

of subiculum, and small, globose to subglobose, hyaline basidiospores. The

original basionym description (Talbot 1956, as Solenia natalensis) indicates

basidiomes up to 2 mm long, but the Brazilian basidiomes are shorter (up

to 1 mm long). Agerer (1973) described the holotype as yellowish, while the

Brazilian material has pale orange external surface and is yellowish only near

the upper part of the basidiome.

Glabrocyphella cyathiformis sp. nov. (Brazil) ... 487

Fic 3. Rectipilus natalensis (PACA 22683). A. Basidiomes on rotten wood; B. Basidiomes in detail;

C. External hyphal tips; D. Basidiospores. Scale bars: A= 1 mm; B= 0.5 mm; C=5 um; D = 10 um.

Agerer (1973) stated that R. confertus (Burt) Agerer and R. fasciculatus

(Pers.) Agerer are similar to R. natalensis, but they lack external hyphae with

twisted or coiled apices, and they present different basidiospore morphologies.

The only Rectipilus species previously reported from Brazil, R. stromatoides

Gorjon, differs from R. natalensis by its basidiomata, which are arranged in a

dense, broadly effused, vivid pink stroma (Gorjon & Jesus 2014).

Acknowledgments

The authors are grateful to Dra. Maria Salete Marchioretto (curator of PACA) for

sending specimens. We sincerely appreciate all valuable comments and suggestions

made by the peer-reviewers: Marcelo A. Sulzbacher (Universidade Federal de

Santa Maria, Brazil), Sergio P. Gorjon (University of Salamanca, Spain), and Mario

Rajchenberg (Centro Forestal CIEFAP, Chubut, Argentina). This study is a partial

result from the post-doctoral research of the first author, with a scholarship provided

by the Brazilian government (CNPq/PDJ, proceeding #150233/2017-5).

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Spegazzini C.L. 1889. Fungi Puiggariani. Pugillus 1. Boletin de la Academia Nacional de Ciencias

en Cérdoba 11(4): 381-622.

Sulzbacher MA, Desjardin DE, Putzke, J. 2008. Calathella columbiana (Basidiomycota): new record

of a cyphelloid fungus from Brazil. Mycotaxon 105: 37-42.

Sulzbacher MA, Coelho G, Cortez VG. 2009. Studies on cyphelloid fungi in Southern Brazil. The

genus Phaeosolenia Speg. Boletin de la Sociedad Micolégica de Madrid 33: 91-96.

Talbot PHB. 1956. The cyphelloid fungi from South Africa. Bothalia 6: 465-487.

MY COTAXON

July-September 2019—Volume 134, pp. 489-516

https://doi.org/10.5248/134.489

New species of Marthamyces and Ramomarthamyces gen. nov.

from New Zealand and the Cook Islands

PETER R. JOHNSTON & DUCKCHUL PARK

Manaaki Whenua - Landcare Research,

Private Bag 92170, Auckland 1142, New Zealand

CORRESPONDENCE TO: johnstonp@landcareresearch.co.nz

ABSTRACT—A leaf-spotting fungus common on Phormium tenax in New Zealand is

described here as Marthamyces harakeke sp. nov. The phylogenetic analysis prepared for

the description of this new species showed Marthamyces to be polyphyletic. To resolve this,

three Marthamyces species from Australia and New Zealand, M. barbatus, M. dracophylli,

and M. gilvus, are recombined in the new genus Ramomarthamyces. Morphologically the

Ramomarthamyces species differ from Marthamyces in having paraphyses distinctly branched,

rather than propoloid. A fungus common on recently fallen leaves of Metrosideros spp. in

New Zealand has been previously referred to as Marthamyces emarginatus, but is recognised

here as a new species, Marthamyces metrosideri. In addition, two new Marthamyces species,

M. maccormackii on Metrosideros collina, and M. renga on Metrosideros collina, Vaccinium

cereum, and Weinmannia samoensis, are described from the Cook Islands, and a new

Ramomarthamyces species, R. tuku on Juncus sp., is described from New Zealand. Finally,

Naemacyclus culmigenus is recombined in Marthamyces.

Key worps—fungi, Leotiomycetes, Marthamycetaceae, New Zealand flax, pathogen

Introduction

A leaf-spotting fungus common on Phormium tenax J.R. Forst. & G. Forst.

(Asphodelaceae) in New Zealand has been tentatively referred to as Propolis

sp. in the literature (McKenzie & al. 1999) and on the NZFungi website

(https://nzfungi2.landcareresearch.co.nz). Macroscopically the fungus

recalls Propolis and Marthamyces in having ascomata immersed in host

tissue, opening by several irregular slits, and the hymenial surface having

490 ... Johnston & Park

a crystalline appearance when the flaps of covering host tissue fold back.

It is distinguished from most Marthamyces species in having bifusiform

to trifusiform ascospores and in being pathogenic, but neither of these

features is unique in the genus. Ascospore shape is variable in these fungi;

e.g., Marthamyces desmoschoeni and M. dendrobii have swollen areas along

their filiform ascospores. A pathogen of Miscanthus (Poaceae), identified as

Naemacyclus culmigenus by Hosoya & al. (2013), is morphologically typical

of Marthamyces, and also matches the genus Marthamyces phylogenetically,

based on the molecular phylogenetic analysis provided in this paper.

DNA sequencing of other Marthamyces specimens from New Zealand,

the Cook Islands, and Australia revealed several other taxonomic issues.

Species placed in Marthamyces by Johnston (1986, 2006) separate into two

distinct clades, one of which contains the type species of Marthamyces,

M. emarginatus. The clade that does not contain M. emarginatus is

segregated here into a newly described genus, Ramomarthamyces. Three

new Marthamyces species are proposed, one for specimens collected

from Metrosideros excelsa Sol. ex Gaertn., M. fulgens Sol. ex Gaertn.,

M. kermadecensis W.R.B. Oliv., M. robusta A. Cunn., and M. umbellata

Cav. (Myrtaceae) in New Zealand, a second from Metrosideros collina

(VJ.R. Forst. & G. Forst.) A. Gray on the Cook Islands, and a third from

M. collina, Vaccinium cereum G. Forst. (Ericaceae) and Weinmannia

samoensis A. Gray (Cunoniaceae) on the Cook Islands. A new

Ramomarthamyces sp. is described from Juncus in New Zealand.

Materials & methods

The Marthamyces specimens studied were collected from native forests in New

Zealand, the Cook Islands, and Australia. Those maintained in culture were grown

from ascospores germinating on water agar plates at the time the specimens were

collected. The fungarium specimens are deposited in PDD, and the living cultures

in the ICMP culture collection, with both collections maintained by Manaaki

Whenua - Landcare Research, Auckland (https://scd.landcareresearch.co.nz/).

DNA extraction, amplification, and sequencing followed the methods of

Johnston & Park (2005). Amplification primers were ITSIF and ITS4 for ITS

(White & al. 1990, Gardes & Bruns 1993), LROR and LR5 for LSU (Bunyard &

al. 1994, Vilgalys & Hester 1990), NS1 and NS4 for SSU (White & al. 1990), and

mrSSU1 and mrSSU3R for mtSSU (Zoller & al. 1999).

DNA sequences from the newly collected specimens were added to those

available for SSU, ITS, LSU, and mtSSU from the Leotiomycetes taxa treated in a

class-wide, 15-gene phylogeny (Johnston & al. 2019) that were not members of the

order Helotiales. Each gene was aligned using MAFFT (Katoh & Standley 2013) as

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Marthamyces spp. nov. & Ramomarthamyces gen. nov. (New Zealand) ... 493

implemented in Geneious 10, the ends were manually trimmed and introns were

removed as necessary, and then the alignments were concatenated. Maximum

likelihood analyses were run with IQ-TREE (Nguyen & al. 2015, Chernomor

& al. 2016) using models selected by ModelFinder using the BIC criterion

(Kalyaanamoorthy 2017) for each partitioned gene; branch support was estimated

using ultrafast bootstrap analysis with 1000 replicates (Hoang & al. 2018). Xylaria

hypoxylon and Neurospora crassa were used as outgroup. We also performed an

ITS analysis, which included additional specimens of each species as well as DNA

sequences from two GenBank accessions of Marthamyces specimens identified

incorrectly as Coccomyces proteae. The ITS analysis used the same methods as

the multi-gene analysis but included only the Marthamycetaceae specimens

together with the Chaetomellaceae specimens treated by Johnston & al. (2019),

Chaetomellaceae being sister to Marthamycetaceae in the Fic. 1 multigene analysis.

The Marthamycetaceae taxa and genes included in the phylogenetic analyses are

provided in TABLE 1.

Phylogeny

The higher-level taxa resolved with our four-locus analysis (SSU, ITS, LSU,

and mtSSU; Fic. 1) matched those in the more data-rich analysis of Johnston

& al. (2019). The monophyletic Marthamycetaceae included Mellitiosporium,

Propolis, Cyclaneusma, Naemacyclus, and Marthamyces. Species previously

named as Marthamyces gilvus, M. barbatus, and M. dracophylli formed a

clade separate from the clade containing the type species of Marthamyces,

and these species are recombined below in a new genus, Ramomarthamyces.

A specimen identified by Hosoya & al. (2013) as Naemacyclus culmigenus

is strongly supported as a member of the Marthamyces sensu stricto clade,

and this species is recombined in Marthamyces below. Two Marthamyces

emarginatus-like specimens collected from Eucalyptus leaves in Australia

are phylogenetically distinct from each other, and from M. emarginatus as

accepted here, but are not formally named as new species because each is

known from only a single specimen. The M. emarginatus-like species from

Metrosideros in New Zealand is recognised as a new species, described

below as M. metrosideri.

The ITS gene tree (Fic. 2) includes additional specimens of several of

the accepted species. It also includes three accessions in GenBank deposited

as Coccomyces proteae that are misidentifications of Marthamyces spp.

The position of Naemacyclus fimbriatus differs between the two trees;

clarification of its relationship requires additional genes or the sampling of

additional species within Naemacyclus.

494 ... Johnston & Park

Marthamyces emarginatus |CMP 22854

Marthamyces desmoschoeni ICMP 17350

Marthamyces quadrifidus ICMP 18329

Marthamyces metrosiden ICMP 17398

Marthamyces harakeke PDD 108765

e Marthamyces cf. emarginatus ICMP 22621

Marthamyces cf. emarginatus ICMP 22628

"i Marthamyces culmigenus TNS F-41728

91 L Marthamyces maccormackii ICMP 16155

Marthamyces renga ICMP 15830 :

Z Naemacyclus fimbriatus CBS 289.61 8

oe Ramomarthamyces gilvus ICMP 22855 E

1 Ramomarthamyces tuku ICMP 22562 Ss

2 = Ramomarthamyces barbatus ICMP 22853

Ramomarthamyces dracophylli |CMP 17381

kets Cyclaneusma minus CBS 496.73

Cyclaneusma minus ICMP 17358

ico Propolis farinosa ICMP 17354

100 Propolis versicolor ILLS 60497

Mellitiosporium versicolor Lantz357

Chaetomellaceae

Rhytismatales

Phacidiales

Thelebolales

“Tympanidaceae’” clade 2

Tympanidaceae sensu stricto

Mniaeciaceae

Leotiales

Xylaria hypoxylon AFTOL Outgroup Sordariomycetes

Neurospora crassa Broad Genome Outgroup Sordariomycetes

Ficure 1. ML tree based on concatenated SSU, ITS, LSU, and mtSSU sequences, with bootstrap

values shown when >90%. Details of the specimens of Marthamycetaceae included in the analysis

are provided in TaBLE 1; the extra-limital taxa in other families and orders are from Johnston et al.

(2019) and their data are available though the Manaaki Whenua - Landcare Research Datastore

(https://doi.org/10.7931/T5Y V-BE95).

Marthamyces spp. nov. & Ramomarthamyces gen. nov. (New Zealand) ... 495

MK599209 Marthamyces emarginatus ICMP 22622

MH921869 Marthamyces emarginatus ICMP 22854

MK599210 Marthamyces emarginatus ICMP 22623

) JN712451 “Coccomyces proteae” CBS 111703

JN712450 “Coccomyces proteae” CBS 111704

KJ606679 Marthamyces desmoschoeni ICMP 17350

MK599211 Marthamyces cf. emarginatus ICMP 22628

MK599218 Marthamyces metrosideri |CMP 22625

MK599220 Marthamyces metrosideri ICMP 22624

MK599219 Marthamyces metrosideri PDD 111232

98 _ MH682227 Marthamyces metrosideri ICMP 17398

MK599212 Marthamyces cf. emarginatus ICMP 22621

MK599213 Marthamyces harakeke PDD 108765

MK599214 Marthamyces quadrifidus ICMP 18329

MK599215 Marthamyces maccormackii ICMP 16155

100_| MK599217 Marthamyces maccormackii ICMP 15829

MK599216 Marthamyces maccormackii ICMP 15831

AB745435 Marthamyces culmigenus TNS-F 41728

97 ) MK599221 Marthamyces renga ICMP 15830

97 MK599222 Marthamyces renga ICMP 15827

JX089385 “Coccomyces proteae” CR252M

MH921868 Ramomarthamyces barbatus ICMP 22853

100 _}* MK599223 Ramomarthamyces barbatus ICMP 22620

MK599224 Ramoarthamyces barbatus ICMP 22627

MH921870 Ramomarthamyces gilvus ICMP 22855

99 108 MK599225 Ramomarthamyces gilvus ICMP 22626

MK599226 Ramomarthamyces tuku ICMP 22562

MH682228 Ramomarthamyces dracophylli ICMP 17381

MH860208 Naemacyclus fimbriatus CBS 440.71

98 KJ606680 Cyclaneusma minus ICMP 17358

NR_153910 Cyclaneusma minus CBS 496.73

100 MH682229 Propolis farinosa ICMP 17354

JQ256425 Propolis versicolor ILLS 60497

100 KF661532 Zoelineria rosarum PDD 102789

99 MH861115 Chaetomella oblonga CBS 110.78

98 NR_119500 Pilidium acerinum CBS 736.68

AY 487097 Pilidium concavum BPI 1107274

KY462816 Xeropilidium dennisii TU 104501

Marthamycetaceae

100

Chaetomellaceae

FiGurE 2. ML tree based on ITS sequences with bootstrap values shown when >90%. Details of the

specimens of Marthamycetaceae included in the analysis are provided in TaBLe 1. Members of the

Chaetomellaceae, sister to Marthamycetaceae in the multi-gene analysis in Fic. 1, are used as the

outgroup; data for the outgroup specimens are available through the Manaaki Whenua - Landcare

Research Datastore (https://doi.org/10.7931/T5Y V-BE95).

496 ... Johnston & Park

Taxonomy

Marthamyces culmigenus (Ellis & Everh.) PR. Johnst., comb. nov.

IF 556322

= Naemacyclus culmigenus Ellis & Everh., Proc. Acad. Nat. Sci. Philadelphia 45: 151, 1893.

Notes: The combination is based on both DNA sequence and morphological

data. The description of Naemacyclus culmigenus from Hosoya & al. (2013) cites

the morphological features typical of Marthamyces with respect to ascomatal

wall structure, the 0-1-septate filiform ascospores, and propoloid paraphyses.

Naemacyclus fimbriatus, a heterotypic synonym of N. pinastri, the type species

of Naemacyclus, differs in having unusually long periphysoids lining the upper

wall of the opened ascoma, and also differs in having multi-septate ascospores

and paraphyses simple at the apex (DiCosmo 1979). A specimen identified as

N. fimbriatus (AFTOL ID-1295, CBS 289.61; Fic. 1) is phylogenetically distinct

from the specimen accepted as N. culmigenus by Hosoya & al. (2013).

Marthamyces emarginatus (Cooke & Massee) Minter, Mycotaxon 87: 51, 2003.

Fic. 3

= Stictis emarginata Cooke & Massee, Grevillea 18: 7, 1889.

= Propolis emarginata (Cooke & Massee) Sherwood, Mycotaxon 5: 323, 1977.

SPECIMENS EXAMINED—AUSTRALIA: Victoria: on Eucalyptus sp., Martin 439 (K

- holotype of Stictis emarginata). TASMANIA: Mt Wellington, on Eucalyptus sp., 4 Oct

1997, coll. PR. Johnston AU97-7 (PDD 81847, duplicate HO, culture ICMP 22621);

Cradle Mountain Lake St Clair National Park, Cynthia Bay-Shadow Lake Track, near

Watersmeet, on Eucalyptus sp., 7 Oct 1997, coll. PR. Johnston AU97-64 (PDD 81844,

duplicate HO, culture ICMP 22623); Cradle Mountain Lake St Clair National Park,

Cynthia Bay—Echo Point Track, on Eucalyptus sp., 8 Oct 1997, coll. PR. Johnston AU97-

84 (PDD 81845, duplicate HO, culture ICMP 22622); Zeehan-Wynyard road, Hellyer

Gorge State Reserve, on Eucalyptus sp., 12 Oct 1997, coll. PR. Johnston AU97-123 (PDD

81846, culture ICMP 22854). NEw SouTH WALEs: New England National Park, Point

Lookout, Lyrebird Walk, on Eucalyptus sp., 31 May 1996, coll. P.R. Johnston AU96-211-2

(PDD 81848, culture ICMP 22628).

Notes: Originally described from fallen Eucalyptus leaves from Australia, this

species has been reported from Eucalyptus from the Canary Islands, North

America, and South Africa (Sherwood 1977, Crous & van der Linde 1993). It

was first reported from New Zealand by Johnston (1986), but on Metrosideros

rather than Eucalyptus, and it has not subsequently been reported on

Eucalyptus from New Zealand (Dick 1990, https://nzfungi2.landcareresearch.

co.nz, accessed 10 October 2018). Here we accept M. emarginatus as a specialist

of Eucalyptus, and the Metrosideros-inhabiting fungus from New Zealand is

described below as a new species.

Marthamyces spp. nov. & Ramomarthamyces gen. nov. (New Zealand) ... 497

FiGuRE 3. Marthamyces emarginatus (A. PDD 81846; B, C. PDD 81845). A. Macroscopic appearance

when dry; B. Asci with rostrate apex; C. Propoloid paraphyses. Scale bars: A = 1 mm; B, C = 10 um.

The Marthamyces specimens examined from fallen Eucalyptus leaves from

Australia are similar in macro- and micro-morphology, but phylogenetically

they form three groups, each probably representing a distinct species. PDD

81844 (ICMP 22623), PDD 81845 (ICMP 22622), and PDD 81846 (ICMP

22854) have almost identical ITS sequences and are morphologically more

or less indistinguishable. PDD 81848 (ICMP 22628) and the three-specimen

498 ... Johnston & Park

Figure 4. Marthamyces cf. emarginatus (A, B. PDD 81848; C, D. PDD 81847). A. Macroscopic

appearance when dry; B. Apex of paraphyses with several long apical branches; C. Macroscopic

appearance when dry; D. Paraphyses broad, sometimes with single narrow branch at apex. Scale

bars: A, C = 0.5 mm; B, D = 10 um.

Marthamyces spp. nov. & Ramomarthamyces gen. nov. (New Zealand) ... 499

group cited above share 92% identical sites across the ITS region; PDD 81848

differs morphologically in having longer, finger-like extensions at the ends

of the propoloid paraphyses (Fic. 4A, B). PDD 81847 (ICMP 22621) and the

three-specimen group cited above share 96% identical sites across the ITS

region; PDD 81847 differs morphologically in having darker ascomata and

paraphyses that are broader near the apex and typically with a single, finger-

like projection (Fic. 4C, D). Here we provisionally accept the three matching

specimens PDD 81844, PDD 81845, and PDD 81846 as M. emarginatus. No

epitype is proposed because no recent specimens are available from Victoria,

the type locality of M. emarginatus.

Marthamyces harakeke PR. Johnst., sp. nov. Fics 5, 6

IF 556323

Differs from Marthamyces emarginatus by its Phormium host preference, by its fruiting

on living leaves, by its ascomata being elongate, and by its broader, bifusiform to

trifusiform ascospores.

Type: New Zealand: Auckland: Waitakere Ranges, Kaitarakihi Beach, on Phormium

tenax living leaves, 20 July 1983, coll. PR. Johnston R328 (holotype PDD 112252).

Etymo.oey: harakeke, Maori name for the host.

Ascomata and conidiomata developing on living leaves within reddish,

elliptic to fusiform, approximately 10-30 x 3-15 mm lesions. Ascomata

more or less cylindric, 0.6-2 x 0.3-0.5 mm, concolorous with host tissue,

immersed, initially with covering host tissue becoming slightly raised,

then the covering host tissue splitting irregularly, the flaps of tissue folding

back to expose the hymenial surface, which has a pale yellow, crystalline

appearance. Undersides of flaps of host tissue also with crystals. Ascomata

subepidermal, in vertical section upper wall up to 40 um thick, comprising

several layers of non-gelatinous tangled hyphae about 4—5 um diam., with

cell walls thin, pale brown, lined with a layer of periphysoids, cylindric,

unbranched, 15-20 x 3-6 um, hyaline. Periphysoids downward projecting

before ascomata open, inward projecting after ascomata open, partly lost

after opening. Lower wall and subhymenium 15-20 um thick, lower wall

comprising tangled hyphae 2-3 um diam., with cell walls thin, pale brown.

Paraphyses 3-4 um diam., simple or with one or more short, tapering,

narrow, 1.5—2 um diam. branches at apex, extending about 10-20 um beyond

asci, small crystals in clumps amongst the parts of the paraphyses above

the asci. Asci 110-180 x 12-14 um, cylindric, apex broadly rounded, wall

undifferentiated or slightly thinner across the apex, 8-spored. Ascospores

55-78 x 3.5-4.5 um (1.5-2.5 um at the constrictions), long-cylindric,

500 ... Johnston & Park

Figure 5. Marthamyces harakeke (A, B. PDD 108765; C, D. PDD 112252). A, B. Macroscopic

appearance when dry; C. Upper wall of ascoma and part of hymenium in vertical section; D. Detail

of upper wall of immature, unopened ascoma showing periphysoid layer. Scale bars: A = 1 cm;

B=1mm;C,D=10 um.

Marthamyces spp. nov. & Ramomarthamyces gen. nov. (New Zealand) ... 50.1

Figure 6. Marthamyces harakeke (A, B. PDD 112252; C-E PDD 108765). A. Conidioma in

vertical section; B. Wall of conidioma, conidiogenous cells, and conidia; C. Asci and ascospores;

D. Apex of paraphyses; E, F Ascospores with small globose gel caps at each end. Scale bars =

10 um.

