Iv ... MYCOTAXON 134(4)

MYCOTAXON

VOLUME ONE HUNDRED THIRTY-FOUR (4) — TABLE OF CONTENTS

134-4: TABLE OF CONTENTS, NOMENCLATURAL UPDATES, PEERS, EDITORIALS

POVIC WONG Perk ols bcs ng ghs_ a tung Sea efor v athe citer sy 3 read RI Us eee Soret get ae vi

Nomernclatural novelties ty pifications soo 4. cok ey tp eras keke eens vii

COPTICONAG RA es Shox. aSecey gen on eau tats eae Este bod Gallente 8a alee g viii

PP ROMISTTCTSAILOT 3, nyc. cess Ses aegy $e stg ents Sea he trv Aes gw cate ce ahh ob ogo eee ix

ZOLO Ss WUMISSIOR DPYOCEAURE? 3: Pe tons Fry Sone Py Ra ver oAd oy Dey ly Yo Sha xi

RESEARCH ARTICLES

Urocystis cumminsii sp. nov., a smut fungus on Themidaceae

from Arizona KYRYLL G. SAVCHENKO, SYLENA R. HARPER,

Lori M. Carrts, Lisa A. CASTLEBURY 591

Leucoagaricus brunneus sp. nov. from Khyber Pakhtunkhwa, Pakistan

Zi1A ULLAH, SANA JABEEN, MUHAMMAD FAISAL,

Hasis AHMAD, ABDUL Nasir KHALID 601

Dictyostelids from Jilin Province, China 3:

new Cavenderia and Dictyostelium records

Pu Liu, SHUNHANG ZHANG, ZHUANG LI*,

YuE Zou, XUEPING KANG, Yu Li 613

Records of Aureobasidium harposporum, Sarcophoma miribelii,

and Stigmina dothideoides from Turkey

MAKBULE ERDOGDU, MERVE ULUKAPI,

ALI IHSAN KARAYEL, ZEKIYE SULUDERE 619

Bactrodesmium pulcherrimum sp. nov. from Ecuador

FERNANDO ESPINOZA, DAYNET SOSA, LIZETTE SERRANO,

ADELA QUEVEDO, FREDDY MAGDAMA, MARCOS VERA,

SIMON PEREZ-MARTINEZ, ELAINE MALOSSO, RAFAEL F. CASTANEDA-RUIZ 627

Dendrographium multiseptatum sp. nov. from China

L1-Guo Ma, YuE-L1 ZHANG, Bo ZHANG,

Kal Qt, CHANG-SONG LI, JUN-SHAN QI 633

Haematomma pluriseptatum sp. nov. from China

CONGCONG MIAO, RONG TANG,

LINLIN DONG, ZHAOJIE REN, ZUNTIAN ZHAO 637

New records of Didymium inconspicuum, D. karstensii, and

D. rugulosporum from China

CHAOFENG YUAN, SHU LI, WAN WANG, SHUWEI WEI, QI WANG, Yu LI 643

OCTOBER-DECEMBER 2019 ... V

Rhomboidia wuliangshanensis gen. & sp. nov.

from southwestern China

TaI-MIN Xu, XIANG-Fu Liu, Yu-Hur CHEN, CHANG-LIN ZHAO 649

Filsoniana lhasanensis sp. nov. from Tibet, China

XuE-MEI! WEN, HurNISA SHAHIDIN, ABDULLA ABBAS 663

Sarcopodium flocculentum, the correct name for S. macalpinei

SHAUN R. PENNYCOOK & PauL M. Kirk) 677

Pteridicolous ascomycetes from a cloud forest in eastern Mexico

ROSARIO MEDEL-ORTIZ, YAJAIRA BAEZA,

FRANCISCO G. LOREA-HERNANDEZ 681

First sexual morph record of Sarcopodium vanillae

NAPALAI CHAIWAN, SAJEEWA S.N. MAHARACHCHIKUMBURA,

DHANUSHKA N. WANASINGHE, MINGKWAN DOILoM,

RUVISHIKA JAYAWARDENA, KEVIN D. HyDE 707

Notes on rust fungi in China 7. Aecidium caulophylli life cycle

inferred from phylogenetic evidence and renamed as

Puccinia caulophylli comb. nov. JiInG-XIn Jt, ZHUANG LI,

Yu Li, MAKOTO KAKISHIMA 719

Exserticlava aquatica sp. nov., a microfungus

from the Brazilian Amazon LUANA TEIXEIRA DO CARMO,

D10GO CARELI DOS SANTOS, CAROLINA RIBEIRO SILVA,

SHEILA MIRANDA LEAO FERREIRA, THAMARA ARAO FELETTI,

Luis FERNANDO PASCHOLATI GuSMAO 731

MycoBioTa (FUNGA) NEW TO THE MYCOTAXON WEBSITE

Ascomycota (lichenized and non-lichenized) on Syagrus coronata

in the Caatinga biome: new and interesting records

for Brazil and South America (suMMARyY)

Mata4ra A.L. Dos SANTOS, NILO G. S. FORTEs,

TAssio E. FE. Sitva, NaApja S. VITORIA 737

Checklist of Bolivian Agaricales. 1:

Species with dark and pink spore prints (suMMaRy)

E. MELGAREJO-ESTRADA, M.E. SUAREZ,

D. RocaBADO, O. MAILLARD, B.E. LECHNER 739

vi ... MYCOTAXON 134(4)

REVIEWERS — VOLUME ONE HUNDRED THIRTY-FOUR (4)

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.

M. Catherine Aime

D. Jayarama Bhat

Lu-Sen Bian

Marcela Eugenia da Silva Caceres

Rafael F. Castahteda-Ruiz

Vagner G. Cortez

Cvetomir M. Denchev

Shouyu Guo

Tom Hsiang

Shah Hussain

Sevda Kirbag

Klaus Kalb

Paul M. Kirk

John C. Landolt

De-Wei Li

José G. Marmolejo

Tom May

Eric H.C. McKenzie

Josiane Santana Monteiro

Karen K. Nakasone

Nicolas Niveiro

Lorelei L. Norvell

Cumali Ozaslan

Shaun R. Pennycook

Marcin Piatek

Meike Piepenbring

Luis Quijada

Scott Redhead

Andrea Irene Romero

Michelline Lins Silvério

Adna Cristina Barbosa de Sousa

Steven L. Stephenson

Jan Vondrak

Xinli Wei

Ze-Fen Yu

Changlin Zhao

OcCTOBER-DECEMBER 2019...

NOMENCLATURAL NOVELTIES AND TYPIFICATIONS

PROPOSED IN MYCOTAXON 134(4)

Bactrodesmium pulcherrimum R.F. Castaneda, F. Espinoza & D. Sosa

[MB 830569], p. 629

Dendrographium multiseptatum L.G. Ma & J.S. Qi

[MB 832847], p. 634

Exserticlava aquatica L.T. Carmo, C.R. Silva, Careli, S.M. Leao, Feletti

& Gusmao [MB 831391], p. 732

Filsoniana lhasanensis X.M. Wen, Shahidin & A. Abbas

[FN 570592], p. 669

Haematomma pluriseptatum R. Tang

[MB 830618], p. 638

Leucoagaricus brunneus Z. Ullah, Jabeen & Khalid

[MB 827985], p. 603

Puccinia caulophylli (Kom.) Jing X. Ji & Kakish.

[MB 830631; epitypified: MBT 386779], p. 726

Rhomboidia C.L. Zhao

[MB 833318], p.654

Rhomboidia wuliangshanensis C.L. Zhao

[MB 833320], p. 656

Urocystis cumminsii Savchenko, Carris & Castl.

[MB 830145], p. 595

VII

vill ... MyCOTAXON 134(4)

CORRIGENDA

VOLUME 134-1

p.174, ACKNOWLEDGMENTS

FoR: The author would like to thank Prof. Dr. Ertugrul Sesli, Prof. Dr. Ibrahim

Tirkekul, and Dr. Shaun Pennycook for their helpful comments and careful

review.

READ: The author would like to thank Yiiziincii Yil University, Coordination of

Scientific Research Projects for financial support (2010-FED-B031 and FYL-

2016-5213) and Prof. Dr. Ertugrul Sesli, Prof. Dr. Ibrahim Tiirkekul, and Dr.

Shaun Pennycook for their helpful comments and careful review.

MYCOTAXON 134-3

p. vii, line 21 FoR: Marthamyces culmigenus (Ellis & Everh.) P.R. Johnst.

READ: Marthamyces culmigenus (Ellis & Langl.) P.R. Johnst.

p.496, lines 3-5

FOR: Marthamyces culmigenus (Ellis & Everh.) P.R. Johnst., comb. nov.

IF 556322

= Naemacyclus culmigenus Ellis & Everh..,

Proc. Acad. Nat. Sci. Philadelphia 45: 151, 1893.

READ: Marthamyces culmigenus (Ellis & Langl.) P.R. Johnst., comb. nov.

IF 556322

= Naemacyclus culmigenus Ellis & Langl., in Ellis & Everhart,

Proc. Acad. Nat. Sci. Philadelphia 45: 151, 1893.

[Semi-bold fonts used above to flag corrected terms. ]

CORRIGENDA IN CURRENT ISSUE (134-4)

Cited below are mistakes or oversights present in approved input files not

detected by authors until after PDF conversion.

p. 613: Pu Liu and fellow coauthors wish to acknowledge the contributions made to

“Dictyostelids from Jilin Province, China 3: new Cavenderia and Dictyostelium

records” (MycoTaxon 134: 613-618) by Prof. Zhuang Li (Shandong Provincial

Key Laboratory for Biology of Vegetable Diseases and Insect Pests, College of Plant

Protection, Shandong Agricultural University, Tai’an 271018, China). The author

sequence originally intended should read “Pu Liu, Shunhang Zhang, Zhuang Li,

Yue Zou, Xueping Kang, Yu Li.”

p-669, line 26 FoR: MK43983

READ: MK439830

OCTOBER-DECEMBER 2019 ... IX

FROM THE EDITOR-IN-CHIEF

DEADLINES, MYCOTAXON & NOMENCLATURE— The most onerous task your esteemed

Editor-in-Chief faces is bringing the year-end issue in ‘on time’ A glance at our

publication history suggests that during her tenure, she has failed miserably. Since

2004, she has met only three of her sixteen December deadlines: New Year's Eve

2004, Boxing Day 2007, 30 December 2009. Given this deplorable 37% success rate,

it is no surprise that, once again, an October-December MycotTaxon will appear

in January. (It could be worse: after adopting electronic submission and new software,

we released the last 2005 volume on 11 May 2006!)

Reasons/excuses for delays are myriad: time-consuming nomenclatural revision

and editorial repair of sloppily prepared submissions (on the misplaced notion that

it is quicker for us to do authors’ work for them), outside research commitments,

severe health problems, uncontrollable press delays. Nonetheless, this year we were

convinced that MycoTaxon 134(4) would be ready for delivery well before 2020.

What we did NoT anticipate was the small number of research papers submitted.

With several authors not sending us their final papers after their 2019 nomenclatural

review, we waited until December 27 before deciding to release the issue with

only 140 pages (the smallest issue EVER since 1974) rather than waiting for final

submissions that have yet to appear. Frustrating, because removal of required page

charges produced a flood of 138 accessions during 2019.

What makes the year-end release date so important to MycoTaxon? With

nomenclatural priority date-based, the date of publication is dictated by the actual

release date and not what is displayed on the cover. That means that taxonomic

names published in periodicals must cite both the actual date of publication and

the ‘nominal’ date printed in the issue, e.g., following the order, Haematomma

pluriseptatum R. Tang, Mycotaxon 134: 638 (2020) but indexed as “Miao, C,

Tang R, Dong L, Ren Z, Zhao Z. 2020 (‘2019’). Haematomma pluriseptatum sp.

nov. from China. Mycotaxon 134: 637-641. https://doi.org/10.5248/134.637 ”

Much less confusing and infinitely more restful to have only ONE date to cite!

MyYcOTAXON 134(4) presents 15 papers by 81 authors (representing 15 countries) as

revised by 34 expert reviewers and the editors.

The 2019 October-December MycoTaxon proposes one new genus (Rhomboidia

from China) and eight new species representing Bactrodesmium from ECUADOR;

Dendrographium, Filsoniana, Haematomma, and Rhomboidia from CHINA;

Exserticlava from BraziL; Leucoagaricus from PAKISTAN; and Urocystis from the

U.S.A. We also offer a new combination in Puccinia and epitypification for Puccinia

caulophylli.

New species range extensions are reported for [ascomycetes] Aureobasidium,

Sarcophoma, Stigmina in TURKEY and new records and hosts of fern-associated ascos

in eastern MExico and [myxomycetes] Cavenderia, Dictyostelium, and Didymium

in CHINA.

x ... MYCOTAXON 134(4)

Two papers on Sarcopodium [1] explain why S. flocculentum is the correct name

for S. macalpinei and [2] discuss the first sexual morph recorded for S. vanillae in

Thailand. Another paper treats the full life cycle for the rust Puccinia (= Aecidium)

caulophylli.

Our small year-end issue closes with the announcements of two mycobiota

[recently posted on www.mycotaxon.com] covering [1] new records of ascomycetes

on Syagrus coronata in Brazil's Caatinga biome and [2] dark- and pink-spored agarics

in Bolivia.

Wishing us all health, happiness, illumination, and PEACE in 2020,

Lorelei L. Norvell (Editor-in-Chief)

8 January 2020

PUBLICATION DATE FOR VOLUME ONE HUNDRED THIRTY-FOUR (3)

MYCOTAXON for JULY-SEPTEMBER 2019 (1—-xIv + 423-590)

was issued on October 2, 2019

OCTOBER-DECEMBER 2019 ... XI

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The Mycotaxon journal publishes four quarterly issues per year. Both open access

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MYCOTAXON

October-December 2019—Volume 134, pp. 591-599

https://doi.org/10.5248/134.591

Urocystis cumminsii sp. nov., a smut fungus

on Themidaceae from Arizona

KYRYLL G. SAVCHENKO"™*, SYLENA R. HARPER’,

Lori M. Carris’, Lisa A. CASTLEBURY?

Department of Biological Sciences, Butler University, Indianapolis, IN 46208

? Department of Plant Pathology, Washington State University, Pullman, WA 99164, USA

> USDA-ARS, Mycology and Nematology Genetic Diversity and Biology Laboratory,

10300 Baltimore Ave, Beltsville, MD 20705, USA

“ CORRESPONDENCE TO: ksavchen@butler.edu

ABSTRACT—The morphology and phylogenetic relationships of a species of Urocystis on

Dichelostemma capitatum (Themidaceae, Asparagales) collected in the Tucson Mountains in

Arizona, United States, were studied using microscopy and ITS rDNA sequence analyses. This

is a first record for smut fungi on hosts from Themidaceae. Molecular phylogenetic analyses

based on ITS sequence data revealed its basal position in relation to species on Poaceae. As

a result, the smut in leaves of Dichelostemma capitatum is described and illustrated here as a

new species, Urocystis cumminsii.

Key worps—plant pathogens, taxonomy Urocystidales

Introduction

Urocystis Rabenh. ex Fuckel contains more than 170 species of plant

pathogenic smut fungi, found all over the world but more common in

temperate areas of both hemispheres (Vanky 2011b). More than 60% of

Urocystis species are found on monocotyledons, with Poaceae serving as a

major monocotyledonous host family, followed by Juncaceae, Hypoxidaceae,

Convallariaceae, Amaryllidaceae, and Hyacinthaceae (Vanky 2011b). The

intra-level phylogenetic relationships of Urocystis species have never been

analysed and thus far only the graminicolous species from triticoid hosts

592 ... Savchenko & al.

have been included in comprehensive phylogenetic analyses (Savchenko

& al. 2017). Most of the Urocystis species described recently—e.g.,

U. achnatheri L. Guo, U. anemones-narcissiflorae Vanky, U. arxanensis

L. Guo, U. beckwithiae Vanky, U. circaeasteri Vanky, U. dunhuangensis

S.H. He & L. Guo, U. glabella Vanky & R. G. Shivas, U. helanensis L. Guo,

U. koeleriae L. Guo, U. phalaridis Vanky, U. puccinelliae L. Guo & H.C.

Zhang, U. rostrariae Piatek, U. sinensis L. Guo, U. skirgielloae Piatek,

U. wangii L. Guo, and U. xilinhotensis L. Guo & H.C. Zhang—were

supported only by morphological and host-specialization data (Guo 2002,

2005, 2006; Guo & Zhang 2004, 2005; He & Guo 2007; Piatek 2006a,

2006b; Vanky 2004, 2005, 2011la; Vanky & Abbasi 2011). The integration

of molecular phylogenetic analyses, host plant taxonomy, and morphology

provides a natural classification for various smut genera (Bauer & al. 2008;

Castlebury & al. 2005; Kruse & al. 2018; Lutz & al. 2008; McTaggart & al.

2012; Savchenko & al. 2013, 2015; Vanky & Lutz 2007), justifying the need

of a phylogenetic study of the genus.

During a survey of Urocystis species diversity in the United States,

we examined a specimen identified as Urocystis sp. on Dichelostemma

capitatum (Benth.) Alph. Wood (Themidaceae) from Arizona in the WSP

herbarium. Previously, no Urocystis species had been recorded on hosts

from this family. The present study aimed to resolve the specific status of

the smut on D. capitatum through morphological analysis and determine

its phylogenetic affinities within Urocystis.

TABLE 1. GenBank sequences used in this study.

SPECIES GENBANK NO. REFERENCE

Antherospora scillae EF653983 Bauer & al. 2008

A. vaillantii EF653988 Bauer & al. 2008

Urocystis bolivarii KX057771 Savchenko & al. 2017

U. colchici DQ839596 Matheny & al. 2006

U. cumminsii MK575496 This study

U. eranthidis JN367299 Kellner & al. 2011

U. fischeri KF668284 Smith & Lutz 2013

U. occulta KX057774, Savchenko & al. 2017

U. trillii HQ239361 Henricot 2010

U. tritici KX057782 Savchenko & al. 2017

Ustilago hordei AY345003 Stoll & al. 2003

Vankya heufleri EF667965 Bauer & al. 2008

V. ornithogali EF635910 Bauer & al. 2008

Urocystis cumminsii sp. nov. (United States) ... 593

Materials & methods

The herbarium specimen is deposited in Washington State University

Mycological Herbarium, Pullman, WA, United States (WSP).

Sorus and spore characteristics were studied using dried herbarium material.

Specimens were examined by light microscopy (LM). Pictures of sori were taken

with a Canon Power Shot G10 camera. For LM, spores were mounted in 90% lactic

acid on a microscope slide, gently heated to boiling point to eliminate air bubbles,

and then examined under a Carl Zeiss Axiostar™ light microscope at 1000x

magnification and photographed with a Canon Power Shot G10 camera. At least 50

spore balls were measured, and the variation is presented as a range with extreme

values given in parentheses. Means and standard deviations (SD) are provided after

the spore size ranges.

For SEM studies, spore balls were attached to metal stubs by double-sided

adhesive tape and coated with gold. Spore surface ornamentation was observed at

15 kV and photographed with a JEOL JSM-6700F scanning electron microscope

with a working distance of c. 12-13 mm.

Sequences from other species of Urocystis and related genera were obtained

from GenBank (TABLE 1). Genomic DNA was isolated from spore balls removed

from the herbarium specimen that had been lysed in 1.5 mL tubes for 1 min using

FastPrep°24. Tubes were incubated in a water bath for 5 hours at 55 °C, and DNA

extracted using DNeasy Plant Mini Kit (QIAGEN) following the manufacturer’s

instructions.

DNA was amplified in 20 ul aliquots on an Applied Biosystems® GeneAmp 9700

thermal cycler using ITS1 as the forward primer and ITS4 as the reverse primer

(White & al. 1990).

Standard cycling parameters with an annealing temperature of 57 °C were used

for amplification. PCR products were purified with USB ExoSAP-IT according

to the manufacturer’s instructions, amplified with the forward and reverse PCR

primers with the BigDye® Terminator v3.1 Cycle Sequencing Kit, and sequenced on

an ABI PRISM? 3100 Genetic Analyzer.

Consensus sequences were assembled, aligned, and edited with Geneious 7.1.8

for MacOS and with MAFFT 6.853 (Katoh & al. 2002, Katoh & Toh 2008) using the

L-INS-i option. Maximum Likelihood (ML) was implemented as a search criterion

in RAxML (Stamatakis 2014). GTR+I+G was specified as the evolution model in

MrModeltest (Nylander & al. 2004). The RAxML analyses were run with a rapid

Bootstrap analysis (command -fa) using a random starting tree and 1000 maximum

likelihood bootstrap replicates. A Markov Chain Monte Carlo (MCMC) search in

a Bayesian analysis (BA) was conducted with MrBayes (Ronquist & Huelsenbeck

2003). Four runs were implemented for 5 million generations. The cold chain was

heated to a temperature of 0.25°C. Substitution model parameters were sampled

every 500 generations and trees were saved every 1000 generations. Convergence

of the Bayesian analysis was confirmed using AWTY (Nylander & al. 2008) and a

594 ... Savchenko & al.

burn-in of 18,000 generations was calculated. The ML and Bayesian analyses were

run three times to test accuracy. The tree was rooted using Ustilago hordei (Pers.)

Lagerh.

Results

The ITS alignment of 13 sequences (including the outgroup Ustilago hordei)

comprised 643 characters including gaps. The different BA and ML analytical

runs yielded consistent topologies in respect to well-supported branches

(a posteriori probability >90% in most cases). The consensus tree of one run of

Bayesian phylogenetic analyses is presented in Fic. 1. The Urocystis sequences

fell into two major clades. ‘The first clade comprised species from Poaceae and

Themidaceae, and the second clade included species from Cyperaceae, Liliaceae

s.l., and Ranunculaceae. The two species from the genus Vankya Ershad

clustered together with the second Urocystis clade.

Ustilago hordei

Antherospora scillae

Anth Jo)

Antherospora vaillantii a IE Ore

Urocystis cumminsii sp. nov.

Urocystis bolivarii

Urocystis 1

Urocystis occulta Oe 7 is,

Urocystis tritici

Urocystis eranthidis

Vankya heufleri

Vankya ornithogali

Urocystis 2

Urocystis colchici rae = 2

Urocystis trillii

Urocystis fischeri

Fic. 1. Bayesian inference of phylogenetic relationships resulting from the analysis of ITS

nucleotide sequence data. Numbers on branches are estimates for PPs from Bayesian inference

(only probabilities >0.8 are shown).

Urocystis cumminsii sp. nov. (United States) ... 595

Fic. 2. Urocystis cumminsii (WSP 68198). A. sori in leaves of Dichelostemma capitatum; B. spore

balls seen by SEM; C, D. spore balls seen by LM. Scale bars: A = 2 mm, B = 5 um, C, D = 20 um.

Taxonomy

Urocystis cumminsii K.G. Savchenko, Carris & Castl., sp. nov. FIG. 2

MB 830145

Differs from Urocystis camassiae by its lighter colored yellowish-brown spores with

thicker walls and its smaller thinner walled sterile cells, and by its host specialization

on Themidaceae.

Type: USA. Arizona, King’s Canyon, 16 km west of Tucson, on Dichelostemma capitatum

(as D. pulchellum), 30.03.1981, leg. G.B. Cummins (Holotype, WSP 68198; GenBank

MK575496).

EryMoLocy: Named after George B. Cummins (1904-2007), an eminent American

mycologist, who collected the holotype specimen.

596 ... Savchenko & al.

Sor! in leaves as slightly elevated, pustular, elongate areas of various size and

shape, sometimes confluent, visible on both sides of the leaf, initially lead-

colored and covered by the epidermis which ruptures exposing the powdery,

black mass of spore balls. SPORE BALLS globose, subglobose, ovoid to irregular,

20-45 um diam., composed of 1-6 (mostly 3) spores and more or less complete

investing layer of sterile cells. Spores subglobose, ovoid, irregularly oblong to

elongated, 11-15 x 12-18 um diam. [mean + SD, 13.3 + 2.6 x 15 + 2.9 um],

medium yellowish brown, wall 1-1.5 um thick, smooth. STERILE CELLS

subglobose, elongated, ovoid, 5-7 x 5-11(-13) um, pale yellow, to almost

hyaline, with smooth, 1 um thick wall.