502 ... Johnston & Park

trifusiform with two constrictions along the length (rarely bifusiform with

a single constriction), a single septum at the upper constriction, small,

globose gelatinous caps at each end, hyaline. Conidiomata developing

within leaf spots prior to the mature ascomata, immersed in host tissue,

host tissue darker around the conidiomata, conidiomata globose, opening

by a small, round pore through stomata, conidiomatal wall about 10 um

thick, comprising 5-6 layers of tangled hyphae 1.5-2.5 um diam. with walls

hyaline, thin. Wall near the opening of the conidioma up to 20 um thick,

comprising hyphae up to 6 um diam., innermost cells lining the ostiole

slightly elongated and periphysoid-like. Conidiomatal wall lined with

conidiogenous layer. Conidiogenous cells 10-17 x 1.5-2 um, cylindric,

tapering slightly towards the barely differentiated apical conidiogenous

locus; several conidiogenous cells held on a conidiophore, cylindric,

8-12 x 2-2.5 um, hyaline. Conidia 16-24 x 0.8-1 um filiform, straight to

irregularly curved, 0-septate, hyaline.

ADDITIONAL SPECIMENS EXAMINED—NEW ZEALAND: AUCKLAND: Hauraki

Gulf, Tiritiri Matangi Is., Ridge Track, on Phormium tenax living leaf, 28 Oct

2000, coll. E-H.C. McKenzie, R.E. Beever REB1978 (PDD 83472); Tiritiri Matangi

Island, on Phormium tenax living leaf, 27 Oct 2000, coll. P.R. Johnston R1017 (PDD

72007); Piha, on Phormium tenax living leaf, 5 Oct 1972, coll. J.M. Dingley (PDD

60494); Piha, on Phormium tenax living leaf, 2 Apr 1956, coll. J.M. Dingley (PDD

60497); Piha, on Phormium tenax living leaf, 5 Sep 1960, coll. J.M. Dingley (PDD

60500); Piha, on Phormium tenax living leaf, Sep 1960, coll. RLER. McNabb (PDD

43290); Piha, on Phormium tenax living leaf, 5 Oct 1972, coll. J.M. Dingley (PDD

43312); Waitakere Swamp, on Phormium tenax living leaf, 3 Apr 1964, coll. J.M.

Dingley (PDD 23268); Karekare, on Phormium tenax living leaf, 27 Apr 1971, coll.

J.M. Dingley (PDD 60495); Anawhata, on Phormium tenax living leaf, 13 May

1972, coll. J.M. Dingley (PDD 60498). CHATHAM IsLANDs: Rekohu, Rangaika

Reserve, on Phormium tenax living leaf, 21 Nov 1992, coll. EH.C. McKenzie, P.R.

Johnston C80 (PDD 62689). COROMANDEL: Little Barrier Island, near East Cape,

on Phormium tenax living leaf, 16 June 1984, coll. R.E. Beever, P.R. Johnston, E.P.

Laracy (PDD 55303). NELSson: Takaka Saddle, on Phormium tenax living leaf, 17

Nov 1970, coll. J.M. Dingley (PDD 60647). NorTHLAND: Maunganui Bluff Scenic

Reserve, on Phormium tenax living leaf, 19 Oct 1987, coll. E.H.C. McKenzie, P.R.

Johnston (PDD 48400); Whangaruru North Head, on Phormium tenax living leaf,

28 Jan 1988, coll. P.R. Johnston (PDD 54127); Waipoua Forest, Lookout Track, on

Phormium tenax living leaf, 11 May 2017, coll. PR. Johnston D2429 (PDD 108765);

Three Kings Islands, Great Island, vic. light beacon, on Phormium tenax living leaf,

27 Nov 1997, coll. R.E. Beever REB 1565-2 (PDD 68427).

Notes: First collected in the 1950s, this fungus is common on living leaves of

Phormium tenax in north New Zealand. Although its host occurs throughout

New Zealand, no specimens have been yet been collected south of Nelson or

the Chatham Islands.

Marthamyces spp. nov. & Ramomarthamyces gen. nov. (New Zealand) ... 503

Marthamyces maccormackii P.R. Johnst., sp. nov. Fic. 7

IF 556324

Differs from M. emarginatus by its darker ascomatal wall and from M. metrosideri by its

rostrate ascus apex.

Type: Cook Islands: Rarotonga: Te Rua Manga, near summit, on Metrosideros collina

fallen leaves, 28 Jun 2005, coll. PR. Johnston RR392 (holotype PDD 112256; ex-type

culture ICMP 15829).

ErymMo.ocy: Named in honour of Gerald McCormack, director of the Cook Islands

Biodiversity and Natural Heritage Trust, who provided the support needed to collect the

Cook Islands specimens cited here.

Ascomata developing on recently fallen leaves, often associated with slightly

paler areas on the leaf, initially appearing as a slightly raised dark spot,

opening initially as a pore, then by three to four radiate slits, the covering

host and fungal tissue folding back as small triangular flaps, the undersides

of which are covered with a layer of white crystals, exposed hymenial surface

with a layer of whitish crystals. Mature ascomata 0.3-0.5 mm diam., more

or less circular in shape, covering tissue normally darkened, pale in some

collections, hymenial surface remaining well below the surface of the host

tissue. Ascomata subepidermal, immature ascomata in vertical section with a

50 um thick covering layer comprising partially decomposed host epidermal

cells and several layers of fungal tissue. The outermost, clypeus-like layer

comprises 3-4 rows of tangled hyphae 2.5-3.5 um diam., with hyphal walls

dark brown, irregularly thickened, but this is lacking near the centre of the

ascoma. The upper wall of the ascoma comprises several layers of globose

to angular cells 3-5 um diam. with walls thin, slightly pigmented, and this

layer of cells is lined with a layer of downward-projecting periphysoids

2-3 um diam., with walls thin, hyaline. In mature ascomata the upper wall

develops clumps of crystals forming amongst the globose to angular cells.

Lower wall of the ascoma 15-20 um thick, comprising several layers of

tangled hyphae 2.5-3.5 um diam. with hyphal walls dark brown, irregularly

thickened. Paraphyses 2 um diam., with several short, tapering branches near

apex 1-1.5 um diam., paraphyses about the same length as the asci. Asci

75-95 x 7.5-9 um, cylindric, tapering suddenly to small, rostrate apex, wall

undifferentiated, 8-spored. Ascospores 50-70 x 1.5-2 um, gelatinous caps at

each end small, flattened, 1-septate, hyaline.

ADDITIONAL SPECIMENS EXAMINED: COOK ISLANDS: RAROTONGA: Cross Island

walk, near Te Rua Manga, on Metrosideros collina, 30 Jun 2005, coll. PR. Johnston RR611

(PDD 112250, culture ICMP 16155); track to Ikurangi, near the top, on M. collina fallen

leaves, 16 Jan 2005, coll. PR. Johnston RR315 (PDD 112258, culture ICMP 15831); track

to Ikurangi, near the top, on M. collina fallen leaves, 4 Jul 2005, coll. PR. Johnston RR635

504 ... Johnston & Park

Marthamyces spp. nov. & Ramomarthamyces gen. nov. (New Zealand) ... 505

(PDD 112260); Te Rua Manga, near summit, on M. collina fallen leaves, 5 Jul 2005, coll.

PR. Johnston RR650 (PDD 112255).

Notes—There are two Metrosideros-inhabiting species of Marthamyces in

the Cook Islands. Marthamyces maccormackii has small, circular ascomata

with a dark upper wall, macroscopically remarkably similar to M. metrosideri

from New Zealand but phylogenetically quite distinct and also different

in having a rostrate rather than rounded ascus apex. A second, sympatric

Marthamyces species from the same host in the Cook Islands has larger,

more angular, less deeply immersed, paler-walled ascomata with a yellow

hymenial surface and is described below as M. renga. Microscopically the

two Cook Island species differ in ascus shape. The two species often form in

lesions adjacent to each other on the same leaves.

Marthamyces metrosideri PR. Johnst., sp. nov. Fia. 8

IF 556325

Differs from Marthamyces emarginatus by its Metrosideros host preference, its darker

upper wall of ascomata, and its ascus tapering gradually to rounded apex.

Type: New Zealand: Wellington: Wainuiomata Waterworks Catchment Area, on

Metrosideros robusta fallen leaves, 19 Nov 1992, coll. P.R. Johnston R965 (holotype PDD

67458; ex-type culture ICMP 22625).

EryMo_ocy: based on host genus name.

Ascomata developing within slightly paler areas on the lower surface of fallen

leaves. Immature ascomata initially with appearance of small, darkened

pustules. Mature ascomata round in outline, 0.3-0.4 mm diam., opening by

several radiates slits, covering layer of host and fungal tissue folding back as

3-4 flaps lined with white crystals, exposing the white, pruinose hymenial

surface. Ascomata subepidermal, in vertical upper wall of immature ascoma

comprising 4-5 layers of angular to globose cells 3-5 um diam., with walls

thin, darkened, lined with layer of downward-projecting periphysoids, 15-30 x

3-3.5 um, cylindric, unbranched, walls thin, hyaline. Lower wall of immature

ascoma 20-35 um thick, of several layers of hyphae 1.5-2.5 um diam., with

walls brown, slightly thickened, forming a loose textura intricata. Mature

ascomata in vertical section with upper wall 70-100 um thick, comprising a

10-30 um wide layer of 3-5 um diam. hyphae with cell walls thin, dark brown,

Figure 7. Marthamyces maccormackii (PDD 112256). A. Macroscopic appearance when dry;

B. mature, opened ascoma in vertical section; C. Immature, unopened ascoma in vertical

section; D. Asci; E. Flattened gel caps at top of ascospores; F. Paraphyses . Scale bars: A = 1 mm;

B, C = 20 um; D-F = 10 um.

506 ... Johnston & Park

and a broad layer of crystals. Upper wall of mature ascomata lined with a layer

of inward-projecting, hyaline, thin-walled periphysoids, individual elements

shorter than in unopened ascomata, up to 10 um long. Paraphyses 2-3 um

diam., with short side-branches 1-1.5 um diam. near apex. Asci 70-105(-120)

x 7-9.5 um, subcylindric to subfusoid, tapering gradually to rounded to broadly

rounded apex, wall undifferentiated at apex, 8-spored. Ascospores 45-85 x

2-3 um, filiform, straight when released, tapering slightly and gradually towards

the base, with small, flattened gelatinous caps at apex and base, 0-1-septate,

hyaline.

ADDITIONAL SPECIMENS EXAMINED: NEW ZEALAND: KERMADEC ISLANDs: Raoul

Island, Denham Bay Track, on Metrosideros kermadecensis, 22 Sep 1988, coll. E.H.C.

McKenzie (PDD 54664); Raoul Island, Blue Lake Track, on M. kermadecensis, 23

Sep 1988, coll. E.H.C. McKenzie (PDD 54666). NORTHLAND: Waipoua State Forest,

Yakas Track, on M. robusta, 20 Oct 1987, coll. PR. Johnston (PDD 48163); vic.

Opononi, Waimamaku River mouth, on M. excelsa, 21 Oct 1987, coll. PR. Johnston

(PDD 53849); Whangaruru North Head, on M. excelsa, 28 Jan 1988, coll. PR.

Johnston (PDD 54117); Ahipara, Shipwreck Bay, on M. excelsa, 8 May 2017, coll. PR.

Johnston D2421 (PDD 111232). AUCKLAND: Waitakere Ranges, Walker Bush, on M.

robusta, 4 Aug 1982, coll. E.H.C. McKenzie, G.J. Samuels, P.R. Johnston R64 (PDD

43951); Huia Dam footbridge, on M. excelsa, 27 Apr 1982, coll. P.R. Johnston R52

(PDD 43952); Kaitarakihi Beach, on M. excelsa, 8 Mar 1983, coll. RR. Johnston (PDD

43953); Piha Valley, Cowan Track, on M. robusta, 27 Apr 1983, coll. P.R. Johnston

R209, G.J. Samuels, R.H. Petersen (PDD 43954); Mt Albert Research Centre, on

Metrosideros sp., May 1983, coll. P.R. Johnston (PDD 43955); Kaitarakihi Beach,

on M. excelsa, 20 Jul 1983, coll. BR. Johnston (PDD 43958); Cornwallis Beach,

on M. excelsa, 13 Apr 1984, coll. PR. Johnston (PDD 49151). COROMANDEL: vic.

Coromandel, 309 Road, Waiau Falls Scenic Reserve, on M. robusta, 25 Apr 1989, coll.

P.R. Johnston (PDD 55341); Thames Coast, between Te Mata and Waikawau, on M.

excelsa, 26 Aug 1999, coll. P.R. Johnston R1003, B.M. Spooner (PDD 70890); Tapu-

Coroglen Road, 2 km Tapu side of summit, on M. robusta, 29 Aug 1984, coll. P.R.

Johnston R552 (PDD 46194); North of Colville, vic. Ohinewai Stream, on M. excelsa,

27 Jan 1985, coll. PR. Johnston (PDD 46775); Waihi, track N of beach, on M. excelsa,

9 May 2003, coll. P.R. Johnston (PDD 78201); Port Charles, between wharf and Big

Sandy Bay, on M. excelsa, 26 Mar 1989, coll. P.R. Johnston (PDD 55293); Awaroa

Stream, on M. fulgens, 12 Jun 1984, coll. PR. Johnston (PDD 55294); Little Barrier

Island, Titoki Point, on M. excelsa, 11 Jun 1984, coll. P.R. Johnston (PDD 45364);

Little Barrier Island, near hut, on M. excelsa, 8 Apr 1988, coll. PR. Johnston R795

(PDD 48768); Little Barrier Island, south of Te Wairere, on M. excelsa, 9 Mar 1974,

coll. W.B. Kendrick (PDD 40728). GISBORNE: Urewera National Park, Ngamoko

FiGureE 8. Marthamyces metrosideri (A-C, E-G. PDD 111232; D. PDD 55341). A. Macroscopic

appearance when dry; B. Mature, opened ascoma in vertical section; C. Immature, unopened

ascoma in vertical section; D. Asci; E. Released ascospore; F. Apex of ascospores showing gel caps;

G. Paraphyses. Scale bars: A = 1 mm; B, C = 20 um; D-G = 10 um.

Marthamyces spp. nov. & Ramomarthamyces gen. nov. (New Zealand) ... 507

508 ... Johnston & Park

Track, on M. robusta, 10 May 2001, coll. P.R. Johnston (PDD 74308); vic. Ruatahuna,

Mangapae, on M. robusta, 24 May 2005, coll. P.R. Johnston R1033, T. Hosoya (PDD

82920). TARANAKI: Mt Egmont, Waiweranui Track, on M. robusta, 25 Apr 1983, coll.

PR. Johnston R274, G.J. Samuels, R.H. Petersen (PDD 43957). WANGANUI: Kai Iwi,

Bushy Park Reserve, on M. robusta, 15 May 1987, coll. P.R. Johnston (PDD 45587).

NELSON: Karamea Bluff, Large Matai Track, on M. robusta, 25 May 1989, coll. P.R.

Johnston, E.M. Gibellini (PDD 57159); Karamea, 1 km S Granite Creek Road, on

M. robusta, 14 Apr 1983, coll. PR. Johnston R262, G.J. Samuels, R.E. Beever (PDD

43956, culture ICMP 22624). BULLER: vic. Greymouth, Point Elizabeth Walkway,

on M. robusta, 1 May 1985, coll. P.R. Johnston R645, G.J. Samuels, L.M. Kohn (PDD

46961). WESTLAND: Haast, Hapuku River mouth, on M. umbellata, 11 May 2002,

coll. S.R. Whitton, T. Atkinson (PDD 75509).

Notes—This common, Metrosideros-inhabiting fungus was previously

identified as M. emarginatus (Johnston 1986, as Propolis emarginata; McKenzie

& al. 1999, as Propolis emarginata). Based on published descriptions (e.g.

Sherwood 1977), examination of the type specimen of M. emarginatus, and the

features of the specimens from Eucalyptus from Australia deposited as PDD

81844, PDD 81845, and PDD 81846, the two species are very similar but slight,

apparently consistent morphological differences (see diagnosis) are matched by

the five transitions across the ITS1 and ITS2 regions.

Marthamyces renga P.R. Johnst., sp. nov. Fic. 9

IF 556326

Differs from Marthamyces emarginatus by its larger, more angular, less deeply immersed

ascomata, and its sharp contrast between the white crystals lining the upper wall and the

yellow crystals on the hymenial surface.

Type: Cook Islands: Rarotonga: Raemaru Track, summit, on Metrosideros collina fallen

leaves, 28 Jun 2005, coll. PR. Johnston RR562 (holotype PDD 112263).

ETyMOLoGy: renga, Cook Island Maori word for yellow, referring to the colour of the

hymenial surface.

Ascomata developing within slightly paler areas on both the upper and lower

surface of recently fallen leaves, initially appear as slightly raised patches, these

splitting open by 3-4(-5) radiate slits, the covering host and fungal tissue

folding back as small flaps, the undersides of which are covered with a layer

of white crystals, the exposed hymenial surface crystalline, yellow. Mature

ascomata 0.4-1.5 x 0.4-0.8 mm, angular, 3-5-sided. Ascomata subepidermal,

in vertical section mature ascomata with a covering layer 50-60 um thick,

FiGuRE 9. Marthamyces renga (A-C. PDD 112263; D-F. PDD 112259). A. Macroscopic appearance

when dry; B. Mature, opened ascoma in vertical section; C. Detail of upper wall of opened ascoma

in vertical section; D. Asci; E. Paraphyses; F. Apex of ascospores with flattened gel cap. Scale bars:

A=1mm; B= 100 um; C-F = 10 um.

Marthamyces spp. nov. & Ramomarthamyces gen. nov. (New Zealand) ... 509

510... Johnston & Park

comprising mostly partially broken-down cells of the host leaf tissue, together

with crystals forming within a thin layer of more-or-less globose fungal cells

with walls thin, hyaline, lined with a layer of broad-cylindric periphysoids,

10-20 x 4-6 um, thin-walled, hyaline, unbranched. Lower wall 10-15 um thick,

comprising a compact layer of 3-4 um diam. hyphae with cell walls hyaline,

slightly thickened. Paraphyses 2-2.5 um diam., with several short, tapering

branches near apex 1-1.5 um diam., intermixed with groups of small crystals,

paraphyses about the same length as the asci. Asci 75-90 x 7-9.5 um, cylindric,

tapering slightly to rounded apex, wall undifferentiated, 8-spored. Ascospores

rarely seen released, 1.5-2 um wide, about 55-70 um long, gelatinous caps at

each end small, flattened, 1-septate, hyaline.

ADDITIONAL SPECIMENS EXAMINED: COOK ISLANDS: RAROTONGA: Te Rua Manga,

on track near summit, on Metrosideros collina fallen leaves, 21 Jan 2005, coll. PR.

Johnston RR393 (PDD 112257; culture ICMP 15830); Te Kou, on track near summit,

on Metrosideros collina fallen leaves, 19 Jan 2005, coll. RR. Johnston RR380 (PDD

112259; culture ICMP 15827). Te Rua Manga, on track near summit, on Vaccinium

cereum fallen leaves, 5 Jul 2005, coll. BR. Johnston RR397 (PDD 112261). Te Rua

Manga, on track near summit, Vaccinium cereum fallen leaves, 5 Jul 2005, coll. P.R.

Johnston RR642 (PDD 112264). Te Rua Manga, on track near summit, on Weinmannia

samoensis fallen leaves, 5 Jul 2005, coll. P.R. Johnston RR641 (PDD 112262).

Notes: A macroscopically distinctive species with a marked contrast in colours

between the white crystals on the underside of the upper wall of the ascoma and

the yellow crystals on the hymenial surface. Although common on Metrosideros

collina, this species is not host-specialized and was found also on recently fallen

leaves of Vaccinium cereum G. Forst. (Ericaceae) and Weinmannia samoensis

A. Gray (Cunoniaceae), both south Polynesian endemic plant species found in

Rarotonga in the same high-altitude plant communities as Metrosideros. See

also notes under M. maccormackii.

Ramomarthamyces P.R. Johnst., gen. nov.

IF 556327

Differs from Marthamyces by its distinctly branched paraphyses.

TYPE SPECIES: Propolis dracophylli P.R. Johnst. [= Ramomarthamyces dracophylli (P.R.

Johnst.) PR. Johnst.]

ETYMOLOGy: ramo = branch; refers to the branched paraphyses in these Marthamyces-

like fungi.

Ascomata immersed in the host tissue, opening by several radiate slits to reveal

the pruinose, crystalline hymenial surface. Ascus wall undifferentiated at the

apex, ascospores filiform, paraphyses branched several times near the apex.

Marthamyces spp. nov. & Ramomarthamyces gen. nov. (New Zealand) ... 511

Notes: A genus in the Marthamycetaceae, but molecular phylogenetic analysis

shows it to be phylogenetically distinct from other genera within the family

(Frc. 1). The genetic difference is supported by a characteristic morphology, all

species known for the genus having distinctly branched, rather than propoloid,

paraphyses. Marthamyces is macro-morphologically similar.

Ramomarthamyces tuku PR. Johnst., sp. nov. Fra. 10

IF 556333

Differs from Ramomarthamyces gilvus by its Juncaceae host, its elongate ascoma, its

ascospores with a swelling above the septum, its undifferentiated ascus wall, and its lack

a well-developed layer of crystals amongst the paraphyses.

Type: New Zealand, Chatham Islands, Rekohu, Tuku Reserve, on Juncus sp., dead leaves,

19 Nov 1992, coll. PR. Johnston C42, R911 & E.H.C. Mckenzie (holotype PDD 62161;

ex-type culture ICMP 22562).

ErymMo oey: tuku, based on type locality.

Ascomata developing on dead leaves, not associated with discoloured areas.

Ascomata 0.5-1 x 0.2-0.3 mm, immersed in the leaf, opening by an elongate slit,

covering layer slightly paler than the surrounding leaf tissue, sometimes with

poorly developed crystalline layer near edge of opening slit, exposed hymenium

translucent yellow, lacking obvious crystals. Ascomata intrahypodermal, in

vertical section immature ascomata with a covering layer 50-70 um thick,

comprising most partly decomposed host cells intermixed with a poorly

developed, clypeus-like layer of hyphae 3-5 um diam. with walls thin, hyaline.

Beneath the host cells is a 10 um thick layer of narrow tangled hyphae with

walls thin, hyaline, this layer lined with broad-cylindric periphysoids 10-15 x

4-5 um, with walls thin, hyaline. Mature ascomata similar in vertical section

but periphysoids more or less lost, and clumps of crystals forming amongst the

hyphae of the upper wall near the opening slit. Lower wall 10-15 um thick,

comprising several rows of narrow, tangled hyphae with walls thin, hyaline.

Paraphyses 1.5-2 um diam., several times branched near the apex, extending

20-25 um beyond asci. Asci 115-125 x 9-10 um, cylindric, apex broadly

rounded, wall undifferentiated at apex, 8-spored. Ascospores 110 x 2-3 um,

1-septate, a small swelling 3.5-4 um diam. immediately above the septum,

otherwise not tapering, more or less globose gelatinous caps at each end, 3-3.5

um diam.

Notes: Although known only from the type specimen, this is a clearly distinct

species, both morphologically and phylogenetically. An inconspicuous

fungus, it is likely to be widespread on Juncus in New Zealand. The closely

Johnston & Park

Slizes

Marthamyces spp. nov. & Ramomarthamyces gen. nov. (New Zealand) ... 513

related R. gilvus is known from several cyperaceous hosts in Australia and it

is possible that with more collections, R. tuku will be found to have a broader

range of hosts. The New Zealand species Marthamyces dendrobii has similar

distinctively shaped ascospores, with a small swelling immediately above the

central septum, but differs in host, its ascoma having a dark upper wall, and its

smaller ascospores.

Ramomarthamyces barbatus (PR. Johnst.) P.R. Johnst., comb. nov.

IF 556329

= Marthamyces barbatus P.R. Johnst., Aust, Syst, Bot, 19: 137, 2006.

NoTE: Recombination based on both DNA sequence and morphological data.

Ramomarthamyces dracophylli (P.R. Johnst.) P.R. Johnst., comb. nov.

IF 556330

= Propolis dracophylli P.R. Johnst., New Zealand J. Bot. 24: 115, 1986.

=Marthamyces dracophylli (P.R. Johnst.) Minter, Mycotaxon 87: 51, 2003.

NoTE: Recombination based on both DNA sequence and morphological data.

Ramomarthamyces gilvus (Rodway) P.R. Johnst., comb. nov.

IF 556331

= Naemacyclus gilvus Rodway, Pap. & Proc. Roy. Soc. Tasmania 1917: 107, 1918.

= Marthamyces gilvus (Rodway) P.R. Johnst., Aust. Syst. Bot. 19: 140, 2006.

NoTE: Recombination based on both DNA sequence and morphological data.

Discussion

Marthamyces harakeke is widespread and common as a leaf spot on

Phormium in New Zealand. The earliest fungarium specimen of the fungus

was collected in 1956 and deposited in PDD as Propolis sp., but with a note

that the identification was uncertain. Later specimens were also referred

to Propolis by several different mycologists. Although macroscopically the

fungus is Propolis-like in having apothecia shallowly immersed in host tissue,

a crystalline hymenial surface, and asci undifferentiated at the apex, there

was always doubt about its identity because of the pathogenic lifestyle. It was

not until DNA sequences were obtained from a specimen collected in 2017

FiGuRE 10. Ramomarthamyces tuku (PDD 62161). A. Macroscopic appearance when dry;

B. Mature, opened ascoma in vertical section; C. Immature, unopened ascoma in vertical section,

showing periphysoids; D. Ascospore with a globose gel cap at each end; E. Apex of asci; F. Apex of

paraphyses. Scale bars: A = 1 mm; B, C = 20 um; D-F = 10 um.

514... Johnston & Park

that its generic position was resolved. Today the genus used for this fungus

and its relatives is Marthamyces, a generic name now used for several species

that had been placed in Propolis in the past.

Both Propolis and Marthamyces were placed in Rhytismataceae at the

time that Lantz & al. (2011) published a molecular phylogeny treating the

Rhytismatales in depth. Their results showed the Rhytismataceae to be

polyphyletic, and they regarded Propolis and Marthamyces as incertae sedis

within the Leotiomycetes. With support from rDNA-based phylogenies, Baral

(2015) placed Propolis and Marthamyces along with several other genera in a

separate family, Marthamycetaceae, and more recently Johnston & al. (2019)

erected the order Marthamycetales, based on a 15-loci analysis. In the present

paper, species placed in the genus Marthamyces have been more intensively

sampled with molecular data, and the genus was shown to be polyphyletic. As

a result we created the genus Ramomarthamyces for species segregated from

Marthamyces sensu stricto.

The phylogeny presented here shows a pathogenic lifestyle to have

evolved more than once in the Marthamycetaceae. Along with M. harakeke,

M. culmigenus (in a clade of species separate from that containing

M. harakeke) and the Cyclaneusma species cause foliar diseases.

Marthamyces emarginatus was originally described from Eucalyptus from

Victoria, Australia. Marthamyces emarginatus-like fungi are common on

recently fallen Eucalyptus leaves in Australia, but the data presented here show

that several closely related but phylogenetically distinct species occur on this

substrate. Insufficient samples are available to determine whether there is a

relationship between this diversity with respect to geography, or to phylogeny

within Eucalyptus. Until the distribution of these fungi in Australian forests is

better understood, a neotype cannot be selected for M. emarginatus.

Two GenBank accessions identified as Coccomyces proteae have ITS

sequences that closely match Marthamyces emarginatus-like species

(CBS 111703 as JN712451; CBS 111704 as JN712450; Fic. 2). Based

on (Marincowitz & al. 2008) the morphology of C. proteae is typical of

Coccomyces. It is possible that the putative C. proteae cultures from which

these sequences were derived represent another, possibly undescribed,

species of Marthamyces that grows on Protea leaves in South Africa. Note

that another GenBank accession identified as Coccomyces proteae (JX089385

ex leaves of Disterigma humboldtii (Ericaceae) ex Puerto Rico) represents

yet another Marthamyces species, this one belonging in the clade containing

M. maccormackii, M. renga, and M. culmigenus (Fic. 2).

Marthamyces spp. nov. & Ramomarthamyces gen. nov. (New Zealand) ... 515

Acknowledgments

The Cook Island specimens were collected with the support of Gerald McCormack

through the Cook Islands Biodiversity and Natural Heritage Trust, under the

conditions of a scientific collecting permit issued by the Cook Islands National

Environment Service. The Australian specimens were collected with the support

of the Participatory Program of the Australian Biological Resources Survey, under

the conditions of permits issued by the New South Wales National Parks and

Wildlife Service, Department of Conservation and Natural Resources—Victoria,

and the Tasmanian Department of Parks, Wildlife and Heritage. The New Zealand

collections were collected under the conditions of permits issued by the New Zealand

Department of Conservation, and through a scientific research agreement with the

Tuhoe Tuawhenua Trust. The authors are supported through the Manaaki Whenua

Systematics portfolio, with funding from the Science and Innovation Group of the

New Zealand Ministry of Business, Innovation and Employment. The authors thank

Danny Haelewaters (Purdue University, West Lafayette IN USA) and Cheng-Lin Hou

(Capital Normal University, Beijing, China) for presubmission review.

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MY COTAXON

July-September 2019—Volume 134, pp. 517-528

https://doi.org/10.5248/134.517

Septobasidium aquilariae sp. nov. from Yunnan, China

Ru1-Linc Ma}, MENG GAO}, XIANG-LIN ZHUANG? , CHANG-LIN ZHAO!”

'Key Laboratory for Forest Resources Conservation and Utilization

in the Southwest Mountains of China, Ministry of Education,

Southwest Forestry University, Kunming 650224, P.R. China

College of Biodiversity Conservation, Southwest Forestry University,

Kunming 650224, P.R. China

College of Forestry, Southwest Forestry University, Kunming 650224, P.R. China

" CORRESPONDENCE TO: fungichanglinz@163.com

ABSTRACT—A new fungal species, Septobasidium aquilariae, is proposed based on

a combination of morphological features and molecular evidence. The species is

characterized by an annual growth habit; a resupinate coriaceous basidiocarp with a

cream to pale brown surface; a monomitic hyphal system with thick-walled generative

hyphae bearing simple septa; reniform, hyaline, thin-walled, smooth basidiospores

measuring 11-19 x 4—7.5 um; and haustoria consisting of irregularly coiled hyphae. The

fungus was found associated with Pseudaulacaspis sp. on Aquilaria sinensis. Sequences of

internal transcribed spacer region (ITS) were analysed maximum likelihood, maximum

parsimony, and Bayesian inference methods. The phylogenies strongly supported

S. aquilariae in a monophyletic lineage (ML = 100%; MP = 100%; PP = 1) and grouped

with “S. cokeri”.

Key worps—felt fungus, Pucciniomycetes, Septobasidiaceae, Septobasidiales, taxonomy

Introduction

Septobasidium Pat. (Septobasidiaceae, Septobasidiales), erected by

Patouillard (1892), is a large, cosmopolitan genus characterized by resupinate

basidiocarps with a white to cream, yellowish brown or brown hymenophore,

a monomitic hyphal system with simple septa, with or without probasidia, 2-4

celled cylindrical, curved, or straight basidia, basidiospores that are hyaline,

thin-walled, smooth, and cylindrical or fusiform, and haustoria consisting

518 ... Ma &al.

of coiled or spindle-shaped hyphae (Patouillard 1892, Couch 1929). About

300 species have been accepted in the genus worldwide (Patouillard 1892;

Bresadola & Saccardo 1897; Burt 1916; Lloyd 1919; Couch 1929, 1935, 1938,

1946; Yamamoto 1956; Gomez & Henk 2004; Henk 2005; Lu & Guo 2009a,b,

2010a,b, 2011; Lu & al. 2010; Chen & Guo 2011, 2012; Li & Guo 2013, 2014).

Molecular systematics have played a powerful role in inferring phylogenies

within fungal groups since the early 1990s (White & al. 1990, Binder &

al. 2013, Dai & al. 2015, Choi & Kim 2017). However, molecular studies

involving Septobasidium are rare (Henk & Vilgalys 2007, Zhao & al. 2017). One

phylogenetic study of a single origin of insect symbiosis in the Pucciniomycetes

suggested that there is little or no support for Septobasidium as a monophyletic

group (Henk & Vilgalys 2007). Zhao & al. (2017) introduced a six-gene

phylogenetic overview of Basidiomycota and allied phyla and confirmed

that S. carestianum Bres. nested within the Septobasidiales and grouped with

Helicobasidium mompa Nobuj. Tanaka and Thanatophytum crocorum (Pers.)

Nees.

During disease investigations on Aquilaria sinensis in southern China, a new

Septobasidium taxon was found that could not be assigned to any described

species. In this study, we expand samplings from previous studies to examine

the taxonomy and ITS phylogeny of this new species within the Septobasidium.

Materials & methods

The specimens studied are deposited at the herbarium of Southwest Forestry

University, Kunming, China (SWFC). Macromorphological descriptions are

based on field notes. Colour terms follow Petersen (1996). Micromorphological

data were obtained from dried specimens observed under a light microscope.

Measurements are presented as: L = mean spore length, W = mean spore width,

Q = range of L/W ratios, n = number of spores/number of specimens.

Genomic DNA was obtained from dried specimens using the EZNA HP Fungal

DNA Kit, according to the manufacturer’s instructions with some modifications.

A small piece of dried fungal specimen (about 30 mg) was ground to powder

with liquid nitrogen. The powder was transferred to a 1.5 ml centrifuge tube,

suspended in 0.4 ml of lysis buffer, and incubated in a 65 °C water bath for 60

min. After that, 0.4 ml phenol-chloroform (24:1) was added to each tube and

the suspension was shaken vigorously. After centrifugation at 13,000 rpm for 5

min, 0.3 ml supernatant was transferred to a new tube and mixed with 0.45 ml

binding buffer. The mixture was then transferred to an adsorbing column (AC) for

centrifugation at 13,000 rpm for 0.5 min. Then, 0.5 ml inhibitor removal fluid was

added in AC for a centrifugation at 12,000 rpm for 0.5 min. After washing twice

with 0.5 ml washing buffer, the AC was transferred to a clean centrifuge tube,

Septobasidium aquilariae sp. nov. from China ... 519

TABLE 1. Species, specimens, and ITS sequences used in Septobasidium molecular

analyses.

SPECIES

Helicobasidium mompa

Pachnocybe ferruginea

Septobasidium alni

S. aquilariae

S. arachnoideum

S. bogoriense

S. broussonetiae

S. burtii

S. carestianum

S. castaneum

S. cavarae

“S. cokeri”

S. fumigatum

S. gomezii

S. grandisporum

S. hainanense

S. kameii

S. maesae

S. marianiae

S. michelianum

S. pallidum

S. pilosum

S. pinicola

S. pseudopedicellatum

S. ramorum

S. septobasidioides

S. taxodii

S. velutinum

S. westonii

S. wilsonianum

SAMPLE

DAH hI

DAH pfl

DAH EP3

CLZhao 6610

CLZhao 6611 [T]

CLZhao 6612

CLZhao 6613

CLZhao 6614

DAH 025

998434

998436

DAH 062

DJM 644

DAH 052

DJM FP1

DAH 061

DAH 005

DAH 031

DAH 065

998437

998432

998433

DAH 283b

DAH EP5

998435

DAH 020

DAH 013

DAH 044

DAH 045a

DAH 032

DAH 194C

DAH 024

DAH FP2001

DAH 037

GENBANK ITS

DQ241472

DQ241473

DQ241441

MK802139

MK802140

MK802141

MK802142

MK802143

DQ241443

HM209414

HM209416

DQ241444

DQ241448

DQ241447

DQ241445

DQ241449

DQ241451

DQ241462

DQ241453

HM209417

HM209412

HM209413

DQ241456

DQ241457

HM209415

DQ241458

DQ241459

DQ241460

DQ241450

DQ241461

DQ241466

DQ241467

DQ241468

DQ241469

REFERENCE

Henk & Vilgalys 2007

Present study

Henk & Vilgalys 2007

Unpublished

Henk & Vilgalys 2007

Unpublished

Henk & Vilgalys 2007

Unpublished

Henk & Vilgalys 2007

520 ... Ma &al.

and 100 ml elution buffer was added to the middle of adsorbed film to elute the

genomic DNA. ITS region was amplified with primer pairs ITS5 and ITS4 (White

& al. 1990). The PCR procedure was as follows: initial denaturation at 95 °C

for 3 min; followed by 35 cycles of 94 °C for 40 s, 58 °C for 45 s, and 72 °C for 1 min;

and a final extension of 72 °C for 10 min. The PCR products were purified and

directly sequenced at Kunming Tsingke Biological Technology Limited Company.

All newly generated sequences were deposited at GenBank (TABLE 1).

Sequencher 4.6 was used to edit the DNA sequence. Sequences were aligned in

MAFFT 6 (Katoh & Toh 2008; http://mafft.cbrc.jp/alignment/server/) using the

“G-INS-I° strategy and manually adjusted in BioEdit (Hall 1999). The sequence

alignment was deposited in TreeBase (submission ID 23722). Sequences of

Helicobasidium mompa and Pachnocybe ferruginea Berk. obtained from GenBank

were used as outgroups to root trees following Henk & Vilgalys (2007) in the ITS

analysis (Fic. 1).

Maximum parsimony analysis was applied to the ITS dataset sequences.

Approaches to phylogenetic analysis followed Zhao & al. (2013) and Song & al.

(2016), and the tree construction procedure was performed in PAUP* version

4.0b10 (Swofford 2002). All characters were equally weighted, and gaps were

treated as missing data. Trees were inferred using the heuristic search option with

TBR+G branch swapping and 1000 random sequence additions. Max-trees were

set to 5000, branches of zero length were collapsed and all parsimonious trees

were saved. Clade robustness was assessed using a bootstrap (BP) analysis with

1000 replicates (Felsenstein 1985). Descriptive tree statistics tree length (TL),

consistency index (CI), retention index (RI), rescaled consistency index (RC),

and homoplasy index (HI) were calculated for each Maximum Parsimonious Tree

(MPT) generated. Sequences were also analyzed using Maximum Likelihood

(ML) with RAxML-HPC2 on Abe through the Cipres Science Gateway (www.

phylo.org; Miller & al. 2009). Branch support (BS) for ML analysis with

GTR+G+I model of site substitution including estimation of Gamma-distributed

rate heterogeneity and a proportion of invariant sites. The branch support was

evaluated with bootstrapping method of 1000 replicates.

MrModeltest 2.3 (Posada & Crandall 1998; Nylander 2004) was used to

determine the best-fit evolution model for each data set for Bayesian inference

(BI). Bayesian inference was calculated with MrBayes3.1.2 with a general time

reversible (GTR+G+I) model of DNA substitution and a gamma distribution rate

variation across sites (Ronquist & Huelsenbeck 2003). Four Markov chains were

run for 2 runs from random starting trees for 3 million generations (Fic. 1) and

trees were sampled every 100 generations. The first one-fourth generations were

discarded as burn-in. A majority rule consensus tree of all remaining trees was

calculated. Branches were considered as significantly supported if they received

maximum likelihood bootstrap (ML)>70%, maximum parsimony bootstrap

(MP) >50%, or Bayesian posterior probabilities (PP) >0.95.

Septobasidium aquilariae sp. nov. from China... 521

S. aquilariae CL Zhao 6612

S. aquilariae CL Zhao 6613

S. aquilariae CL Zhao 6614

100/93/0.98 S. aquilariae CL Zhao 6611 [T]

S. aquilariae CL Zhao 6610

“S. coker?’ DAH 061

S. hainanense 998437

S. maesae 998433

S. pseudopedicellatum DAH 044

S. cavarae DAH FP1

100/100/1

-/66/ -

wilh S. pallidum 998435

S. carestianum DIM 644

70/671 - ~~" S. alni DAH FP3

S. kameii 998432

S. marianiae DAH 283b

S. velutinum DAH 024

S. ramorum DAH 045a

S. pinicola DAH 013

S. michelianum DAH FP5

-1551 - S. taxodii DAH 194C

S. bogoriense 998434

100/100/1 S. broussonetiae 998436

usioors S. wilsonianum DAH 037

S. pilosum DAH 020

pielie 100/98/1 L's grachnoideum DAH 025

S. burtii DAH 062

S. castaneum DAH 052

*)661- S. septobasidioides DAH 032

eLpE IS 100/99/1 S. grandisporum DAH 065

S. fumigatum DAH 005

S. gomezii DAH 031

99/94/0.98 S. westonii DAH FP2001

Helicobasidium mompa DAH hl

Pachnocybe ferruginea DAH pfl

“4

Fic. 1. Maximum Parsimony strict consensus tree illustrating the phylogeny of Septobasidium

aquilariae and related species in Septobasidium based on ITS sequences, with Helicobasidium

mompa and Pachnocybe ferruginea as outgroup. Branches are labeled with maximum likelihood

bootstrap >70%, maximum parsimony bootstrap >50% and Bayesian posterior probabilities >0.95.

Phylogeny

The ITS dataset included sequences from 34 fungal specimens representing

29 species. The dataset had an aligned length of 617 characters, of which

315 are constant, 106 are variable and parsimony-uninformative, and 196

are parsimony-informative. Maximum parsimony analysis yielded 12

522 ... Ma &al.

equally parsimonious trees (TL = 1143, CI = 0.434, HI = 0.566, RI = 0.454,

RC = 0.197). Best model for the ITS dataset estimated and applied in the

Bayesian analysis: GIR+1+G, lset nst = 6, rates = invgamma; prset statefreqpr

= dirichlet (1,1,1,1). Bayesian analysis and ML analysis resulted in a similar

topology as MP analysis, with an average standard deviation of split

frequencies = 0.008724 (BI).

The Maximum Parsimony strict consensus tree (Fic. 1) inferred from ITS

sequences within Septobasidium demonstrated that the new species formed a

monophyletic lineage with strong support (ML = 100%; MP = 100%; PP = 1)

and formed a sister clade with “S. cokeri” Couch [nom. inval.] and forming a

group with S. hainanense C.X. Lu & L. Guo and S. maesae C.X. Lu & L. Guo.

Taxonomy

Septobasidium aquilariae C.L. Zhao, sp. nov. Figs 2, 3

MB 830332

Differs from “Septobasidium cokeri” by its slightly brown to brown hymenial surface

upon drying and its growth on Aquilaria sinensis.

Type: China. Yunnan Province: Xishuangbanna, Jinghong, Menghai County, in

association with Pseudaulacaspis sp. on Aquilaria sinensis, 31 May 2018, CLZhao 6611

(Holotype, SWFC 006611; MK802140).

ErymMo.oey: The specific epithet aquilariae (Lat.) refers to the plant genus on which all

of the specimens were collected.

BASIDIOMATA on branches, annual, resupinate, hard to separate from substrate,

becoming coriaceous upon drying, up to 8 cm long, up to 4 cm wide, up to

1 mm thick. Hymenial surface smooth, cream to pale brown when fresh, slightly

brown to brown upon drying. Sterile margin indeterminate, white to cream.

HYPHAL STRUCTURE monomitic; generative hyphae with simple septa, pale

brown, thick-walled. In section 800-1000 um thick; subiculum pale brown,

50-150 um thick; pillars brown, 300-400 um high, 50-100 um wide; hyphal

layer pale brown, 150-350 um thick; hymenium hyaline or brown, 70-100 um

thick.

Basip1a arising directly from the hyphae without a probasidial cell,

cylindrical, curved, or straight, hyaline, 2-4-celled, 15-26.5 x 4-6 um.

BASIDIOSPORES reniform, hyaline, thin-walled, smooth, (10-)11-19(-20.5)

x (3.5-)4-7.5(-8) um, L = 14.15 um, W = 6.12 um, Q = 1.81-2.52 (n = 150/5).

Haustoria consisting of irregularly coiled hyphae.

Fig. 2. Septobasidium aquilariae (holotype, SWFC 006611). A, B. Basidiomata on branch;

C. Sections of basidiomata; D. Basidiospore; E. Basidia; F. Hyphae; G. Haustoria. Scale bars:

a=5cm;b=2 cm; c-g= 10 um.

Septobasidium aquilariae sp. nov. from China ... 523

524 ... Ma &al.

Fic. 3. Septobasidium aquilariae (holotype, SWFC 006611).

A. Basidiospores; B. Generative hyphae from hyphal layer; C. Basidia. Scale bars = 10 um.

HABITAT AND DISTRIBUTION growing in association with Pseudaulacaspis

sp. on Aquilaria sinensis (Lour.) Spreng. (Thymelaeaceae).

Septobasidium aquilariae sp. nov. from China... 525

ADDITIONAL SPECIMENS EXAMINED: CHINA. YUNNAN PROVINCE. Xishuangbanna:

Jinghong, Menghai County, in association with Pseudaulacaspis sp. on Aquilaria

sinensis, 31 May 2018, CLZhao 6610 (SWFC 006610; MK802139); CLZhao 6612 (SWFC

006612; MK802141); CLZhao 6613 (SWFC 006613; MK802142); CLZhao 6614 (SWFC

006614; MK802143).