Discussion

Dichelostemma Kunth is a North American genus of wild hyacinths, closely

related to Brodiaea Sm., from the family Themidaceae. Plants from this family

are native to Central America and western North America, from British

Columbia to Guatemala (Pires & Sytsma 2002, Stevens 2018). No members of

Themidaceae were previously known to be parasitized by smut fungi (Vanky

2011b). Our molecular phylogenetic analyses and morphological data have

helped resolve the systematic position of Urocystis on D. capitatum.

The only possible close relative to U. cumminsii might be another native

North American species, U. camassiae Vanky, found on Camassia Lindl.

(Agavaceae; Fay & Chase 1996, Pires & Sytsma 2002). However, U. camassiae

is distinguished by its darker colored reddish-brown spores with thinner spore

walls (0.5-1 um) and its larger (5-17 um) sterile cells with thicker walls (1-2

(-3) um; Vanky 1994).

ITS phylogenetic analysis infers that U. cumminsii is sister to the clade of

Urocystis species on grasses and separate from species found on hosts from

families more closely related to Themidaceae, such as U. colchici (Schltdl.)

Rabenh. ex A.A. Fisch. Waldh. and U. trillii H.S. Jacks., indicating multiple

inter-family host jumps during the evolution of Urocystis species, similar to

those in the closely related genus Thecaphora Fingerh. (Vasighzadeh & al.

2014). Interestingly, Vankya heufleri (Fuckel) Ershad and V. ornithogali (J.C.

Schmidt & Kunze) Ershad also clustered within the Urocystis lineage (Fic. 1).

Most sequences of Urocystis in GenBank are derived from the LSU region.

Unfortunately, our preliminary analysis showed that LSU is not informative for

Urocystis phylogenetics. Hence, we based our current phylogeny on ITS data,

utilizing the limited number of Urocystis ITS sequences available in GenBank.

Additional sequencing is needed to resolve the taxonomy and evolutionary

Urocystis cumminsii sp. nov. (United States) ... 597

relationships within Urocystis, as well as the phylogenetic affiliation of closely

related genera, such as Vankya.

Urocystis cumminsii expands the occurrence of Urocystis species to the

host family Themidaceae. Future combined molecular phylogenetic and

morphological analyses may reveal higher diversity among Urocystis species in

North America.

Acknowledgments

The authors are grateful to Mary Catherine Aime (Purdue University, Lafayette IN,

US.A.) and Marcin Piatek (W. Szafer Institute of Botany, Polish Academy of Sciences,

Krakow) for peer reviewing the manuscript and Shaun Pennycook for his valuable

comments. Funding for this work was provided by USDA-APHIS 2017 Farm Bill

Projects 3.0245.01 and 3.0245.02.

Literature cited

Bauer R, Lutz M, Begerow D, Piatek M, Vanky K, Bacigalova K, Oberwinkler F. 2008. Anther

smut fungi on monocots. Mycological Research 112: 1297-1306.

https://doi.org/10.1016/j.mycres.2008.06.002

Castlebury LA, Carris LM, Vanky K. 2005. Phylogenetic analysis of Tilletia and allied genera

in order Tilletiales (Ustilaginomycetes; Exobasidiomycetidae) based on large subunit nuclear

rDNA sequences. Mycologia 97: 888-900. https://doi.org/10.1080/15572536.2006.11832780

Fay ME, Chase MW. 1996. Resurrection of Themidaceae for the Brodiaea alliance, and

recircumscription of Alliaceae, Amaryllidaceae and Agapanthoideae. Taxon 45: 441-451.

https://doi.org/10.2307/1224136

Guo L. 2002. Two new species of Urocystis and a Urocystis (Ustilaginales) new to China.

Mycotaxon 81: 431-434.

Guo L. 2005. Two new species of Urocystis (Urocystales) from China. Mycotaxon 92: 269-272.

Guo L. 2006. Urocystis wangii (Urocystales), a new species from China. Mycosystema 25:

364-365.

Guo L, Zhang H. 2004. A new species and two new records of Ustilaginomycetes from China.

Mycotaxon 90: 387-390.

Guo L, Zhang H. 2005. Two new species of Urocystis (Urocystales) from China. Nova Hedwigia

81: 199-204. https://doi.org/10.1127/0029-5035/2005/0081-0199

He SH, Guo L. 2007. Two new species of Urocystales from China. Mycotaxon 101: 1-4.

Henricot B. 2010. New diseases of Trillium in the UK caused by Colletotrichum lineola and

Urocystis fischeri. New Disease Reports 22: 32.

https://doi.org/10.5197/j.2044-0588.2010.022.032

Katoh K, Toh H. 2008. Recent developments in the MAFFT multiple sequence alignment

program. Briefs in Bioinformatics 9: 286-298. https://doi.org/10.1093/bib/bbn013

Katoh K, Misawa K, Kuma K, Miyata T. 2002. MAFFT: a novel method for rapid multiple

sequence alignment based on fast Fourier transform. Nucleic Acid Research 30: 3059-3066.

https://doi.org/10.1093/nar/gkf436

Kellner R, Vollmeister E, Feldbrugge M. Begerow D. 2011. Interspecific sex in grass smuts and

genetic diversity of their pheromone-receptor system. PLoS Genetics 7: e1002436 [17 p.].

https://doi.org/10.1371/journal.pgen.1002436

598 ... Savchenko & al.

Kruse J, Piatek M, Lutz M, Thines M. 2018. Broad host range species in specialised pathogen

groups should be treated with suspicion - a case study on Entyloma infecting Ranunculus.

Persoonia 41: 175-201. https://doi.org/10.3767/persoonia.2018.41.09

Lutz M, Piatek M, Kemler M, Chlebicki A, Oberwinkler F. 2008. Anther smuts of Caryophyllaceae:

Molecular analyses reveal further new species. Mycological Research 112: 1280-1296.

https://doi.org/10.1016/j.mycres.2008.04.010

Matheny PB, Gossman JA, Zalar P, Kumar TKA, Hibbett DS. 2006. Resolving the phylogenetic

position of Wallemiomycetes: an enigmatic major lineage of Basidiomycota. Canadian

Journal of Botany 84: 1794-1805. https://doi.org/10.1139/b06-128

McTaggart AR, Shivas RG, Geering ADW, Callaghan B, Vanky K, Sharaschkin T. 2012.

Soral synapomorphies are significant for the systematics of the Ustilago-Sporisorium-

Macalpinomyces complex (Ustilaginaceae). Persoonia 29: 63-77.

https://doi.org/10.3767/003158512X660562

Nylander JA, Ronquist F, Huelsenbeck JP, Nieves-Aldrej JL. 2004. Bayesian phylogenetic analysis

of combined data. Systematic Biology 53: 47-67. https://doi.org/10.1080/10635 150490264699

Nylander JA, Wilgenbusch JC, Warren DL, Swofford DL. 2008. AWTY (are we there yet?):

A system for graphical exploration of MCMC convergence in Bayesian phylogenetics.

Bioinformatics 24: 581-583. https://doi.org/10.1093/bioinformatics/btm388

Piatek M. 2006a. Urocystis rostrariae, a new species of smut fungi on Rostraria from Jordan.

Mycotaxon 97: 119-124.

Piatek M. 2006b. Urocystis skirgielloi, a new graminicolous smut fungus infecting Heteropogon

contortus in India. Acta Mycologica 41: 7-10. https://doi.org/10.5586/am.2006.002

Pires JC, Sytsma KJ. 2002. A phylogenetic evaluation of a biosystematics framework: Brodiaea

and related petaloid monocots (Themidaceae). American Journal of Botany 89: 1342-1359.

https://doi.org/10.3732/ajb.89.8.1342

Ronquist F, Huelsenbeck JP. 2003. MrBayes 3: Bayesian phylogenetic inference under mixed

models. Bioinformatics 19: 1572-1574. https://doi.org/10.1093/bioinformatics/btg180

Savchenko KG, Lutz M, Piatek M, Heluta VP, Nevo E. 2013. Anthracoidea caricis-meadii is a

new North American smut fungus on Carex sect. Paniceae. Mycologia 105: 181-193.

https://doi.org/10.3852/12-074

Savchenko KG, Carris LM, Castlebury LA, Heluta VP, Wasser SP, Nevo E. 2015. Entyloma

scandicis, a new smut fungus on Scandix verna from Mediterranean forests of Israel.

Mycotaxon 130: 1061-1071. https://doi.org/10.5248/130.1061

Savchenko KG, Carris LM, Demers J, Manamgoda DS, Castlebury LA. 2017. What causes flag

smut of wheat? Plant Pathology 66: 1139-1148. https://doi.org/10.1111/ppa.12657

Smith PA, Lutz M. 2013. The rare smut fungus Urocystis fischeri (Urocystidales, Ustilaginomycotina)

from the Outer Hebrides, Scotland, with notes on its systematic position. Glasgow Naturalist

26: 112-114.

Stamatakis A. 2014. RAxML Version 8: a tool for phylogenetic analysis and post-analysis of large

phylogenies. Bioinformatics 30: 1312-1313. https://doi.org/10.1093/bioinformatics/btu033

Stevens, PF. 2018 [continuously updated] Angiosperm phylogeny website. Version 14, July 2017.

http://www.mobot.org/MOBOT/research/APweb [accessed August 2018].

Stoll M, Piepenbring M, Begerow D, Oberwinkler F. 2003. Molecular phylogeny of Sporisorium

and Ustilago species (Basidiomycota, Ustilaginales) based on internal transcribed spacer

(ITS) sequences. Canadian Journal of Botany 81: 976-984. https://doi.org/10.1139/b03-094

Vanky K. 1994. Taxonomical studies on Ustilaginales. XI. Mycotaxon 51: 153-174.

Vanky K. 2004. Taxonomic studies on Ustilaginomycetes — 24. Mycotaxon 89: 55-118.

Vanky K. 2005. Taxonomic studies on Ustilaginomycetes —- 25. Mycotaxon 91: 217-272.

Urocystis cumminsii sp. nov. (United States) ... 599

Vanky K. 201 1a. Seven new species of smut fungi (Ustilaginomycotina). Mycologia Balcanica 8:

97-104.

Vanky K. 2011b [“2012”]. Smut fungi of the world. St. Paul, APS Press.

Vanky K, Abbasi M. 2011.Urocystis phalaridis sp. nov. on Phalaris sp. from Iran. Rostaniha 12:

187-190.

Vanky K, Lutz M. 2007. Revision of some Thecaphora species (Ustilaginomycotina) on

Caryophyllaceae. Mycological Research 111: 1207-1219.

https://doi.org/10.1016/j.mycres.2007.06.007

Vasighzadeh A, Zafari D, Selcuk E, Huseyin E, Kursat M, Lutz M, Piatek M. 2014. Discovery

of Thecaphora schwarzmaniana on Rheum ribes in Iran and Turkey: implications for the

diversity and phylogeny of leaf smuts on rhubarbs. Mycological Progress 13: 881-892.

https://doi.org/10.1007/s11557-014-0972-4

White TJ, Bruns T, Lee S, Taylor J. 1990. Amplification and direct sequencing of fungal

ribosomal RNA sequences for phylogenetics. 315-322, in: MA Innis & al. (eds).

PCR protocols: a guide to methods and applications. San Diego, Academic Press.

https://doi.org/10.1016/B978-0-12-372180-8.50042-1

MY COTAXON

October-December 2019—Volume 134, pp. 601-611

https://doi.org/10.5248/134.601

Leucoagaricus brunneus sp. nov.

from Khyber Pakhtunkhwa, Pakistan

ZIA ULLAH"*?, SANA JABEEN?®3, MUHAMMAD FAISAL’,

HaBiB AHMAD‘, ABDUL NASIR KHALID”

' Department of Microbiology and Molecular Genetics & "Department of Botany:

University of the Punjab,

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

*Department of Botany, Division of Science & Technology,

University of Education, Township, Lahore, Pakistan

“Islamia College, University Peshawar, Peshawar, Pakistan

* CORRESPONDENCE TO: Ziaullah.phd.mmg@pu.edu.pk

ABsTRACT—A field survey of macrofungi from the Hindu Kush range of District Swat

revealed a fungus morphologically similar to Leucoagaricus but representing a new species,

here named L. brunneus. A detailed description and comparison with closely allied taxa are

provided. Maximum likelihood analysis based on the internal transcribed spacer rDNA

region (ITS) confirm its placement in Leucoagaricus sect. Rubrotincti.

Key worps—Agaricaceae, Agaricales, Agaricomycetes, phylogeny, polyphyletic

Introduction

Leucoagaricus Locq. ex Singer, assigned to the family Agaricaceae (Singer

1986, Vellinga 2004), is represented by over 100 species in the northern and

southern hemispheres (Kumar & Manimohan 2009; Ge 2010; Liang & al. 2010;

Vellinga 2010; Vellinga & Balsley 2010; Vellinga & al. 2010; Mufioz & al. 2012,

2014; Kumari & Atri 2013; Malysheva & al. 2013; Ye & al. 2014; Ge & al. 2015;

Qasim &al. 2015; Dovana & al. 2017; Hussain & al. 2018; Usman & Khalid 2018).

From Pakistan only ten Leucoagaricus species have been reported previously:

L. asiaticus Qasim & al., L. badius S. Hussain & al., L. lahorensiformis S. Hussain

602 ... Ullah & al.

& al., L. lahorensis Qasim & al., L. leucothites (Vittad.) Wasser, L. pabbiensis

Usman & Khalid, L. pakistaniensis Jabeen & Khalid, L. serenus (Fr.) Bon &

Boiftard. L. sultanii S. Hussain & al., and L. umbonatus S. Hussain & al. (Ahmad

& al. 1997, Ge & al. 2015, Qasim & al. 2015, Hussain & al. 2018, Usman &

Khalid 2018). The genus is characterized morphologically by small- to medium-

sized basidiomata with entire pileus margins, free lamellae, a central, equal to

bulbous stipe, a membranous, persistent annulus, a white, cream, or pink spore

print white, usually dextrinoid basidiospores, the presence of cheilocystidia,

rare occurrence of pleurocystidia, and absence of clamp connections (Singer

1986).

Several molecular phylogenies indicate that Leucoagaricus is polyphyletic

(Johnson & Vilgalys 1998, Johnson 1999, Vellinga 2004). Vellinga (2003,

2004) showed that Leucoagaricus and Leucocoprinus species cluster together

phylogenetically but intermix within a single clade. Because of the large number

of species in the clade and relatively limited molecular data, the taxonomic

and phylogenetic relationships among many Leucoagaricus and Leucocoprinus

species have not yet been resolved.

During the present investigation, one Leucoagaricus specimen collected

from the Shawar Valley (district Swat, Khyber Pakhtunkhwa, Pakistan) is

proposed here as a new species, L. brunneus, supported by our morphological

and phylogenetic analyses.

Material & methods

The mushroom was collected during a forest foray in Shawar valley, Khyber

Pakhtunkhwa, Pakistan, during the 2017 monsoon season and photographed at the

sampling site. Macrocharacters were recorded from the fresh basidioma, with colors

coded according to Munsell’s Soil Color Charts (1975). The specimen was air dried for

further analyses and deposited in the Herbarium of University of the Punjab, Quaid-

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

For microscopic studies, fruiting body tissues were mounted in 2% KOH and

Congo red (to increase contrast). Basidia, basidiospores, cheilocystidia, and pellicular

elements were measured using Scopelmage 9.0. DNA extraction followed Bruns

(1995). The internal transcribed spacer region (ITS1-5.8S-ITS2 = ITS) was amplified

with the ITS1F/ITS4 primer combination (White & al. 1990, Gardes & Bruns 1993).

PCR products were purified and both strands were sequenced by Beijing Genomic

Institute (BGI).

For phylogenetic analysis, an ITS consensus sequence was generated in BioEdit

(Hall 1999) and BLAST searched at NCBI (https://www.ncbi.nlm.nih.gov/);

closely related sequences in L. sect. Rubrotincti were retrieved from both database

and literature (Hussain & al. 2018). Lepiota subgracilis Kihner was chosen as

outgroup (Liang & al. 2010). All sequences were aligned online using PRANK tool

Leucoagaricus brunneus sp. nov. (Pakistan) ... 603

(https://www.ebi.ac.uk/goldman-srv/webprank/). Gaps were treated as missing data.

The final aligned ITS dataset was phylogenetically analyzed through MEGA 6.0

software. Maximum Likelihood (ML) analysis was performed by selecting Kimura

2-parameter with Gamma Distributed model using Best-Fit Substitution Model

approach in MEGA 6.0 (Tamura & al. 2013).

Taxonomy

Leucoagaricus brunneus Z. Ullah, Jabeen & Khalid, sp. nov. FIGS 1, 2

MB 827985

Differs from Leucoagaricus truncatus by its smaller size, smooth shiny non-squamulose

pileus with brown striations, and its longer and narrower elongate to cylindrical

basidiospores.

Type: Pakistan. Khyber Pakhtunkhwa Province, Swat district, Lower Shawar, under

Quercus oblongata D. Don (Fagaceae), 8 July 2017, Zia Ullah LS4 (Holotype, LAH35862;

GenBank MH990662).

EryMotoey: brunneus (Latin) refers to the brown coloration of the pileus fibrils.

Figure. 1. Leucoagaricus brunneus (holotype, LAH35862). Basidioma. Scale bars = 0.5 cm.

604 ... Ullah & al.

Figure. 2. Leucoagaricus brunneus (holotype, LAH35862). A. Basidiospores; B. Basidia;

C. Cheilocystidia; D. Pileipellis; E. Stipitipellis. Scale bars = 10 um. (Drawing by Sana Jabeen.)

PiLteEus 35 mm diam, plane with incurved margins and umbonate center,

surface smooth and shiny, dark brown (10YR1/2) from center to lighter brown

(10YR2/4) towards margin in the form of radial striations, context cream.

Leucoagaricus brunneus sp. nov. (Pakistan) ... 605

LAMELLAE free, close to crowded, margins entire, cream. LAMELLULAE

absent. STIPE central, 9 x 4-8 mm, narrower towards the apex and wider

(<8 mm) towards the base, then again narrowing (<6 mm) at the base,

cream with grayish brown (10YR4/4) patches over the central part; annulus

superior (at the center of the upper half), white.

BasIDIOSPORES [30/1/1] (8.0—)8.2-10.5(-10.6) x (4-)4.1-4.9(-5.1) um,

Q = (1.8-)1.9-2.1(-2.2), avQ = 2, elongate to cylindrical in face view,

amygdaliform in side view, smooth, germ pore lacking, dextrinoid in

Melzer’s reagent, hyaline in KOH. Basip1a 17.3-19 x 7.4—-8.1 um, clavate,

smooth, hyaline, 4-spored. CHEILOCYSTIDIA 29.7-32.8 x 8.5-12.7 um,

clavate to subclavate, hyaline, without crystals. PLEUROCysTIDIA absent.

PILEIPELLIS a cutis with slightly clavate to cylindrical elements, 4.1-6.7 um

diam, hyaline in KOH. Stiprripexuis cylindric hyphae, 3.0-5.2 um diam,

hyaline in KOH. CLAMP CONNECTIONS absent in all tissues.

EcoLtocy & DISTRIBUTION—Saprobic and solitary on humus-rich

soil under Quercus oblongata [= Q. incana Roxb., nom. illeg.], at 1200 m

elevation, in moist temperate Quercus vegetation.

Phylogenetic analysis

The 72 ITS sequence dataset (TABLE 1, Fic. 3) comprises 757 positions.

Maximum likelihood analysis clusters the local collection in a sister clade with

L. truncatus Z.W. Ge & Zhu L. Yang and L. purpureolilacinus Huijsman with a

bootstrap support of 72% (Fia. 3).

Discussion

Leucoagaricus brunneus can be distinguished from L. truncatus based on

basidioma size. Leucoagaricus truncatus produces medium to large (40-80

mm) basidiomata (Ge & al. 2015) and is further separated by the orange-white

to gray-orange furfuraceous squamules (Ge & al. 2015) on its pileus surface

in contrast to the smooth shiny pileus with brown striations that distinguish

L. brunneus. Leucoagaricus truncatus is further separated microscopically by

its more broadly ovoid and broadly amygdaliform basidiospores (Ge & al.

2015). The ITS sequence analysis also provides strong bootstrap support for

L. brunneus as an independent taxon.

Leucoagaricus brunneus differs from L. purpureolilacinus, which is

characterized by a pinkish brown pileus and presence of crystalliferous

cheilocystidia (Vellinga 2001).

Pileus morphology also separates Leucoagaricus brunneus with its dark

brown umbo and brownish striations running towards the margin from

606 ... Ullah & al.

the center from L. serenus and L. crystallifer Vellinga, both characterized

by whitish basidiomata and pilei with white-to cream obtuse umbos with

obvious striations near the margins. Leucoagaricus serenus and L. crystallifer

are further distinguished by their ovoid and broadly amygdaliform

TABLE 1. Leucoagaricus species and specimens, and Lepiota subgracilis outgroup, used

for ITS phylogenetic analyses.

SPECIES VOUCHER COUNTRY GENBANK NO.

L. americanus Vellinga 2454 (UCB) USA AY 176407

JRH091509-1 (TENN) USA MF773593

L. asiaticus LAH5872011 Pakistan KP 164972

LAH10012012 Pakistan KP 164971

L. badius LAH SH210 Pakistan KU647734

LAH SH148 Pakistan KU647736

L. bresadolae Bas7981 USA AF295929

MCVE:756 Italy GQ329047

CCBAS802 Czech Rep. LN714565

L. brunneus LAH35862 [T] Pakistan MH990662

L. crystallifer Huijser (L) Germany AF482863

SFC 1010003-02 Spain KY350216

L. dyscritus Vellinga 3532B (UC) USA GU136181

Vellinga 3956 (UC) USA GU136180

L. gaillardii MCVE:16517 Italy GQ329064

MCVE:736 Italy GQ329042

L. griseodiscus MCVE:13719 Italy GQ329059

L. japonicus J. Li221 China KY039572

L. jubilaei Guinberteau 99101101 France AY243635

10115A USA KX258658

L. lacrymans Zhang 599 China KY039574

L. lahorensiformiS FH-SHL2 Pakistan KU647730

LAH SHL2 Pakistan KU647729

L. littoralis MCVE:856 Italy GQ329051

MCVE:702 Italy GQ329041

MCVE:13721 Italy GQ329060

L. cf. majusculus MFLU 09-0164 Thailand HM488764

L. medioflavoides MCVE:2324 Italy GQ329055

L. meleagris IMG 1671 USA KY680786

Vellinga 2095 (L) Netherlands AF482867

Vellinga 1990 (L) Netherlands AY176419

CAW-9 India GQ249888

L. menieri

L. nivalis

L. pakistaniensis

L. purpureolilacinus

L. rubroconfusus

L. rubrotinctus

L. sardous

Leucoagaricus sp.

L. subcrystallifer

L. sublittoralis

L. subpurpureolilacinus

L. subvolvatus

L. sultanii

L. truncatus

L. umbonatus

L. vassiljevae

L. viscidulus

L. volvatus

Lepiota subgracilis

herb. Huijser

Yang 5792

LAH SJF13

LAH SJF23

MCVE:754

Vellinga 2291 (L)

ZT13003 (ZT)

gr0557

gr157

KUN:HKAS 54317

KUN:HKAS 54240

AC5195

Thiele 2646

JZB2115002

BAB 4737

Ge 97

Yang 3972

Vellinga 2561

TENN:070790

FLAS-F-60259

M.M. Rogers (UCB)

Vellinga 2484 (UC)

Ge 878

Ecv2235

Yang 3959

Ge 406

Brand s.n. (L)

LAH SH115b

LAH SH115

Ge 793

LAH SHL1

LAH SHL8

LE 10350

LE 289338

LE 289432

AC4187

AC1785

Vellinga 1783 (L)

HKAS 5802

Netherlands

China

Pakistan

Italy

Netherlands

USA

China

Spain

Australia

China

India

China

USA

China

Netherlands

China

USA

Pakistan

China

Pakistan

Russia

Spain

Netherlands

China

Leucoagaricus brunneus sp. nov. (Pakistan) ... 607

KP300879

KY039573

KU647727

KU647728

GQ329045

AF482869

KP300875

KP300877

KP300876

JN944082

JN944081

JX827166

KT992149

AY176432

JN907015

KR154966

KP096237

KP096238

AY176430

MF686514

MF153051

AY176434

GU136182

KP096236

AY176442

KP096234

KP096233

KP300878

KU647732

KU647733

KP096235

KU647737

KU647738

JX133169

JX133170

JX896447

KT992148

KT992150

AY176490

EU416290

608 ... Ullah & al.