Discussion

In the present study, a new species, Septobasidium aquilariae, is described

based on phylogenetic analyses and morphological characters.

Phylogenetically, S. aquilariae is closely related to “S. cokeri,

S. hainanense, and S. maesae (Fic. 1). But morphologically “S. cokeri” differs

from S. aquilariae by its pure white hymenial surface and restricted growth

on Quercus rubra (Gomez & Henk 2004); S. hainanense differs in its purple

hymenial surface and larger (25-36 x 7-13 um) basidia (Lu & Guo 2010a);

and S. maesae differs by its perennial basidiocarps peeled off after maturity

and larger (28-55 x 7.5-11.5 um) basidia (Lu & Guo 2009a).

Several species found in China are morphologically similar to S. aquilariae.

Septobasidium broussonetiae C.X. Lu & al. is distinguished by its cracking

basidiocarps and growth on Broussonetia papyrifera (Lu & al. 2010);

S. brunneum Wei Li bis & L. Guo differs in its purple-brown hymenial

surface with many cracks and growth on Eurya sp. (Li & Guo 2014);

S. capparis S.Z. Chen & L. Guo differs by its thicker (<2 mm thick) section

and larger (45-56 x 8-12 um) basidia (Chen & Guo 2012); S. euryae-groffii

C.X. Lu & L. Guo is distinguished by its cinnamon to chestnut brown

hymenium and growth on Eurya groffii (Lu & Guo 2010b); S. fissuratum

Wei Li bis & L. Guo differs in its larger (32-45 x 6-9 um) basidia and growth

on Castanea sp. (Li & Guo 2013); S. gaoligongense C.X. Lu & L. Guo differs

in its dark brown hymenium and thinner (260-580 um) section (Lu & Guo

2010b).

Septobasidium guangxiense Wei Li bis & L. Guo differs from S. aquilariae

in its yellowish brown hymenium with numerous fissures at maturity and

larger (27-38 x 5-10 um) basidia (Li & Guo 2014); S. hoveniae Wei Li bis

& al. differs in its cinnamon-brown hymenium and growth on Hovenia

acerba (Li & al. 2013); S. polygoni C.X. Lu & L. Guo differs in its white

to cinnamon-brown hymenium and growth on Polygonum campanulatum

(Lu & Guo 2010b); S. reevesiae S.Z. Chen & L. Guo differs in its thicker

(1.65-2.20 mm) section and larger (37-55 x 8-13 um) basidia and growth

on Reevesia longipetiolata (Chen & Guo 2012); and S. transversum Wei Li

bis & L. Guo differs in its cinnamon-brown basidiomata, its transverse

526 ... Ma &al.

layer at the pillar bases, and larger (42-60 x 9-16 um) basidia (Li & Guo

2014).

The diversity of Septobasidium is rich in China and 57 species have been

reported, especially in subtropics and tropics (Lu & Guo 2009a,b, 2010a,b,

2011; Lu & al. 2010; Chen & Guo 2011, 2012; Li & Guo 2013, 2014). Several

Septobasidium species have been described from Yunnan Province (Lu &

Guo 2010b, 2011; Li & Guo 2013), and our new species, S. aquilariae, is

also from Yunnan Province. It is expected that new taxa will be found after

further research.

Acknowledgments

Special thanks are due to Dr. Sana Jabeen (Division of Science and Technology,

University of Education, Lahore, Pakistan) and Dr. Jie Song (Research Institute

of Tropical Forestry, Chinese Academy of Forestry, P.R. China) who reviewed the

manuscript. The research is supported by the National Natural Science Foundation

of China (31700023), Yunnan Agricultural Foundation Projects (2017FG001-042),

Science Foundation of Southwest Forestry University (Project No. 111715) and the

Science Foundation of Education Department in Yunnan (2018JS326).

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MY COTAXON

July-September 2019—Volume 134, pp. 529-534

https://doi.org/10.5248/134.529

Limacella bangladeshana,

first record of the genus in Thailand

JATURONG KUMLA’, NAKARIN SUWANNARACH’, SAISAMORN LUMYONG**

' Department of Biology, Faculty of Science & Center of Excellence in Microbial Diversity and

Sustainable Utilization, Chiang Mai University, Chiang Mai, 50200, Thailand

? The Royal Society of Thailand, Academy of Science, Bangkok 10300, Thailand

* CORRESPONDENCE TO: saisamorn.l@cmu.ac.th

ABSTRACT—Mushroom specimens collected in Thailand were identified as Limacella

bangladeshana, based on morphology and LSU and ITS sequence analyses. A full description

and illustration are provided. This is the first record of the genus Limacella from Thailand.

Key worps—Agaricales, Amanitaceae, gill fungus, taxonomy

Introduction

Limacella (Amanitaceae, Agaricales) was described by Earle (1909) with

L. delicata (Fr.) Earle ex Konrad & Maubl. as the type species. This genus is

mainly distributed in Europe and America, but some species are found in Asia

(Smith 1945; Hongo 1978; Pegler 1983, 1986; Yang & Chou 2002; Neville &

Poumarat 2004; Sato & al 2010; Ferreira & al. 2013; Kumari & al. 2013; Yang

2015; Hosen & Li 2017; Nascimento & Wartchow 2018; Yang & al. 2018).

Approximately 47 taxa of Limacella are known (http://www.indexfungorum),

but no Limacella species have been reported from Thailand (Chandrasrikul

& al. 2011). During a taxonomic survey of macrofungi collected in northern

Thailand, we found specimens that corresponded to the description of

L. bangladeshana, previously reported from Bangladesh (Hosen & Li 2017).

Herein we describe and illustrate the morphological characteristics of the

Thai material and provide supporting evidence based on LSU and ITS gene

sequence matches in GenBank.

530 ... Kumla, Suwannarach, Lumyong

Materials & methods

Morphology

Basidiomata were collected from the campus of Chiang Mai University,

Chiang Mai Province, Thailand, and kept in plastic collection boxes until

transported back to the laboratory. Macroscopic descriptions are based on

observations of the fresh basidiomata. Color names and codes follow Kornerup

& Wanscher (1978). The specimens were dried at 40—45 °C in an electric

food dryer. The micromorphological data were derived from dried specimens

mounted in 95% ethanol and rehydrated in distilled water, 3% KOH, or Melzer’s

reagent. Dimensions of anatomical features were calculated from at least 50

measurements of each structure. For basidiospores, Q represents the range of

length/width ratios from individual spores and is the mean + standard deviation

of the ratios from all measured spores. Dried specimens were deposited in the

herbarium of Research Laboratory for Excellence in Sustainable Development

of the Biological Resources, Faculty of Science, Chiang Mai University, Thailand

(SDBR-CMU).

Sequence analyses

Genomic DNA was extracted from fresh specimens using a Favorgen DNA

Extraction Mini Kit following the manufacturer's instructions. The large subunit

(LSU) region of ribosomal DNA (rDNA) was amplified by polymerase chain

reaction (PCR) with LROR/LRS primers, and the rDNA internal transcribed spacer

(ITS) region was amplified with ITS4/ITSIF primers, under the following thermal

conditions: 94 °C for 2 min; 35 cycles of 95 °C for 30 s, 52 °C for 30 s, 72 °C for 1 min;

and 72 °C for 10 min. PCR products were checked under UV light on 1% agarose

gels stained with ethidium bromide and purified using a NucleoSpin Gel and PCR

Clean-up Kit following the Macherey-Nagel protocol. The purified PCR products

were directly sequenced according to the Sanger method by 1°" Base Company in

Kembangan, Malaysia, using the same PCR primers cited above. Sequences were

used to query GenBank database via BLAST (http://blast.ncbi.nlm.nih.gov/Blast.cgi).

Taxonomy

Limacella bangladeshana Hosen, Phytotaxa 332: 281 (2017). FIG. 1

Pileus 20-50 mm diam., subconical to sub-hemispherical, becoming

planoconvex, surface dry, orange white (5A2) to brown (7E4), slightly

darker at the centre; margin non-striate, with brownish orange (6C4) to

light orange (5A4) at edge, covered with white velvety to cottony squamules.

Context 5-8 mm thick at the center of the pileus, white, unchanging when

cut or bruised. Lamellae 22—30 x 7-10 mm, free, white to brownish orange

(6C4) or brown (7E4) to dark brown (7F4) when dried. Stipe 25-50 x 5-12

mm, cylindrical to subcylindrical, slight swollen at base, dry, not slimy,

Limacella bangladeshana new to Thailand ... 531

Fic. 1. Limacella bangladeshana (SDBR-CMU-JK0146).

A. basidiomata; B. basidiospores; C. basidia; D. pileipellis.

Scale bars: A = 10 mm; B, C=5 um; D = 10 um.

surface covered with fibrillose or floccose squamules, white to brownish

orange (6C4). Annulus absent.

Basidiospores 3—5 x 3—4.5 um, Q = 1.00—-1.11, = 1.04 + 0.06, globose

to (rarely) subglobose, hyaline, inamyloid, thin-walled, smooth; apiculus

cylindrical to subcylindrical <1.2 um long. Basidia 20-30 x 5—8 um,

clavate, hyaline, thin-walled, 4-spored, with sterigmata <4 um long, with

basal clamp connections. Subhymenium <35 um thick, with 2—3 layers

of inflated to subglobose cells, thin-walled, hyaline. Hymenophore trama

532 ... Kumla, Suwannarach, Lumyong

bilateral, filamentous hyphae 3—9 um diam. mixed with inflated cells 15-32

x 10-20 um. Lamellar edge fertile. Cheilocystidia and pleurocystidia absent.

Pileipellis an ixo-trichodermium composed of more or less erect hyphae

embedded in a gelatinized matrix, hyphae septate, hyaline, cylindrical,

branched, with terminal cell 25—75 x 5—8 um, clamp connections present.

Stipe surface hyphae interwoven, filamentous, hyaline, thin-walled 3—5 um

diam., cells not inflated; stipe trama hyphae longitudinally arranged, 4—8

um diam., mixed with acrophysalidic cells 10-16 um diam. thin-walled,

hyaline to pale yellow, clamp connections rare.

SPECIMENS EXAMINED—THAILAND, CHIANG MAI PROVINCE, Muang

District, Chiang Mai University, 18°48’03”N 98°57’21”E, elevation 334 m, on

soil, 2 June 2018, J. Kumla & N. Suwannarach (SDBR-CMU-NK0356; GenBank

MK288133, MK290392); 3 June 2018, J. Kumla (SDBR-CMU-JK0146, GenBank

MK290393, MK290394).

Molecular analysis

The LSU and ITS sequences of specimens SDBR-CMU-NK0357 and

SDBR-CMU-JK0146 were deposited in GenBank. Our LSU and ITS sequences

were 100% similar to L. bangladeshana ex-type sequences KR816866 and

KR816865.

Discussion

The agaricoid basidiomata, slightly glutinous pileus, a fertile lamellar edge,

lack of annulus, a stipe that is not viscid, and a trichodermium pileipellis

embedded in a gelatinous matrix support the placement of our collections

in Limacella (Singer 1986, Tulloss & al. 2016). According to the descriptions

provided by Singer (1986), this genus is divided into two sections based mainly

on the stipe characters. There are L. sect. Lubricae (viscid or glutinous stipe)

and L. sect. Limacella (dry stipe). Based on its dry stipe, L. bangladeshana

belongs to L. sect. Limacella (Hosen & Li 2017).

Thirteen Limacella species have been reported from Asia: L. anomologa

(Berk. & Broome) Pegler, L. asperulospora Corner, L. bangladeshana,

L. glioderma (Fr.) Maire, L. illinita (Fr.) Maire, L. magna B. Kumari &

R.C. Upadhyay, L. myxodictyon (Berk. & Broome) Pegler, L. ochraceolutea

P.D. Orton, L. olivaceobrunnea Hongo, L. quilonensis Sathe & J.T. Daniel,

L. singaporeana Corner, L. subglischra (S. Imai) S. Ito, and L. taiwanensis Zhu

L. Yang & W.N. Chou (Corner 1994, Hosen & Li 2017, Imai 1938, Kumari &

al. 2013, Pegler 1986, Sathe & Daniel 1981, Sato & al. 2010, Yang 2015, Yang &

Chou 2002). Limacella asperulospora, L. glioderma, L. illinita, L. myxodictyon,

Limacella bangladeshana new to Thailand ... 533

L. ochraceolutea, L. olivaceobrunnea, and L. singaporeana_ with buff white,

dark cinnamon to reddish brown, pale brown, dark reddish brown, dark

yellowish brown colors and larger (3.5—12.5 x 3—7 um) basidiospores with

higher (1.05—1.2) values (Hongo 1978, Pegler 1986, Corner 1994, Yang

2015) are distinguished from L. bangladeshana, with orange white to brown

basidiomata and globose basidiospores (3—5 x 3—4.5 um) with a lower value

(1.04) (Hosen & Li 2017). The yellowish to pale orange basidiomata distinguish

L. subglischra (Imai 1938) from L. bangladeshana. Shorter basidiospores

separate L. bangladeshana from L. anomologa (4—5.7 x 2.7—3.3 um), L. magna

(7-12.5 x 5-7 um), and L. quilonensis (7-10 x 3—4 um) (Sathe & Daniel

1981, Pegler 1986, Kumari & al. 2013), while the subglobose basidiospores of

L. taiwanensis (3—4.5 x 3—3.5 um; Yang & Chou 2002) with a verruculose surface

clearly distinguish that species from L. bangladeshana. The LSU molecular

analysis supports the morphological differences between L. bangladeshana and

other Limacella species (Hosen & Li 2017).

The LSU and ITS sequence matches in GenBank confirmed that the two

Thai specimens belong to L. bangladeshana. Therefore, the combination of

morphological and molecular characteristics supports the identification of

L. bangladeshana, a new record in Thailand.

Acknowledgments

This work was supported by grants from Chiang Mai University and Center of

Excellence in Microbial Diversity and Sustainable Utilization, Chiang Mai University.

We also thank Dr Steve L. Stephenson (University of Arkansas, Fayetteville AR USA)

and Dr Eric H.C. McKenzie (Manaaki Whenua Landcare Research, Auckland NZ) for

their helpful comments and careful review of this manuscript.

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description. Phytotaxa 332: 280-286. https://doi.org/10.11646/phytotaxa.332.3.4

534 ... Kumla, Suwannarach, Lumyong

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MY COTAXON

July-September 2019—Volume 134, pp. 535-543

https://doi.org/10.5248/134.535

New records of Acarospora and Psora from China

CHUN-XIAO WANG*, CHUAN-FENG ZHENG*, ZUN-TIAN ZHAO’

Key Laboratory of Plant Stress Research, College of Life Sciences,

Shandong Normal University, Jinan, 250014, P. R. China

“CORRESPONDENCE TO: chunxiao6wang@163.com

ABSTRACT—Acarospora impressula, A. umbilicata, Psora gresinonis, and P. himalayana are

reported for the first time from China and Psora decipiens for the first time from Qinghai

Province. Descriptions, illustrations, and distributions are given for each species.

Keyworps —Acarosporaceae, East Asia, lichenized fungi, Psoraceae, taxonomy

Introduction

Acarospora (Acarosporaceae, Acarosporales) was established by

Massalongo (1852). The genus is characterized mainly by crustose (often

distinctly areolate or subsquamulose) thalli, 16-100+-spored asci with

non-amyloid apical domes, and hymenium typically over 100 um thick

(Knudsen 2007, Fletcher & al. 2009). The genus contains about 200 species

(Licking & al. 2016), usually on siliceous rocks, rarely on calcareous

rocks, wood, and soil, often in arid habitats (Knudsen 2007, Fletcher &

al. 2009, Ovstedal & al. 2018). In China, 35 species have been reported

(Dongling & al. 2017, Knudsen 2007, Wei 1991, Nurtai & al. 2019).

Psora (Psoraceae, Lecanorales) comprises 30 species growing on soil and

rock, mainly in arid areas, in arctic to subtropical regions (Timdal 2002).

The genus is characterized mainly by squamulose thalli, a hypothecium

containing calcium oxalate crystals, porpidia-type asci, laminal pycnidia,

and acrogenous bacilliform pycnoconidia. In China, 11 species have been

reported (Abbas 1998, Mamut & Abbas 2016, Wei 1991).

* CHUN-XIAO WANG & CHUAN-FENG ZHENG contributed equally to this manuscript.

536 ... Wang, Zheng, Zhao

In this paper we provide valuable information on Acarospora and Psora

in China in preparation for a revised edition of Wei’s (1991) book, AN

ENUMERATION OF LICHENS IN CHINA. Here, we identify four species new

to the country—Acarospora impressula, A. umbilicata, Psora gresinonis,

P. himalayana—and report P. decipiens for the first time from Qinghai

Province.

Materials & methods

The specimens studied were collected in Inner Mongolia Autonomous Region

(Neimenggu) and Qinghai Province, China, and are preserved in Lichen Section

of the Botanical Herbarium, Shandong Normal University, Jinan, China (SDNU).

The specimens were examined morphologically under a COIC XTL7045B2

stereomicroscope and Olympus CX41 polarizing microscope. Thallus and medulla

were tested for identification with K (10% potassium hydroxide aqueous solution),

C (saturated aqueous sodium hypochlorite solution), and I (Lugol’s iodine). The

lichen substances were identified using standardized thin layer chromatography

techniques (TLC) with system C (Orange & al. 2001). The lichens were

photographed using an Olympus DP72 camera attached to Olympus SZX16 and

BX61 microscopes.

Taxonomy

Acarospora impressula Th. Fr., Lich. Scand. 1: 214 (1871) Fie. 1

THALLUS areolate, rugulose, with cracks between areoles, 0.3-0.7 mm

wide, 0.3-0.5 mm high. Upper surface brown, slightly white pruinose.

Epinecral layer 20 um. Cortex 22.5-37.5 um thick. Algal layer uneven,

interrupted by hyphal bundles, <175 um thick, algal cells 10-17.5 um in

diam. Medulla white, plectenchymatous, <300 um thick. APOTHECIA

0.36—0.41 mm wide, disc dark brown. Exciple hyaline, distinct, 2.5-125 um

wide. Epihymenium light yellow, 12.5-17.5 um tall. Hymenium hyaline,

45—75 um tall; paraphyses hyaline, simple or slightly branched, conglutinate.

Asci hyaline, 62.5—75 x 15-17.5 um, 100-200-spored; ascospores simple,

ellipsoid, 2-3 x 5-6 um. Subhymenium hyaline, 12.5-52.5 um tall.

Hypothecium hyaline, indistinct. Algal layer uneven, interrupted by hyphal

bundles, <150 um thick, algal cells 7.5-15 um in diam. Medulla containing

many crystals.

CHEMISTRY—Thallus K-, C-, KC-—, hymenium I+ blue; no secondary

metabolites detected by TLC.

SPECIMEN EXAMINED—CHINA. NEIMENGGU, Hu He Ba Shi Ge, alt. 1600 m, 16 Aug.

2011, D.B. Tong 20123540 (SDNU).

Acarospora & Psora species new for China ... 537

Fic. 1. Acarospora impressula (Tong 20123540, SDNU). A, B. Apothecia section; C. Thallus and

apothecia; D. Apothecia; E. Asci; F. Ellipsoid ascospores.

ECOLOGY & DISTRIBUTION—On siliceous or calcareous rock. Europe, Asia,

North Africa, Greenland, North America, Australia (Temina & al. 2005,

Ovstedal & Gremmen 2014). New to China.

CoMMENTS— Through studying Acarospora from China, it was found that

A. brevilobata H. Magn. and A. gobiensis H. Magn. have similar ascospores

to A. impressula but differ in their thallus, which is circular in A. brevilobata

and yellow in A. gobiensis (Abbas 1998, Temina & al. 2005). Our specimen

538 ... Wang, Zheng, Zhao

closely matches the modern description of A. impressula in Ovstedal &

Gremmen (2014).

Acarospora umbilicata Bagl., Mém. R. Accad. Sci. Torino,

Ser. 2, 17: 397 (1857) Fig?2

THALLUS squamulose, convex, 0.4-1.4 mm wide, 0.1-0.3 mm high. Upper

surface grey-brown, densely white-pruinose. Epinecral layer 20-25 um tall.

Cortex 22.5 um thick, with crystals. Algal layer even, 30-112.5 um thick,

algal cells 7.5-17.5 um in diam. Medulla white, plectenchymatous, scattered

to contiguous with crystals in longitudinal sections. ApoTHecta black,

disc depression 0.19-1.28 mm. Exciple hyaline, distinct, 25-38 um wide.

Epihymenium brown, 20-30 um tall. Hymenium hyaline, 37.5-100 um tall,

paraphyses hyaline, simple, apex expanded, conglutinate. Asci 87.5-95 x

20—22.5 um, 100-200-spored; ascospores simple to 1-septate, 4—5 x 1.5—2 um,

ellipsoid. Subhymenium hyaline, 25-40 um tall. Hypothecium hyaline, 35-60

um tall. Algal layer uneven, interrupted by hyphal bundles, contains crystals,

<100 um thick, algal cells 7.5-12.5 um in diam. Medulla containing many

crystals, 50—162.5 um tall.

ate sir

tall I 44

MIE

i (Uy

Ocm1

aos oe / ie v4 254nm = 365nm light

before before after acid

Fic. 2. Acarospora umbilicata (Tong 20123044, SDNU). A. Thallus and apothecia; B, C. Thallus

section; D, E. Apothecia section; F. Asci; G. Ascospores; H. Asci I+ blue; I. TLC chromatograms.

Acarospora & Psora species new for China ... 539

CHEMISTRY— Thallus K-, C+ red, KC+ red; hymenium I+ blue. Secondary

metabolites: gyrophoric acid.

SPECIMEN EXAMINED — CHINA. NEIMENGGU, Ho-lan Mountains, alt. 1500 m, 19 Aug.

2011, D.B. Tong 20123044 (SDNU).

ECOLOGY & DISTRIBUTION—On siliceous rock. Europe, Asia, North Africa,

Canary Islands, Madeira Islands (Temina & al. 2005, Galloway 2007). New to

China.

CoMMENTS—Acarospora fulva N.S. Golubk., A. fuscata (Nyl.) Th. Fr.,

A. glypholecioides H. Magn., and A. jenisejensis H. Magn. are similar

to A. umbilicata by the C+ red reaction indicating gyrophoric acid as

secondary metabolites. However their thalli differ, lacking pruina in

A. fuscata, colored yellow in A. fulva, and wider in A. glypholecioides and

A. jenisejensis. Acarospora glypholecioides and A. jenisejensis are further

distinguished by their shorter ascospores (A. glypholecioides, 3—3.5 x 1.7

um; A. jenisejensis, 3—3.5 x 2.5-3 um; Abbas 1998, Temina & al. 2005,

Galloway 2007, Knudsen 2007). Our specimen agrees closely with modern

descriptions of A. umbilicata (Timdal 1987, Galloway 2007).

Psora decipiens (Hedw.) Hoffm., Descr. Pl. Cl. Crypt. 2: 68 (1794) Fic. 3A-D

SQUAMULES <6 mm wide, adnate, scattered to contiguous, concave to

convex; upper surface bright red, epruinose or partly to entirely white pruinose,

sparingly fissured; margin white, slightly up-turned, soon becoming crenulate;

upper cortex <150 um thick, containing crystals of calcium oxalate; medulla,

containing calcium oxalate crystals; lower cortex poorly developed or absent.

APOTHECIA $2.5 mm diam., marginal, immarginate, black, epruinose. Asc

8-spored, ascospores ellipsoid, simple, hyaline, 10-15 x 6-9 um. PYCNIDIA not

seen.

CHEMISTRY— thallus K-, C-, KC-, hymenium I+ blue; no secondary

metabolites detected.

SPECIMEN EXAMINED: CHINA. QinGHAI, Haibeizhan Fengxiakou alpine meadow, alt.

3280 m, 6 Aug. 2007, C. Yuan 20070787 (SDNU).

ECOLOGY & DISTRIBUTION—On soil. Africa, Asia, Australia, Europe, New

Zealand, and North and South America (Timdal 2002, Galloway 2007). New

to Qinghai Province.

CoMMENTS—Psora decipiens is characterized by its bright red squamules,

marginal apothecia, white margin, and absence of chemical substances. It is

closely related to P. crenata, which differs by its broader and thicker (10 mm)

540 ... Wang, Zheng, Zhao

squamules, usually with a regular central depression, and a down-turned, more

or less entire margin. Our specimen is closely similar to modern descriptions

of P. decipiens (Timdal 1987, Galloway 2007).

Psora gresinonis B. de Lesd., Bull. Soc. Bot. France 77: 614 (1930) Fic. 3E-H

SQUAMULES discrete, attached to the substrate, concave, 2—3 mm wide;

upper surface dark brown; margin darker with upper side; upper cortex <60

uum thick, lacking calcium oxalate crystals; lower cortex poorly developed.

APOTHECIA $1.4 mm diam., laminal or submarginal, immarginate, black. Asc1

8-spored, ascospores ellipsoid, simple, hyaline, 10—15 x 7.5-10 um. PYCNIDIA

not seen.

CHEMISTRY—Cortex K—, C-, medulla C—, K+ red, KC+ red; norstictic acid

detected by TLC.

SPECIMEN EXAMINED: CHINA. QiNGHAI, Haibeizhan Fengxiakou alpine meadow. alt.

3250 m, 6 Aug. 2007, Z.S. Sun 20070741 (SDNU).

ECOLOGY & DISTRIBUTION — On soil. Italy, France (Timdal 1984). New to

China.

CoMMENTS—Psora gresinonis is characterized by its dark brown squamules,

black margin, and norstictic acid. The species is closely related to P. vallesiaca

(Schaer.) Timdal and P. tenuifolia Timdal, which are distinguished by their

reddish brown squamules and white margin (Timdal & Zhurbenko 2004).

Our specimen agrees with the original description of P. gresinonis by Bouly de

Lesdain (1930).

Psora himalayana (C. Bab.) Timdal, Bryologist 89: 262 (1987 [“1986”]) Fic. 31-L

SQUAMULES <4 mm wide, ascending and imbricate; upper surface medium

brown or reddish brown to dark brown, epruinose or more rarely faintly white

pruinose (especially along the margin); margin white, slightly up-turned,

underside brown in center, white near the margin; upper cortex <150 um thick,

sometimes containing calcium oxalate crystals; medulla sometimes containing

calcium oxalate crystals; lower cortex not sharply delimited from the medulla,

containing calcium oxalate crystals. APOTHECIA <2 mm diam., laminal,

immarginate, black, epruinose. Ascr 8-spored, ascospores ellipsoid, simple,

hyaline, 12.5-17 x 5-9 um. PYCNIDIA not seen.

CHEMISTRY— thallus K-, C-, KC-, hymenium I+ blue; no substances

detected by TLC.

SPECIMEN EXAMINED: CHINA. QinGual, National Highway 109, Mt. Xiangpi, alt. 3520

m, 15 Aug. 2007, Y.D. Du & X.L. Shi 20071940 (SDNU).

Acarospora & Psora species new for China... 541

Fic. 3. Psora decipiens (Yuan 20070787, SDNU). A. Thallus and apothecia; B. Apothecia section;

C. Asci; D. Ascospores. Psora gresinonis (Sun 20070741, SDNU). E. Thallus and apothecia;

F. Apothecia section; G. Asci; H. Ascospores. Psora himalayana (Du & Shi 20071940, SDNU).

I. Thallus and apothecia; J. Apothecia section; K. Asci; L. Ascospores.

ECOLOGY & DISTRIBUTION—On soil. Asia, Europe, and western North America

(Timdal 2002). New to China.

ComMENtTS—Psora himalayana is characterized by squamules distinctly white

pruinose along the margin and calcium oxalate crystals present in the medulla

and/or the lower cortex. The species is closely related to P globifera (Ach.)

A. Massal., which differs by its pruina distributed mainly on the upper side

near the squamule margins, and its pale to medium brown lower side near the

542 ... Wang, Zheng, Zhao

squamule margins (Timdal 1987). Psora vallesiaca differs from P. himalayana

mainly in having norstictic acid (Timdal & al. 2016). Our specimen agrees with

the original description of P himalayana by Timdal (1987).

Acknowledgments

The authors thank Dr. Kerry Knudsen (Dept. of Ecology, Czech University of Life

Sciences, Prague) and Dr. Shou-Yu Guo (State Key Laboratory of Mycology, Institute

of Microbiology, CAS, Beijing) for presubmission reviews. We thank Dr. Einar

Timdal (Natural History Museum, University of Oslo, Norway) for providing great

help during the study. This work was supported by Emergency Management Project

of National Natural Science Foundation of China (31750001), the National Natural

Science Foundation of China (31400015), the National Natural Science Foundation of

China Youth Science Foundation (31600100) and the Scientific Research Foundation

of Graduate School of Shandong Normal University (SCX201945).

Literature cited

Abbas A, Wu J, Wu J. 1998. Lichens of Xinjiang. Urumdi, Sci-Tech & Hygiene Publishing House,

124 p.

Bouly de Lesdain M. 1930. Notes lichénologiques. XXIV. Bulletin de la Société Botanique de France

77: 612-615. https://doi.org/10.1080/00378941.1930.10837199

Dongling N, Xinyu W, Yufei B, Lifang H. 2017. A revision of the lichen genus Pleopsidium in China.

Acta Botanica Boreali-Occidentalia Sinica 37(1): 191-195.

Fletcher A, James PW, Purvis OW. 2009. Acarospora. 125-132, in: CW Smith & al. (eds). The

Lichens of Great Britain and Ireland. The British Lichen Society, Natural History Museum

Publications, United Kingdom.

Galloway DJ. 2007. Flora of New Zealand: lichens, revised second edition, including lichen-

forming and lichenicolous fungi. Volumes 1 and 2. Manaaki Whenua Press, Landcare Research.

Knudsen K. 2007. Acarospora. 1-38, in: TH Nash III & al. (eds). Lichen Flora of the Greater Sonoran

Desert Region. Volume 3. Lichens Unlimited, Arizona State University, Tempe, Arizona.

Liicking R, Hodkinson BP, Leavitt SD. 2016. The 2016 classification of lichenized fungi in the

Ascomycota and Basidiomycota - approaching one thousand genera. Bryologist 119: 361-416.

https://doi.org/10.1639/0007-2745-119.4.361

Mamut R, Abbas A. 2016. Preliminary study on the lichen genus Psora in Xinjiang, China. Acta

Botanica Boreali-Occidentalia Sinica 36(7): 1482-1485,

Massalongo AB. 1852. Ricerche sull’ autonomia dei licheni crostosi e materiali pella loro naturale

ordinazione. Verona, Dalla Tipografia di A. Frizierio. 207 p.

Nurtai L, Knudsen K, Abbas A. 2019 [“2018”]. A preliminary study of the yellow Acarospora of

China. Mycotaxon 133(4): 681-701. https://doi-org/10.5248/133.681

Orange A, James PW, White FJ. 2001. Microchemical methods for the identification of lichens.

British Lichen Society, London. 101 p.

@vstedal DO, Gremmen NJM. 2014. Additional lichen records from Subantarctica III. Marion

Island. Australasian Lichenology 74: 2-7.

@Ovstedal DO, Lindblom L, Knudsen K, Fryday AM. 2018. A new species of Acarospora

(Acarosporaceae, Acarosporales, lichenized Ascomycota) from the Falkland Islands (Islas

Malvinas). Phytotaxa 340(1), 86-92. https://doi.org/10.11646/phytotaxa.340.1.7

Acarospora & Psora species new for China ... 543

Temina M, Nevo E, Wasser SP. 2005. The lichen genus Acarospora in Israel and its vicinity. Nova

Hedwigia 80: 433-452. https://doi.org/10.1080/00378941.1930.10837199

Timdal E. 1984. The delimitation of Psora (Lecideaceae) and related genera, with notes on some

species. Nordic Journal of Botany 4: 525-540.

https://doi.org/10.1080/00378941.1930.10837199

Timdal E. 1987 [“1986”]. A revision of Psora (Lecideaceae) in North America. Bryologist 89:

253-275. https://doi.org/10.2307/3243197

Timdal E. 2002. Psora. 418-430, in: TH Nash III & al. (eds). Lichen Flora of the Greater Sonoran

Desert Region. Volume 1. Lichens Unlimited, Arizona State University, Tempe, Arizona.

Timdal E, Zhurbenko MP. 2004. Psora indigirkae, a new species from eastern Siberia, with key to

the Asian species of Psora. Symbolae Botanicae Upsalienses 34(1): 25-29.

Timdal E, Obermayer W, Bendiksby M. 2016. Psora altotibetica (Psoraceae, Lecanorales),

a new lichen species from the Tibetan part of the Himalayas. MycoKeys 13: 35-48.

https://doi.org/10.3897/mycokeys.13.8824

Wei JC. 1991. An enumeration of lichens in China. Beijing, International Academic Publishers.

MY COTAXON

July-September 2019—Volume 134, pp. 545-553

https://doi.org/10.5248/134.545

Phylogenetic affinity and taxonomic reassessment of

Pseudoidium kalanchoes

MONIKA GOTz’, ELKE IDCZAK’', UWE BRAUN**

"Institute for Plant Protection in Horticulture and Forests, JKI, Julius Kiihn Institute,

Federal Research Centre for Cultivated Plants,

Messeweg 11/12, 38104 Braunschweig, Germany

? Martin Luther University, Institute of Biology, Department of Geobotany, Herbarium,

Neuwerk 21, 06099 Halle, Germany

* CORRESPONDENCE TO: uwe.braun@botanik.uni-halle.de

AsBstTRACT—Pseudoidium kalanchoes, retrieved from powdery mildew on Kalanchoe spp. in

Asia, has been re-examined and reassessed based on phylogenetic analyses of the ITS and 28S

rDNA regions. The results confirmed that sequences retrieved from Asian Kalanchoe powdery

mildew have 100% congruence with sequences obtained from Erysiphe sedi on Sedum spp.

A German collection of P kalanchoes on Kalanchoe blossfeldiana was morphologically and

genetically examined and analyzed and is designated as neotype with an ex-neotype reference

sequence. Morphologically, the asexual morph on Kalanchoe spp. agrees with the asexual

morph of E. sedi. Consequently, P. kalanchoes is reduced to synonymy with E. sedi.

Key worps —Erysiphales, “Oidium calanchoeae,’ phylogeny

Introduction

Lustner (1935) introduced the name “Oidium calanchoeae” for an asexual

powdery mildew on Kalanchoe sp. (very probably K. blossfeldiana Poelln.)

from Germany. He failed to present the Latin description or diagnosis

required by ICN (Shenzhen) Art. 39.1, making his binomial invalid; and

his epithet “calanchoeae” required a twofold correction, to kalanchoes, an

orthography based on the accepted name of the host genus (ICN Art. F.9.1),

Kalanchoe Adans. 1763 (instead of Calanchoe Pers. 1805), and use of the

correct genitive case ending (ICN Art. 32.2). Braun (1987: 602) validated

546 ... Gotz, Idczak, Braun

Oidium kalanchoes by adding a Latin description and an effective typification

based on Listner’s original material (although this could not be traced).

The corrected orthography was first published in Braun & al. (2003), and

Braun & Cook (2012) later combined this species in Pseudoidium Y.S. Paul &

J.N. Kapoor.

Pseudoidium kalanchoes has been reported (sometimes under synonyms

or misdeterminations) from Asia, Australasia, Europe, and North America on

crassulacean hosts: Crassula ovata, Kalanchoe blossfeldiana, K. (Bryophyllum)

daigremontiana, K. (Bryophyllum) delagoensis, K. (Bryophyllum) pinnata,

Kalanchoe spp., and Sedum praealtum (Adhikari 2017; Amano 1986;

Boesewinkel 1980; Braun 1987, 1995; Braun & Cook 2012; Braun & al.

2003; Cho & al. 2012; Fang & al. 2017; Ialongo 1992; Jage & al. 2010; Liu

2010; Nagy 1975; Tang & al. 2016; see also TaBLE 1). Chinese collections on

K. blossfeldiana and K. pinnata were assigned to Erysiphe sedi on the basis of

ITS analyses (Tang & al. 2016; Fang & al. 2017). The three hosts classified in

Kalanchoe sect. Bryophyllum are retained in Kalanchoe (rather than in genus

Bryophyllum) on the basis of phylogenetic analyses (Gehrig & al. 2001, Mort

& al. 2010).

The present examinations aimed at re-examining and analyzing German

collections through morphological and molecular methods in order to

clarify the identity of Oidium kalanchoes and to stabilize the application of

this name by neotypification with an ex-neotype reference sequence.

Materials & methods

Collection and maintenance of powdery mildew isolates

The powdery mildew isolate used for sequence analyses was collected from

naturally infected Kalanchoe blossfeldiana plants from Germany in January 2018.

Morphological characterization and molecular analysis described below were

performed using the single spore isolate JKI-GFZ-C20-EM25A, obtained by

transferring a single conidium onto washed cut-off Kalanchoe leaves. Inoculated

cuttings were incubated in plastic boxes at room temperature and natural day-night

periods for two to three weeks until colonies could be detected. To ensure a pure

isolate, four subsequent sub-cultures were performed. Uninoculated leaves served as

controls. Artificially infected leaves were deposited at the Herbarium of the Institute of

Biology, Department of Geobotany and Botanical Garden, Martin Luther University

Halle-Wittenberg, Germany (HAL) as voucher HAL 3298 F.

Morphological characterization

For morphological characterization, fresh powdery mildew structures were

stripped off the leaf with sticky tape and mounted in water. Herbarium samples

Pseudoidium kalanchoes: a synonym of Erysiphe sedi ... 547

were mounted in lactic acid and gently heated. Mycelia from the upper and lower

leaf surfaces were inspected. Morphological characteristics covering size and shape

of conidia (n 2100) and conidiophores (n =50), position of the basal septum, shape

and position of hyphal appressoria, and presence or absence of fibrosin bodies were

assessed. Conidial length and width were analyzed following Frank (1990) with

modifications: the applied values indicate the minimum, lower limit, upper limit, and

maximum values; lower and upper limits indicate the range of 90% of all values (not

95% as described by Frank 1990).

All morphological characteristics were examined using a Zeiss® Axio Imager.Al

standard light microscope equipped with an AxioCam MRc5 camera and differential

interference contrast at 200x, 400x, and 1000x. Measurements were made and images

were taken with the calibrated Axiovision software rel. 4.8, and the images were

processed using Adobe® Photoshop CS4 software version 11.0.

Conidial germination patterns were assessed following the method of Zaracovitis

(1965) with slight modifications. Preferably young conidia (~ 24 h) were dusted onto

microscope slides and incubated in a moist chamber at 20 + 1 °C in the dark for 24-48

h and analysed and documented as above. The patterns were compared with those

described in Cook & Braun (2009).

Molecular characterization

Total DNA was extracted from conidia and mycelia using the Qiagen DNeasy

plant mini kit following the manufacturer's instructions with slight modifications.

The internal transcribed spacer (ITS) was amplified using the primers ITS5 and PM6

(Takamatsu & Kano 2001), and the 28S rRNA was amplified using PM5 (Takamatsu

& Kano 2001) and NLP2 (Hirose & al. 2005). The reaction mix contained Hot FirePol

Mastermix (containing 7.5 mM MgCl ), 0.2 uM of each primer, and 4 ul template DNA

in a total volume of 50 ul. PCR was performed in a MyCycler thermal cycler with an

initial denaturation step at 95 °C for 13 min; followed by 40 cycles of 30 s at 94 °C for

denaturation, 30 s/50 s (ITS/28S rRNA) at 52 °C for annealing, and 30 s/50 s (ITS/28S

rRNA) at 72 °C for extension; and a final extension for 10 min at 72 °C. PCR products

were purified using a MSB Spin PCRapace Kit. Fragments were bidirectionally

sequenced twice with the same primers used for PCR by LGC Genomics GmbH. A

contig was generated and edited using CLC Main Workbench 8.1 following the EPPO

recommendations for sequence analysis (OEPP/EPPO 2016). The consensus sequence

was deposited in GenBank under the accession number MK411006.

The sequence obtained for the isolate JKI-GFZ-C20-EM25A was aligned to

sequences of three Erysiphe sedi isolates and 12 additional isolates from the Erysiphe

aquilegia clade (see Takamatsu & al. 2015 for further details of the sequences

concerned) spanning the same region using CLC Main Workbench 8.1 with the

settings gap open cost 10.0 and gap extension cost 1.0 and the very accurate alignment

mode. Erysiphe linderae (LC010067; Abasova & al. 2018) was chosen as outgroup

taxon. The alignment was manually refined using CLC Main Workbench.

A phylogenetic tree was constructed using the Neighbor-Joining method and

the nucleotide substitution model Kimura’s (1980) two parameter model with a

548 ... Gotz, Idczak, Braun

transition/transversion ratio of 2.0 (Fic. 1). The strength of the internal branches of

the resulting tree was tested with bootstrap analysis using 1000 replications (CLC

Main Workbench 8.1).

Results

Phylogenetic analysis

Sequence data including genes for ITS1, 5.88 rRNA, ITS2, 28S rRNA

(partial and complete) obtained from the isolate JKI-GFZ-C20-EM25A

(MK411006) showed 100% identity to three sequences of E. sedi (LC010045,

LC010046, LC010047) spanning the same region deposited in GenBank

(Fic. 1). They were also identical to sequences of E. hommae (LC009926)

and Pseudoidium hortensiae (LC009915). All sequences belong to the

Erysiphe aquilegiae clade (see Takamatsu & al. 2015), which is characterized

by insufficient resolution on species level, indicating that the resolution of

the region analyzed is not high enough to distinguish all members of this

clade. However, both morphological characteristics and host range support

assignment of JKI-GFZ-C20-EM25A to E. sedi.

Erysiphe sedi LC010047

Erysiphe sedi LC010045

7i¢ Pseudoidium pedaliacearum LC342967

Pseudoidium pedaliacearum LC342968

Erysiphe hommae LC009926

Erysiphe sedi LC010046

Pseudoidium neolycopersici LC009912

Pseudoidium hortensiae LC009915

Erysiphe aquilegiae LC009942

Erysiphe chloranthi LC009931

Erysiphe circaeae LC010044

Erysiphe knautiae LC010042

Erysiphe caricae-papayae _LC228614

Erysiphe caricae-papayae LC228613

Erysiphe euphorbiae LC010073

Erysiphe linderae LC010067

0,018

Fic. 1. Phylogenetic analysis (combined data of ITS1, 5.88 rRNA, ITS2, and 28S rRNA

complete and partial sequences) for the isolate JKI-GFZ-C20-EM25A (framed in red)

including 15 selected sequences of the Erysiphe aquilegiae clade. The bootstrap support value

for maximum likelihood >70% is presented above the node. The tree is rooted to Erysiphe

linderae (LC0O10067).

Morphology

The following description is based on collections on Kalanchoe

spp. (Eliade 1990, Bolay 2005, Liu 2010, Braun & Cook 2012, present

examinations):

Pseudoidium kalanchoes: a synonym of Erysiphe sedi ... 549

Erysiphe sedi U. Braun, Feddes Repert. 92(7-8): 502, Sept. 1981. FIG. 2

= Erysiphe sedi R.Y. Zheng & G.Q. Chen, Sydowia 34: 253, 31 Dec. 1981, nom. illeg.

= Oidium kalanchoes Listner ex U. Braun, Beih. Nova Hedwigia 89: 602, 1987, as

“calanchoeae” [validation of “Oidium calanchoeae” Listner, Nachrichtenbl. Deutsch.

Pflanzenschutzdienst 15(4): 41, 1935, nom. inval., ICN (Shenzhen) Art. 39.1].

TypPE: Germany, on leaves of Kalanchoe blossfeldiana, 1935, G. Lustner (holotype;

not preserved). Germany, Niedersachsen, Braunschweig, Julius Kiihn Institute,

greenhouse, on living leaves of Kalanchoe blossfeldiana, Jan. 2018, M. Gotz (neotype,

HAL 3298 FE, designated here MBT 385509; GenBank MK411006.)

= Pseudoidium kalanchoes (Liistner ex U. Braun) U. Braun &

R.T.A. Cook, Taxon. Man. Erysiphales: 608, 2012.

Mycelium amphigenous, also caulicolous, forming white or grayish

white patches, sometimes confluent or effuse, persistent. HyPHAE straight

to flexuous-sinuous, branched, (2—)4—8(-10) um wide, septate, hyaline,

thin-walled, smooth. HyPHAL AppRESSORIA Solitary or in opposite pairs,

1-4 per cell, almost nipple-shaped to distinctly lobed, 5-10 um diam.