69 KP300877 L. rubrotinctus

AY 176442 L. sublittoralis

KP300876 L. rubrotinctus

69 | JX133169 L. vassiljevae

KY 039572 L. japonicus

JX133170 L. vassilievae

JX896447 L. vassilievae

96 |75 JN944082 L. rubrotinctus

GQ329045 L. purpureolilacinus

99 KU647730 L. lahorensiformis

KU647729 L. lahorensiformis

4 JN944081 L. rubrotinctus

V1 KP300875 L. rubroconfusus

50 JX827166 L. rubrotinctus

63 AY 176432 Leucoagaricus sp.

99 | KP096234 L. subpurpureolilacinus

KP096233 L. subpurpureolilacinus

93 | KU647727 L. pakistaniensis

KU647728 L. pakistaniensis

92| P24 JN907015 Leucoagaricus sp.

99 KR154966 Leucoagaricus sp.

98 /KU647734 L. badius

KU647736 L. badius

99 — KP164972 L. asiaticus

KP164971 L. asiaticus

KT992148 L. viscidulus

90 99 KU647732 L. sultanii

66 50 KU647733 L. sultanii

I 99 — AF482863 L. crystallifer

3p KY350216 L. crystallifer

hg -— KP300879 L. menieri

g, KP300878 L. subvolvatus

4 KT992150 L. volvatus

46 KP096236 L. subcrystallifer

KT992149 L. sardous

99 | GQ329051 L. littoralis

GQ329041 L. littoralis

27 GQ329060 L. littoralis

95 | KP096237 Leucoagaricus e

45 Q329059 L. griseodiscus

KP096238 Leucoagaricus sp.

KY 039573 L. nivalis

78 99 KU647737 L. umbonatus

53 KU647738 L. umbonatus

52 AY 176430 Leucoagaricus sp.

99 |MF686514 Leucoagaricus sp.

MF 153051 Leucoagaricus sp.

51 AF482869 L. purpureolilacinus

Se ? KP096235 L. truncatus

@ MH990662 L. brunneus

AY 176434 Leucoagaricus sp.

99 AY243635 L. jubilaei

18 KX258658 L. jubilaei

is [197 ,GU136181 L. yee

53 GU136180 L. dyscritus

GU 136182 Leucoagaricus sp.

99 ,GQ329064 L. gaillardii

56 GQ329042 L. gaillardii

GQ329055 L. medioflavoides

“4 KY039574 L. lacrymans

4 og | KY680786 L. meleagris

AF 482867 L. meleagris

50 AY 176419 L. meleagris

GQ249888 L. meleagris

84 HM488764 L. cf. majusculus

AY 176407 L. americanus

81 | _ AF295929 L. bresadolae

56} GQ329047 L. bresadolae

MF773593 L. americanus

LN714565 L. bresadolae

99 ,AY176490 Lepiota subgracilis

—| £U416290 Lepiota subgracilis

-——XH

0.02

Ficure. 3. Molecular phylogenetic analysis of ITS sequences of Leucoagaricus species, with Lepiota

subgracilis outgroup. The evolutionary history was inferred by using the Maximum Likelihood

method based on the Tamura 3-parameter model. The analysis involved 72 nucleotide sequences.

All positions containing gaps and missing data were eliminated. There were a total of 448 positions

in the final dataset. Our new species is marked with @.

Leucoagaricus brunneus sp. nov. (Pakistan) ... 609

basidiospores, contrasting with the elongated to cylindric and amygdaliform

basidiospores of L. brunneus. In addition, in L. crystallifer the cheilocystidia

have obvious crystals on the surface (Vellinga 2000, 2001) unlike the smooth

cheilocystidia of L. brunneus.

In comparison with Pakistani Leucoagaricus species, L. brunneus differs

from L. asiaticus characterized by oblong to ellipsoid spores and cheilocystidia

with crystals at the apex (Ge & al. 2015). Morphologically, its pileus color

distinguishes L. brunneus (brown) from L. badius (red), L. pakistaniensis

(whitish), L. lahorensis (dark reddish brown), L. lahorensiformis (light

orange), L. umbonatus (yellowish pink), and L. sultanii (dark yellow) (Qasim

& al. 2015, Hussain & al. 2018). Basidiospore morphology also separates

these species from L. brunneus (Qasim & al. 2015, Hussain & al. 2018). All

the earlier named Pakistani species have smaller spores except L. lahorensis,

whose spores measure more or less the same as L. brunneus (Qasim &

al. 2015), and none except L. brunneus present the same basidiospore

shape—elongate to cylindrical in face view and amygdaliform in profile:

amygdaliform to oblong in L. badius (6.5-7.5 x 4-5 um), ellipsoid to rarely

amygdaliform in L. pakistaniensis (7.5-8 x 4.5-5 um), broadly ellipsoid to

ellipsoid in L. lahorensis (8-10.6 x 6.4—7.6 um), fusiform to amygdaliform

in L. lahorensiformis (6.5-7.5 x 3.5-4 um), amygdaliform to ellipsoid

in L. umbonatus (5.5-6.5 x 3.5-4 um), and amygdaliform to ellipsoid in

L, sultanii (5.5-7 x 3.5-4.5 um) (Qasim & al. 2015, Hussain & al. 2018). The

crystals on the cheilocystidial apices in L. lahorensiformis and L. sultanii also

separate both species from L. brunneus.

Therefore a combination of morphological differences and a phylogenetic

bootstrap support of 72% (Fic. 3) provide convincing support for our new

species, Leucoagaricus brunneus.

Acknowledgments

The work is funded by HEC Research Project number 20-3383/HEC/R&D/14/184.

Sincere thanks to Dr. Chang-Lin Zhao (Forestry College, Southwest Forestry

University, Yunnan, P.R. China) and Dr. Shah Hussain (Center for Plant Sciences and

Biodiversity, University of Swat, Pakistan) for presubmission review of the manuscript.

Their comments and suggestions greatly helped to improve the document. We are

thankful to Sheraz Khan, Numan Fazal, and Niaz Ali for their help in sampling.

Literature cited

Ahmad S., Iqbal SH., Khalid AN. 1997. Fungi of Pakistan. Sultan Ahmad Mycological Society of

Pakistan, Department of Botany, University of the Punjab, Quaid-e-Azam campus, Lahore.

610 ... Ullah & al.

Bruns TD. 1995. Thoughts on the processes that maintain local species diversity of ectomycorrhizal

fungi. 63-73, in: HP Collins & al. (eds). The significance and regulation of soil biodiversity.

Springer, Netherlands. https://doi.org/10.1007/978-94-011-0479-1_5

Dovana F, Contu M, Angeli P, Brandi A, Mucciarelli M. 2017. Leucoagaricus ariminensis sp. nov.,

a lilac species from Italy. Mycotaxon 132: 205-216. https://doi.org/10.5248/132.205

Gardes M., Bruns TD. 1993. ITS primers with enhanced specificity of basidiomycetes:

application to the identification of mycorrhizae and rusts. Molecular Ecology 2: 113-118.

https://doi.org/10.1111/j.1365-294X.1993.tb00005.x

Ge ZW. 2010. Leucoagaricus orientiflavus, a new yellow lepiotoid species from southwestern

China. Mycotaxon 111: 121-126. https://doi.org/10.5248/111.121

Ge ZW, Yang ZL, Qasim T, Nawaz R, Khalid AN, Vellinga EC. 2015. Four new species in

Leucoagaricus (Agaricaceae, Basidiomycota) from Asia. Mycologia 107(5): 1033-1044.

https://doi.org/10.3852/14-351

Hussain S, Jabeen S, Khalid AN, Ahmad H, Afshan NUS, Sher H, Pfister DH. 2018.

Underexplored regions of Pakistan yield five new species of Leucoagaricus. Mycologia 1-14.

https://doi.org/10.1080/00275514.2018.1439651

Johnson J. 1999. Phylogenetic relationships within Lepiota sensu lato based on morphological

and molecular data. Mycologia 91: 443-458. https://doi.org/10.2307/3761345

Johnson J, Vilgalys R. 1998. Phylogenetic systematics of Lepiota sensu lato based on nuclear

large subunit rDNA evidence. Mycologia 90: 971-979.

https://doi.org/10.1080/00275514.1998.12026994

Kumar TKA, Manimohan P. 2009. The genera Leucoagaricus and Leucocoprinus (Agaricales,

Basidiomycota) in Kerala state, India. Mycotaxon 108: 385-428.

https://doi.org/10.5248/108.385

Kumari B, Atri NS. 2013. New additions of basidiomycetous fungi in Indian mycoflora.

Mycosphere 4: 53-59. https://doi.org/10.5943/mycosphere/4/1/4

Liang JF, Yang ZL, Xu J, Ge ZW. 2010. Two new unusual Leucoagaricus species (Agaricaceae)

from tropical China with blue-green staining reactions. Mycologia 102: 1141-1152.

https://doi.org/10.3852/09-021

Locquin M. 1943a. Etude du développement des spores du genre Leucocoprinus Pat. (Troisiéme

partie) suivie de la description d'une espéce nouvelle et dune espéce critique. Bulletin Mensuel

de la Société Linnéenne de Lyon 12(5): 75-80. https://doi.org/10.3406/linly.1943.9740

Locquin M. 1943b. Etude du développement des spores du genre Leucocoprinus Pat. (Troisiéme

partie) suivie de la description d'une espéce nouvelle et dune espéce critique. Bulletin Mensuel

de la Société Linnéenne de Lyon 12(6): 91-96. https://doi.org/10.3406/linly.1943.9747

Malysheva EF, Svetasheva TY, Bulakh EM. 2013. Fungi in the Russian Far East. I. Leucoagaricus

lateritiopurpureus and new species of Leucoagaricus (Agaricaceae) with reddish brown

basidiocarps. Mikologiya i Fitopatologiya 47: 169-179.

Mufioz G, Caballero A, Contu M, Vizzini A. 2012. A new Leucoagaricus species of section Piloselli

(Agaricales, Agaricaceae) from Spain. IMA Fungus 2: 117-123.

Mufioz G, Caballero A, Contu M, Ercole E, Vizzini A. 2014. Leucoagaricus croceobasis (Agaricales,

Agaricaceae), a new species of section Piloselli from Spain. Mycological Progress 13(3):

649-655. https://doi.org/10.1007/s11557-013-0947-x

Munsell Soil Color Charts. 1975. Munsell Color Co, Baltimore, MD.

Qasim T, Amir T, Nawaz R, Niazi AR, Khalid AN. 2015. Leucoagaricus lahorensis, a new species of

L. sect. Rubrotincti. Mycotaxon 130(2): 533-541. https://doi.org/10.5248/130.533

Singer R. 1948. Diagnoses fungorum novorum agaricalium. Sydowia 2: 35-36.

Leucoagaricus brunneus sp. nov. (Pakistan) ... 611

Singer R. 1986. The Agaricales in modern taxonomy. 4th ed. Koenigstein: Koeltz Scientific

Books. 450 p.

Tamura K, Stecher G, Peterson D, Filipski A, Kumar S. 2013. MEGA6: Molecular Evolutionary

Genetics Analysis version 6.0. Molecular Biology and Evolution 30: 2725-2729.

https://doi.org/10.1093/molbev/mst197

Usman M., Khalid AN. 2018. Leucoagaricus pabbiensis sp. nov. from Punjab, Pakistan.

Mycotaxon 133(2): 354-363. https://doi.org/10.5248/133.355

Vellinga EC. 2000. Notes on Lepiota and Leucoagaricus. Type studies on Lepiota magnispora,

Lepiota barssii and Leucoagaricus americanus. Mycotaxon 76: 429-438.

Vellinga EC. 2001la. Leucoagaricus. 85-108, in: ME Noordeloos & al. (eds). Flora Agaricina

Neerlandica, Vol. 5.

Vellinga EC. 2001b. Notulae ad floram agaricinam neerlandicam—XXXVIII. Leucoagaricus

subgenus Sericeomyces. Persoonial7(3): 473-480.

Vellinga EC. 2003. Phylogeny and taxonomy of lepiotaceous fungi [Doctoral dissertation].

Leiden: Univ Leiden Press. 259 p.

Vellinga EC. 2004. Genera in the family Agaricaceae: evidence from nrITS and nrLSU sequences.

Mycological Research 108: 354-377. https://doi.org/10.1017/S0953756204009700

Vellinga, EC. 2010. Lepiotaceous fungi in California, U.S.A. Leucoagaricus sect. Piloselli.

Mycotaxon 112: 393-444. https://doi.org/10.5248/112.393

Vellinga EC, Balsley RB. 2010. Leucoagaricus dacrytus — a new species from New Jersey, USA.

Mycotaxon 113: 73-80. https://doi.org/10.5248/113.73

Vellinga EC, Contu M., Vizzini A. 2010. Leucoagaricus decipiens and L. erythrophaeus, a new

species pair in sect. Piloselli. Mycologia 102: 447-454. https://doi.org/10.3852/09-164

Vellinga E, Sysouphanthong P, Hyde KD. 2011. The family Agaricaceae: phylogenies and two

new white-spored genera. Mycologia 103: 494-509. https://doi.org/10.3852/10-204

White TJ, Bruns T, Lee S, Taylor J. 1990. Amplification and direct sequencing of fungal

ribosomal RNA genes for phylogenetics. 315-322, in: MA Innis & al. (eds). PCR

protocols: a guide to methods and applications. Academic Press, San Diego, pp. 315-322.

https://doi.org/10.1016/B978-0-12-372180-8.50042-1

Ye Y, Li YK., Liang JF. 2014. Leucoagaricus tangerinus, a new species with drops from Southern

China. Mycological Progress 13: 893-898. https://doi.org/10.1007/s11557-014-0974-2

MY COTAXON

October-December 2019—Volume 134, pp. 613-618

https://doi.org/10.5248/134.613

Dictyostelids from Jilin Province, China 3:

new Cavenderia and Dictyostelium records

Pu Liu', SHUNHANG ZHANG', YUE ZOU’, XUEPING KANG’, Yu LI"

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

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

?Yanbian Academy of Agricultural Sciences, Yanji 133001, P. R. China

* CORRESPONDENCE TO: fungi966@126.com

ABSTRACT—Cavenderia parvispora, Dictyostelium vermiforme, and D. dimigraforme, isolated

from samples of soil collected in Jilin Province, China, represent new records from China.

Descriptions and illustrations based on these isolates are provided.

Key worps—Cavenderiaceae, cellular slime molds, Dictyosteliaceae, Mycetozoa, taxonomy

Introduction

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

(protozoan) and fungus-like characteristics, are primarily inhabitants of the

soil and leaf litter/humus zone of fields and forests, along with animal dung,

where they feed mostly on bacteria. These organisms play an essential role in

maintaining the balance that exists between bacteria and other soil organisms

(Singh 1947; Cavender & Raper 1965a,b; Raper 1973, 1984). Traditional

morphological classifications distributed dictyostelids among four genera

based on differences in sorophore structure and branching patterns (Kirk &

al. 2008). However, a new classification based on unique 18S rRNA sequence

signatures was proposed by Sheikh & al. (2018).

Jilin Province is situated in the temperate zone of China and is characterized

by a monsoon climate (Liu & Li 2014). Previously, a total of 15 species, including

two new dictyostelid species, have been reported from Jilin Province (He & Li

614 ... Liu & al.

2008; Liu & Li 2014, 2017). We report another three species in Cavenderia and

Dictyostelium for the first time from China.

Materials & methods

SAMPLING, ISOLATION, CULTIVATION—Samples of soil were collected from three

localities in Jilin Province in 2008 and 2009. Each 10-20 g soil sample was placed in a

sterile whirl-pack plastic bag for isolation according to Cavender & Raper (1965a). Each

sample was weighed and enough ddH,O added for an initial dilution of 1:10. A 0.5 mL

aliquot of this dilution was added to each of five duplicate culture plates prepared with

hay infusion agar (Raper 1984). After c. 0.4 mL of a heavy suspension of the bacterium

Escherichia coli was added to each culture plate, the plates were incubated at 23 °C with

a 12h light : 12 h dark cycle. Each plate was examined at least once daily for two weeks

after the appearance of initial aggregations. Each isolate recovered from one of the plates

was purified and cultivated for taxonomic studies. Spores from these isolates were frozen

in HL 5 media (Cocucci & Sussman 1970) and stored at —80 °C in the herbarium of the

Mycological Institute of Jilin Agricultural University, Changchun, China (HMJAU).

OBSERVATION—The isolates were identified using morphological descriptions

(Raper 1984) and molecular characteristics (Sheikh & al. 2018). After we marked the

location of each early aggregating clone and sorocarp in a plate, we observed life cycle

stages under a Zeiss Axio Zoom V16 dissecting microscope with a 1.5x objective

and a 10x ocular. Sorocarps were mounted in water on slides for observation and

measurements of spores, sorophores, and sorocarps using a Zeiss Axio Imager A2 light

microscope mounted with 10x ocular and 10, 40, and 100x (oil) objectives. Photographs

were taken with Zeiss Axiocam 506 color microscope camera.

Taxonomy

Cavenderia parvispora (H. Hagiw.) S. Baldauf, S. Sheikh & Thulin,

Protist 169(1): 20. 2018. PLATE 1A-D

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

gregarious or solitary, unbranched or sparsely irregularly branched, sometimes

prostrate, phototropic, normally 0.2-4.4 mm long. Sorophore colorless, sinuose,

slender, tapering from bases to tips, consisting of one tier of cells except for

the in base and tip, bases clavate, tips clavate. Sori white, globose, commonly

30-150 um diam. Spores hyaline, elliptical, 3.7-6.0 x 1.9-2.9 um, with a few

conspicuous polar granules. Aggregations with radiate streams or mound-like.

SPECIMEN EXAMINED: CHINA, JILIN PROVINCE, Zuojjia, isolated from soil ($0425) from

a broadleaf forest, 9 Sep. 2008 (HMJAU MR056).

COMMENTS: Cavenderia parvispora was first isolated from forest soil sampled in

Japan (Hagiwara 1989). Its medium-sized sorocarps, thin delicate sorophores

with clavate tips, and small elliptical spores readily distinguish this species

from morphologically similar dictyostelids.

Cavenderia and Dictyostelium spp. new from Jilin (China) ... 615

198 @ OVE t.

PEGI @ o® =

a” ee cy “Fry “aK A/ QO see ee

if : _ pee “| ee! Cy =e : .

~ off *

PLATE 1. Cavenderia parvispora (HMJAU MR056): A. Sorocarps; B. Sorophore base;

C. Sorophore tip; D. Spores. Dictyostelium vermiforme (HMJAU MR058): E. Sorocarps;

F, G. Vermiform and curved sorogens; H. Sorophore base; I. Sorophore tip; J. Spores. Scale bars:

A, E= 2 mm; B, C, H, 1 =5 um; D,J = 10 um; KF G=1 um.

616 ... Liu & al.

Dictyostelium vermiforme Vadell & Cavender, Mycologia 99: 118. 2007. PLATE 1E-J

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

clustered or solitary, normally prostrate, 1.0-2.0 mm long. Sorophore colorless,

tapering from bases to tips, consisting of one tier of cells except for the base

and tip, bases small round or expanded, tips capitate or expanded. Sori white,

globose, commonly 50-130 um diam. Spores hyaline, oblong to elliptical,

4.7-7.0 x 2.9-3.9 um, with sparse or prominent polar granules. Aggregations

with flattened streams, forming mound-like early sorogens. Sorogens

developing rapidly, becoming vermiform and curved, migrating freely to form

sinuous and curled late sorogens with prostrate sorophores.

SPECIMEN EXAMINED: CHINA, JILIN PROVINCE, National Nature Reserve, isolated

from soil (S0083-1) from a broadleaf forest, 10 Jun. 2009 (HMJAU MR058).

ComMENTs: Dictyostelium vermiforme was originally isolated from soil/litter

sampled in Iguazu Regional Park, Argentina (Vadell & Cavender 2007). This

species is characterized by its vermiform and curved early migrating sorogens.

Dictyostelium dimigraforme Cavender, J. Gen. Microbiol. 62:115.1970. | PLATE 2

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

normally solitary, unbranched, erect or inclined, 3.5-10.5 mm long. Sorophore

colorless or slightly yellow, tapering from bases to tips, consisting of one tier of

cells except for the base and tip, bases round or slightly enlarged, tips capitate.

Sori globose, off-white to yellow, 220-380 um diam. Spores hyaline, oblong

to elliptical, 7.7-12.0 x 2.9-4.9 um, without polar granules. Aggregations

radiating.

SPECIMEN EXAMINED: CHINA, JILIN PROVINCE, Chanchung, Jingyuetan National

Natural Park, isolated from soil (S0446) from a broadleaf forest, 10 Sep. 2008 (HMJAU

MRO059).

ComMENTs: Dictyostelium dimigraforme was isolated the first time from surface

soil sampled in a tropical forest in the West Indies (Raper 1984). This species is

characterized by its yellow sori and round bases, and can be distinguished from

D. discoideum by its lack of basal disk.

Discussion

Our study has raised to 18 the number of dictyostelid species known from

Jilin Province. Jilin Province is located in the temperate zone which has rich

forests optimal for dictyostelids. The three dictyostelid species (Cavenderia

parvispora, Dictyostelium vermiforme, and D. dimigraforme) reported here do

not appear to be distributed worldwide (Hagiwara 1989, Vadell & Cavender

2007, Raper 1984). In particular, Dictyostelium vermiforme (originally isolated

Cavenderia and Dictyostelium spp. new from Jilin (China) ... 617

ra €

PLATE 2. Dictyostelium dimigraforme (HMJAU MR059): A, B. Sorocarps; C. Sorophore base;

D. Sorophore tip; E. Spores. Scale bars: A, B = 2 mm; C, D =5 um; E= 10 um.

618 ... Liu & al.

from c. 26°S in South America) and D. dimigraforme (originally isolated from

c. 11°N in the West Indies) have now been isolated from c. 44°N in Asia. Clearly,

the dictyostelids of Jilin Province need to be investigated in more detail.

Acknowledgments

We wish to express our appreciation to peer reviewers Prof. Steven L. Stephenson

(University of Arkansas, U.S.A.) and Prof. John C. Landolt (Shepherd University, U.S.A.)

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. 1965a. The Acrasieae in nature. I. Isolation. American Journal of

Botany 52: 294-296. https://doi.org/10.2307/2439943

Cavender JC, Raper KB. 1965b. The Acrasieae in nature. II. Forest soil as a primary habitat.

American Journal of Botany 52: 297-302. https://doi.org/10.2307/2439944

Cocucci SM, Sussman M. 1970. RNA in cytoplasmic and nuclear fractions of cellular slime

mold amoebas. Journal of Cell Biology 45: 399-407. https://doi.org/10.1083/jcb.45.2.399

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

Science Museum, Tokyo.

He XL, Li Y. 2008. A new species of Dictyostelium. Mycotaxon 106: 379-383.

Kirk PM, Cannon PF, Minter DW, Stalpers JA. 2008. Dictionary of the fungi, 10th edition.

CABI, UK.

Liu P, Li Y. 2014. Dictyostelids from Jilin Province, China. I. Phytotaxa 183(4): 279-283.

https://doi.org/10.11646/phytotaxa.183.4.7

Liu P, Li Y. 2017. Dictyostelids from Jilin Province, China. II. Phytotaxa 323(1): 77-82.

https://doi.org/10.11646/phytotaxa.323.1.6

Raper KB. 1973. Acrasiomycetes. 9-36, in: GC Ainsworth &al. (eds). The Fungi, vol. IVB.

Academic Press, Inc. New York.

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

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): 1-28.

https://doi.org/10.1016/j.protist.2017.11.001.

Singh BN. 1947. Studies on soil Acrasieae. 1. Distribution of species of Dictyostelium in soils

of Great Britain and the effects of bacteria on their development. Journal of General

Microbiology 28: 417-429. https://doi.org/10.1099/00221287-1-1-11

Vadell EM, Cavender JC. 2007. Dictyostelids living in the soils of the Atlantic Forest, Iguazu

Region, Misiones, Argentina: description of new species. Mycologia 99(1): 112-124.

https://doi.org/10.1080/15572536.2007.11832606

MYCOTAXON

October-December 2019—Volume 134, pp. 619-625

https://doi.org/10.5248/134.619

Records of Aureobasidium harposporum, Sarcophoma miribelii,

and Stigmina dothideoides from Turkey

MAKBULE ERDOGDU’, MERVE ULUKAPT?,

ALI IHSAN KARAYEL?, ZEKIYE SULUDERE?