CONIDIOPHORES arising from the upper surface of mother cells, erect,

40-200 um long. Foot-ce.ts cylindrical, straight, (15-)18-50(-60) x

6-11 um, followed by (1-)2(-3) cells, usually about as long as the foot-cell,

shorter or even longer, occasionally followed by a very long cell up to 90 um,

forming conidia singly. Conrp14 cylindrical to ellipsoid, (22—)30-55(-85) x

(10.5—)14—22(-25) um, length/width ratio (1.5—)1.7-3.2(-3.8). GERM TUBES

Pseudoidium type, occasionally in a longitubus pattern, usually with a single

perihilar germ tube, rarely with 2-3 germ tubes, usually short to moderately

long, straight, unbranched and aseptate, occasionally with a single septum,

very rarely branched, apex usually with a lobed appressorium, sometimes

only swollen and unlobed.

SPECIMENS EXAMINED—GERMANY. HEssen: sine loco: on living leaves of Kalanchoe

blossfeldiana, 1.1V.1998, J. Dalchow (HAL 405 F); Wiesbaden, on living leaves of Crassula

ovata, 12.V.2003, S. Krause (HAL, s.n.). NIEDERSACHSEN: Braunschweig, on living

leaves of Crassula ovata, 1.2003, U. Brielmaier-Liebetanz (HAL, s.n.); Braunschweig,

Julius Kuhn Institute, greenhouse, on living leaves of Kalanchoe blossfeldiana, 1.2018, M.

Gotz (HAL 3298 F, neotype).

Discussion

Until the application of molecular methods for the purpose of

identification of powdery mildew species, the names Pseudoidium/

Oidium kalanchoes were widely used for an asexual morph occurring

on Kalanchoe blossfeldiana and other Kalanchoe species (including

Bryophyllum) as well as Crassula ovata (Eliade 1990, Braun 1995, Braun

& al. 2003, Bolay 2005, Liu 2010, Braun & Cook 2012). First molecular

550 ... Gotz, Idezak, Braun

Fic. 2. Pseudoidium kalanchoes on Kalanchoe blossfeldiana (neotype, HAL 3298 F):

A-D. Conidiophores; E-H. Conidia; I-L. Conidial germination types in vitro [I. Longitubus

pattern; J-L. Pseudoidium type]; M-P. Hyphal appressoria (background removed with Adobe

Photoshop to clarify relevant features of the conidiophores). Scale bars = 10 um.

Pseudoidium kalanchoes: a synonym of Erysiphe sedi ... 551

examinations of this powdery mildew in Asia on Kalanchoe spp. raised

serious doubts as to the taxonomic status and true affinity of this

anamorph-typified powdery mildew species (Cho & al. 2012, Tang

& al. 2016, Fang & al. 2017). On the basis of analyses of ITS data and

corresponding morphological traits, these authors identified Chinese and

Korean powdery mildew specimens on Kalanchoe spp. as Erysiphe sedi.

Therefore, it was logical to assume that P kalanchoes must be considered

a heterotypic synonym of E. sedi. In order to confirm this assumption,

it was necessary to neotypify P kalanchoes (type not preserved) with a

German collection on K. blossfeldiana and to retrieve an ex-neotype

reference sequence. A corresponding neotype has been designated and

morphologically as well as phylogenetically examined. The morphology of

the neotype material coincided with the asexual morph of E. sedion Sedum

spp. and the ex-neotype ITS1-5.8S rRNA-ITS2-28S rRNA sequence shows

a congruence of 100% with sequences retrieved from E. sedi on Sedum

and Kalanchoe spp. The results of a combination of morphological and

phylogenetic analyses justify reducing P kalanchoes to synonymy with

E. sedi, while the limited resolution of ITS data on species level within the

E. aquilegiae clade (see Takamatsu & al. 2015) may not be concealed.

Multilocus analyses of this clade are necessary but not yet available.

TABLE 1. Host range and distribution of Erysiphe sedi on Kalanchoes spp.

Host

Crassula ovata

Kalanchoe blossfeldiana

Kalanchoe daigremontiana;

= Bryophyllum daigremontianum

Kalanchoe delagoensis;

= Bryophyllum delagoense;

= Kalanchoe tubiflora

Kalanchoe pinnata;

= Bryophyllum pinnatum

Kalanchoe spp. [incl. hybrids]

Sedum praealtum

COUNTRY

Germany

South Korea; Australia;

Germany, Hungary,

Norway, Romania,

Switzerland; USA

China, Nepal

Germany; New Zealand

China

Italy, Netherlands; Canada

New Zealand

REFERENCE

Braun & al. 2003

von Arx 1952, Nagy 1975,

Spencer 1978, Amano 1986,

Talongo 1992, Braun 1995,

Jage & al. 2010

Liu 2010

Adhikari 2017

Braun & al. 2003;

Boesewinkel 1980, as

“Microsphaera polonica’

Tang & al. 2016

Amano 1986

Boesewinkel 1980, as

“Microsphaera polonica’

552 ... Gotz, Idcezak, Braun

Acknowledgments

The authors thank Roger T.A. Cook (York, UK) and Susumu Takamatsu (Mie

University, Graduate School of Bioresources, Tsu, Japan) for presubmission reviews

of this manuscript and Elvira Drefler and Petra Mitschke for excellent technical

assistance.

Literature cited

Abasova LV, Aghayeva DN, Takamatsu S. 2018. Erysiphe azerbaijanica and E. linderae: two new

powdery mildew species (Erysiphales) belonging to the Microsphaera lineage of Erysiphe.

Mycoscience 59: 181-187. https://doi.org/10.1016/j.myc.2017.10.002

Adhikari MA. 2017. Research on the Nepalese mycoflora-3: Erysiphales from Nepal. Published

by the author, Kathmandu.

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https://doi.org/10.1016/s0007-1536(65)80031-6

MY COTAXON

July-September 2019—Volume 134, pp. 555-560

https://doi.org/10.5248/134.555

Beltraniomyces panthericolor and B. pulcher spp. nov.

from Brazil

FLAVIA RODRIGUES BARBOSA’, PATRICIA OLIVEIRA FIUZA’,

GLEYSON CRISTIANO KORPAN BARBOSA’, LOISE ARAUJO COSTA3,

GABRIEL GINANE BARRETO‘*, LUIS FERNANDO PASCHOLATI GUSMAO’,

RAFAEL FELIPE CASTANEDA-RUIZ°

"Instituto de Ciéncias Naturais, Humanas e Sociais, Universidade Federal de Mato Grosso,

Ay. Alexandre Ferronato, 1200, 78557-267, Sinop, Brazil

? Programa de Pés-graduacdo em Sistemdtica e Evolucao,

Universidade Federal do Rio Grande do Norte, Campus Universitario,

Av. Senador Salgado Filho, 3000, Lagoa Nova, Natal-RN, 59078-970, Brazil

* Departamento de Ciéncias Biologicas, Laboratorio de Microbiologia,

Centro de Ciéncias Agrarias, Universidade Federal da Paraiba,

Rodovia PB-079, Areia, Brazil

‘Departamento de Micologia, Universidade Federal de Pernambuco,

Avenida da engenharia, Cidade Universitaria 50740-570, Recife, Brazil

° Departamento de Ciéncias Biologicas, Laboratorio de Micologia,

Universidade Estadual de Feira de Santana,

BR116 KMO03, 44031-460, Feira de Santana, Brazil

° Instituto de Investigaciones Fundamentales en Agricultura Tropical (INIFAT)

Alejandro de Humboldt, Académico Titular de la Academia de Ciencias de Cuba,

Calle 1 Esq. 2, Santiago de Las Vegas, C. Habana, Cuba, C.P. 17200

“CORRESPONDENCE TO: faurb10@yahoo.com.br

ABSTRACT—Two new Beltraniomyces species found on decaying leaves and wood in Brazil

are described and illustrated. Beltraniomyces panthericolor is distinguished by biconic, brown

conidia with multiple, irregular, dispersed, smoky to gray-brown spots that resemble leopard

spots. Beltraniomyces pulcher is characterized by biconic conidia with rounded to subobtuse

apices.

Key worps—biodiversity, conidial fungi, hyphomycetes, Pezizomycotina, taxonomy

556 ... Barbosa & al.

Introduction

Asexual fungi belong primarily to Ascomycota and are abundant in nature

as decomposers of litter (Seifert & al. 2011). In Brazil, research involving

asexual ascomycetous decomposers has concentrated in the Caatinga

and Mata Atlantica biomes (Santa Izabel & Gusmao 2018, Barbosa & al.

2013). The monotypic genus Beltraniomyces Manohar. & al. was originally

described from dead unidentified twigs in India and is characterized

by macronematous, mononematous conidiophores with polyblastic,

sympodial conidiogenous cells producing biconic conidia with central

hyaline transverse bands (Manoharachary & al. 2003). The genus is typified

by B. lignicola Manohar. & al., and no additional species have been described

(Wijayawardene & al. 2017).

Two undescribed species resembling Beltraniomyces that were found

during a survey of asexual fungi on decaying plants from Brazil are

described and illustrated here. A key for the three Beltraniomyces species is

also presented.

Materials & methods

Expeditions were made in Fazenda Sao Nicolau (Cotriguacu, Mato Grosso),

Parque Estadual Cristalino (Novo Mundo, Mato Grosso), and Brejo Paraibano

(Areia, Paraiba) to collect leaf litter and wood debris. Samples were placed in

paper bags, taken to the laboratory, and processed as described in Castafieda-Ruiz

& al. (2016). Reproductive structures were placed on slide prepared with PVL

resin (polyvinyl alcohol, lactic acid, and phenol), measured, and photographed

using a microscope equipped with phase contrast. The type specimens were

deposited in the Herbario Centro-Norte Mato-Grossense, Sinop, Mato Grosso,

Brazil (CNMT), and Herbario Jayme Coelho de Moraes, Areia, Paraiba, Brazil

(EAN).

Taxonomy

Beltraniomyces panthericolor L.A. Costa, G.G. Barreto, ER. Barbosa,

Fiuza, R.F. Castafieda, sp. nov. Fig. 1

MB 830313

Differs from Beltraniomyces lignicola by its longer, brown conidia with multiple smoky

or gray-brown spots covering the conidial body.

Type: Brazil, Paraiba State, Areia, Arboreto Jayme Coelho de Moraes, 7°02’04”S

35°48’28’”W, on dead leaves of Clusia sp. (Clusiaceae), 20.1X.2018, coll. L.A. Costa

(Holotype, EAN 26.231).

EryMo_oey: Latin, panthericolor, referring to the brown conidia with multiple smoky

or gray-brown spots, similar to the skin patterning of the leopard (Panthera pardus).

Beltraniomyces panthericolor & B. pulcher spp. nov. (Brazil) ... 557

0000!

Fic. 1. Beltraniomyces panthericolor (holotype, EAN 26.231). A-D. Conidia; E. Conidiogenous cell

and conidium; F. Conidiogenous cells and conidium; G. Conidiophore, conidiogenous cells, and

conidia; H. Conidiophores in group, conidiogenous cells, and conidia. Scale bars: A-D = 10 um;

E= 20 um; F-H = 40 um.

COLONIES scattered, dark brown. Mycelium superficial and immersed,

composed of slightly branched, septate, smooth, pale brown to brown

hyphae. CONIDIOPHORES macronematous, mononematous, erect, straight

or flexuous, unbranched, geniculate, 5-25-septate, brown, smooth, 83-522

558 ... Barbosa & al.

x 4-10 um. CONIDIOGENOUS CELLS polyblastic, integrated, terminal and

intercalary, brown to pale brown, with indeterminate, sympodial, percurrent

extensions, <390 um long. Conidial secession schizolytic. Conrp1a solitary,

biconic, subulate to mucronate apex, cuneiform and truncate at the base,

brown, with multiple, irregular, smoky to gray-brown spots covering the

conidial body; median transversal band subhyaline, smooth, dry, 31-62(-45)

x 15-22 um.

ComMMENTS—Beltraniomyces lignicola is distinguished by smooth biconic

conidia that are acute at the apex and truncate at the base, subhyaline to

pale brown, with a hyaline median transverse band, 31-38.5 x 19-24 um

(Manoharachary & al. 2003). Beltraniomyces panthericolor is easily distinguished

from B. lignicola by conidial shape, pigmentation, and size.

Beltraniomyces pulcher ER. Barbosa, Fiuza, G.C.K. Barbosa,

R.F. Castafieda & Gusmao, sp. nov. Fic. 2

MB 830314

Differs from Beltraniomyces lignicola by its rounded to subobtuse conidial apex.

Type: Brazil, Mato Grosso State, Cotriguagu, 9°52’24”S 58°13’17”W, on decaying wood,

15.XII.2017, coll. G.C.K. Barbosa (Holotype, CNMTf 77).

Erymo oey: Latin, pulcher, meaning beautiful.

COLONIES scattered, dark brown. Mycelium superficial and immersed,

composed of slightly branched, septate, smooth, pale brown to brown hyphae.

CONIDIOPHORES macronematous, mononematous, erect, straight or flexuous,

unbranched, geniculate, 10-25-septate, brown, smooth, 450-750 x 6-12 um.

CONIDIOGENOUS CELLS polyblastic, integrated, terminal and intercalary, with

indeterminate, sympodial, percurrent extensions, brown to pale brown, <300

um long. Conidial secession schizolytic. Conip1 solitary, biconic, rounded to

subobtuse apex, truncate at the base, brown with a hyaline median transversal

band, smooth, dry, 30-45 x 15-23 um.

ADDITIONAL SPECIMENS EXAMINED: BRAZIL, Mato Grosso STATE, Novo Mundo,

9°28'06"S 55°50’36”W, on decaying wood, 29.XI.2016, coll. G.C.K. Barbosa (CNMTf

78); 3.X1I.2015, coll. G.C.K. Barbosa (CNMTf 79).

COMMENTS—Beltraniomyces pulcher is separated from B. lignicola and

B. panthericolor by its apically rounded or subobtuse conidia and larger

conidiophores, with the conidia distinguished by acute apices in B. lignicola

(Manoharachary & al. 2003) and subulate to mucronate in B. panthericolor.

Beltraniomyces pulcher also lacks the distinctive leopard-like coloration found

in B. panthericolor.

Beltraniomyces panthericolor & B. pulcher spp. nov. (Brazil) ... 559

Fic. 2. Beltraniomyces pulcher (holotype, CNMTf 77). A-D. Conidia; E. Conidiogenous cell and

conidium; F. Conidiogenous cells; G. Conidiophore. Scale bars: A-E = 10 um; F = 25 um; G = 90 um.

Key to Beltraniomyces species

1. Conidia with multiple smoky or gray-brown spots, subulate to mucronate apex,

CUNETOriT base ie, vce he oie hd cd ca Mee Stee eee Boe B. panthericolor

TrEOniGid NOU AS A DOY ease, 2 ietiey ouetee ener cncetiee ta et ed sates Scat ee gran than. as thant 2

2. Conidia with acute apex, 31-38.5 x 19-24 um ........... eee eee eee B. lignicola

2. Conidia with rounded to subobtuse apex, 30-45 x 15-23 um ........... B. pulcher

560 ... Barbosa & al.

Acknowledgments

The authors express their sincere gratitude to Dr. Huzefa A. Raja and

Dr. De- Wei Li for their critical review of the manuscript. ER. Barbosa thanks CNPQ

(Proc. 445245/2014-0), Programa de Pesquisa em Biodiversidade da Amazonia

(Proc. 558225/2009-8, 569382/2008-4) for financial support and Programa de

Pés-Graduacao em Ciéncias Ambientais (PPGCAM/UFMT). Patricia Fiuza thanks

Coordenac¢ao de Aperfeigoamento de Pessoal de Nivel Superior —- Programa Nacional

de Pés-doutorado (CAPES-PNPD) for scholarship (Proc. 88882.306016/2018-01).

Dr. Lorelei Norvell’s editorial review and Dr. Shaun Pennycook’s nomenclature

review are greatly appreciated.

Literature cited

Barbosa FR, Raja HA, Shearer CA, Gusmao LFP. 2013. Some freshwater fungi from the Brazilian

semi-arid region, including two new species of hyphomycetes. Cryptogamie, Mycologie

34(3): 243-258. https://doi.org/10.7872/crym.v34.iss2.2013.243

Castafieda-Ruiz RF, Heredia G, Gusmao LFP, Li DW. 2016. Fungal diversity of Central and

South America. 197-217, in: DW Li (ed.). Biology of Microfungi. Springer, Switzerland.

https://doi.org/10.1007/978-3-319-29137-6_9

Manoharachary C, Agarwal DK, Rao NK. 2003. Beltraniomyces, a new genus of dematiaceous

hyphomycetes from India. Indian Phytopathology 56(4): 418-421.

Santa Izabel TS, Gusmao LE. 2018. Richness and diversity of conidial fungi associated with plant

debris in three enclaves of Atlantic Forest in the Caatinga biome of Brazil. Plant Ecology and

Evolution 151(1): 35-47. https://doi.org/10.5091/plecevo.2018.1332

Seifert K, Morgan-Jones G, Gams W, Kendrick B. 2011. The genera of hyphomycetes. CBS

Biodiversity Series 9. 997 p.

Wijayawardene NN, Hyde KD, Rajeshkumar KC, Hawksworth DL, Madri H & al. 2017. Notes

for genera: Ascomycota. Fungal Diversity 86. 594 p. https://doi.org/10.1007/s13225-017-0386-0

MY COTAXON

July-September 2019—Volume 134, pp. 561-576

https://doi.org/10.5248/134.561

Coenogonium hainanense sp. nov.

and new records from China

XIAO-HAN Wu’, WEI-CHENG WANG”?,

MING-ZHU Dou’, ZE-FENG JIA‘ ~

"College of Life Sciences, Liaocheng University, Liaocheng, China

State Key Laboratory of Mycology, Institute of Microbiology,

Chinese Academy of Sciences, Beijing 100101, China

> University of Chinese Academy of Sciences, Beijing 100049, China

" CORRESPONDENCE TO: Zffia2008@163.com

ABSTRACT—A new species, Coenogonium hainanense, is proposed, characterized by

its small slightly transparent pale yellow to brownish apothecia with plane to concave

discs; slightly thick, not prominent, denticulate margins with short dense hairs; and

uniseriate or irregularly biseriate, ellipsoid 1-septate ascospores (10-11.25 x 2.5-3.75

um). Eight additional Coenogonium species reported as new to China are C. coronatum,

C. curvulum, C. fallaciosum, C. geralense, C. hypophyllum, C. moniliforme, C. siquirrense,

and C. usambarense. Descriptions and distributions of the new species and records are

presented, and a key to the 20 Coenogonium species known from China is provided.

KEY woRDS—Coenogoniaceae, Lecanoromycetes, lichens, Ostropales, taxonomy

Introduction

Coenogonium Ehrenb. (Ostropales: Coenogoniaceae), comprising more

than 90 species worldwide, has a tropical and subtropical distribution and

usually occurs on organic substrata, including bark, bryophytes, and leaves

(Baloch & al. 2010, Ferraro & Michlig 2013, Joshi & al. 2015, Licking &

al. 2017, Kantvilas & al. 2018). The genus is characterized by a crustose or

filamentous thallus with yellowish, orange, or brown biatorine apothecia and

rounded, plane, or concave discs, usually distinct margins, a well-developed

excipulum, simple paraphyses, narrowly cylindrical 8-spored asci with thin

562 ... Wu &al.

TABLE 1. Distributions of Coenogonium species previously reported from China.

SPECIES LOCALITY: REFERENCE

C. dilucidum GuaNGx!I & HAINAN: Wu & al. 2018. Hone Kone: Aptroot & Seaward 1999, as Dimerella

dilucida; Aptroot & Sipman 2001, as D. dilucida.

C. disjunctum ‘TAIWAN: Wu & al. 2018.

C. interplexcum TAIWAN: Wang-Yang 1972; Wang-Yang & Lai 1973; Wu & al. 2018. YUNNAN: Wu & al. 2018.

C. isidiatum SICHUAN & XIZANG: Obermayer 2004, as D. isidiata.

C. leprieurii Fujian: Zahlbruckner 1930; Wu & al. 2018. YUNNAN: Wu & al. 2018.

C. linkii GUANGXI & HAINAN: Wu & al. 2018. Hone Kone: Aptroot & Seaward 1999; Seaward &

Aptroot 2005, as C. cf. linkii; Pfister 1978, as C. disjunctum; Aptroot & Sipman 2001. Tarwan:

Zahlbruckner 1933, as C. subvirescens; Asahina 1939, as C. boninense; Wang-Yang 1972;

Wang-Yang & Lai 1973; Yoshimura 1974, as C. boninense; Wu 1987, as C. boninense.

C. luteum ZHEJIANG: Wu & Qian 1989, D. lutea. ANHUI: Wu & al. 2018. Fujian: Zahlbruckner 1930,

as Microphiale lutea; Wu & al. 2018. Huse: Chen & al. 1989, as D. lutea. HAINAN: Wu &

al. 2018. Hone Kone: Aptroot & Seaward 1999, as D. lutea; Seaward & Aptroot 2005, as D.

lutea; Pfister 1978, as Gyalecta lutea; Aptroot & Sipman 2001, as D. lutea.

C. minimum GUANGXI: Wang & Wei 2018.

C. pineti Honc Kone: Aptroot & Sipman 2001, as D. pineti. Tarwan: Aptroot & Sparrius 2003, as D.

pineti.

C. subluteum Hone Kone: Aptroot & Seaward 1999, as D. epiphylla; Aptroot & Sipman 2001, as D.

epiphylla. TAIWAN: Chung 1995, as D. epiphylla; Aptroot & al. 2003. YUNNAN: Zahlbruckner

1930, as M. brachyspora, M. epiphylla, and M. lutea f. foliicola; Santesson 1952, as D. epiphylla;

Wei & Jiang 1991; Aptroot & al. 2003; Wu & al. 2018.

C. zonatum YUNNAN: Aptroot & al. 2003.

walls and unthickened apices lacking internal amyloid structures; typically

(0—)1-septate ellipsoid to fusiform hyaline ascospores; the absence of lichen

secondary substances; and the presence of a trentepohliod photobiont

(Licking & Kalb 2000, Rivas Plata & al. 2006, Liicking 2008).

Eleven Coenogonium species have been reported from China: C.

dilucidum (Kremp.) Kalb & Lticking, C. disjunctum Nyl., C. interplexum

Nyl., C. isidiatum (G. Thor & Vézda) Licking & al., C. leprieurii (Mont.)

Nyl., C. linkii Ehrenb., C. luteum (Dicks.) Kalb & Liicking, C. minimum

(Mull. Arg.) Licking, C. pineti (Ach.) Liicking & Lumbsch, C. subluteum

(Rehm) Kalb & Lticking, and C. zonatum (Mill. Arg.) Kalb & Liicking

(TaBLE 1). Our current research on Coenogonium revealed one new species

and eight species records for China.

Materials & methods

The specimens are deposited in the Herbarium Mycologicum Academiae Sinicae-

Lichenes, Beijing, China (HMAS-L), and the Fungarium, College of Life Sciences,

Coenogonium hainanense sp. nov. (China) ... 563

Liaocheng University, China (LCUF). An Olympus SZX16 dissecting microscope and

Olympus BX53 light microscope were used for the morphological and anatomical

studies. Measurements were taken from manual cross sections of fruit bodies in water.

Amyloidity of the ascospores was tested using Lugol’s solution. The thallus surface

and thin thallus sections were spot tested with 20% KOH. The lichen substances were

detected and identified by thin-layer chromatography (Culberson & Kristinsson 1970,

Culberson 1972, White & James 1985, Jia & Wei 2016).

Taxonomy

Coenogonium hainanense X.H. Wu & Z.E Jia, sp. nov. FIG. 1

EN 570605

Differs from Coenogonium lisowskii by its larger apothecia and ascospores.

Type: CHINA. HAINAN PROVINCE, Changjiang County, Bawang Ridge, 19°16’N

109°03’E, alt. 700 m, on leaves, 5 Sep. 2008, B. Gao HNF056 (Holotype, HMAS-L

126438).

ErymMo.ocy: The specific epithet refers to the type locality, Hainan Island.

THALLUS crustose, foliicolous, very thin, smooth, pale green to grey green,

slightly shiny. APOTHECIA epiphyllous, sessile, rounded, 0.1-0.3 mm

Fic. 1. Coenogonium hainanense (holotype, HMAS-L 126438). A. Thallus with apothecia;

B. Apothecia; C. Cross section of apothecium; D. Ascus with ascospores. Scale bars: A = 1 mm;

B =0.5 mm; C = 100 um; D = 20 um.

564 ... Wu &al.

diam.; disc plane to concave, pale yellow to brownish, slightly transparent;

margin slightly thick, not prominent, denticulate, with short dense hairs

of fungal hyphae, wax-colored; ExciPLE colorless, 20-35 um broad;

HYMENIUM 60-70 um high, colorless; HyPOTHECIUM 15-20 um high,

colorless to pale brownish. Asci 8-spored, 50-70 x 7—9 um; ASCOSPORES

uniseriate or irregularly biseriate, ellipsoid, 1-septate, 10-11.25 x 2.5-3.75

um, 3-4 times as long as broad. Pycnip1a not observed.

CHEMISTRY: No lichen substances detected by TLC.

ECOLOGY & DISTRIBUTION: On the surface of living leaves in humid,

semi-exposed forests of Hainan Island, south China.

REMARKS: Coenogonium hainanense is characterized by its small pale

yellow to brownish, slightly transparent apothecia with plane to concave

discs; slightly thick, not prominent, and denticulate margins covered by

short dense hairs; ellipsoid ascospores, 1-septate, 10-11.25 x 2.5-3.75

um. The new species is very similar to C. lisowskii (Vézda) Licking and

C. dilucidum, but C. lisowskii has smaller apothecia (0.05-0.15 mm diam.)

with smooth hairless margins and smaller ascospores (7-9 x 1.8-2.5 um),

and C. dilucidum has broader ascospores (3.75-5 um diam.; Licking

2008).

Coenogonium coronatum G. Neuwirth & Stock.-Worg.,

Bryologist 117: 161 (2014) Fic. 2

THALLUS crustose, foliicolous, smooth, yellowish green to pale green.

APOTHECIA epiphyllous, sessile, rounded, 0.15-0.35 mm diam.; discs

plane (concave in young apothecia), pale yellow to creamy; margins

slightly thick, densely pilose, cream-colored, hairs consisting of fungal

hyphae. Asci 8-spored, 35-50 x 6-8 um; ASCOSPORES irregularly biseriate,

ellipsoid, 1-septate, 10-12.5 x 3-3.75 um, 2.7—3.3 times as long as broad.

SPECIMEN EXAMINED: CHINA. HAINAN PROVINCE, Ledong County, Jianfeng

Ridge, Mingfeng Valley, alt. 960 m, 6 Sep. 2017, W.C. Wang HN20170170 (HMAS-L

139527).

ECOLOGY & DISTRIBUTION: On leaves. South-East Asia: Thailand (Neuwirth

& al. 2014, Kalb & al. 2016). New record for China.

REMARKS: Coenogonium coronatum is characterized by its small and

pale-yellow apothecia with concave disc, and slightly thick, densely pilose

margin. The species resembles C. subluteum, which differs in having cilia

on the outer excipulum (Kalb & al. 2016).

Coenogonium hainanense sp. nov. (China) ... 565

cia eer fe OF ee ee See

Fic. 2. Coenogonium coronatum (HMAS-L 139527). A. Thallus with apothecia; B. Apothecia;

C. Cross section of apothecium; D. Ascus with ascospores; Scale bars: A, B = 0.5 mm; C = 100

um; D = 50 um.

Coenogonium curvulum Zahlbr., Ann. Cryptog. Exot. 1: 164 (1928) FIG. 3

THALLUS filamentous, foliicolous, green to bright green, forming a thin

crust of irregular filaments attached to the substrate from which a short

and dense layer of erect filaments emerge. APOTHECIA sessile, rounded,

0.2-0.45 mm diam., immersed in a loose algal carpet, pale yellow to

yellow; discs plane to slightly concave; margins glabrous, smooth, cream-

colored. Asci 8-spored; AscosPoREs curved, colorless, biseriate, ellipsoid,

1-septate, 6.25—7.5 x 2.5 um, 2.5-3 times as long as broad.

SPECIMENS EXAMINED: CHINA. HAINAN PROVINCE, Wuzhishan City, Wuzhishan

Tropical Rainforest Scenic Area, alt. 600 m, 9 Sep. 2017, W.C. Wang HN20170360

(HMAS-L 139783); Wuzhishan National Forest Park, alt. 800 m, 8 Sep. 2017, W.C.

Wang HN20170335 (HMAS-L 139428); Changjiang County, Bawang Ridge,

Baishitan, alt. 700 m, 9 Sep. 2017, W.C. Wang HN20170039 (HMAS-L 139418).

ECOLOGY & DISTRIBUTION: On leaves. Neotropics and eastern Paleotropics:

Indonesia (Rivas Plata & al. 2006). New record for China.

REMARKS: Coenogonium curvulum is characterized by its filamentous,

foliicolous thallus with short, upright hair-like algal threads and

566 ... Wu &al.

Fic. 3. Coenogonium curvulum (HMAS-L 139783). A. Thallus with apothecia; B. Apothecia;

C. Cross section of apothecium; D. Ascus with ascospores. Scale bars: A, B = 0.5 mm; C = 100 um;

D=50 um.

curved ellipsoid ascospores. It resembles C. epiphyllum Vain., which

is distinguished by larger (0.3-0.8 mm diam.) apothecia and straight

ascospores.

Coenogonium fallaciosum (Miill. Arg.) Kalb & Liicking,

Bot. Jahrb. Syst. 122: 32 (2000) Fic. 4

THALLUS crustose, foliicolous, pale green to grey green. APOTHECIA mostly

marginally hypophyllous on mycelium-free algae, sessile, rounded, 0.3-1(-1.5)

mm diam.; disc plane, pale yellow to yellow orange; margin very thin, smooth,

concolorous with disc or slightly paler. Asci 8-spored; Ascospores colorless,

uniseriate or irregularly biseriate, ellipsoid, 1-septate, 7-10 x 2.5-3.5 um,

2.5-3.5 times as long as broad.

SPECIMENS EXAMINED: CHINA. HAINAN PROVINCE, Changjiang County, Bawang

Ridge, Baishitan, alt. 700 m, 4 Sep. 2017, W.C. Wang HN20170026 (HMAS-L 139458),

HN20170031 (HMAS-L 139463), HN20170036 (HMAS-L 139468), HN20170038

(HMAS-L 139470), HN20170042 (HMAS-L 139473); Baoting County, Qixian Ridge

National Forest Park, alt. 350 m, 7 Sep. 2017, W.C. Wang HN20170239 (HMAS-L

139553), HN20170274 (HMAS-L 139494), HN20170275 (HMAS-L_ 139495),

HN20170276 (HMAS-L 139496), HN20170277 (HMAS-L 139497), S.H. Jiang

Coenogonium hainanense sp. nov. (China) ... 567

Fic. 4. Coenogonium fallaciosum (HMAS-L 139775). A. Thallus with apothecia; B. Cross section of

apothecium; C. Ascospores uniseriate D. Ascospores irregularly biseriate. Scale bars : A = 2 mm;

B= 100 um; C, D = 20 um.

HN20171262 (HMAS-L 144212); Wuzhishan City, Wuzhishan National Forest Park,

alt. 800 m, 8 Sep. 2017, W.C. Wang HN20170328 (HMAS-L 139770), HN20170333

(HMAS-L 139775); YUNNAN PROVINCE, Xishuangbanna, Mengla County, Tropical

Botanical Garden of Chinese Academy of Sciences, alt. 529 m, 25 Jan 2018, W.C. Wang

20180225 (HMAS-L 144228).

ECOLOGY & DISTRIBUTION: On leaves. Pantropical: Brazil (Rivas Plata & al.

2006). New record for China.

REMARKS: Coenogonium fallaciosum is characterized by its apothecia

mostly marginally hypophyllous on mycelium-free algae and very thin,

smooth margins. It resembles C. geralense, which differs in smaller

apothecia (0.4-1 mm diam.), smaller ascospores (6.25-8.75 x 1.8-2.5 um),

and pale yellow to bright yellow (never orange) apothecia (Lticking 2008).

Coenogonium geralense (Henn.) Licking,

Fl. Neotrop., Monogr. 103: 579 (2008) FIG. 5

THALLUS crustose, foliicolous, thin, yellowish green to grey green.

APOTHECIA epiphyllous or marginally hypophyllous, sessile, rounded,

568 ... Wu &al.

Fic. 5. Coenogonium geralense (HMAS-L 144268). A. Thallus with apothecia; B. Cross section

of apothecium; C. Ascus with ascospores; D. Ascospores. Scale bars: A = 1 mm; B = 100 um;

C = 50 um; D = 20 um.

0.4-1 mm diam.; disc plane to concave, pale to bright yellow; margins

thin, smooth, concolorous to disc or paler. Asci 8-spored, 45-55 x 6-7

lum; ASCOSPORES uniseriate or irregularly biseriate, narrowly ellipsoid,

1-septate, 6.25-8.75 x 1.8-2.5 um, 3-4 times as long as broad.

SPECIMENS EXAMINED: CHINA. HAINAN PROVINCE, Qiongzhong, Mt. Limu,

alt. 800 m, 18 Dec. 2000, M.R. Huang 458 (HMAS-L 108841); Lingshui County,

Mt. Diaoluo, alt. 1050 m, 14 Sep. 2000, M.R. Huang 337 (HMAS-L 112801);

Ledong County, Jianfeng Ridge, Mingfeng Valley, alt. 960 m, 6 Sep. 2017, W.C.

Wang HN20170168 (HMAS-L 139525), HN20170169 (HMAS-L 139526),

HN20170171 (HMAS-L 139434); YUNNAN PROVINCE, Mengla County, Tropical

Botanical Garden of Chinese Academy of Sciences, Greenstone Forest Scenic

Area, 614 m alt., 26 Jan. 2018, W.C. Wang 20180236 (HMAS-L 144268).

ECOLOGY & DISTRIBUTION: On leaves. Pantropical: Brazil (Rivas Plata &

al. 2006). New record for China.

REMARKS: Coenogonium geralense is characterized by its crustose,

foliicolous, smooth, yellowish green to grey green thallus and ellipsoid,

Coenogonium hainanense sp. nov. (China) ... 569

1-septate ascospores. It is similar to C. dilucidum, which is distinguished

by larger ascospores (6-12 x 2.5-3.5 um; Kalb & al. 2016).

Coenogonium hypophyllum (Vézda) Kalb & Liicking,

Bot. Jahrb. Syst. 122: 32 (2000) FIG. 6

THALLUS crustose, foliicolous, hypophyllous, thin, smooth, pale greenish

grey to yellow brownish. ApoTHEcIA hypophyllous, sessile, rounded,

0.25-0.35 mm diam.; discs plane to concave, pale yellow; margins thin,

smooth to minutely dentate, wax-colored. Ascr 8-spored, 45-60 x 6-8 um;

ASCOSPORES irregularly biseriate, fusiform, 1-septate, 11.25-15 x 2.5 um,

5-6 times as long as broad.

SPECIMEN EXAMINED: CHINA. HAINAN PROVINCE, Ledong County, Jianfeng

Ridge, alt. 900 m, 3 Apr. 1993, Y.M. Jiang & S.Y. Guo H-0695 (HMAS-L 138640).

ECOLOGY & DISTRIBUTION: On leaves. Pantropical: Tanzania (Rivas Plata &

al. 2006). New record for China.

REMARKS: Coenogonium hypophyllum is characterized by its foliicolous

thallus and regularly hypophyllous apothecia with smooth to minutely

Fic. 6. Coenogonium hypophyllum (HMAS-L 138640). A. Thallus with apothecia; B. Cross section

of apothecium; C. Ascus with ascospores; D. Ascospores. Scale bars: A = 0.5 mm; B = 100 um;

C, D = 20 um.

570 ... Wu &al.

dentate margins. It resembles C. subzonatum (Licking) Lticking & Kalb,

which differs in its apothecia marginally hypophyllous on a mycelium free

of algae and smaller ascospores (7-10 x 2.5-3.5 um; Lticking 2008).

Coenogonium moniliforme Tuck., Proc. Amer. Acad. Arts 5: 416 (1862) FIG. 7

THALLUS filamentous with densely woven filaments forming closely

appressed mats, moderately thick, pale to dark green or brownish green.

APOTHECIA sessile, rounded, 0.2-0.35 mm diam.; discs plane to concave,

pale to dark orange; margins thin, smooth, wax-colored. Asci 8-spored,

45-55 x 6-8 um; ASCOSPORES irregularly biseriate, fusiform, 1-septate,

8.75-11.25 x 2.5-3.75 um, 2.3-4.5 times as long as broad.

SPECIMENS EXAMINED: CHINA. HAINAN PROVINCE, Ledong County, Jianfeng

Ridge, Mingfeng Valley, alt. 960 m, 6 Sep. 2017, W.C. Wang HN20170218 (HMAS-L

139421), S.H. Jiang HN20171048 (HMAS-L 144213).

ECOLOGY & DISTRIBUTION: On leaves. Pantropical: Cuba (Rivas Plata & al.

2006). New record for China.

REMARKS: Coenogonium moniliforme is characterized by its filamentous thallus

with the densely woven filaments forming closely appressed mats and plane to

a7 , : ‘er |

Fic. 7. Coenogonium moniliforme (HMAS-L 144213). A. Thallus with apothecia; B. Apothecia;

C. Cross section of apothecium; D. Ascus with ascospores. Scale bars: A = 1 mm; B = 0.5 mm;

C = 100 um; D = 30 um.

Coenogonium hainanense sp. nov. (China) ... 571

concave, pale to dark orange discs. It closely resembles C. flavoviride M. Caceres

& Licking, which is distinguished by its much smaller ascospores (4-6 x

1.8-2 um; Liicking 2008).

Fic. 8. Coenogonium siquirrense (HMAS-L 139552). A. Thallus with apothecia; B. Cross section of

apothecium; C. Ascospores irregularly biseriate; D. Ascospores uniseriate. Scale bars: A = 1 mm;

B = 100 um; C, D = 20 um.

Coenogonium siquirrense (Liicking) Licking,

Fl. Neotrop., Monogr. 103: 580 (2008) Fic. 8

THALLUS crustose, foliicolous, continuous, thin, smooth, green to dark

green. APOTHECIA mostly epiphyllous, sessile, rounded, 0.5-0.8 mm diam.;

discs plane, orange with darker marginal zone and paler centre; margins thin,

smooth, pale orange. Asci 8-spored, 30-45 x 3-4 um; ASCOSPORES uniseriate

or irregularly biseriate, narrowly ellipsoid, 1-septate, 7.5-10 x 2-2.5 um,

3-4 times as long as broad.

SPECIMEN EXAMINED: CHINA. HAINAN PROVINCE, Baoting County, Qixian Ridge

National Forest Park, 350 m alt., 7 Sep. 2017, W.C. Wang HN20170238 (HMAS-L

139552).

ECOLOGY & DISTRIBUTION: On leaves. Neotropics: Costa Rica (Rivas Plata & al.

2006). New record for China.

572... Wu &al.

REMARKS: Coenogonium siquirrense is characterized by its crustose,

foliicolous thallus, mostly epiphyllous sessile 0.5-0.8 mm diam. apothecia,

and narrowly ellipsoid ascospores. The closely similar C. geralense differs in

its pale to bright yellow often marginally hypophyllous apothecia with plane

to concave discs (Lticking 2008). The also similar C. luteum is distinguished

by its larger (0.5-2 mm diam.), more prominent apothecia and broader

ascospores (2.5-3.75 um diam.; Liicking 2008).

Fic. 9. Coenogonium usambarense (HMAS-L 139777). A. Thallus with apothecia; B. Cross section

of apothecium; C. Ascus with ascospores; D. Ascospores. Scale bars: A = 2 mm; B = 200 um;

C=50 um, D= 10 um.

Coenogonium usambarense (Vézda & Farkas) Liicking & Kalb,

Biblioth. Lichenol. 78: 255 (2001) FIG. 9

THALLuS crustose, foliicolous, epiphyllous, thin, smooth, green to

bright green. APOTHECIA sessile, rounded, 0.4-0.8 mm diam.; discs plane

to slightly convex, pale yellow to orange; margins smooth, cream-colored.

Ascli 8-spored, 40-45 x 4-6 um; ASCOSPORES irregularly biseriate, ellipsoid,

1-septate, 8.5-11.25 x 2.5-3 um, 4-5 times long as broad.

SPECIMENS EXAMINED: CHINA. HAINAN PROVINCE, Wuzhishan City, Wuzhishan

National Forest Park, 800 m alt., 8 Sep. 2017, W.C. Wang HN20170336 (HMAS-L

Coenogonium hainanense sp. nov. (China) ... 573

139777); YUNNAN PROVINCE, Mengla County, Tropical Botanical Garden of Chinese

Academy of Sciences, 549 m alt., 25 Jan. 2018, X.H. Wu YN18082 (LCU); Greenstone

Forest Scenic Area, Dabangen, 672 m alt., 26 Jan. 2018, X.H. Wu YN18160 (LCU).

ECOLOGY & DISTRIBUTION: On leaves. African and eastern Paleotropics:

Tanzania (Rivas Plata & al. 2006). New record for China.

REMARKS: Coenogonium usambarense is characterized by a crustose,

foliicolous, epiphyllous, thin, smooth, and green or bright green thallus

and ellipsoid ascospores (8.5-11.25 x 2.5-3 um). The similar C. epiphyllum

produces much shorter and broader ascospores (6-8 x 2.5-3.75 um; Vézda &

Farkas 1988).

Key to all the known species of Coenogonium in China

Dei i til aeeTTEO US: “i We tata it Geeta te tee Ste le teen Cheon Lawes Fees Eee ey 2

A, dehval lurstendietoGe. scutes. suite os studs c. xeudla sh ites boeuilg ste dnvudl a te deed! act, ould poli ts peu dee 7

2. Thallus distinctly filamentous, in loose, erect mats or

horizontally projecting, shelf-like structures ............ 00... c cee eee eee 3

2. Thallus indistinctly filamentous, in thin densely woven mats or

with well-developed upright short filaments ................... 0c eee eee eee 6

3) ASCOSPOTES Sipe, 5-8. 2S LUN oy. 6' ila holed Pw ele SG Me ahve e' C. leprieurii

OT ASCOSP ORCS TESEPLAlee «dele tue thiy Banting Boor sne iin eas Bais oe Louk one Podepere Podkapers Bowes enh Spa 4

4, Thallus dimidiate or shelf-like; ascospores 6-9 x 2-3 UM ............000- C. linkii

4, Thallus‘prostrate, irresularly-woven: +. 2.024206. s%eues sans yes yen ween Megs 5

DAASCOSPOLES 7 0; M28 UU pu soya we see hye be seek a, be nok ate Se nod ate boned ate: Sonal a C. interplexum

Sy ASCOSpOtesaL 0 '4iKe 2 SUH e bree Sistee SL La La a C. disjunctum

6. Ascospores:8:/5=1 1.25% 25-3775 path... wvins ngien sagen saga emees C. moniliforme

G.ASCOSWORES.O: 75 lo eR SUE cls oe Le oa A ta ME ota ora Roa C. curvulum

7. Thallus with isidia; ascospores 9-14 x 3-4Um ..... eee eee eee eee C. isidiatum

oo Pal bet SSAC RAST IAG 1 5 ore ha We oem ga Spc a Ha esa WL Fea o peda We dreds bs bres hts fovea ts reo 8

8. Thallus and apothecia regularly hypophyllous;

ascospores 1425-15 425.um, fusiform, fost fie deat fe toe ie C. hypophyllum

8. Thallus and apothecia epiphyllous or apothecia often marginally hypophyllous ... 9

Ov mpotnecia, Seryssiitalls OvE On 3 Tam afi wh re Sct are hh ae eb a PL tet peat cna tte 10

D>: Apothects larger O25 —2 Ms ote cacw sean low nate od awe asaled atmo oe Moet snl 11

10. Apothecia light brown to yellow brown with strongly concave discs

and thick margins; ascospores 10-14 x 3-4um ................. C. minimum

10. Apothecia wax-colored to pale yellow with flat to slightly concave discs;

aSCOSpOres1 0-2 SoS FOS. Se oes alee Bad oe Bai n-p heads gold C. dilucidum

11. Apothecia margin densely pilose or slightly denticulate ..................... 1

PieApothecamarein Smooth -s.f.7%, sus .s6uls nbastd able ables ablwa were: wholes) 13

574A ... Wu &al.