' Department of Landscape Architects, Faculty of Agriculture &

? Graduate School of Natural and Applied Sciences:

Ahi Evran University, Bagbas1, Kirsehir, Turkey

* Department of Biology, Faculty of Science, Gazi University,

Teknikokullar, Ankara, Turkey

* CORRESPONDENCE TO: merdogdu@ahievran.edu.tr

ABsTRACT—During field studies on the microfungi in Mucur District of Kirsehir Province,

Aureobasidium harposporum, and Stigmina dothideoides were revealed as new records for

Turkey; and Sarcophoma miribelii, previously recorded from northeastern Turkey, is reported

from central Turkey. Distinguishing morphological characters of these species are described,

and their photographs are provided.

Key worps—Ascomycota, Dothideales, leaf pathogenic fungi, Mycosphaerellales, SEM

Introduction

Aureobasidium Viala & G. Boyer (Saccotheciaceae, Dothideales)

contains 24 species (Index Fungorum 2018). The genus is characterized by

synchronous conidium production on hyaline conidiogenous cells. Species of

Aureobasidium produce variously shaped one-celled conidia from terminal,

lateral, or intercalary hyaline conidiogenous cells. Members of this genus

occur as saprophytes on a variety of substrates or as parasites on phanerogams

(Hermanides-Nijhof 1977).

Sarcophoma Hohn. (Dothioraceae, Dothideales) was proposed by Hohnel

in 1906 as a monotypic genus, but four species are now accepted (Index

Fungorum 2018).

620 ... Erdogdu & al.

Stigmina Sacc. (Mycosphaerellaceae, Mycosphaerellales) was established

by Saccardo in 1880; c. 80 species are currently accepted in the genus (Index

Fungorum 2018). In Turkey, the Stigmina species are poorly known and have

not yet been intensively studied; of the four species reported from Turkey—

S. carpophila, S. compacta, S. obtecta, S. platani—(Gdobelez 1964, Hiiseyin &

al. 2003, Cimen & Ertugrul 2007, Erdogdu & Huseyin 2008), two have been

transferred to other genera: S. compacta [= Thyrostroma compactum] and

S. platani [= Pseudocercospora platanigena].

Materials & methods

Host specimens were prepared following conventional herbarium techniques.

Host plants were identified using the Flora of Turkey and East Aegean Islands (Davis

1965-85). Thin fungal sections prepared from host tissue were examined under a

Leica DM E light microscope and measured from mounts in tap water. Infected

host surfaces were photographed using a Leica EZ4D stereomicroscope. The fungi

were identified using relevant literature (for Aureobasidium: Saccardo 1892 (as

Gloeosporium), Hermanides-Nijhof 1977; for Sarcophoma: Saccardo 1884 (under

Phoma), Morgan-Jones 1971, Aa 1975; for Stigmina: Ellis 1959, Shoemaker & Egger

1982). All examined specimens were deposited in the Mycology Laboratory of Ahi

Evran University, Department of Biology, Kirsehir, Turkey (AEUT).

For scanning electron microscopy (SEM), 8-10 mm square pieces of infected

leaf were mounted on aluminium stubs with double-sided adhesive tape. They were

coated with gold using Polaron SC 502 Sputter Coater and were examined with

Jeol JSM 6060 scanning electron microscope operated at 5-10 kV in the Electron

Microscopy Unit, Faculty of Science, Gazi University, Ankara, Turkey.

Taxonomy

Aureobasidium harposporum (Bres. & Sacc.) Herm.-Nijh.,

Stud. Mycol. 15: 151 (1977) PLATE 1

Spots visible on both sides of leaves and twigs, generally leaves beginning

to dry from the tip, brown on the upper surface, fuscous chestnut on the lower

surface, spots margin dark brown; infected leaves dropping prematurely from

the shrub. Conrp1omarta acervular, subepidermal, yellowish, broadly elliptical,

196-330 um diam. CONIDIOGENOUS CELLS erect, densely packed, clavate to

subcylindrical, 10-20 x 3.5-6.5 um, forming conidia simultaneously at the

apex. Conip1 one-celled, falcate, sometimes elliptical or slightly curved, oil

droplet, smooth-walled, 13.7-19.3(-21.8) x 4.4-5.4 um, hyaline.

SPECIMEN EXAMINED—TURKEY, Kirsenrr, Mucur District, Tekken Village,

39°02'13”N, 34°13’08”E, 977 m asl, on living leaves and stems of Viscum album L. subsp.

album (Santalaceae), 19.06.2013, M. Ulukap1 (AEUT MU1050).

Aureobasidium, Sarcophoma, Stigmina spp. in Turkey ... 621

10k \ xS"6Ge— ) Sam GUFEF | | {OkU ~~ x4,388 Sum GuFEF

PLATE 1. Aureobasidium harposporum (AEUT MU1050). A. leaf spots; B. acervuli on leaf;

C. acervuli on leaf (SEM); D, E. acervulus on leaf (SEM); E acervulus, vertical section; G. conidia;

H, I. conidia (SEM).

Notes: Viscum album (European mistletoe) is an evergreen, perennial,

epiphytic, hemiparasitic shrub that lives on a wide range of woody plant species

(Zuber 2004). European mistletoes can affect their host trees in many ways.

Known effects from mistletoe infection include lowering the vigor of the host,

inducing premature mortality, reducing quality and quantity of wood grown,

reducing fruit production, and predisposing trees to secondary infection by

other agents, such as insects or decay fungi (Hawksworth 1983).

Biological control of parasites by using plant pathogens has gained

acceptance as a practical, safe, and environmentally beneficial management

method applicable to agro-ecosystems (Charudattan 2001). The use of biological

control agents for weed control has attracted increased attention (Ozaslan &

al. 2013; Ozaslan 2016), and diagnosing host weeds and their natural enemies

is the foundation stone of a successful biological control program (Ozaslan

2016). Control of European mistletoe is an important consideration for the

forest service in Turkey (Yiiksel & al. 2005). Over 20 microscopic fungi live on

European mistletoe, but only a few of them cause major damage to the plant

622 ... Erdogdu & al.

(Karadzic & al. 2004). Of these, Aureobasidium harposporum, which causes leaf

spot disease of European mistletoe, appears to have potential as a biological

control agent against of this semi-parasite. In this study, Aureobasidium

harposporum on living leaves and stems of Viscum album subsp. album is

reported as new to the mycobiota of Turkey.

The Turkish specimen agrees with other reports of Aureobasidium

harposporum in conidiomata and conidia morphology, the only observable

difference being the smaller dimensions of acervuli and conidia. Saccardo

(1892) describes 200 um diam. acervuli and 18-20 x 4-5 um conidia, while

Hermanides-Nijhof (1977) cites 250-500 um acervuli and (16-)17-21(-23) x

3.5-5 um conidia.

XZ66 160)tmh

X7> 566 Zharn

Aureobasidium, Sarcophoma, Stigmina spp. in Turkey ... 623

Sarcophoma miribelii (Fr.) Héhn., Hedwigia 60: 133 (1918) PLATE 2

= Macrophoma miribelii (Fr.) Berl. & Voglino, Atti Soc.

Veneto-Trent. Sci. Nat.10(1): 179 (1886)

PYcNIDIA numerous, hypophyllous (rarely epiphyllous), at first covered

by the epidermis, later becoming erumpent, scattered, 143-289 um diam.,

yellowish or brown. Conrp1a single-celled, ovoid, rounded at apex, attenuate

at the base, smooth-walled, (8.4—)10-13.6(-14.3) x 6.2-8.7 um, hyaline,

with granular contents.

SPECIMEN EXAMINED—TURKEY, Kirseuir, Mucur District, Seyfegolti, 39°06’58’N,

34°12'11”E, 1135 m asl, on living leaves and stems of Buxus sempervirens L. (Buxaceae),

16.04.2014, M. Ulukapi (AEUT MU1099).

Notes: Our Turkish specimen is morphologically similar to Sarcophoma

miribelii specimens described in the literature (Saccardo 1884, Morgan-Jones

1971, Aa 1975). Sarcophoma miribelii has previously been reported (without

description or illustration) from Rize Province in coastal northeastern

Turkey (Gobelez 1964, Hiiseyin & al. 2005; both as Macrophoma mirbelii

[sic]), whereas our new record is from Kirsehir Province in central Turkey at

c. 1100 m asl. Sarcophoma miribelii is found worldwide, wherever the Buxus

host occurs (Aa 1975).

Stigmina dothideoides (Ellis & Everh.) M.B. Ellis, Mycol. Pap. 72: 53 (1959) PLATE 3

SPORODOCHIA erumpent through epidermis, pustulate, 1-4 mm

diam., circular to elliptical, blackish. CoNIDIOPHORES densely crowded,

arising from stromatic hyphae, pale brown to medium brown, cylindrical

to lageniform, 1-2- septate, often branched at the base. Conip1a blastic,

solitary and terminal, elliptical to obovoid, at first one-celled, becoming

3-septate, non-constricted or slightly constricted at the septum, rounded

at apex, truncated to obtusely rounded at the base, smooth-walled, 25-38 x

11.8-13.9(-14.5) um, golden brown.

SPECIMEN EXAMINED— TURKEY, Kirsenir, Mucur District, Gumtiskiimbet Village,

39°05'52”N, 34°12’26”E, 1180 m asl, on stems of Artemisia sp. (Asteraceae), 16.04.2014,

M. Ulukapi (AEUT MU1087).

Notes: The species was first described by Ellis & Everhart (1891, as

Clasterosporium dothideoides), who observed symptoms on stems of

Artemisia cana and Shepherdia argentea in Montana, USA; and it has

PLATE 2 (left). Sarcophoma miribelii (AEUT MU1099). A. pycnidia on leaf; B. pycnidia on

leaf (SEM); C. pycnidium on leaf (SEM); D. pycnidium, vertical section; E. conidia; F. conidia

(SEM).

624 ... Erdogdu & al.

15 um

Cp - ls

PLATE 3. Stigmina dothideoides (AEUT MU1087). Top Lert. conidia and conidiophores; Top

RIGHT. conidium and conidiophore; BoTTom. conidia.

been recorded on Gaillardia aristata further north in Alberta, Canada

(Shoemaker & Egger 1982). Stigmina dothideoides on branches of

Artemisia sp. is reported as new to the mycobiota of Turkey.

Although our Turkish specimen of Stigmina dothideoides is

morphologically similar to specimens described in the literature,

it differs slightly in having shorter and narrower 3-septate conidia,

compared with 25-46 x 13-19 um 3-septate conidia in Ellis (1959) and

30-38(-42) x (12.3-)14-16.8 um 3(—4)-septate conidia in Shoemaker

& Egger (1982).

Acknowledgments

The authors thank Sevda Kirbag (Department of Biology, Firat University, Elazig,

Turkey) and Cumali Ozaslan (Department of Plant Protection, Dicle University,

Diyarbakir, Turkey) for pre-submission review. This work was supported by the

Ahi Evran University Scientific Research Projects Coordination Unit. (Project

Number: PYO-Fen.4003.13.005 and FEF.A4.17.006).

Aureobasidium, Sarcophoma, Stigmina spp. in Turkey ... 625

Literature cited

Aa HA van der. 1975. The perfect state of Sarcophoma miribelii. Persoonia 8(3): 283-289.

Charudattan R. 2001. Biological control of weeds by means of plant pathogens: significance

for integrated weed management in modern agro-ecology. Biocontrol 46(2): 229-260.

https://doi.org/10.1023/A:101147753

Cimen I, Ertugrul BB. 2007. Determination of mycoflora in almond plantations under drought

conditions in Southeastern Anatolia project Region, Turkey. Plant Pathology Journal 6(1):

82-86. https://doi.org/10.3923/ppj.2007.82.86

Davis PH (ed.). 1965-85. Flora of Turkey and East Aegean Islands. Vols 1-9. Edinburgh

University Press, Edinburgh.

Ellis MB. 1959. Clasterosporium and some allied dematiaceae—phragmosporae. II. Mycological

Papers 72.75 p.

Ellis JB, Everhart BM. 1891. New species of fungi from various localities. Proceedings of the

Academy of Natural Sciences of Philadelphia 43: 76-93.

Erdogdu M, Huseyin E. 2008. Microfungi of Kurtbogazi1 Dam (Ankara) and its environment. Ot

Sistematik Botanik Dergisi 14(1): 131-150.

Gébelez M. 1964. La mycoflore de Turguie. (List of fungi of Turkey). II. Mycopathologia et

Mycologia Applicata 23(1): 47-67. https://doi.org/10.1007/BF02049185

Hawksworth FG. 1983. Mistletoes as forest parasites. 317-333, in: M Calder, P Bernhardt (eds).

The biology of mistletoes. Australia, Academic Press.

Hermanides-Nijhof EJ. 1977. Aureobasidium and allied genera. Studies in Mycology. 15: 141-177.

Huseyin E, Selcuk FE, Gaffaroglu M. 2003. Some materials on mitosporic fungi from Turkey I.

Hyphomycetes. Botanica Lithuanica 9(2): 151-160.

Hiiseyin E, Selcuk F, Gaffaroglu M. 2005. Materials on the micromycetes on box tree (Buxus) and

Rhododendron from Turkey. 62-68, in: Proceedings of the XVI Symposium of Mycologists

and Lichenologists of Baltic States. 21-25 September, 62-68, Cesis, Latvia.

Index Fungorum 2018: http://www.indexfungorum.org. [accessed September 2018].

Karadzi¢ D, Lazarev V, Milenkovi¢ M. 2004. The most significant parasitic and saprophytic

fungi on common mistletoe (Viscum album L.) and their potential application in biocontrol.

Bulletin Faculty of Forestry, University of Bajna Luka, Serbia 89: 115-126.

Morgan-Jones G. 1971. Conidium ontogeny in coelomycetes. I. Some amerosporous

species which possess annellides. Canadian Journal of Botany 49(11): 1921-1929.

https://doi.org/10.1139/b71-267

Ozaslan C, Hiiseyin E, Erdogdu M. 2013. Microfungi species on the weeds of agro-ecosystem

(wheat ecosystem) in Adtyaman City. Mantar Dergisi 4(2): 10-18.

Ozaslan C. 2016. Downy mildews species on the weeds of lentil fields in Diyarbakir in Turkey.

Scientific Papers. Series A. Agronomy 59: 365-367.

Saccardo PA. 1884. Sylloge Spheropsidearum et Melanconiarum. Sylloge Fungorum 3. 860 p.

Saccardo PA. 1892. Supplementum universale, pars II. Discomyceteae-Hyphomyceteae. Sylloge

Fungorum 10. 964 p.

Shoemaker RA, Egger KN. 1982. Stigmina dothideoides. Fungi Canadenses 212. 2 p.

Yuksel B, Akbulut S, Keten A. 2005. The damage, biology and control of pine mistletoes (Viscum

album ssp. austriacum (Wiesb.) Vollman). Turkish Journal of Forestry 2: 111-124.

Zuber D. 2004. Biological flora of Central Europe: Viscum album L. Flora 199(3): 181-203.

https://doi.org/10.1078/0367-2530-00147

MY COTAXON

October-December 2019—Volume 134, pp. 627-632

https://doi.org/10.5248/134.627

Bactrodesmium pulcherrimum sp. nov.

from Ecuador

FERNANDO ESPINOZA’, DAYNET SOSA’, LIZETTE SERRANO’,

ADELA QUEVEDO’, FREDDY MAGDAMA’, MARCOS VERA’,

SIMON PEREZ-MARTINEZ’, ELAINE MALOSSO},

RAFAEL F, CASTANEDA-Rut1Iz?*

' Escuela Superior Politécnica del Litoral, ESPOL, (CIBE),

Campus Gustavo Galindo Km. 30.5 Via Perimetral,

PO. Box 09-01-5863, Guayaquil, Ecuador

? Universidad Estatal de Milagro (UNEMI), Facultad de Ingenieria,

Cdla. Universitaria Km. 1.5 via Milagro-Km26. Milagro 091706, Guayas, Ecuador

* Centro de Biociéncias, Departamento de Micologica, Universidade Federal de Pernambuco,

Avenida da Engenharia, s/n Cidade Universitaria, Recife, PE, 50.740-600, Brazil

‘Instituto de Investigaciones Fundamentales en Agricultura (INIFAT),

Tropical ‘Alejandro de Humboldt’, OSDE, Grupo Agricola,

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

* CORRESPONDENCE TO: dasosa@espol.edu.ec

ABSTRACT—A new species Bactrodesmium pulcherrimum, found on decaying wood

in Ecuador, is described and illustrated. The fungus is distinguished by subnapiform to

broadly subturbinate (maize kernel-shaped) 1-septate asymmetrical ochreous-brown

conidia. Illustrations of Bactrodesmium novageronense and B. simile and a comparison

table of bicellular Bactrodesmium species are also provided.

KEY worDs—asexual fungi, freshwater, hyphomycetes, taxonomy, tropics

Introduction

Saprobic dematiaceous hyphomycetes are highly diverse on plant

materials in Central and South American tropical forests, where many

new genera and species have recently been discovered (Castafieda-Ruiz &

628 ... Espinoza & al.

Fic. 1. Cacao plantation, Guayas Province, Ecuador.

Bactrodesmium pulcherrimum sp. nov. (Ecuador) ... 629

al. 2016). During a survey of hyphomycetes associated with plant debris of

cacao plantations in the Naranjal, Guayas province, southern Ecuador (Fic.

1), we collected a Bactrodesmium specimen that differs remarkably from all

previously described species (Hernandez-Restrepo & al. 2013, Arias & al.

2016). It is described here as a new species, Bactrodesmium pulcherrimum.

Materials & methods

Samples of decaying plant materials were collected and placed in plastic bags for

transport to the laboratory, where they were washed, treated according to Castaneda-

Ruiz & al. (2016), and placed in humid chambers. Several attempts to obtain this

species in pure culture were unsuccessful after using a flamed needle to transfer

conidia to corn meal agar mixed 1:1 with carrot extract and incubating at 25 °C.

Mounts were prepared in PVL (polyvinyl alcohol, lactic acid) and measurements

were made at 1000x magnification. Microphotographs were obtained with an

Olympus BX51 microscope equipped with bright field and Nomarski interference

optics. The type specimen is deposited in the Herbarium of Universidade Federal de

Pernambuco, Recife, Brazil (URM).

Taxonomy

Bactrodesmium pulcherrimum RF. Castafieda, F. Espinoza & D. Sosa,

sp. nov. Pre;

MB 830569

Differs from Bactrodesmium simile by its hemispherical conidial basal cells and maize

kernel-shaped conidia.

Type: Ecuador, Guayas Province, Guayaquil, Naranjal, 02°41’26”S 79°36’46”W, on

decaying wood of Theobroma cacao L. (Malvaceae), 7 March 2018, E. Espinoza & M.

Vera (Holotype, URM 91287).

Erymo oey: Latin, pulcherrimum, meaning most beautiful.

CONIDIOMATA on the natural substrate sporodochial, scattered, pulvinate

ochreous to brown, <20 um diam. Mycelium superficial and immersed composed

of septate, branched, hyaline, smooth hyphae, 2-3 um diam. CONIDIOPHORES

macronematous, mononematous, unbranched or with short lateral branches

scorpioid after 1-2 sympodial elongations of the conidiogenous cells, erect,

flexuous, 2-3-septate, hyaline, smooth, 30-80 x 4-5 um. CONIDIOGENOUS

CELLS monoblastic, integrated or discrete, cylindrical or slightly clavate,

sometimes inflated near the conidiogenous loci, determinate or indeterminate

with sympodial extensions, hyaline, 10-18 x 4-7 um. Conidial secession

rhexolytic. Conrp1A solitary, acrogenous, sometimes acropleurogenous,

broadly obovoid to subnapiform, 1-septate, ochreous-brown, smooth, 18-24 x

630 ... Espinoza & al.

Fic. 2. Bactrodesmium pulcherrimum (holotype, URM 91287): A, B. Conidia with a basal

frill after rhexolytic conidial secession and a detached conidium with a lateral extension of

the conidiogenous cell; C. Conidiogenous cells with lateral sympodial extensions, attached

conidia; D. Detached conidium and conidiogenous loci indicated by arrows; E. Conidiophores,

conidiogenous cells, and conidia; F. Sporodochium. Scale bars: A-E = 10 um; F = 20 um.

Bactrodesmium pulcherrimum sp. nov. (Ecuador) ... 631

13-19 um, asymmetrical, with a basal cell hemispherical, 5-7 x 10-12 um, and

an apical cell semi-elliptical or subglobose, 12-17 x13-17 um, with markedly

pale ochreous-brown lumina.

Fic. 3. Bactrodesmium simile (holotype, XAL CB1689): A. Conidia with a basal frill after

rhexolytic conidial secession. Bactrodesmium novageronense (holotype, INIFAT C84/101-2):

B. Conidia with a basal frill after rhexolytic conidial secession. Scale bars = 10 um.

Note: Bactrodesmium simile Arias & al. (Fic. 3A, TABLE 1) is similar to

B. pulcherrimum in the conidial sizes, but B. simile has broadly pyriform,

obovoid, golden brown to pale olivaceous-brown conidia, with a basal

TABLE 1. Comparison of the 1-septate Bactrodesmium species.

SPECIES

B. novageronense

B. pulcherrimum

B. simile

COLOR

Pale brown

Ochreous-brown

Golden brown to

pale olivaceous

brown

CONIDIAL MORPHOLOGY

SHAPE & SIZE

(um)

Obovoid

to subglobose

8-13 x 7-10

Broadly obovoid

to subnapiform

18-24 x 13-19

Broadly pyriform

to obovoid

19-24 x 12-16

BASAL CELL

(um)

Hemispherical

3-3.5 x 5-7

Hemispherical

5-7 x 10-12

Cuneiform or

obconical

7.5-11 x 5-8.5

REFERENCE

Castafieda-Ruiz

1985

This paper

Arias & al.

2016

632 ... Espinoza & al.

cell cuneiform, 7.5-11 x 5-8.5 um (Arias & al. 2016). Bactrodesmium

novageronense R.F. Castaneda (Fic. 3B, TABLE 1) is similar to B. pulcherrimum

but can be distinguished by its smaller conidia (8-13 x 7-10 um;

Castanieda-Ruiz 1985).

Acknowledgments

We are indebted to Dr. Josiane Santana Monteiro (Museu Paraense Emilio

Goeldi, Belém, Brazil) and Dr. De-Wei Li (The Connecticut Agricultural

Experiment Station Valley Laboratory, Windsor, U.S.A.) for their critical reviews.

The authors thank Mr. Jairo Gonzalez for providing permission to collect samples.

The authors are grateful to Escuela Superior del Litoral (ESPOL), CIBE, for financial

support and the International Society for Fungal Conservation for facilities. RFCR

is grateful to the Cuban Ministry of Agriculture. We acknowledge the websites

provided by Dr. P.M. Kirk (Index Fungorum) and Dr. K. Bensch (MycoBank).

Dr. Lorelei Norvell’s editorial review and Dr. Shaun Pennycook’s nomenclature

review are greatly appreciated.

Literature cited

Arias RM, Heredia G, Castafieda-Ruiz RF. 2016. Two new species of Bactrodesmium and

Dictyoaquaphila from Mexico. Mycotaxon. 131: 291-295. https://doi.org/10.5248/131.291

Castafeda-Ruiz RE 1985. Deuteromycotina de Cuba. Hyphomycetes 2. Instituto de

Investigaciones Fundamentales en Agricultura Tropical, Cuba. 23 p.

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 International

Publishing. https://doi.org/10.1007/978-3-319-29137-6_9

Hernandez-Restrepo M, Mena-Portales J, Gené J, Cano J, Guarro J. 2013. New

Bactrodesmiastrum and Bactrodesmium from decaying wood in Spain. Mycologia 105:

172-180. https://doi.org/10.3852/12-004

MY COTAXON

October-December 2019—Volume 134, pp. 633-635

https://doi.org/10.5248/134.633

Dendrographium multiseptatum sp. nov.

from China

Li-Guo Ma, YUE-LI ZHANG, Bo ZHANG,

Kal QI, CHANG-SONG LI, JUN-SHAN QT

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

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

“CORRESPONDENCE TO: gi999@163.com

ABSTRACT—A new species, Dendrographium multiseptatum, is described and illustrated from

specimens collected on rotten twigs in China. This fungus is characterized by synnematous

conidiophores with polytretic conidiogenous cells producing obclavate, 6-9-distoseptate,

pale brown conidia.

KEY wWoRDS—asexual morph, microfungi, hyphomycete, taxonomy

Introduction

Dendrographium Massee was established by Massee (1892) to

accommodate the Brazilian type species D. atrum Massee. The genus is

characterized by synnematous, macronematous, unbranched conidiophores

with polytretic, integrated, terminal (becoming intercalary), sympodial,

cylindrical conidiogenous cells that produce solitary, acropleurogenous,

simple, distoseptate conidia (Ellis 1971, Seifert & al. 2011). Species

assignment in Dendrographium is based primarily on conidial morphology,

including shape, septal number, size, pigmentation, and ornamentation. Six

additional species have been described (all from India), but Dendrographium

kamatii V.G. Rao with monotretic conidiogenous cells has been recombined

as Corynespora kamatii (V.G. Rao) M.B. Ellis (Rao 1963, Ellis 1976). Ghosh

& al. (1977) provided a comparison table for the six Dendrographium species

(including D. kamatii) reported from India.

634 ... Ma &al.

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

Dendrographium species was found. As this species did not match any of the

currently described species in the genus, it is proposed here as new. Specimens

are deposited in the Herbarium of Institute of Plant Protection, Shandong

Academy of Agricultural Sciences, Jinan, Shandong, China (HSAAS) and

the Mycological Herbarium, Institute of Microbiology, Chinese Academy of

Sciences, Beijing, China (HMAS).

Taxonomy

Dendrographium multiseptatum L.G. Ma & J.S. Qi, sp. nov. FIG. 1

MB 832847

Differs from Dendrographium atrum by its narrower conidiophores and its shorter,

broader conidia with fewer distosepta and from D. mitteri by its longer synnemata, and

solitary, shorter, 6-9-distoseptate conidia.

Type: China, Shandong Province, Taian City, Taishan District, on rotten twigs of an

unidentified plant, 22 May 2018, L.G. Ma (Holotype, HSAAS 0415; isotype, HMAS

146095).

ETyMOLoGy: multiseptatum, referring to the numerous conidial septa.

CoLoNnIEs on the natural substratum effuse, black to dark brown, tufted.

Mycelium mostly immersed in the substratum. Synnemata large, black,

erect, tapered towards the apex, with dark brown stalks, consisting of

parallel conidiophores, 520-1380 um long, 25-85 um diam. at the base.

CONIDIOPHORES macronematous, synnematous, mostly adpressed along

lower part of the length, divergent towards the upper part, unbranched,

erect, straight or flexuous, cylindrical, septate, dark brown, smooth,

thick-walled, brown towards the apex, <1400 um long, 3.5-6.5 um diam.

CONIDIOGENOUS CELLS integrated, terminal becoming intercalary,

polytretic, cylindrical, brown, smooth, thick-walled, 8-24 x 5.0-7.5 um.

Conipi1A solitary, acropleurogenous, simple, obclavate, 6—9-distoseptate,

smooth, thick-walled, pale brown, 32-66 x 8-11.5 um, tapered towards the

apex, with a basal scar, truncate at the base, 2.5-3.5 um diam.

CoMMENTS—Dendrographium multiseptatum most closely resembles

D. atrum and D. mitteri Syd. in conidial shape. However, D. atrum has

broader (6-10 um) conidiophores and longer and narrower conidia

with more distosepta (40-130 x 7-9 um, 6-15 distosepta; Massee 1892).

Dendrographium mitteri differs by its shorter synnemata (170-250 um) and

occasionally catenate, longer (35-110 um), 3-10-septate conidia (Sydow &

Mitter 1933).

Dendrographium multiseptatum sp. nov. (China) ... 635

Fic. 1. Dendrographium multiseptatum (holotype, HSAAS 0415): A-D. Synnematous

conidiophores, conidiogenous cells, and conidia; E. Synnematous conidiophores with

conidiogenous cells and conidium; F. Conidia.

Acknowledgments

The authors express gratitude to Dr. De-Wei Li (The Connecticut Agricultural

Experiment Station Valley Laboratory, Windsor, U.S.A.) and Dr. Ze-Fen Yu

(Laboratory for Conservation and Utilization of Bio-resources, Yunnan University,

Kunming, P.R. China) for serving as pre-submission reviewers and to Dr. Shaun

Pennycook for nomenclatural review. This project was supported by Young Talents

Training Program of Shandong Academy of Agricultural Sciences (CXGC2018E04),

National Key R & D Program of China (2016YFD0300700, 2017YFD0201700), and

National Natural Science Foundation of China (31400019).

Literature cited

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

Surrey, England.

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

Kew, Surrey, England.

Ghosh RN, Pathak NC, Singh MS. 1977. Two new records of stilbaceous fungi from India.

Norwegian Journal of Botany 24: 79-81.

Massee GE. 1892. Notes on exotic fungi in the Royal Herbarium, Kew. Grevillea 21: 1-6.

Rao VG. 1963. A new species of Dendrographium from India. Current Science 32: 473-474.

Seifert KA, Morgan-Jones G, Gams W, Kendrick B. 2011. The genera of Hyphomycetes.

CBS Biodiversity Series 9. 997 p.

Sydow H, Mitter JH. 1933. Fungi indici. I. Annales Mycologici 31: 84-97.

MY COTAXON

October-December 2019—Volume 134, pp. 637-641

https://doi.org/10.5248/134.637

Haematomma pluriseptatum sp. nov.

from China

CONGCONG MIAO *!, RONG TANG **4,

LINLIN DoNG?, ZHAOJIE REN?, ZUNTIAN ZHAO **

"Key Laboratory of Plant Stress Research, College of Life Sciences &

? Institute of Environment and Ecology:

Shandong Normal University, Jinan, 250014, PR. China

* Shandong Museum, Jinan, 250014, P. R. China

‘The First High School of Liangshan, Shandong Province, 272600, P. R. China

“CORRESPONDENCE TO: ccmjy123@163.com

ABSTRACT—A new species, Haematomma pluriseptatum, is described from southwestern

China. This corticolous species is characterized by large ascospores with (19-)20-26(-27)

septa and by the presence of atranorin, russulone, and pseudoplacodiolic acid. A detailed

description of its morphology, high resolution photographs, chemistry, comments, and

distribution are provided. Related lichen taxa are discussed, and a key to the species of

Haematomma from China is also provided.

Key worps—East Asia, Haematommataceae, Lecanorales, lichenized fungi, taxonomy

Introduction

The crustose Haematomma lichens, with their small but brilliant-red

lecanorine apothecia, are often conspicuous on smooth-barked trees in

warm-temperate to tropical regions of the world (Staiger & Kalb 1995,

Brodo & al. 2008). Haematomma was proposed originally to house two

species, H. vulgare A. Massal. and H. ventosum (L.) A. Massal. (Massalongo

1852). The genus is characterized by Haematomma-type asci, blood-red

lecanorine apothecia, and transversely septate, submuriform to muriform

*CONGCONG MIAO & RonG TANG contributed equally to this work.

638 ... Miao, Tang & al.

ascospores (Brodo & al. 2001, Elix 2004, Nelsen & al. 2006, Nash & al. 2004,

Smith & al. 2009). Apothecial pigmentation and secondary chemistry play

important roles for species delimitation in this genus (Messuti & de la Rosa

2009). The number of spore septa is also used as an important character

in separating species (Nelsen & al. 2006). Of the forty Haematomma

species reported in the world, twelve have been reported in China (Brodo

& al. 2001, Elix 2004, Nelsen & al. 2006, Nash & al. 2004, Smith & al.

2009, Tang & al. 2018). For this study, we examined about two hundred

specimens of Haematomma morphologically and chemically, describe a

new species, H. pluriseptatum, from China, and present a key to the species

of Haematomma from China.

Materials & methods

Four collections are deposited in the Lichen Section of the Botanical Herbarium,

Shandong Normal University, Jinan, China (SDNU), and one is deposited in the

Lichen Herbarium of the Kunming Institute of Botany, Kunming, China (KUN).

The specimens were examined morphologically and anatomically morphological

under an Olympus SZ51 stereomicroscope and Olympus CX21 polarizing

microscope. Both thallus and medulla were tested with K (a 10% KOH aqueous

solution) and C (a saturated aqueous NaOCl solution). The lichen substances were

identified using standardized thin layer chromatography techniques (TLC) with

solvent system A and C (Orange & al. 2010). The specimens were photographed

using a DP72 camera on Olympus SZX16 and BX61 microscopes.

Taxonomy

Haematomma pluriseptatum R. Tang, sp. nov. Fie. 1

MB 830618

Differs from Haematomma rufidulum by its ascospores with more septa and its

production of pseudoplacodiolic acid.

Type: China. Yunnan, Diging Co., Shangri-La, Tianbao Mt., 27°36’37”N 99°53’37”E,

alt. 3790 m, on bark of living trees, 17 Aug. 2018, Wang Chunxiao 20181135 (Holotype,

SDNU).

EryMo.ocy: The epithet pluriseptatum refers to the morphology of the ascospores with

many septa.

Thallus crustose, corticolous, pale greyish to greenish grey, rugose rimose-

areolate to areolate, continuous, thickness 0.6 mm. Soredia and isidia

absent. Apothecia scattered, 0.3-0.95(-1.0) mm diam., sessile, convex,

constricted at the base, smooth when young, red. Disc orange-yellow to

brownish yellow, white pruina present when young but absent when mature;

Thalline margin well developed; amphithecium present, 70-90 um thick.

Haematomma pluriseptatum sp. nov. (China) ... 639

Fic. 1. Haematomma pluriseptatum (holotype, SDNU (Wang Chunxiao 20181135)). A. Thallus

and apothecia; B. Apothecium section; C. K+ reaction (red) of epihymenium and outer exciple;

D. Ascus; E. Ascospores. Scale bars: A = 200 um; B, C = 100 um; D = 5 um; E = 10 um.

Epihymenium orange, K+ red, 9-20 um thick. Hymenium hyaline, 70-95

um high. Hypothecium hyaline or slightly brown. Paraphyses branched

and anastomosing. Asci clavate, containing 8 spores. Ascospores twisted in

ascus, filiform, straight or slightly curved, (19-)20-26(-27)-septate, 70-95 x

5-6.5(—7) um. Conidia absent.

CHEMISTRY—Cortex K+ yellow, C-, KC-. Atranorin, russulone, and

pseudoplacodiolic acid detected by TLC.

ADDITIONAL SPECIMENS EXAMINED: CHINA. YUNNAN, Diging Co., Shangri-La,

Tianbao Mt., 27°36’37”N 99°53’37’E, alt. 3790 m, on bark of living trees, 17 Aug.

2018, Wang Chunxiao 20181175, 20181209, 20181126 (SDNU); Deqin Co., Yubeng

Village, Xiaonong Base Camp. 28°23.92’N 98°46.16’E, alt. 3500 m, on bark of living

trees, 14 Sep. 2012, Niu Dongling 12-36322 (KUN-L).

COMMENTS—Haematomma pluriseptatum is characterized by the large

number of septa (20-27) in each ascospore and the presence of atranorin,

russulone and pseudoplacodiolic acid in the thallus.

640 ... Miao, Tang & al.

The new ascomycete is morphologically similar to H. rufidulum (Fée)

A. Massal., which differs by the occurrence of slightly irregular, white,

convex to hemispherical soredia and elongate-fusiform, 8-25-celled,

50-72(-85) x (3.5—)4-6(-—7) um ascospores (Brodo & al. 2008). Secondary

metabolites also differ: H. pluriseptatum contains pseudoplacodiolic acid,

while H. rufidulum contains placodiolic acid.

Haematomma pluriseptatum may also be confused with H. dolichosporum

(B. de Lesd.) Kalb & Staiger, which also produces twisted spores in the

asci, but H. dolichosporum is readily distinguished by its shorter (42-62

um) spores with fewer (7—9(-11)) septa and the production of placodiolic

acid (Staiger & Kalb 1995; Brodo & al. 2008).

Key to the species of Haematomma in China

1. Epihymenium with russulone, K+ Ted 25 soe sien seston nie Sn olew slew ales 2

1. Epihymenium with haematommone, K+ purple (entirely dissipating) ........... 8

2 siallastcomtaimi ne UgMte BOR | nse ne hms nadie Rate ree Sad een Bade ea ade eek ae H. fauriei

2. Uhalluslackine-usnic acid.) 25. ebay. Bey ee tie ee tk el MEE in ee ae De geee ts mes 3

SeOMTOCKE 5h nds Awe ade eH eH lng W eiagee nN efegeed ehogee me Ong mee H. fenzlianum

Pe EU Dial Raters, onye- dag, nyee apa > aeons teen otis #o genie key ie sn gh mesa ean eM S ees ote ee -:

4, Thallus containing placodiolic acid ............. 0... e eee eee eee H. rufidulum

#. balls ekino-placOd iol Ge: dGid. cna ee Ms aren neers Basten Paks ea ede oe Sly or Rodgers Bootes 5

>) thallus lacking sphderophotin ts, atac. eitore a ara eee nea ee hee Paes Pee 5 6

5. Thallus containing sphaerophorin,

Sporesto= IA seclledanw to, We M rim oe meh tans Rens Rees CAR a eRe ne ee 7

6. Thallus containing pseudoplacodiolic acid,

spores’ 20=27—celled oo n.cey vin eee Reed ee oad Bee EE Be oeS H. pluriseptatum

6. Thallus lacking pseudoplacodiolic acid,

spores S=l6scelled) .. is. ese eee ae ee eee es H. puniceum subsp. pacificum

Fc SPOVesOa7 ACE 6 x 4 bsg kag t rhea y eAGY 4S MENS MEE EAR H en tie Dy nae H. persoonii

7 Spores 01a xcelled. 28 Anh thal fatter Ma nat Ane nak Met May H. collatum

SeSPOTEs MOE SUDEIUTIIOLI, 5 oF 58 ue «Aste eg. ike recom ga-rhc om gro mG aaa oni arec phe e cn eget wie 9

SPS Pores Su DHMITMOLN a ac, tila kn eas Cees earn ies dee tten aeeeeen Rete ss hae H. wattii

Re ApOecia: Sorediatee.. M.S ath. a sgeh ese h ead ven tak wonton moe ete H. caperaticum

9 Apothecranorsorediate +5. 2565 28 ls oh Besos eed bs ee hee ema ee et Metgs oe eins 10

10. Thallus containing isoplacodiolic, isopseudoplacodiolic acids ...... H. flexuosum

10. Thallus lacking isoplacodiolic, isopseudoplacodiolic acids ................... 11

Ines poreg celled 7 itr eh Fut i oP RNa aI H. accolens

I Sporess eas ceed ie is carte Merde Bevery Rebstes Roden Sods ee Babee hon H. africanum

Haematomma pluriseptatum sp. nov. (China) ... 641

Acknowledgments

We thank Dr. Klaus Kalb (Lichenological Institute Neumarkt, Germany) and

Dr. Shou-Yu Guo (State Key Laboratory of Mycology, Institute of Microbiology,

Chinese Academy of Sciences, Beijing) for presubmission review. The authors would

also like to thank Lisong Wang and Xinyu Wang (Kunming Institute of Botany, CAS,

China) for assistance during this study. This work was supported by the National

Natural Science Foundation of China Youth Science Foundation (31600100), and

Emergency management project of National Natural Science Foundation of China

(31750001).

Literature cited

Brodo IM, Sharnoff SD, Sharnoff S. 2001. Lichens of North America. Yale University Press, New

Haven and London, 795 p.

Brodo IM, Culberson WL, Culberson CE. 2008. Haematomma (Lecanoraceae) in North and

Central America. Bryologist 111: 363-423.

https://doi.org/10.1639/0007—2745(2008) 111[363:hlinac]2.0.co;2

Elix JA. 2004. Haematommataceae. 4-10, in: Flora of Australia 56A—Lichens 4. Australian

Biological Resources Study/CSIRO Publishing. 222 p.

Massalongo AB. 1852. Ricerche sull’ autonomia dei licheni crostosi. Verona: Friziero. 207 p.

Messuti MI, de la Rosa IN. 2009. Notes on the genus Haematomma (Ascomycota, Lecanoraceae)

in Argentina. Darwiniana 47(2): 297-308.

Nash TH, Ryan BD, Diederich P, Gries C, Bungartz F. 2004. Lichen flora of the Greater Sonoran

Desert Region, vol. 2. Tempe, Lichens Unlimited.

Nelsen MP, Liicking R, Chaves JL, Sipman HJM, Umafa L, Navarro E. 2006. A first assessment

of the Ticolichen biodiversity inventory in Costa Rica: the genus Haematomma (Lecanorales:

Lecanoraceae). Lichenologist 38: 251-262. https://doi.org/10.1017/s0024282906005573

Orange A, James PW, White FJ. 2010. Microchemical methods for the identification of lichens.

2™ edition. London, British Lichen Society.

Smith CW, Aptroot A, Coppins BJ, Fletcher A, Gilbert OL, James PW, Wolseley PA. 2009.

The lichens of Great Britain and Ireland. London, British Lichen Society.

Staiger B, Kalb K. 1995. Haematomma-Studien: I. Die Flechtengattung Haematomma. Bibliotheca

Lichenologica 59. 198 p.

Tang R, Yan SK, Sun MJ, Zhang LL. 2018. New records of Haematomma and Ophioparma from

China. Mycotaxon 133(1): 175-181. https://doi-org/10.5248/133.175

MY COTAXON

October-December 2019—Volume 134, pp. 643-648

https://doi.org/10.5248/134.643

New records of Didymium inconspicuum,

D. karstensii, and D. rugulosporum

from China

CHAOFENG YUAN, SHU LI, WAN WANG,

SHUWEI WEI, Qi WANG, YU LI

Engineering Research Center of Chinese Ministry of Education

for Edible and Medicinal Fungi, Jilin Agricultural University,

Changchun, Jilin 130118 China

* CORRESPONDENCE TO: giwang@jiau.edu.cn

ABSTRACT—Didymium inconspicuum, D. karstensii, and D. rugulosporum are reported as

new records for China. Morphological characteristics were observed and described by light

microscopy and scanning electron microscopy and are discussed and compared with similar

species.

Key worps—Didymiaceae, Myxogastrea, taxonomy

Introduction

Didymium was introduced by Schrader (Schrader 1797) and characterized

by sporophores with a peridium covered with stellate lime crystals. In

contrast, the capillitium is almost always limeless. The lime crystal nature on

the peridium separates Didymium from Diderma, in which the lime occurs

as amorphous granules (Martin & Alexopoulos 1969). Currently Didymium

includes approximately 94 species around the world (Kirk & al. 2008, Lado

2018), of which 35 have been recorded in China (Li & al. 1996, Liu & Chen

1998, Chen 1999, Li 2008, Liu & Chang 2011, Gao & al. 2018). The objectives

of this work were to examine some putative Didymium samples, characterize

them by light and scanning electron microscopy, and compare them with

similar species.

644 ... Yuan & al.

Materials & methods

Specimen collection and moist chamber culture

Didymium rugulosporum was collected from Huzhong Nature Reserve,

Heilongjiang Province in 2016. Didymium karstensii was collected from Changbai

Mountain, Jilin Province in 2018. Didymium inconspicuum was collected from the

desert transition zone of Suwu Township, Mingin County, Gansu Province in 2016.

The specimens are deposited in the Herbarium of the Mycological Institute of Jilin

Agricultural University, Changchun, China (HMJAU).

Morphological identification

The morphological characteristics of the sporocarps were observed by light

microscope and scanning electron microscope. Each sporocarp was placed on a

slide and one drop of 4% KOH added. The slide cover was placed on top and gently

tapped, and filter paper used to remove excess solution. The capillitium, calcareous

crystals, and spores were observed under a Leica DM 2000 light microscope

(LM). Up to 20 spores were randomly selected and their diameters measured.

The ultrastructure was observed under the Hitachi SU8010 scanning electron

microscope (SEM) operating at 5kV (Martin & Alexopoulos 1969, Zhu & al. 2012,

Li & al. 2017).

Taxonomy

Didymium inconspicuum Nann.-Bremek. & D.W. Mitch.,

Proc. Kon. Ned. Akad. Wetensch., C 92: 508. 1989 Fic. 1

Sporocarps sessile, scattered, occasionally mingled with — short

plasmodiocarps, small and inconspicuous. 0.5-1 mm long, about 0.5 mm high,

ovoid, often depressed in the center, grayish white. Columella absent. Peridium

membranous, thin, coated with grayish white lime crystals. Capillitium profuse,

threads slender and curved, pale brown, 1-1.5 um diam., often branching and

anastomosing to an incomplete network, sometimes with flattened and darker

colored sections on the branches. Spores dark brown in mass, brown by LM,

ovoid to globose, 8-11 um diam., evenly slightly warted.

SPECIMEN EXAMINED: CHINA, GANSU PROVINCE, Mingin County, Suwu Township,

38°16'42”N 103°06’17’E, 1360 m, on leaf by moist chamber culture, 16 September 2016,

Chaofeng Yuan & Shuwei Wei (HMJAU 10650).

CoMMENTs: Didymium inconspicuum is similar to Didymium obducens

P. Karst. [= D. fulvum Sturgis] and Didymium ochroideum G. Lister. The spore

of D. obducens is tuberculate on one side, while the other side is smooth and

the color is pale, but the spore of D. inconspicuum has relatively uniform

small warts. The spore of D. ochroideum is 6-8 um diam. whereas the spore

of D. inconspicuum is 8-11 1m diam.

Didymium spp. new to China... 645

Fic 1. Didymium inconspicuum. A- C. Sporocarps; D. Capillitium (LM); E. Spores (LM);

F. Capillitium (SEM); G. Lime crystal (SEM); H. Spores (SEM). Scale bars: A- C = 500 um;

D, F= 10 pm; E, G, H=5 pm.

Nannenga-Bremekamp (1989) described D. inconspicuum as having white

or pale yellow-brown tiny lime crystals, with a coarse capillitium and spores

that are 12-14(-15) um diam.; Schnittler & Novozhilov (2000) described this

species as having 3-6 um long lime crystals smaller than spores, a pale to pale

brown, 1-2(-2.5) um diam. capillitium, and (9.5—)10.5-11.5(-12.5) um diam.

spores. Our specimen has white lime crystals about 5-8 um diam., a light

brown 1-1.5 um diam. capillitium, and spores 8-11 um diam.; other features

are consistent with the published descriptions.

Didymium karstensii Nann.-Bremek., Acta Bot. Neerl. 13: 247. 1964 FIG. 2

Sporocarps gregarious, sessile or with a short stalk, white, subglobose,

1 mm wide, up to 0.5 mm high, stalk (when present) calcareous, yellowish

white, about 0.2 mm long. Peridium double, outer layer white, with stellate

lime crystals about 5 um diam. forming the wrinkled shell; the inner layer

membranous and colorless. Columella white, calcareous, slightly thickened at

646 ... Yuan & al.

Fic 2. Didymium karstensii. A. Sporocarps; B. Capillitium and swelling (LM); C. Spores

(LM); D. Lime crystals (SEM); E. Capillitium (SEM); F. Spores (SEM). Scale bars: A = 1 mm;

B-E = 10 um; F=5 um.

the base. Capillitium pale brown, threads slender, branching and anastomosing

to a network, with dark swellings, 2-3 um diam., attached to the peridium and

to the base of the sporocarp. Spores dark brown in mass, brown by LM, densely

warted and ridged, 7.5-10 um diam.

SPECIMEN EXAMINED: CHINA, JILIN PROVINCE, Changbai Mountain, 42°22’18”N

128°00'14”E, 828 m, on the leaf, 5 September 2018, Chaofeng Yuan & Shuwei Wei

(HMJAU 10651).

Comments: Didymium karstensii is similar to Didymium squamulosum (Alb. &

Schwein.) Fr., but D. karstensii is inconspicuously umbilicate at the base, whereas

D. squamulosum is deeply umbilicate at the base. Nannenga-Bremekamp

(1964) described stalks of D. karstensii as generally not exceeding half the

diameter of the sporocarps, and spinulose spores, 10-12 um diam. Demirel &

Kasik (2012) described stalks of D. karstensii generally not exceeding 0.2 mm

long, and warted spores, 10-12 um diam. Our specimen had stalks about 0.2

mm long and warted spores, 7.5-10 um across; other features are consistent

with the published descriptions.

Didymium rugulosporum Kowalski, Mycologia 61: 636. 1969 Fic. 3

Sporocarps sessile, scattered to clustered, globose to subglobose, 1-2.5 mm

diam., white. Peridium double, the outer layer thick and fragile, white smooth

calcareous shell, stellate lime crystals; the inner layer membranous, thin,

colorless and transparent. Columella absent. Capillitium dense, rigid, brown,

Didymium spp. new to China ... 647

Fic 3. Didymium rugulosporum. A, B. Sporocarps; C. Capillitium and spores (LM); D. Capillitium

and peridium (SEM); E. Lime crystal (SEM); FE. Spores (SEM). Scale bars: A, B = 1 mm; C = 20 um;

D = 10 um; E, F=5 um.