12. Apothecia pale yellow to creamy, margins cream-colored,

densely pilose (sometimes abraded in older apothecia);

ascospores: 10S12.5 3 SSA. 75 eB hog oe ee he ena ela eet ee ey C. coronatum

12. Apothecia pale yellow to brownish, slightly transparent,

margins denticulate; ascospores 10-11.25 x 2.5-3.75 um ........ C. hainanense

13. Ascospores narrowly ellipsoid to oblong (1.8-2.5 um broad),

Lista yeintes ulachysbiseP ates 20 aie. tila bleibs Mee conic ds beegissterh ape oensease ene see ee 14

13. Ascospores broadly ellipsoid (2.5-5 um broad), usually uniseriate .............. 15

14. Apothecia pale to bright yellow with pale margins;

ASCOSPOLCE:O57.9=9.7 D2 LO =2 SUE AAA Ns at es tah Pe aceall ie ot C. geralense

14. Apothecia orange with darker marginal zones and paler centres;

ASCOSD CECE he LOI B= 7 5 UII a ana RA ell Peete te Me C. siquirrense

15. Apothecia 1-2 mm, bright yellow to yellow orange;

ASCOSPOLCSIID=5.75:0-Z, 525: FOI: 6 Siem, sou sn eg dm peu die pede pelea C. luteum

15. Apothecia 0.25-1 mm, wax-colored to pale orange ............... 0.0.0 eee 16

16. Apothecia usually marginally hypophyllous, 0.3-1(-1.5) mm, pale yellow

to yellow orange; margins very thin, smooth, concolorous with disc

or slightly paler; ascospores 7-10 X 2.5-3.5 um .............0.. C. fallaciosum

lorpotheciaustal hyepiphy lots ry. ih rth ee Ms wa yeh ak, ahha Melek atom h a iy

P7, Apotheciarwax=colored to pale Grains wl PE os steae o aigeae Pps digheg bir Seb prasad ps 18

17. Apothecia white to pale yellow or orange ....... 0.0... eee cece eee 19

18. Prothallus present, white; ascospores 7.5-10 x 2.5-4um ............ C. zonatum

18. Prothallus absent; ascospores 10-12.5 x 2.5-3.75 um .............. C. subluteum

19. Apothecia white to pinkish (rarely orange tinged), 0.2-0.5 mm,

margins smooth; ascospores 9-14 x 2.3-4.5 UM ...... ee ee eee ee eee C. pineti

19. Apothecia pale yellow to orange, margins smooth, cream-colored;

ASCOSPOECS SISAL ASK 2S SAAT gw seas ya te: sake abe Sead ant: brake lee C. usambarense

Acknowledgments

This study was supported by the National Natural Science Foundation of China

(31750001, 31270066 & 31470149). The authors are grateful to the presubmission

reviewers Dr. Wei Xin-li (State Key Laboratory of Mycology, Institute of

Microbiology, Chinese Academy of Sciences, Beijing) and Prof. Qiang Ren (College

of Life Sciences, Shandong Normal University, China) for reading and improving

the manuscript.

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https://doi.org/10.5248/134.577

Cystostereum sirmaurense sp. nov. from India

RAMANDEEP KAurR, AVNEET P. SINGH”, G.S. DHINGRA

Department of Botany, Punjabi University, Patiala 147002, India

*CORRESPONDENCE TO: avneetbot@gmail.com

ABSTRACT—A new corticioid species, Cystostereum sirmaurense, is described from Himachal

Pradesh, India.

Key worps—Agaricales, Agaricomycetes, Basidiomycota, Cystostereaceae, Himalaya

Introduction

During the fungal forays conducted in August 2015 in the Nahan area of

district Sirmaur, Himachal Pradesh, India, Ramandeep & Avneet collected

a corticioid specimen growing in association with a living angiospermous

tree in a mixed forest. Based on its macro and micromorphological details

and comparison with the literature (Rattan 1977, Eriksson & Ryvarden 1975,

Bernicchia & Gorjon 2010, Prasher & Ashok 2013, Dhingra & al. 2014,

Sanyal 2014, Mycobank 2019) it has been placed in Cystostereum Pouzar

(Cystostereaceae), close to the type species C. murrayi (Berk. & M.A. Curtis)

Pouzar. A portion of the basidiocarp was sent for expert comment to Prof. Nils

Hallenberg (University of Gothenburg, Sweden), who confirmed the findings

and supported the proposal of a new species, described here as Cystostereum

sirmaurense.

Materials & methods

The collection was sampled during a field trip conducted in the Nahan forest

division of the Sirmaur district (Himachal Pradesh, India) during August 2015. Colour

standards follow Kornerup & Wanscher (1978). The dried specimen was deposited

at the Herbarium, Department of Botany, Punjabi University, Patiala, India (PUN).

578 ... Kaur, Singh, Dhingra

PLATE 1. Cystostereum sirmaurense (holotype, PUN (Kaur 10097)). 1. Basidiocarp showing

hymenial surface (fresh); 2. Basidiocarp showing hymenial surface (dry). Scales in mm.

Crush mounts and free hand sections were used to study the morphology of hyphae,

cystidia, basidia, basidiospores, etc. in water and 3%, 5%, and 10% KOH solutions, and

stained in cotton blue (1% in lactophenol), Congo red (1% in distilled water), phloxine

(1% in distilled water), sulphovanillin (0.5gm vanillin + 4.0ml conc. sulphuric acid +

2.0ml distilled water), and Melzer’s reagent (0.5gm iodine + 1.5gm KI + 20gm chloral

hydrate + 20ml distilled water). Line diagrams of various microscopic structures were

drawn using a camera lucida.

Taxonomy

Cystostereum sirmaurense R. Kaur, Avn.P. Singh & Dhingra, PLATES 1,2

MB 829964

Differs from Cystostereum murrayi by its almost smooth hymenium with few scattered

tubercles and more broadly ellipsoid basidiospores.

Type: India, Himachal Pradesh: Sirmaur, Nahan, 2 km from Ambwala towards Nahan,

on a living angiospermous tree, 23 August 2015, Ramandeep Kaur & Avneet Pal Singh

10097 (, PUN).

ErymMo.ocy: The epithet refers to the district where the collection was made.

Basidiocarp resupinate, effused, adnate, <340 um thick in section; hymenial

surface smooth but with minute scattered tubercles, white when fresh,

becoming yellowish white to orange white on drying; margins thinning

or indeterminate, paler concolorous. Hyphal system dimitic. Generative

hyphae branched, with clamps, thin-walled, subhyaline up to 2.8 um wide.

Skeletal hyphae unbranched, aseptate, thick-walled, subhyaline up to 5.2

um diam. Cystidia 26-44 x 7-8.5 um, variably shaped but generally with an

obtuse apex, thin- to somewhat thick-walled, with basal clamp, embedded

in the hymenium and subhymenium, not staining in sulphovanillin.

Basidia 15-20 x 3.8-4.7 um, subclavate to clavate, 4-sterigmate, with basal

clamps; sterigmata <3.3 um long. Basidiospores 3.3-4.2 x 2.3-3.3 um,

ellipsoid to broadly ellipsoid, apiculate, thin-walled, smooth, inamyloid,

acyanophilous.

Cystostereum sirmaurense sp. nov. (India) ... 579

UN (Kaur 10097)). 3. Basidiospores;

PLATE 2. Cystostereum sirmaurense (holotype, P

s; 5. Skeletal hypha; 6. Generative hypha.

4. Reconstruction of hymenial and subhymenial zone

580 ... Kaur, Singh, Dhingra

Discussion

Pouzar (1959) proposed Cystostereum on the basis of unique combination

of numerous gloeocystidia, dimitic hyphal system with very scarce light-

couriered skeletal hyphae, hard consistency of trama, and inamyloid

basidiospores. With the addition of C. sirmaurense, eight species are now

represented in Cystostereum (MycoBank 2019). From India, previously

only C. murrayi has been described, from Himachal Pradesh (Rattan 1977;

Kaur 2018) and Uttarakhand (Sanyal 2014). Cystostereum murrayi differs

from C. sirmaurense in having a tuberculate hymenial surface and narrower

basidiospores.

Acknowledgments

The authors thank Head (Department of Botany, Punjabi University, Patiala)

for providing research facilities, Dr. Nils Hallenberg (Professor Emeritus in Botany,

University of Gothenberg) for expert comments and peer review, Prof. B.M. Sharma

(Department of Plant Pathology, COA, CSKHPAU, Palampur, India) for peer review,

and University Grants Commission (New Delhi, India) for financial assistance under

DSA-I scheme.

Literature cited

Bernicchia A, Gorjon SP. 2010. Corticiaceae s.l. Fungi Europaei 12. Edizioni Candusso. Alassio.

Italia. 1008 p.

Dhingra GS, Singh AP, Kaur J, Priyanka, Kaur H, Rani M, Sood S, Singla N, Kaur H, Jain N,

Gupta S, Kaur M, Sharma J, Rajnish, Kaur G. 2014. A checklist of resupinate, non-poroid

agaricomycetous fungi from Himachal Pradesh, India. Synopsis Fungorum 32: 8-37.

Eriksson J, Ryvarden L. 1975. The Corticiaceae of North Europe. Volume 3. Coronicium - Hyphoderma.

Fungiflora, Oslo, Norway. pp. 287-546.

Kaur M. 2018. Systematic studies on resupinate non-poroid hymenomycetous fungi from district

Shimla (H.P.) and evaluation of selected taxa for ligninolytic activity. Ph.D. Thesis. Punjabi

University, Patiala. 436 p.

Kornerup A, Wanscher JH. 1978. Methuen handbook of colour, 3rd ed. Methuen and Co. Ltd.

London. 252 p.

Mycobank. 2019. Fungal databases. Nomenclature and species banks. [Accessed: 02/02/2019]

http://www.mycobank.org/

Pouzar Z. 1959. New genera of higher fungi III. Ceska Mykologie 13(1): 10-19.

Prasher IB, Ashok D. 2013. A checklist of wood rotting fungi (non-gilled Agaricomycotina) from

Himachal Pradesh. Journal on New Biological Reports 2(2): 71-98.

Rattan SS. 1977. The resupinate Aphyllophorales of the North Western Himalaya. Bibliotheca

Mycologica 60. Cramer, Germany. 427 p.

Sanyal SK. 2014. Taxonomic studies on resupinate Polyporales of Uttarakhand. Ph.D. Thesis.

Punjabi University, Patiala. 300 p.

MYCOTAXON

July-September 2019—Volume 134, pp. 581

https://doi.org/10.5248/134.581

Regional annotated mycobiota new to the Mycotaxon website

ABSTRACT—MycotTaxon is pleased to add a new annotated species distribution list to our

133 previously posted free-access fungae. The 9-page “Diversity of macrofungi in Yushan,

Jiangsu, China” by Bing Xu, Haoyu Lu, Donglei Zhao, Wei Wang, and Hong Ji may be

downloaded from our website via http://www.mycotaxon.com/mycobiota/index.html

ASIA

China

BinG Xu, Haoyu Lu, DONGLE! ZHAO, WEI WANG, Hone Ji. Diversity

of macrofungi in Yushan, Jiangsu, China. 9 p.

AxBstRAcT—A checklist is presented of 84 taxa of macrofungi collected from

Yushan National Forest Park, Changshu city, Jiangsu province, China. The taxa

represent 48 genera and 28 families, with taxa in Russulaceae, Agaricaceae,

Amanitaceae, Inocybaceae, and Psathyrellaceae comprising 48.8% of the total

collected. The food and medicinal potentials of the taxa are indicated.

Key Worps—Ascomycota, Basidiomycota, natural resources

MYCOTAXON

July-September 2019—Volume 134, pp. 583-590

https://doi.org/10.5248/134.583

BOOK REVIEWS AND NOTICES:

ELSE C. VELLINGA

861 Keeler Avenue, Berkeley CA 94708 USA

ABSTRACT— Books reviewed include: An introduction to the edible, poisonous and

medicinal fungi of northern Laos by Lessee, Sparre Pedersen, and Sysouphanthong

(2019), Mushrooms of the Gulf Coast States (A field guide to Texas, Louisiana,

Mississippi, Alabama, and Florida) by Bessette, Bessette, and Lewis (2019), and

Fungi of temperate Europe (volumes | & 2) by Leessoe & Petersen (2019).

An introduction to the edible, poisonous |

and medicinal fungi of northern Laos.

By T. Lessee, O. Sparre Pedersen, Ph.

Sysouphanthong, 2019. — Svampetryk, |

Kornblomsvej 6, 8381 Tilst, Denmark;

svampetryk@svampe.dk 192p. 330 photos. This

is an open-access publication, distributed under

the terms of the Creative Commons Attribution

Licence, which permits unrestricted use,

distribution, and reproduction in any medium,

provided it is for non-commercial purpose and

the original source and the three authors are

credited. It does not have an ISBN number.

vase wich

w a ai

monnuta, Wuiio wax duels

luwaniwie suv aro

An Introduction to the

Edible, Poisonous and Medicinal

Fungi of Northern Laos

Thomas Lzessoe, Ole Sparre Pedersen and Phongeun Sysouphanthong

This book is one result from a Darwin Initiative grant to the James

Hutton Institute (UK) for the purpose of “building mycological capacity

* Book reviews or books for consideration for coverage in this column should be sent to the

EDITOR-IN-CHIEF <editor@mycotaxon.com> 6720 NW Skyline, Portland OR 97229 USA.

584 ... MyCOTAXON 134

for sustainable resource management in Lao PDR®” Training of Laotian

students in a laboratory was also part of this project.

This ‘introduction provides an overview of macrofungi from Xieng

Khouang province in northern Laos. The focus is on those fungi that are

considered either beneficial (edible or medicinal) or poisonous for people,

and all text is in both English and Laotian.

The vegetation and mycoflora of Xieng Khouang province resemble

those of northern Thailand, Yunnan province in China, and Japan—all

three regions where mycology has seen a huge outpouring of publications

on macrofungi in the last 15 years.

The book starts with a brief introduction to mushrooms, the different

nutritional modes of fungi (including a paragraph on termite fungi), a page

on classification and morphology, and another page addressing the various

adverse reactions to and toxins of the poisonous fungi.

The fungi are organized systematically within the edible, potentially

edible, poisonous, potentially poisonous, and medicinal categories, with

Microporus xanthopus as the only representative of a fungus that is used for

decoration.

Each species gets its own page, with one to several excellent photos, a

description, and notes on use in Laos. All photos, with the exception of two

species, were taken in Laos—this type of information is very welcome.

Many species are identified as ‘close to X’ or ‘resembling Y, as Southeast

Asia is still extraordinarily rich in undescribed fungal species, and the

identification of the Laotian species was based solely on morphology.

An extensive reference list and an index complete the book.

This is the first book I know of that deals with Laotian fungi. In any

case, information on the mycoflora of Laos is exceedingly rare. As in

neighbouring countries, wild mushrooms form a huge part of the local

diet and are a source of income as well; some species (for instance several

species of red russulas) are dried and exported.

I like the cautious approach to the naming, and the efforts to link the

local Laotian names to a scientific name. As I have been mushrooming in

northern Thailand and Yunnan province, this book evokes happy memories

of the forests and the visits to the markets and roadside mushroom sellers.

Book Reviews ... 585

Mushrooms of the Gulf Coast States. A field f :

guide to Texas, Louisiana, Mississippi,

Alabama, and Florida. By A.E. Bessette, Pe BK ir

A.R. Bessette, and D.P Lewis, 2019. | he

PELs of Texas Press, utpress.utexas.edu § {> | Gulf Coast

https://doi,.org/10.7560/318157 ISBN 978-1- - oe S

4773-1815-7. 614 pp, 667 colour photos, $ 39.95 =a SSS

Paperback [also available as an e-book].

It is always exciting to open a mushroom @&

field guide for a region one does not know. «

And indeed, the mycoflora of the Gulf Coast [paateiseasnrns

states of the USA is quite different from that eeoratin

of California where I live. |

This guide provides a description and photo for around 650 species,

each species on its own page. The species are organized by group, and

alphabetically within each group. The same artificial categories are used

as in the earlier field guide for the southeastern USA (Bessette et al. 2007),

which works well for a field guide. The alphabetical approach within a

group, however, means that species that look alike and/or are closely

related do not end up together.

A short introduction to the geographical area, the lifestyle of mushrooms,

collecting methods, how to use the book, and pictorial keys to the main

groups precede the descriptions and illustrations. The same type of photos

as those serving to illustrate the species is used in the keys, but printed

very small. Appendices at the back of the book provide information on

microscopy and the chemicals used for identification, and there is a chapter

devoted to mushrooms for the table. A glossary, a short of list of references

and resources, and indices to common and scientific names complete the

volume.

As noted, the bulk of the book is dedicated to the illustration and

description of the mushroom species. The emphasis is on the more

conspicuous species, so that only one Inocybe species is included and

the two Psathyrella species feature only in the picture keys, but don't get

their own page. Unfortunately, the introduction does not state how the

descriptions were made: are they based on observations of the authors on

material from the Gulf Coast states? Or, are they based on the literature?

586 ... MYCOTAXON 134

There is a list of photographers, but there is no information whether the

photographed material has been preserved in a herbarium or where the

photos were taken. Unfortunately, many photos are too dark.

Some names that are used have not been officially published, such as

“Amanita amerifulva” for the North American relative of the European

A. fulva; in other cases, a European name is used, even when it is clear

that the American species is different (e.g., Macrolepiota procera does not

occur in North America, but the North American species have not yet

been described). Only in a few cases are such taxonomic issues indicated

by adding ‘group’ or ‘complex’ to a name; in my opinion this should have

been done in many more cases. Pluteus cervinus is one such a case where

more than one species occurs in the area. Synonyms are given for a number

of species, but not for all, so one wonders how that selection was made;

why is Agaricus alboviolaceus listed, but not Agaricus conigenoides? Some

identification mistakes are obvious—the photo of Tremella mesenterica

shows T: aurantia with Stereum. Tremella mesenterica parasitizes

Peniophora species, and Dacrymyces chrysospermus (compared in the

remarks to the Tremella), growing on conifer wood, is a wood decayer, not

a fungal parasite. The photo of Pluteus atromarginatus shows a different

species, and the text and the photo of Mycena epipterygia var. viscosa do

not match, as the photo shows pruinose stipes, and the text calls the stipe

‘smooth:

One element that I sorely miss is a description of the main types of habitat

the species were found in. What do these forests look like? Where are they?

When and where to go if one would like to see these mushrooms with one's

own eyes? Which species are really rare, and are there conservation plans

for them or their habitats?

Also, the volume is probably too heavy to carry around in a backpack.

These problems aside, the book gives a wealth of information on

southeastern North American species and will form a basis for much more

in-depth work.

Bessette AE, Roody WC, Bessette AR, Dunaway DL. 2007. Mushrooms of the

southeastern United States. Syracuse University Press.

Book Reviews ... 587

of Temperate Europe of Temperate Europe

VOLUME 1 VQ Mig

Fungi of temperate Europe (volumes 1 & 2). T. Lzessoe, J.H. Petersen.

2019. Princeton University Press. https://press.princeton.edu/titles/13983.html

$110.00. ISBN 978-0-691-18037-3. 1708p. 7 1/2 x 11 1/2 inches, 7,000 color illus.

This translation of the Danish NORDEUROPAS SVAMPE gives descriptions and

illustrations of around 2800 species of fungi occurring in northern temperate

Europe.

This is not your standard guidebook. The whole book is extremely well

thought out, with ease of identification as the most important criterion for lay-

out and presentation. And that alone makes it worth buying.

The two authors are extremely experienced broadly interested field

mycologists, educators, excellent photographers, and Jens Petersen has proven

his skills in presenting data in the general mycology/coffee table book THE

KINGDOM OF FunaI (Petersen 2013). This book builds on that experience and

on the digital identification tool Mycoxey by the same two authors.

The two volumes (one for the introduction and covering most agarics and

boletes, and a second one for crusts, polypores, ascomycetes and other fungi)

enthuse, and make you want to look much deeper and in much more detail to

find all those beautiful fungi.

Identification tools are not in the form of dichotomous keys but as so-

called ‘fungal wheels’—pie charts with the identifying characters as drawings

and photos in the outer rings, where one starts in the middle and works one’s

way outwards to find the right species or group of species. I'd like to try these

588 ... MyCcOTAXON 134

x « Lasieboes

pope 10 toe speees pape 47 Tsoecien pope 1366

bape TAT hoo

with students and people who dont have any experience with fungi to see

how effective these are. These wheels are free for download from the Mycokey

website and can be used by everybody, whether you have the books or not:

( www.mycokey.com/Downloads/FungiOfTemperateEurope_Wheels.pdf )

The text of the descriptions is in the form of a narrative, which always starts

with the name of the species (no extras like author names or synonyms), gives

the essential characters, followed by the habitat and distribution, and some

remarks on other similar species. Conservation status of species, IUCN red

data listings are given as well.

Space is not wasted in this book: the inside of the covers is used for the index

and explanation of symbols, and a one-page explanation of often-used terms.

What are some of the other characters that make this book stand out?

—Inclusion of many species of crust fungi, small and inconspicuous

ascomycetes, rusts, smuts, slime molds and fungi that do not form

macroscopic fruitbodies but are often encountered (e.g. on rotten fruit).

Also the astounding numbers of small Agaricales that is included is unique:

e.g., five species of Pseudobaeospora, four Mycenella species, and 28 pages of

inocybes. Even Ceratellopsis aculeata is presented (a tiny, tiny awl-shaped

fruitbody on Cladium mariscus leaves). A total of 2,800 species! It must

be said that this book is not an identification guide for the plant parasites

(rusts, smuts etc.) and slime molds, but as they are conspicuous and show

up at observation sites such as iNaturalist, their inclusion is definitely

warranted.

—The attention to detail—the insets with close ups of the important parts

of the mushrooms;

Book Reviews ... 589

—An introduction of almost 40 pages that cover many different aspects of

mushrooms, the study of mushrooms, and the scope, geographic coverage,

and lay-out of the book. Estimates are given for the species representation

per group; e.g., an estimated 50% of gilled mushrooms from the study area

is included (for small genera all species are included, but only a fifth of the

Cortinarius species are represented).

—Group names on the top of the pages so that one knows where one is,

without having to move back and forth through the book. An example of

such a group: “large, dry + colourful cortinarias with red-brown to orange-

brown gills.”

—Scale bars in the photos, no guessing about size anymore, and no need to

give sizes in the descriptions.

—Because the species are grouped by shape not all species of a genus will

be automatically together; this is obvious for truffle-shaped species and

their agaricoid relatives but is also the case for genera with agaricoid and

cyphelloid forms.

The only place I did not agree with the identification is for Leucoagaricus

americanus—the photo shows L. meleagris.

An endearing character of these books is the narrow green ribbon that

comes in handy to mark the pages with the identification wheels.

This is not a book to put in your backpack on your hike, as it is much too

heavy. But it will be used back home, in the lab, with a microscope at hand.

Not only for people in Europe is this the best guidebook to get, it also is

extremely useful for people in other parts of the world. It is a splendid example

how to present the multitude of forms in a way that makes identification

possible and fun, while at the same time showing the beauty and diversity of

fungi.

Petersen, J.H., 2013. The kingdom of Fungi. Princeton University Press,

Princeton. 265 pp.