1-1.5 um diam., weakly attached to the base and apex of the sporocarps, brown,

branched with many colorless swellings. Spores dark brown in mass, purple

brown by LM, globose, 17-19 um diam., with large dense warts about 1.5 um

high and with some ridges forming a reticulum.

SPECIMEN EXAMINED: CHINA, HEILONGJIANG PROVINCE, Huzhong Nature Reserve,

52°06'02”N 123°20'38”E, 168 m, on the bark, 24 September 2016, Wan Wang (HMJAU

10652).

Comments: Didymium rugulosporum is similar to Didymium trachysporum

G. Lister and Didymium quitense (Pat.) Torrend, which differ by their smaller

spores (D. trachysporum 9-10 um diam.; D. quitense is 12-15 um diameter).

Kowalski (1969) described spores of D. rugulosporum as (16—)18-20(-22) um

diam. Our specimen had spores 17-19 um diam.; other characteristics are

consistent with the original description.

Acknowledgments

We would like to thank Tom Hsiang (University of Guelph, Canada) and Paul

M. Kirk (RBG Kew, UK) for presubmission reviews of this manuscript. This work

was financed by the National Natural Science Foundation of China (NO.31770011,

NO.31370065).

Literature cited

Chen SL. 1999. Fungal flora of tropical Guangxi, China: a survey of myxomycetes from southwestern

Guangxi. Mycotaxon 72: 393-401.

648 ... Yuan & al.

Demirel G, Kasik G. 2012. Four new records for Physarales from Turkey. Turkish Journal of

Botany 36: 95-100. https://doi.org/10.3906/bot-1010-12

Gao Y, Yan SZ, Wang GW, Chen SL. 2018. Two new species and two new records of myxomycetes

from subtropical forests in China. Phytotaxa 350(1): 51-63.

https://doi.org/10.11646/phytotaxa.350.1.6

Kirk PM, Cannon PF, Minter DW, Stalpers JA. 2008. Ainsworth & Bisby’s dictionary of the

fungi. 10th edition. CAB International Publishing, Wallingford. 771 p.

Kowalski DT. 1969. A new coprophillous species of Didymium. Mycologia 61(3): 635-639.

https://doi.org/10.2307/3757253

Lado C. (2018). An online nomenclatural information system of Eumycetozoa. Real Jardin

Botanico, CSIC. Madrid, Spain. http://www.nomen.eumycetozoa.com (Accessed:

September 2018).

Li Y. 2008. Flora myxomycetes sinicorum. Science Press.

Li HZ, Li Y, Wang Q. 1996 [“1995-96”]. Myxomycetes from China XIII: a new species of

Didymium. Mycosystema 8-9: 173-175.

Li S, Wang W, Wang W, Wang Q, Li Y. 2017. Morphology and life-cycle of Fuligo leviderma,

a newly recorded myxomycete species of China. Mycosystema 36(4): 528-531.

https://doi.org/10.13346/j.mycosystema.160122

Liu CH, Chang JH. 2011. Myxomycetes of Taiwan XXIII. The genera Diachea and Didymium.

Taiwania 56: 287-294. https://doi.org/10.6165/tai.2011.56(4).287

Liu CH, Chen YE. 1998. Myxomycetes of Taiwan X. Three new records of Didymium. Taiwania

43: 177-184. https://doi.org/10.6165/tai.1998.43(3).177

Martin GW, Alexopoulos CJ. 1969. The myxomycetes. University of Iowa Press, Iowa City.

560 p.

Nannenga-Bremekamp NE. 1964. Notes on myxomycetes VIII. A new species of

Didymium from the Netherlands. Acta Botanica Neerlandica 13: 246-249.

https://doi.org/10.1111/j.1438-8677.1964.tb00155.x

Nannenga-Bremekamp NE. 1989. Notes on myxomycetes XXIII. Seven new species of

myxomycetes. Proceedings, Koninklijke Nederlandse Akademie van Wetenschappen,

Series C, 92: 505-515.

Schnittler M, Novozhilov YK. 2000. Myxomycetes of the winter-cold desert in western

Kazakhstan. Mycotaxon 74(2): 267-285.

Schrader HA. 1797. Nova genera plantarum. Pars prima, Lipsiae.

Zhu H, Song XX, Li $, Wang Q, Li Y. 2012. Three new recorded species of myxomycetes in

China. Mycosystema 31(6): 947-951. https://doi.org/10.13346/j.mycosystema.2012.06.021

MY COTAXON

October-December 2019—Volume 134, pp. 649-662

https://doi.org/10.5248/134.649

Rhomboidia wuliangshanensis gen. & sp. nov.

from southwestern China

TaI-MIN XU?”, XIANG-Fu Liu3, YuU-HuI CHEN’, CHANG-LIN ZHAO?

"Yunnan Provincial Innovation Team on Kapok Fiber Industrial Plantation;

? College of Life Sciences; * College of Biodiversity Conservation:

Southwest Forestry University, Kunming 650224, PR. China

" CORRESPONDENCE TO: fungichanglinz@163.com

ABSTRACT—A new, white-rot, poroid, wood-inhabiting fungal genus, Rhomboidia,

typified by R. wuliangshanensis, is proposed based on morphological and molecular

evidence. Collected from subtropical Yunnan Province in southwest China, Rhomboidia is

characterized by annual, stipitate basidiomes with rhomboid pileus, a monomitic hyphal

system with thick-walled generative hyphae bearing clamp connections, and broadly

ellipsoid basidiospores with thin, hyaline, smooth walls. Phylogenetic analyses of ITS and

LSU nuclear RNA gene regions showed that Rhomboidia is in Steccherinaceae and formed

as distinct, monophyletic lineage within a subclade that includes Nigroporus, Trullella, and

Flabellophora.

Key worps—Polyporales, residual polyporoid clade, taxonomy, wood-rotting fungi

Introduction

Polyporales Gaum. is one of the most intensively studied groups of fungi

with many species of interest to fungal ecologists and applied scientists (Justo

& al. 2017). Species of wood-inhabiting fungi in Polyporales are important

as saprobes and pathogens in forest ecosystems and in their application in

biomedical engineering and biodegradation systems (Dai & al. 2009, Levin

& al. 2016). With roughly 1800 described species, Polyporales comprise about

1.5% of all known species of Fungi (Kirk & al. 2008). Currently, there are

46 genomes of polyporalean taxa available from the Joint Genome Institute

MycoCosm portal (Grigoriev & al. 2013).

650 ... Xu & al.

TABLE 1. Species and sequences used in the phylogenetic analyses

SPECIES

Abortiporus biennis

Antrodiella americana

A. semisupina

Antrodiella sp.

Climacocystis borealis

Diplomitoporus flavescens

Elaphroporia ailaoshanensis

Flabellophora sp.

Flaviporus brownii

E liebmannii

Frantisekia mentschulensis

Hypochnicium bombycinum

H. lyndoniae

Irpex lacteus

Ischnoderma benzoinum

I. resinosum

Junghuhnia crustacea

J. micropora

Loweomyces fractipes

Mycorrhaphium adustum

SAMPLE

TFRI 274

Gothenburg 3161

FCUG 960

X 418

KH 13318

X 84

CLZhao 595

CLZhao 596

X340

X 1216

X 251

X 249

X 666

BRNM 710170

1377

MA 15305

NL 041031

CBS 431.48

DO 421/951208

KHL 12099

FD-328

X 1127

X 262

Spirin 2652

X 1149

X 1253

X 1250

8024

Dai 10173

GENBANK ACCESSION NO.

ITS

EU232187

JN710509

EU232182

JN710523

JQ031126

FN907908

MG231568

MG231572

JN710534

JN710537

JN710541

JN710539

JN710540

FJ496728

JN710544

FN552537

JX124704

MH856423

JX109852

JX109841

KP135303

JN710554

JN710553

JN710559

JN710570

JN710569

JN710568

JN710573

KC485537

LSU

EU232235

JN710509

EU232266

JN710523

JQ031126

MG231568

MG231572

JN710534

JN710537

JN710541

JN710539

JN710540

JN710544

JX124704

MH867969

JX109852

JX109841

KP135225

JN710554

JN710553

JN710559

JN710570

JN710569

JN710568

JN710573

KC485554

REFERENCES

Larsson 2007

Miettinen & al. 2012

Binder & al. 2005

Miettinen & al. 2012

Binder & al. 2013

Miettinen & al. 2012

Wu & al. 2018

Miettinen & al. 2012

TomSsovsky & al. 2010

Miettinen & al. 2012

Binder & al. 2013

Binder & al. 2005

Vu & al. 2019

Binder & al. 2013

Floudas & Hibbett

2015

Miettinen & al. 2012

Miettinen et al. 2012

Miettinen & al. 2012

Nigroporus vinosus

Panus conchatus

P strigellus

Physisporinus

sanguinolentus

P. vitreus

Podoscypha venustula

Pseudolagarobasidium

acaciicola

P. belizense

Rhomboidia

wuliangshanensis

Skeletocutis

novae-zelandiae

Spongipellis spumeus

Steccherinum fimbriatum

S. ochraceum

Trullella dentipora

T. duracina

T. polyporoides

Xanthoporus syringae

Rhomboidia wuliangshanensis gen. & sp. nov. (China) ... 651

X 839

8182

BHS2008- 100

X 1234

INPA 243940

BRNM 699576

3163

KHL11959

CBS 486.72

CBS 65684

CBS 115543

CBS 115544

CFMR DLC 04-31

CLZhao 4406 [T]

CLZhao 4411

Ryvarden 38641

PRM 891931

BRNM 712630

BRNM 734877

KHL 11905

Ryberg s.n.

KHL 11902

X 200

X 290

X 510

X 339

Cui 2177

Gothenburg 1488

N710576

JN710728

JX109857

JN710579

JQ955725

FJ496671

JN710580

JQ031129

MH860538

JN649367

DQ517883

DQ517882

JQ070173

MK860715

MK860716

JN710582

HQ728287

HQ728288

HQ728283

EU118668

EU118669

JQ031130

JN710512

JN710513

JN710602

JN710606

DQ789395

JN710607

N710576

JN710728

JX109857

JN710579

JQ955732

FJ496725

JN710580

JQ031129

MH872244

JN649367

MK860710

MK860711

JN710582

HQ729021

HQ728288

HQ728283

EU118668

EU118670

JQ031130

JN710512

JN710513

JN710602

JN710606

JN710607

Miettinen & al. (012

Miettinen & al. 2012

Binder & al. 2013

Miettinen & al. 2012

Binder & al. 2013

TomSsovsky & al. 2010

Miettinen & al. 2012

Sjokvist & al. 2012

Vu & al. 2019

Binder & al. 2013

Miettinen &

Rajchenberg 2012

Miettinen &

Rajchenberg 2012

Miettinen &

Rajchenberg 2012

Present study

Miettinen & al. 2012

Tomsovsky & al. 2010

Larsson 2007

Sjokvist & al. 2012

Miettinen & al. 2012

652 ... Xu & al.

Systematics of the Polyporales has benefitted from numerous molecular

phylogenetic studies (e.g., Binder & al. 2005, 2013; Larsson 2007; Miettinen

& al. 2012; Dai & al. 2015; Choi & Kim 2017). Steccherinaceae Parmasto, one

of 18 families recognized in Polyporales (Justo & al. 2017), has been included

in several molecular studies (e.g., Binder & al. 2005, 2013; Miettinen & al.

2012; Miettinen & Ryvarden 2016; Justo & al. 2017; Westphalen & al. 2018).

Miettinen & al. (2012) published a multigene, molecular phylogenetic study

of Steccherinum and allied taxa that clearly delineated Steccherinaceae. They

uncovered surprising morphological diversity and plasticity in this family,

requiring revision of generic concepts and 15 new genera to accommodate

existing and new species. Subsequently, Miettinen & Ryvarden (2016)

introduced five new genera, revised one genus, and described two new

species that had been identified earlier by Miettinen & al. (2012). Justo &

al. (2017), who revised family-level classification in Polyporales, confirmed

Steccherinaceae as a distinct lineage in Polyporales that grouped with

Cerrenaceae Miettinen & al. and Panaceae Miettinen & al. Ina morphological

and molecular study of Neotropical taxa of Junghuhnia and Steccherinum,

Westphalen & al. (2018) uncovered a new genus and several new species and

reclassified four taxa.

Cosmopolitan in distribution, Steccherinaceae has a rich diversity because

it is found in boreal, temperate, subtropical, and tropical ecosystems (Nunez

& Ryvarden 2001, Dai 2012, Ryvarden & Melo 2014, Dai & al. 2015, Zhou

& al. 2016). Many new species in Polyporales have been described from

southern, subtropical China (e.g., Li & Cui 2010, Zhao & Wu 2017, Zhao

& Ma 2019). Recently, we collected an undescribed taxon from Yunnan

Province that could not be assigned to any described genus. We present

morphological and molecular phylogenetic evidence that support the

recognition of a new monotypic genus in Steccherinaceae—Rhomboidia,

typified by R. wuliangshanensis.

Materials & methods

The specimens studied are deposited at the herbarium of Southwest Forestry

University, Kunming, China (SWFC). Macromorphological descriptions are based

on field notes. Special colour terms follow Petersen (1996). Micromorphological

data were obtained from the dried specimens and observed under a light microscope

following Dai (2010). The following abbreviations are used: KOH = 5% potassium

hydroxide; CB = cotton blue; CB- = acyanophilous; IKI = Melzer’s reagent; IKI- =

non-amyloid and non-dextrinoid; L = mean spore length (arithmetic average of all

spores); W = mean spore width (arithmetic average of all spores); Q = variation in

Rhomboidia wuliangshanensis gen. & sp. nov. (China) ... 653

the L/W ratios between the specimens studied; (n = a/b) = number of spores (a)

from number of specimens (b).

HiPure Fungal DNA Mini Kit II was used to obtain genomic DNA from dried

specimens, according to the manufacturer’s instructions with some modifications.

A small piece (about 30 mg) of dried fungal material 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, and 100 ml elution buffer was added to

the middle of adsorbed film to elute the genomic DNA. The internal transcribed

spacer region (ITS) was amplified with primer pairs ITS5 and ITS4 (White & al.

1990). The nuclear large subunit region (LSU) was amplified with primer pairs

LROR and LR7 (https://sites.duke.edu/vilgalyslab/rdna_primers_for_fungi/). The

PCR procedure for ITS was: 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 procedure for LSU was: initial denaturation at 94 °C for 1

min, followed by 35 cycles of 94 °C for 30 s, 48 °C 1 min, and 72 °C for 1.5 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 24216). Xanthoporus syringae

(Parmasto) Audet obtained from GenBank was used as an outgroup to root trees

following Miettinen & al. (2012) in the ITS+LSU analyses (Fic. 1).

The ITS+LSU sequences were analyzed phylogenetically using maximum

parsimony, maximum likelihood, and Bayesian inference methods. Maximum

parsimony (MP) analyses followed Zhao & Wu (2017), and tree construction 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 branch swapping and 1000 random sequence

additions. Max-trees was set to 5000, branches of zero length were collapsed and

all parsimonious trees were saved. Clade robustness was assessed using bootstrap

(BP) analysis with 1000 replicates (Felsenstein 1985). 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 generated. Sequences were analyzed using Maximum Likelihood (ML) with

654 ... Xu & al.

RAxML-HPC2 through the Cipres Science Gateway (www.phylo.org; Miller & al.

2009). Branch support (BS) for ML analysis was determined by 1000 bootstrap

replicates.

MrModeltest 2.3 (Posada & Crandall 1998; Nylander 2004) was used to

determine the best-fit evolution model for data set for Bayesian inference (BI).

Bayesian inference was calculated with MrBayes_3.1.2 using a general time

reversible (GTR+G) 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 4 million generations (ITS+LSU) in Fic.

1 and trees were sampled every 100 generations. The first 25% of the generations

were discarded as burn-in. A majority rule consensus tree of all remaining trees was

calculated. Branches that received bootstrap support for maximum likelihood (BS)

275%, maximum parsimony (BP) 275%, and Bayesian posterior probabilities (BPP)

20.95 were considered significantly supported.

Molecular phylogenetic results

The ITS+LSU (Fic. 1) dataset comprised sequences from 55 fungal

specimens representing 34 taxa, including the outgroup taxon. The dataset

had an aligned length of 2296 of which 1395 were constant, 201 parsimony-

uninformative, and 700 parsimony-informative. MP analysis yielded two

equally parsimonious trees (TL = 3897, CI = 0.376, HI = 0.624, RI = 0.627, RC

= 0.236). The best-fit model for ITS+LSU alignment estimated and applied

in the BI was GTR+I+G, lIset nst = 6, rates = invgamma; prset statefreqpr =

dirichlet (1,1,1,1). BI resulted in a similar topology with an average standard

deviation of split frequencies equal to 0.006862.

Rhomboidia wuliangshanensis forms a monophyletic lineage with strong

support (BS = 100%, BP = 100%, BPP = 1) and is sister to the Nigroporus-—

Trullella clade (Fic. 1).

Taxonomy

Rhomboidia C.L. Zhao, gen. nov.

MB 833318

Differs from Nigroporus and Trullella by its stipitate to substipitate basidiomata, its

orange-brown to reddish brown surface, and its monomitic hyphal system in both

context and trama.

Fic. 1. Maximum parsimony strict consensus tree illustrating the phylogeny of Rhomboidia

wuliangshanensis and related species in the residual polyporoid clade based on ITS+nLSU

sequences. Branches are labeled with maximum likelihood bootstrap >70%, parsimony bootstrap

proportions >50% and Bayesian posterior probabilities >0.95.

Rhomboidia wuliangshanensis gen. & sp. nov. (China) ... 655

G Cerrenaceae

+ Hyphodermataceae

@ Meripilaceae 95/95/1

A Meruliaceae -/-/l

& Panaceae - /81/0.95

84/81/1

100/100/1

Physisporinus vitreus CBS 486.72 @

Irpex lacteus CBS 431.48 @

Irpex lacteus DO 421/951208 @

Ischnoderma benzoinum KHL 12099

Ischnoderma resinosum FD-328 ©

Diplomitoporus flavescens X 84 *

Climacocystis borealis KH 13318 @®

Panus strigellus TNPA 243940 &

Panus conchatus X 1234 &

100/100/1

| Steccherinaceae

@ > Fomitopsidaceae

WME P, Leo} 0/100/1|Pseudolagarobasidium acaciicola CBS 115543 @

ee ae ae Pseudolagarobasidium belizense CFMR DLC 04-31 @

x Polyporaceae Oe Pseudolagarobasidium acaciicola CBS 115544 @©

Hypochnicium lyndoniae NL 041031 +

Hypochnicium bombycinum MA 15305 +

95/63/ -| Spongipellis spumeus BRNM 712630 ©

S782 100/100/1

100/100/1

100/91/1

97/61/1

100/100/1

92/94/1

100/88 1! 1 o9/10071

100/58/0.98

100/100/1

Spongipellis spumeus BRNM 734877 &

Spongipellis spumeus PRM 891931 @&

100/99/1)Physisporinus vitreus KHL 11959 (GB) @

Physisporinus vitreus 3163 @

Physisporinus sanguinolentus BRNM 699576 @

Podoscypha venustula CBS 65684 pea

Abortiporus biennis TFRI 274 Ht

100/63/ - |Loweomyces fractipes X 1253 na

10010011 roweomves fractipes X 1250 [jj

Loweomyces fractipes X 1149 [J

100/100/1) Junghuhnia crustacea X 262

Junghuhnia crustacea X 1127 Jj

Flaviporus brownii X 1216

Flaviporus liebmannii X 666 Ij

Flaviporus liebmannii X 251 fj

100/100/I' Flaviporus liebmannii X 249 fj

100/100/1 p Bigpirapers ailaoshanensis CLZhao 595 [ij

Elaphroporia ailaoshanensis CLZhao 596 |

95/100/1- Trullella duracina X290

Trullella polyporoides X510

Trullella dentipora X 200 B

Nigroporus vinosus BHS2008-100 [ij

Nigroporus vinosus 8182 Q

Nigroporus vinosus X 839 fj

100/100/1; Rhomboidia wuliangshanensis CLZhao 4406 BH

Rhomboidia wuliangshanensis CLZhao 441 Ij

Flabellophora sp.X340

Mycorrhaphium adustum Dai 10173 [i

Mycorrhaphium adustum 8024 [fj

100/99/1 - Junghuhnia micropora Spirin 2652 fj

100/100/1 | l Antrodiella sp. X 418 Pil

100/100/1 Skeletocutis novae-zelandiae Ryvarden 38641 ij

Frantisekia mentschulensis BRNM 710170 i |

00/100/1 Fy antisekia mentschulensis 1377 |

Steccherinum fimbriatum KHL 11905 ia

Antrodiella semisupina FCUG 960 [ij

Antrodiella americana Gothenburg 3161 [fj

Steccherinum ochraceum KHL 11902

100/100/l' steecherinum ochraceum Rybergs.n. [i

Xanthoporus syringae X 339 Oo

an Xanthoporus syringae Cui 2177

Xanthoporus syringae Gothenburg 1488 [J

100/100/1

——i

656 ... Xu & al.

TYPE SPECIES: Rhomboidia wuliangshanensis C.L. Zhao

EryMoLoGy: Rhomboidia (Lat.): referring to the rhomboid pileus of the basidiocarp

with the poroid hymenophore.

BASIDIOMATA annual, stipitate. Pileus rhomboid, arising from a multiple

branched stipe. Pores angular, small, dissepiments thin, entire. Hyphal system

monomitic; generative hyphae thick-walled bearing clamp connections,

IKI-, CB-; tissues unchanged in KOH. Cystidia absent, fusoid cystidioles

numerous; hyphal ends numerous. Basidia barrel-shaped to clavate.

Basidiospores broadly ellipsoid, hyaline, thin-walled, smooth, IKI-, CB-.

TYPE OF ROT: white rot.

Rhomboidia wuliangshanensis C.L. Zhao, sp. nov. Fics 2, 3

MB 833320

Differs from Nigroporus vinosus by its stipitate to substipitate basidiomata and

monomitic hyphal structure.

Type: China. Yunnan Province: Puer, Jingdong County, Wuliangshan National Nature

Reserve, on angiosperm trunk, 6 Oct 2017, CLZhao 4406 (Holotype, SWFC 0004406;

GenBank MK860715, MK860710).

EryMo.Locy: The specific epithet wuliangshanensis (Lat.) refers to the type locality,

Wuliangshan.

BASIDIOMATA annual, stipitate to substipitate. Pileus rhomboid, arising from

a multiple branched stipe, edges curling slightly inward, 3.5 cm from the base

to margin, 4 cm wide, up to 3 mm thick; pileus surface radially striate, slightly

brown to orange brown when fresh, drying brown to reddish; the margin

acute, entire. Pore surface white when fresh, cream to buff upon drying. Pores

angular, 7-9 per mm, dissepiments thin, entire. Context corky, white, thin, up to

0.5 mm thick. Tubes concolorous with pore surface, corky, up to 2.5 mm long.

- TYPE OF ROT: white rot.

ADDITIONAL SPECIMEN EXAMINED: CHINA. YUNNAN PROVINCE. Puer:

Jingdong County, Wuliangshan National Nature Reserve, on angiosperm trunk,

6 Oct 2017, CLZhao 4411 (SWFC 004411; GenBank MK860716, MK860711).

Discussion

Rhomboidia is supported as a new genus by phylogenetic analyses and

morphological characters. It is embedded in Steccherinaceae with strong

support. Phylogenetically, Rhomboidia is closely related to Flabellophora

G. Cunn., Nigroporus Murrill, and Trullella Zmitr. based on ITS+LSU nuclear

RNA gene analyses (Fic. 1), which is similar to the previous multigene sequence-

based study (Miettinen & al. 2012). The genera closely related to Rhomboidia

Rhomboidia wuliangshanensis gen. & sp. nov. (China) ... 657

Fic. 2. Rhomboidia wuliangshanensis (holotype, SWFC 0004406). Scale bars = 5 mm.

658 ... Xu & al.

Leciiiian

G (a)

Fic. 3. Rhomboidia wuliangshanensis (drawn from the holotype, SWFC 0004406). A. Basidiospores;

B. Basidia and basidioles;

subiculum. Scale bars: a

C. Cystidioles; D. Hyphal ends; E. Hyphae from trama; F. Hyphae from

= 5 um; b-f = 10 um.

Rhomboidia wuliangshanensis gen. & sp. nov. (China) ... 659

are easily separated morphologically: In Flabellophora basidiomata arise

from a submerged pseudosclerotium and develop unilateral pilei with a

crust and a coriaceous context (Nufez & Ryvarden 2001). Nigroporus differs

from Rhomboidia by developing resupinate to pileate basidiocarps with

vinaceous brown to pink or violet pore surface and a dimitic hyphal system

(Gilbertson & Ryvarden 1987). In Trullella basidiomata are spathulate and

light-coloured, with a monomitic hyphal system in the context but dimitic

in the trama (Miettinen & al. 2012, Zmitrovich 2018).

Rhomboidia resembles other stipitate genera in Polyporales such as

Abortiporus Murrill, Jahnoporus Nuss, and Polyporus P. Micheli ex Adans.

Abortiporus, however, has a duplex structure and thick-walled basidiospores

(Nunez & Ryvarden 2001). Jahnoporus is characterized by large spindle-

shaped basidiospores (Gilbertson & Ryvarden 1987), and Polyporus has a

dimitic hyphal system (Bernicchia & Gorjon 2010).

Polypores are extensively studied and well-known in North America

(Gilbertson & Ryvarden 1987, Zhou & al. 2016) and Eurasia (Nufez &

Ryvarden 2001, Bernicchia & Gorjén 2010, Dai 2012, Ryvarden & Melo

2014, Dai & al. 2015), but Chinese polypore diversity is still being explored,

especially in subtropical and tropical areas. Rhomboidia wuliangshanensis

was collected from Yunnan Province, where many new taxa in Polyporales

and Hymenochaetales have been described (e.g., Li & Cui 2010, He &

Li 2011, Yu & al. 2013, Yang & He 2014, Zhao & Wu 2017, Zhao & Ma

2019). We anticipate that additional, undescribed polypore taxa will be

discovered throughout China after extensive collections are analyzed both

morphologically and molecularly.

Acknowledgments

Special thanks are due to Drs. Karen K. Nakasone (Center for Forest Mycology

Research, Northern Research Station, U.S. Forest Service, USA) and Lu-Sen Bian

(Experiment Center of Forestry in North China, Chinese Academy of Forestry,

P.R. China) who reviewed the manuscript. The research is supported by the

National Natural Science Foundation of China (Project No. 31700023) and Yunnan

Agricultural Foundation Projects (2017FG001-042) and the Yunnan Provincial

Innovation Team on Kapok Fiber Industrial Plantation (2018HC014) and the

Science Foundation of Education Department in Yunnan (2018JS326).

Literature cited

Bernicchia A, Gorjon SP. 2010. Fungi Europaei 12: Corticiaceae |. Edizioni Candusso, Lomazzo.

1007 p.

660 ... Xu & al.

Binder M, Hibbett DS, Larsson KH, Larsson E, Langer E, Langer G. 2005. The phylogenetic

distribution of resupinate forms across the major clades of mushroom-forming fungi

(Homobasidiomycetes). Systematics and Biodiversity 3: 113-157. https://doi.org/10.1017/

$1477200005001623

Binder M, Justo A, Riley R, Salamov A, Lépez-Giraldez F, Sjékvist E, Copeland A, Foster B,

Sun H, Larsson E, Larsson KH, Townsend J, Grigoriev IV, Hibbett DS. 2013. Phylogenetic

and phylogenomic overview of the Polyporales. Mycologia 105: 1350-1373.

https://doi.org/10.3852/13-003

Choi JJ, Kim SH. 2017. A genome Tree of life for the fungi kingdom. Proceedings of the

National Academy of Sciences of the United States of America 114: 9391-9396.

https://doi.org/10.1073/pnas.1711939114

Dai YC. 2010. Hymenochaetaceae (Basidiomycota) in China. Fungal Diversity 45: 131-343.

https://doi.org/10.1007/s13225-010-0066-9

Dai YC. 2012. Polypore diversity in China with an annotated checklist of Chinese polypores.

Mycoscience 53: 49-80. https://doi.org/10.1007/s10267-011-0134-3

Dai YC, Yang ZL, Cui BK, Yu CJ, Zhou LW. 2009. Species diversity and utilization of medicinal

mushrooms and fungi in China (Review). International Journal of Medicinal Mushrooms

11: 287-302. https://doi.org10.1615/Int) MedMushr.v1 1.13.80

Dai YC, Cui BK, Si J, He SH, Hyde KD, Yuan HS, Lui XY, Zhou LW. 2015. Dynamics of

the worldwide number of fungi with emphasis on fungal diversity in China. Mycological

Progress 14(62). [9 p.] https://doi.org/10.1007/s11557-015-1084-5

Felsenstein J. 1985. Confidence intervals on phylogenetics: an approach using bootstrap.

Evolution 39: 783-791. https://doi.org/10.1111/j.1558-5646.1985.tb00420.x

Floudas D, Hibbett DS. 2015. Revisiting the taxonomy of Phanerochaete (Polyporales,

Basidiomycota) using a four gene dataset and extensive ITS sampling. Fungal Biology 119:

679-719. https://doi.org/10.1016/j.funbio.2015.04.003

Gilbertson RL, Ryvarden L. 1987. North American polypores 2. Megasporoporia-

Wrightoporia. Fungiflora, Oslo.

Grigoriev IV, Nikitin R, Haridas S$, Kuo A, Ohm R, Otillar R, Riley R, Salamov A, Zhao X,

Korzeniewski F, Smirnova T. 2013 [“2014”]. Mycocosm portal: gearing up for 1000 fungal

genomes. Nucleic Acids Research 42(D1): D699-D704.

https://doi.org/10.1093/nar/gkt1183

Hall TA. 1999. BioEdit: a user-friendly biological sequence alignment editor and analysis

program for Windows 95/98/NT. Nucleic Acids Symposium Series 41: 95-98.

He SH, Li HJ. 2011. Hymenochaete in China. 2. A new species and three new records from

Yunnan Province. Mycotaxon 118: 411-422. https://doi.org/10.5248/118.411

Justo A, Miettinen O, Floudas D, Ortiz-Santana B, Sjokvist E, Lindner D, Nakasone

K, Niemela T, Larsson KH, Ryvarden L, Hibbett DS. 2017. A revised family-level

classification of the Polyporales (Basidiomycota). Fungal Biology 121: 798-824.

https://doi.org/10.1016/j.funbio.2017.05.010

KatohK, TohH. 2008. Recentdevelopmentsinthe MAFFT multiplesequencealignmentprogram.

Briefings in Bioinformatics 9: 286-298. https://doi.org/10.3767/003158514X681828

Kirk PM, Cannon PF, David JC, Minter DW, Stalpers JA. 2008. Ainsworth and Bisby’s

dictionary of the fungi. 10th ed. Wallingford, Oxon, UK: CAB International Press.

https://doi.org/10.1079/9780851998268.0000

Larsson KH. 2007. Re-thinking the classification of corticioid fungi. Mycological Research

111: 1040-1063. https://doi.org/10.1016/j.mycres.2007.08.001

Rhomboidia wuliangshanensis gen. & sp. nov. (China) ... 661

Levin L, Maira C, Martin H, Rene U. 2016. Degradation of 4-nitrophenol by the white-

rot polypore Trametes versicolor. International Biodeterioration & Biodegradation 107:

174-179. https://doi.org/10.1016/j.ibiod.2015.11.023

Li HJ, Cui BK. 2010. A new Trametes species from Southwest China. Mycotaxon 113:

263-267. https://doi.org/10.5248/113.263

Miettinen O, Rajchenberg M. 2012. Obba and Sebipora, new polypore genera related to

Cinereomyces and Gelatoporia (Polyporales, Basidiomycota). Mycological Progress 11:

131-147. https://doi.org/10.1007/s11557-010-0736-8

Miettinen O, Ryvarden L. 2016. Polypore genera Antella, Austeria, Butyrea, Citripora,

Metuloidea and Trulla (Steccherinaceae, Polyporales). Annales Botanici Fennici 53:

157-172. https://doi.org/10.5735/085.053.0403

Miettinen O, Larsson KH, Sjékvist E, Larsson KL. 2012. Comprehensive taxon sampling

reveals unaccounted diversity and morphological plasticity in a group of dimitic polypores

(Polyporales, Basidiomycota). Cladistics 28: 251-270.

https://doi.org/10.1111/j.1096-0031.2011.00380.x

Miller MA, Holder MT, Vos R, Midford PE, Liebowitz T, Chan L, Hoover P, Warnow T. 2009.

The CIPRES Portals. CIPRES. URL: http://www.phylo.org/sub_sections/portal. 2009-08-04.

(Archived by WebCite(r) at http://www.webcitation.org/5imQlJeQa).

Nunez M, Ryvarden L. 2001. East Asian polypores 2. Polyporaceae s. lato. Synopsis Fungorum

14: 165-522.

Nylander JAA. 2004. MrModeltest v2. Program distributed by the author. Evolutionary

Biology Centre, Uppsala University.

Petersen JH. 1996. Farvekort. The Danish Mycological Society’s colour-chart. Foreningen til

Svampekundskabens Fremme, Greve.

Posada D, Crandall KA. 1998. Modeltest: testing the model of DNA substitution. Bioinformatics

14: 817-818. https://doi.org/10.1093/bioinformatics/14.9.817

Ronquist F, Huelsenbeck JP. 2003. MrBayes 3: Bayesian phylogenetic inference under mixed

models. Bioinformatics 19: 1572-1574. https://doi.org/10.1093/bioinformatics/btg180

Ryvarden L, Melo I. 2014. Poroid fungi of Europe. Synopsis Fungorum 31. 455 p.

Sjokvist E, Larsson E, Eberhardt U, Ryvarden L, Larsson KH. 2012. Stipitate stereoid basidiocarps

have evolved multiple times. Mycologia 104: 1046-1055. https://doi.org/10.3852/11-174

Swofford DL. 2002. PAUP*: phylogenetic analysis using parsimony (*and other methods).

Version 4.0b10. Sinauer Associates, Massachusetts.

Tomsovsky M, Menkis A, Vasaitis R. 2010. Phylogenetic relationships in European

Ceriporiopsis species inferred from nuclear and mitochondrial ribosomal DNA sequences.

Fungal Biology 114: 350-358. https://doi.org/10.1016/j.funbio.2010.02.004

Vu D, Groenewald M, De VM, Gehrmann T, Stielow B, Eberhardt U, Al-Hatmi A, Groenewald

JZ, Cardinali G, Houbraken J, Boekhout T, Crous PW, Robert V, Verkley GJM. 2019.

Large-scale generation and analysis of filamentous fungal DNA barcodes boosts coverage

for kingdom fungi and reveals thresholds for fungal species and higher taxon delimitation.

Studies in Mycology 92: 135-154. https://doi.org/10.1016/j.simyco.2018.05.001

Westphalen MC, Rajchenberg M, Tomsovsky M, Gugliotta AM. 2018. A re-evaluation of

Neotropical Junghuhnia s. lat. (Polyporales, Basidiomycota) based on morphological and

multigene analyses. Persoonia 41: 130-141. https://doi.org/10.3767/persoonia.2018.41.07

White TJ, Bruns T, Lee S, Taylor J. 1990. Amplification and direct sequencing of fungal

ribosomal RNA genes for phylogenetics. 315-322, in: MA Innis & al. (eds). PCR protocols:

a guide to methods and applications. Academic Press, San Diego.

https://doi.org/10.1016/B978-0-12-372180-8.50042-1

662 ... Xu & al.

Wu ZQ, Xu TM, Shen S, Liu XF, Luo KY, Zhao CL. 2018. Elaphroporia ailaoshanensis gen. et

sp nov in Polyporales (Basidiomycota). MycoKeys 29: 81-95.

https://doi.org/10.3897/mycokeys.29.22086

Yang J, He SH. 2014. Hymenochaete in China. 8. H. biformisetosa sp. nov. with a key to species

with denticulate setae. Mycotaxon 128: 137-144. https://doi.org/10.5248/128.137

Yu HY, Zhao CL, Dai YC. 2013. Inonotus niveomarginatus and I. tenuissimus spp. nov.

(Hymenochaetales), resupinate species from tropical China. Mycotaxon 124: 61-68.

https://doi.org/10.5248/124.61

Zhao CL, Ma X. 2019. Perenniporia mopanshanensis sp. nov. from China. Mycotaxon 134:

125-137. https://doi.org/10.5248/134.125

Zhao CL, Wu ZQ. 2017. Ceriporiopsis kunmingensis sp. nov. (Polyporales, Basidiomycota)

evidenced by morphological characters and phylogenetic analysis. Mycological Progress

16: 93-100. https://doi.org/10.1007/s11557-016-1259-8

Zhou LW, Nakasone KK, Burdsall Jr. HH, Ginns J, Vlasak J, Miettinen O, Spirin V, Niemela

T, Yuan HS, He SH, Cui BK, Xing JH, Dai YC. 2016. Polypore diversity in North America

with an annotated checklist. Mycological Progress15: 771-790.

https://doi.org/10.1007/s11557-016-1207-7

Zmitrovich IV. 2018. Conspectus systematis Polyporacearum v. 1.0. Folia Cryptogamica

Petropolitana 6: 3-145.

MY COTAXON

October-December 2019—Volume 134, pp. 663-675

https://doi.org/10.5248/134.663

Filsoniana lhasanensis sp. nov.

from Tibet, China

XuE-MEI WEN”?, HURNISA SHAHIDIN'?, ABDULLA ABBAS!

‘College of Life Science and Technology, Xinjiang University,

Urumai, 830046, P. R. China

*Tibet Plateau Institute of Biology, Lhasa, 850001, P. R. China

* CORRESPONDENCE TO: hurnisa_xju@sina.com, zxg_lichen@163.com

ABSTRACT—A new species, Filsoniana lhasanensis, is described from Tibet, China, where it

grows on rocks at altitudes 4200-4432 m. The lichen is characterised by a poorly developed or

absent dark orange areolate thallus, lecanorine to zeorine orange to deep-orange aggregated

apothecia, and richly branched and anastomosed paraphyses. DNA was obtained for three

gene loci—nuclear internal transcribed spacer (ITS), nuclear large subunit (LSU), and

mitochondrial small subunit (SSU). Phylogenetic analyses support the taxon as a new species

of Filsoniana.

Key worps—Himalaya, phylogeny, taxonomy, Teloschistoideae

Introduction

Teloschistaceae is a widespread and well-delimited family of lichenized

fungi, estimated at over 1000 species (Arup & al. 2013). Most species

contain anthraquinone (which confers a yellow to orange color) and

grow under sun-exposed conditions (Gaya & al. 2015). Recent molecular

phylogenetic research supports subdivision of the Teloschistaceae into three

subfamilies: Xanthorioideae, Caloplacoideae, and Teloschistoideae (Arup &

al. 2013; Gaya & al. 2015). Kondratyuk & al. (2013, 2015), who evaluated

Teloschistoideae based on three gene loci (ITS, LSU, SSU), proposed an

additional subfamily Brownlielloideae; however this analysis was based

on a hybrid sequence dataset comprising ITS and LSU data derived from

664 ... Wen, Shahidin, Abbas

the lichen mycobiont and the SSU derived from a fungal contaminant

(Vondrak & al. 2018). Many small monophyletic genera established in

Brownlielloideae were based solely on DNA data without any distinguishing

phenotypic characters, and these genera remain within the Teloschistoideae

as proposed by Arup & al. (2013).

During our study on Chinese Teloschistaceae, we collected some

interesting samples from the suburbs of Lhasa and Shannan in Tibet. The

Tibetan plateau is a unique biogeographical region with varied landscapes,

altitudinal belts, and alpine ecosystems and is considered a world center of

species formation (Zhang & al. 2002). Teloschistaceae in this region have

been inadequately studied, with only 15 species reported (Obermayer 2004;

Wei 1991, 2017; Poelt & Petutschnig 1992; Poelt & Hinteregger 1993; Wei

& Jiang 1986). More species are known from the neighboring Himalayan

regions: India, where >74 species of Teloschistaceae have been recorded

(Joshi & Upreti 2006, 2011; Joshi & al. 2008, 2009, 2014; Kondratyuk & al.

2018) and Nepal, where 47 species have been discovered (Olley & Sharma

2013). According to available literature, there are still large underexplored

areas in the Tibet region, and additional undescribed lichenized fungi

species are expected to be discovered in this poorly studied region.

Most of crustose Teloschistaceae species have few taxonomically

significant morphological characters. Traditional species concepts based in

morphology and chemistry are overly simplified for the crustose species

in Teloschistaceae. Modern classification methods are essential for accurate

taxonomic placement of rare and undescribed species. We combined

sequence analyses of three gene loci—ITS, LSU, SsU—with morphological

and chemical methods to describe a new species, Filsoniana lhasanensis.

This is the first report of the genus Filsoniana in China.

Materials & methods

Morphology & chemistry

Specimens collected from Lhasa and Shannan, Tibet, China, were preserved in

Tibet Plateau Institute of Biology, Lhasa, China (XZ) and Lichen Research Center

in Arid Zone of Northwest China, Xinjiang University, Urumqi, China (XJU-L).

The specimens were examined morphologically using a Shunyu SZM45 dissecting

microscope and Nikon Eclipse E100 compound microscope. Colors follow RAL

color system (https://www.ralcolorchart.com/) Sections for anatomical details and

measurements were made manually and mounted in water. Chemical constituents

were identified by thin-layer chromatography using solvent systems C (Orange

& al. 2010). Thallus morphological structures were photographed with a Nikon

Filsoniana lhasanensis sp. nov. (China) ... 665

TABLE 1. Primers and cycling parameters used for PCR.

Locus PRIMERS PCR SETTINGS

ITS ITS1F (Gardes & Bruns 1993) 94°C-5 min; 6 cycles of: 94°C-45 sec, 57-52°C

1 ITS4 (White & al. 1990) : (decreasing 1°C per cycle) -55 sec, 72°C-1 min; 30

:_ cycles of: 94°C-45 sec, 51°C-55sec, 72°C-1 min; and

| 72°C-7 min

LSU { ALIR (Déring & al. 2000) | 94°C-10 min; 6 cycles of: 94°C-1 min, 57-52°C

| LR6 (Vilgalys & Hester 1990) 1 (decreasing 1°C per cycle) -1 min, 72°C-1.2 min; 30

| cycles of: 94°C-1 min, 51°C-1 min, 72°C-1.2 min;

' and 72°C-2 min

ssu mrSSU1 (Zoller & al. 1999) 94°C-10 min; 6 cycles of: 94°C-45 sec, 57-52°C

mrSSU7 (Zhou & Stanosz 2001) : (decreasing 1°C per cycle) —55 sec, 72°C-1 min; 30

1 cycles of: 94°C-45 sec, 51°C-55sec, 72°C-1 min; and

72°C-7 min

DS-Fi2 Digital Camera using a Nikon MZ25 and Nikon Eclipse Ci stereomicroscope.

All measurements were made using the Nikon Nis elements (D). Morphology and

anatomy were described according to Vondrak & al. (2013).

DNA extraction, PCR amplification, DNA sequencing

Total genomic DNA was extracted using the Fungi Genomic DNA Extraction

Kit, following manufacturer's instructions. Molecular data were generated for three

loci: the nuclear internal transcribed spacer (ITS), nuclear large subunit (LSU), and

mitochondrial small subunit (SSU). Primers and PCR cycling parameters used for

amplification are summarized in TABLE 1.

Sequence analysis

The newly generated sequences were compared to GenBank database sequences

using BLASTN search (http://www.ncbi.nlm.nih.gov/BLAST/). All sequences were

aligned with sequences of selected representatives of Teloschistoideae obtained

from GenBank (TABLE 2) using MAFFT version 7 (Katoh & Standley 2013) with

G-INS-i method.

Phylogenies were generated using Bayesian inference and Maximum Likelihood

(ML) methods. Bayesian inference used the best fitting model as inferred by j Model

Test v2.1.6 (Darriba & al. 2012), implemented in MrBayes v3.2.6 (Ronquist & al.

2012) on the Cipres Scientific gateway (Miller & al. 2010), TrNef+I+G for ITS and

LSU and HKY+I+G for SSU. Two parallel MCMCMC runs were executed, each

using eight chains and 10,000,000 generations, with trees sampled every 1000th

generation.

Maximum likelihood (RAxML) analyses were performed using RAxMLHPC

v.8 on XSEDE (Stamatakis 2014) under the GTRGAMMA model on CIPRES

Science Gateway (Miller & al. 2010). Rapid bootstrap analyses were performed

with 1000 bootstrap replicates. The phylogenetic tree was drawn using FigTree ver.

1.4.3 (http://tree.bio.ed.ac.uk/software/figtree/).

666 ... Wen, Shahidin, Abbas

TABLE 2. Sequences used in the phylogenetic analyses.

(Sequences generated for this study are in bold.)

SPECIES VOUCHER ITS LSU SSU

Brownliella aequata | SK838 [4] | KF264627 | KF264662 { KF264688

! SK831 [4] ! KF264626 ! KF264661 ! KF264687

B. kobeana ! 130318 [5] ! KT456214 ! KT456229 ! KT456244

! 120032 [5] ! KT456212 ! KT456227 ! KT456242

B. montisfracti ! SK231 [4] ! KF264625 ! KF264660 ! KF264686

! SK230 [4] ! KF264624 ! KF264659 ! _

Browniliella sp. SK2013a [4] KF264628 KF264663 ! —

Caloplaca sp. ! Casan 242 [3] ! KT291463 ! KT291558 ! KT291508

! Gaya 110 [3] ! KT291456 ! KT291552 ! KT291499

Elixjohnia bermaguiana ! Kondratyuk 20487 [1] ! KC179299 ! KC179245 ! KC179584

E. gallowayi ! Karnefelt 997504 [1] ! KC179301 ! KC179247 ! KC179586

E. jackelixii ! U1402 [1] ! KC179303 ! KC179248 ! KC179587

! SK910 [4] ! KF264655 ! KF264683 ! KF264707

Filsoniana australiensis ! SK851 [4] ! KF264631 ! KF264665 ! KF264691

! SK850 [4] ! KF264632 ! KF264666 ! KF264692

EF. lhasanensis 224, Wen 20110910-05 MK153161 MK439835 MK439831

! Z19, Wen 20110901-29 ! MK153160 ! MK439834 ! MK439830

E rexfilsonii ! SK859 [4] ! KF264638 ! KF264671 ! —

! SK861 [4] ! KF264636 ! KF264670 ! =

Follmannia orthoclada ! Frodén 1772 [1] ! KC179291 ! KC179191 ! _

Haloplaca sp. ! 6-UA2013 [1] ! KC179295 ! KC179203 ! KC179537

Harusavskia elenkinianoides ! SK997 [6] ! KY614404 ! KY614452 ! KY614485

! SK269 [6] ! KY614405 ! KY614453 ! KY614486

Josefpoeltia parva ! Frédén 1671 [1] ! KC179296 ! KC179204 ! KC179539

J. sorediosa ! Frodén 1593 [1] ! KC179297 ! KC179205 ! KC179540

! SK992 [4] ! KF264646 ! KF264674 ! KF264697

! SK991 [4] ! KF264645 ! KF264673 ! KF264696

Kaernefia albocrenulata ! SK245 [4] ! KF264647 ! KF264675 ! KF264698

! SK246 [4] ! KF264648 ! KF264676 ! KF264699

K. gilfillaniorum ! SK999 [4] ! KF264650 ! KF264678 ! KF264701

! SK253 [4] ! KF264649 ! KF264677 ! KF264700

K. kaernefeltii ! SK919 [4] ! KF264651 ! KF264679 ! KF264702

! SK921 [4] ! KF264652 ! KF264680 ! KF264703

Nevilleiella lateritia | SK 261 [6] 1 KY614427 | KY614464 | KY614502

Filsoniana lhasanensis sp. nov. (China) ... 667

SPECIES VOUCHER ITS LSU SSU

| SK 878 [6] | KY614426 | KY614463 | KY614501

N. marchantii ! SK D18 [6] ! KY614425 ! KY614462 ! KY614500

Scutaria andina ! PF209 [1] ! KC179298 ! KC179242 ! KC179581

Sirenophila cliffwetmorei ! SK A93 [6] ! KY614438 ! KY614471 ! KY614513

S. eos ! U1408 [1] ! KC179300 ! KC179246 ! KC179585

S. gintarasii ! SK D17 [6] ! KY614437 ! KY614470 ! KY614512

S. maccarthyi ! Karnefelt 977801 [1] ! KC179304 ! KC179249 ! KC179588

Solitaria chrysophthalma ! Cchr157 [3] ! KT291446 ! KT291537 ! KT291484

Stellarangia elegantissima ! PF 81 [1] ! KC179310 ! KC179254 ! KC179593

! Cele 75 [3] ! KT291454 ! KT291541 ! KT291488

Teloschistes exilis ! Gaya 12 [2] ! JQ301630 ! JQ301577 ! JQ301519

T. flavicans ! AFTOL-ID 315 [2] ! JQ301578 ! JQ301631 ! JQ301520

! Tflav103 [3] ! KT291472 ! KT291565 ! KT291523

T. hosseusianus Gaya 50 [2] KC179318 ! JQ301579 ! JQ301521

T. hypoglaucus ! PF68 [1] ! KC179319 ! KC179256 ! KC179595

T. sieberianus ! Tsie423 [3] ! EU639655 ! pez ! KT291525

Teloschistopsis bonae-spei ! PF82 [1] ! KC179322 ! KC179257 ! KC179596

Ts. eudoxa ! PF72 [1] ! KC179324 ! KC179258 ! KC179597

Thelliana pseudokiamae SK925 [4] KF264633 KF264667 —

! SK926 [4] ! KF264634 ! KF264668 ! =

! SK 925 [5] ! KT456225 ! KT456240 ! _—

! SK 926 [5] ! KT456226 ! KT456241 ! _—

Villophora isidioclada ! USE563 [1] ! KC179325 ! KC179266 ! KC179606

Villophora sp. 50 ! U1342 [1] ! KC179330 ! KC179268 ! —

Villophora sp. 40 ! USE574 [1] ! KC179328 ! KC179267 ! KC179607

Wetmoreana decipioides Cadec188 [3] KT291453 KT291540 KT291487

! SK689 [4] ! KF264644 ! — ! KF264695

W. texana ! SK536 [4] ! KF264658 ! = ! KF264711

! USE527 [1] ! KC179337 ! KC179273 ! KC179612

W. sp. 52 ! Frédén 1645 [1] ! KC179334 ! KC179270 ! KC179609

W. sp. 54 ! Frédén 1521 [1] ! KC179336 ! KC179272 ! KC179611

Xanthomendoza fallax ! Gaya 33 [2] ! JQ301687 ! JQ301580 ! JQ301633

Xa. ulophyllodes ! T11, Shahidin 145507 ! MK414784 ! MK439836 ! MK439832

! T22, Shahidin 145449 ! MK414785 ! MK439837 ! MK439833

Xanthoria sp. ! Gaya 31 [2] ! JQ301692 ! JQ301591 ! JQ301532

Sources: [1] Arup & al. (2013); [2] Gaya & al. (2012); [3] Gaya & al. (2015);

[4] Kondratyuk & al. (2013); [5] Kondratyuk & al. (2015); [6] Kondratyuk & al. (2017).

668 ... Wen, Shahidin, Abbas

Xanthomendoza ulophyllodes

Xanthomendoza fallax

1/100) Villophora isidioclada

1/100 illophora sp. 50

Villophora sp. 40

Josefpoeltia parva

q- Josefpoeltia sorediosa

Josefpoeltia sorediosa

1/80 0.99/99L Josefpoeltia sorediosa

1/100 Teloschistes exilis

1/100 , Teloschistes flavicans

1/94 Teloschistes flavicans

Teloschistes sieberianus

oor- Teloschistes hosseusianus

1/100 5 Teloschistes hypoglaucus

Teloschistes chrysophthalmus

1/100 1/100

Caloplaca sp.

+ ~ 219 Filsoniana lhasanenses

SK861 Filsoniana rexfilsonii

-/88} SK925 Thelliana pseudokiamae

SK925 Thelliana pseudokiamae

SK926 Thelliana pseudokiamae

SK859 Filsoniana rexfilsonii

0.97/86 Filsoniana australiensis

1/100 SK850 Filsoniana australiensis

iy Nevilleiella marchantii

1/9 Nevilleiella lateritia

1/95) 1/99 Nevilleiella lateritia

1/100 Brownliella sp.

1/100 Brownliella aequata

Brownliella aequata

1/100, 8rownliella kobeana

Brownliella kobeana

1/100 Brownliella montisfracti

1/73 Brownliella montisfracti

1/100; Keernefia gilfillaniorum

Kaernefia gilfillaniorum

1/100

1/93

thi

1/94

001

1/100 », Kaernefia kaernefeltii

SL Kaernefia kaernefeltii

O— Kaernefia albocrenulata

99 1/99 Kaernefia albocrenulata

/ Haloplaca sp.

Elixjohnia bermaguiana

1 1/100 as ; R

1/94 1/100 Elixjohnia gallowayi

1/100 Elixjohnia jackelixii

1/90 AGk Bie a

1/1004 Elixjohnia jackelixii

1/99 Sirenophila eos

1/95 Sirenophila maccarthyi

Sirenophila gintarasii

-/95 1/100 . labbod

enophila cliffwetmor

1/100 Teloschistopsis bonae-spei

Teloschistopsis eudoxa

1/100 Stellarangia elegantissima

1/70 Stellarangia elegantissima

1/100 Wetmoreana decipioides

Wetmoreana decipioides

1/100 1/100 Wetmoreana texana

1/100} Wetmoreana texana

1/95 Wetmoreana sp. 54

-94/90]

0.949 Wetmoreana sp. 52

1/85) 0.95/9 Caloplaca sp.

Solitaria chrysophthaima

1/100 Xanthoria sp.

Scutaria andina

1/100 , Harusavskia elenkinianoides

1/99 Harusavskia elenkinianoides

Follmannia orthoclada

0.95/

ae O09

Fic. 1. The ML tree based on a concatenated 3-loci data matrix. The numbers above each node

represent posterior probability (PP) and bootstrap support (BS) values. Only PP values >0.90

and BS values >70% are shown. The species described in this paper is shown in blue font.

Results & discussion

New ITS, LSU, and SSU sequences were generated in this study. A

total of 70 ITS sequences, 67 LSU sequences, and 58 SSU sequences were

included in the analyses (Fic. 1). Trees of similar topologies were also

generated using the maximum likelihood method and Bayesian approach,

Filsoniana lhasanensis sp. nov. (China) ... 669

with Xanthomendoza ulophyllodes and X. fallax as outgroup. Our combined

analyses grouped our new species with Filsoniana rexfilsonii and Thelliana

pseudokiamae in a monophyletic clade with high support value (BS = 93,

PP = 1). The new species is closely related to Thelliana pseudokiamae

(S.Y. Kondr. & Karnefelt) S.-Y. Kondr.& al. and E rexfilsonii (S.Y. Kondr.

& Karnefelt) S.Y. Kondr. & al.. Almost all other members of Filsoniana

have well-developed thalli and occur in Australia. The sequences listed

by Kondratyuk & al. (2013) under the name “Squamulea kiamae” were

misdetermined; they were derived from specimen Karnefelt 994101 [LD

1101337], which was resequenced by Kondratyuk &al. (2015) and designated

as the holotype of a new genus and species, Thelliana pseudokiamae S.Y.

Kondr. & al. Therefore all four of the “Squamulea kiamae” sequences

represent Th. pseudokiamae, and authentic sequences of S. kiamae are not

included in our analyses. Filsoniana kiamae and Th. pseudokiamae differ in

the sorelia and lobe size (Kondratyuk & al. 2007, 2015). Our results conflict

with Kondratyuk & al. (2015), whose ITS-LSU-SSU combined analyses

placed Filsoniana in Brownlielloideae; our analyses place most members of

Brownlielloideae in the Teloschistoideae.

Taxonomy

Filsoniana lhasanensis X.M. Wen, Shahidin & A. Abbas sp. nov. Fic. 2

FN 570592

Differs from Filsoniana kiamae and E rexfilsonii by its reduced thallus, its aggregated

adnate apothecia, and its Asian distribution.

Type: China. Tibet, Lhasa City, Maizhuokunggar county, Riduo Village, 29°42’35”N

92°05’10’E, alt. 4200 m, 1 September 2011, X. M. Wen 20110901-29 (Holotype, XZ;

isotype, XJU-L; GenBank MK153160, MK439834, MK43983).

EryMo.ocy: The epithet /hasanensis refers to the region in which the type specimen

was collected.

THALLUS crustose, areolate, poorly developed or absent, usually confined

to the apothecia, 0.1-0.2 mm diam., orange to deep orange [RAL 2011].

CorTEX paraplectenchymatous, 12.9-67.1 um high, composed of thin-

walled cells, (2.38-)4.8-7.4(-10.5) um diam. ALGAL LAYER + continuous,

49.5-97.9 um thick. MEDULLA loosely interwoven by thin-walled hyphae

under the apothecia, 59.8-86.1 um high.

APOTHECIA lecanorine to zeorine, numerous, adnate, always

aggregated (2-7) or crowded together, disc margins present, smooth,

entire, concolorous with thallus, sun yellow [RAL 1037] to deep orange

670 ... Wen, Shahidin, Abbas

[RAL 2011], disc round to irregular, yellow-orange [RAL 2000] to signal

orange [RAL 2010], flat to concave, 0.3-0.7 mm diam. EXxcIPLE 9.6-14.8

um thick, consisting of 2-4 layers of cells, upper part of cells spherical,

3.2-6.0 um diam, cells of lower part elongated, 3.5-10.6 x 1.4-3.6 um.

HypoTuHEcium thin, 22.3-34.7 um high, composed of several lines of

paraplectenchymatous cells, 2.9-5.2 x 2.3-4.6 um. HYMENIUM 52.2-89.5

um high, colorless. PARAPHYSES anastomosed, generally dichotomously

branched or trigeminally branched at tips, the 3-4 terminal cells gradually

enlarging towards tips, the widened cells nonuniform in size, the second

cells 3.5-6.7 um diam., and the third ones 3.2-5.3 um diam.

Ascr_ teloschistes-type, 8-spored. Ascospores colorless, ellipsoid,

(11.2-)11.7-17.5(-4.7) x 5.8-8.4(-9.3) um, septum 3.3-6.5 um diam.

SPOT TESTS—K+ red.

CHEMISTRY—Parietin, emodin, and two unknown anthraquinones.

EcoLtocy—Filsoniana lhasanensis grows on rocks in open areas

and associates with species of Aspicilia, Lecanora, Rusavskia, Physcia,

Rinodina, Rhizoplaca, and Acarospora. The new lichen is distributed

in arid or semiarid areas of Lhasa city and Shannan city at elevations of

4200-4432 m.

ADDITIONAL SPECIMEN EXAMINED: CHINA. TIBET, Shannan City, Nagarze county,

Yamzho Yumco, 29°09’34”N 90°30'19’E, alt. 4432 m, 10 September 2011, X.M. Wen

20110910-05 (XZ; GenBank MK153161, MK439835, MK439831).

REMARKS: Filsoniana lhasanensis is characterized by a reduced, dark orange

to brownish orange thallus and aggregated apothecia. It is morphologically

similar to Xanthocarpia ferrarii (Bagl.) Frodén & al., which differs by its

narrower septum, larger ascospores, well developed true exciple, zeorine

to biatorine apothecia, and classification in Xanthorioideae (Arup & al.

2013).

Filsoniana australiensis, F. rexfilsonii, and F. lhasanensis have similar

paraplectenchymatous cortex and broad paraphyses (Kondratyuk & al.

2007). However, F. australiensis and E rexfilsonii have well-developed thalli

(E australiensis has a placodioid thallus, and FE rexfilsonii has a squamulose

thallus) with scattered or rare apothecia, whereas F [hasanensis has a poorly

developed thallus consisting of only a few scattered areoles.

A summary of the characteristics of FE [hasanensis and morphologically

similar species is provided in TABLE 3.

Filsoniana lhasanensis sp. nov. (China) ... 671

Fic. 2. Filsoniana lhasanensis (XZ (Wen 20110901-29)): A. Thallus and apothecia; B. Cortex

of thallus; C. Cross section of the apothecium; D. Hypothecium; E. Paraphyses branched and

enlarged at the tips; F. Paraphyses branched and anastomosed; G. Ascus containing eight

ascospores. Scale bars: A = 1 mm; B-F = 10 um; G=5 um.

672 ... Wen, Shahidin, Abbas

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Filsoniana lhasanensis sp. nov. (China) ... 673

Acknowledgments

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

(31093440, 31260008) and Natural Science Foundation of Xizang Autonomous

Region, China (2016ZR-QY-05). The authors are grateful to Dr. Jan Vondrak

(Institute of Botany, Academy of Sciences of the Czech Republic, Prihonice) and

Prof. Xinli Wei (State Key Laboratory of Mycology, Institute of Microbiology,

Chinese Academy of Sciences, Beijing) for peer review.

Literature cited

Arup U, Sechting U, Frédén P. 2013. A new taxonomy of the family Teloschistaceae. Nordic

Journal of Botany 31: 16-83. https://doi.org/10.1111/j.1756-1051.2013.00062.x

Darriba D, Taboada GL, Doallo R, Posada D. 2012. jModelTest 2: more models, new heuristics

and parallel computing. Nature Methods 9(8): 772. https://doi.org/10.1038/nmeth.2109

Doring H, Clerc P, Grube M, Wedin M. 2000. Mycobiont specific PCR primers for the

amplification of nuclear ITS and LSU rDNA from lichenised ascomycetes. Lichenologist

32: 200-204. https://doi.org/10.1006/lich.1999.0250

Gardes M, Bruns TD. 1993. ITS primers with enhanced specificity for basidiomycetes

-application to theidentification ofmycorrhizaeandrusts. Molecular Ecology2:113-118.

https://doi.org/10.1111/j.1365-294X.1993.tb00005.x

Gaya E, Hégnabba F, Holguin A, Molnar K, Fernandez-Brime S, Stenroos S, Arup

U, Sochting U, Boom PV, Liicking R, Sipman HJ, Lutzoni F. 2012. Implementing a

cumulative approach for a comprehensive phylogenetic study of the Teloschistales

(Pezizomycotina, Ascomycota). Molecular Phylogenetics and Evolution 63(2): 374-387.

https://doi.org/10.1016/j.ympev.2012.01.012

Gaya E, Fernandez-Brime S, Vargas R, Lachlan RF, Gueidan C, Ramirez-Mejia M,

Lutzoni F. 2015. The adaptive radiation of lichen-forming Teloschistaceae is associated

with sunscreening pigments and a bark-to-rock substrate shift. Proceedings of the

National Academy of Sciences of the United States of America 112(37): 11600-11605.

https://doi.org/10.1073/pnas.1507072112

Joshi Y, Upreti DK. 2006. Caloplaca amarkantakana, a new species in the Caloplaca sideritis group

from India. Lichenologist 38(6): 537-540. https://doi.org/10.1017/S0024282906006360

Joshi Y, Upreti DK. 2011. Four new records of Caloplaca (lichenized Ascomycetes) from India.

Mycotaxon 116: 53-60. https://doi.org/10.5248/116.53

Joshi Y, Upreti DK, Sati SC. 2008. Three new species of Caloplaca from India. Lichenologist

40(6): 535-541. https://doi.org/10.1017/S0024282908007652

Joshi Y, Upreti DK, Sati SC. 2009. Caloplaca himalayana, a new epiphytic lichen from the Indian

subcontinent. Lichenologist 41(3): 249-255. https://doi.org/10.1017/S0024282909008214

Joshi Y, Ram TAMJ, Sinha GPPS. 2014. Caloplaca indica, a new lichenized Ascomycetes

(Teloschistaceae) from Eastern Himalaya, India. National Academy Science Letters 37(6):

517-519. https://doi.org/10.1007/s40009-014-0270-3

Katoh K, Standley DM. 2013. MAFFT multiple sequence alignment software version

7: improvements in performance and usability. Mol. Biol. Evol 30(4): 772-780.

https://doi.org/10.1093/molbev/mst010

Kondratyuk SY, Karnefelt I, Elix JA, Thell A. 2007. New species of the genus Caloplaca in

Australia. Bibliotheca Lichenologica 95: 341-386.

674 ... Wen, Shahidin, Abbas

Kondratyuk SY, Jeong MH, Yu NH, Karnefelt I, Thell A., Elix JA, Kim J, Kondratyuk AS,

Hur JS. 2013. Four new genera of teloschistoid lichens (Teloschistaceae, Ascomycota)

based on molecular phylogeny. Acta Botanica Hungarica 55(3-4): 251-274.

https://doi.org/10.1556/ABot.55.2013.3-4.8

Kondratyuk SY, Karnefelt I, Thell A, Elix JA, Kim J, Kondratiuk AS, Hur JS. 2015.

Brownlielloideae, a new subfamily in the Teloschistaceae (Lecanoromycetes, Ascomycota).

Acta Botanica Hungarica 57(3-4): 321-343. https://doi.org/10.1556/034.57.2015.3-4.6

Kondratyuk SY, Lékés L, Upreti DK, Nayaka S, Mishra GK, Ravera S, Jeong MH, Jang SH,

Park JS. Hur JS. 2017. New monophyletic branches of the Teloschistaceae (lichen-forming

Ascomycota) proved by three gene phylogeny. Acta Botanica Hungarica 59(1-2): 71-136.

https://doi.org/10.1556/034.59.2017.1-2.6

Kondratyuk SY, Persson PE, Hansson M, Mishra GK, Nayaka S, Liu D, Hur JS, Thell A. 2018.

Upretia, a new caloplacoid lichen genus (Teloschistaceae, lichen-forming Ascomycota)

from India. Cryptogam Biodiversity and Assessment, Special Volume: 22-31.

https://doi.org/10.21756/cab.esp5

Miller MA, Pfeiffer W, Schwartz T. 2010. Creating the CIPRES Science Gateway

for inference of large phylogenetic trees. 1-8, in: Proceedings of the Gateway

Computing Environments Workshop (GCE), 14 Nov. 2010, New Orleans, LA, USA.

https://doi.org/10.1109/GCE.2010.5676129

Obermayer W. 2004. Additions to the lichen flora of the Tibetan region. Bibliotheca

Lichenologica 88: 479-526.

Olley L, Sharma LR. 2013. A provisional checklist of the lichens of Nepal. Journal of the

Department of Plant Resources Nepal 35: 18-21.

Orange A, James PW, White FJ. 2010. Microchemical methods for the identification of lichens.

2nd edn. London, British Lichen Society. 101 p.

Poelt J, Hinteregger E. 1993. Beitrage zur Kenntnis der Flechtenflora des Himalaya. VII. Die

Gattungen Caloplaca, Fulgensia und Ioplaca. Berlin, Stuttgart. 247 p.

Poelt J, Petutschnig W. 1992. Beitrage zur Kenntnis der Flechtenflora des Himalaya. IV. Die

Gattungen Xanthoria und Teloschistes zugleich Versucheiner Revision der Xanthoria

candelaria-Gruppe. Nova Hedwigia 54: 1-36.

Ronquist F, Teslenko M, Mark P, Ayres DL, Darling A, Hohna S, Larget B, Liu L, Suchard

MA, Huelsenbeck JP. 2012. MrBayes 3.2: efficient Bayesian phylogenetic inference and

model choice across a large model space. Systematic Biology 61: 539-542.

https://doi.org/10.1093/sysbio/sys029

Stamatakis A. 2014. RAxML version 8: a tool for phylogenetic analysis and post-analysis of

large phylogenies. Bioinformatics 30(9): 1312-1313.

https://doi.org/10.1093/bioinformatics/btu033

Vilgalys R, Hester M. 1990. Rapid genetic identification and mapping of enzymatically

amplified ribosomal DNA from several Cryptococcus species. Journal of Bacteriology

1990(8): 4238-4246. https://doi.org/10.1128/jb.172.8.4238-4246.1990

Vondrak J, Frolov I, Arup U, Khodosovtsev A. 2013. Methods for phenotypic evaluation of

crustose lichens with emphasis on Teloschistaceae. Chernomorski Botanical Journal 9(3):

382-405. https://doi.org/10.14255/2308-9628/13.93/6

Vondrak J, Shahidin H, Moniri MH, Hali G, KoSnar J. 2018. Taxonomic and functional diversity

in Calogaya (lichenised Ascomycota) in dry continental Asia. Mycological Progress 17(8):

897-916. https://doi.org/10.1007/s11557-018-1402-9

Filsoniana lhasanensis sp. nov. (China) ... 675

Wei JC. 1991. An enumeration of lichens in China. Beijing, International Academic Publishers.

278 p.

Wei JC. 2017. An enumeration of lichenized and lichenicolous fungi in China. Beijing, China

Forestry Publishing House. 596 p.

Wei JC, Jiang YM. 1986. Lichens of Xizang. Beijing, Science Press. 130 p.

White TJ, Bruns T, Lee S, Taylor J. 1990. Amplification and direct sequencing of fungal

ribosomal RNA genes for phylogenetics. 315-322, in: MA Innis & al. (eds). PCR

protocols: a guide to methods and applications. New York, NY: Academic Press.

https://doi.org/10.1016/B978-0-12-372180-8.50042-1

Zhang BP, Chen XD, Li BL, Yao YH. 2002. Biodiversity and conservation in the Tibetan Plateau.

Journal of Geographical Sciences 12(2): 135-143.

Zhou S, Stanosz GR. 2001. Primers for amplification of mtSSU rDNA, and a phylogenetic

study of Botryosphaeria and associated anamorphic fungi. Mycological Research 105(9):

1033-1044. https://doi.org/10.1016/S0953-7562(08)61965-6

Zoller S, Scheidegger C, Sperisen C. 1999. PCR primers for the amplification of mitochondrial

small subunit ribosomal DNA of lichen-forming ascomycetes. Lichenologist 31(5):

511-516. https://doi.org/10.1006/lich.1999.0220

MY COTAXON

October-December 2019— Volume 134, pp. 677-679

https://doi.org/10.5248/134.677

Sarcopodium flocculentum,

the correct name for S. macalpinei

SHAUN R. PENNYCOOK" & PAUL M. KIRK?

'Manaaki Whenua - Landcare Research

Private Bag 92170, Auckland 1072, New Zealand

? Royal Botanic Gardens

Kew, Surrey, TW9 3DS, UK

* CORRESPONDENCE TO: PennycookS@LandcareResearch.co.nz

ABSTRACT— The nomenclatural background of the species treated as Sarcopodium macalpinei

is briefly outlined, and the requirement to supersede that name with a combination based on

Nectriella flocculenta [= Lanatonectria flocculenta] is explained.

Key worps—Actinostilbe flocculenta, Actinostilbe macalpinei, Kutilakesopsis macalpinei,

ICN, code-compliance

In a morphological study, Sutton (1981) synonymised Actinostilbe Petch

and Kutilakesopsis Agnihothr. & G.C.S. Barua with the earlier genus Sarco-

podium Ehrenb. and published a new combination Sarcopodium macalpinei

for the type of Kutilakesopsis.

Rossman & al. (1999) proposed Lanatonectria Samuels & Rossman,

typified by a sexual morph for which they published the new combination

L. flocculenta, additionally proposing a new combination Actinostilbe

macalpinei for the conspecific asexual morph.

In a multi-gene analysis, Lombard & al. (2015) included sequences from

the type species of Sarcopodium (S. circinatum), Actinostilbe (A. vanillae =

S. vanillae), and Kutilakesopsis (K. macalpinei = S. macalpinei), demonstrating

that they were all included within a monophyletic Sarcopodium clade,

thereby confirming the synonymy of these three genera. They also included

678 ... Pennycook & Kirk

Lanatonectria as a synonym based on the established synonymy of its

type (L. flocculenta) with Sarcopodium macalpinei; consequently, they

recombined four of the five Lanatonectria species in Sarcopodium but did

not recombine L. flocculenta (presumably because of its existing heterotypic

Sarcopodium synonym).

Rossman & al. (2016) recommended Sarcopodium as the accepted

generic name in preference to Lanatonectria and Actinostilbe, a decision

based on priority, widespread use, and the greater number of existing

names; but they continued to cite L. flocculenta as S. macalpinei.

However, Sarcopodium macalpinei is not the correct name for this

species, since it fails to use the epithet of the earliest available legitimate

name, as required by ICN (Shenzhen) Art. 11.4 (Turland & al. 2018). The

correct name has been proposed by Pennycook & Kirk (2019):

Sarcopodium flocculentum (Henn. & E. Nyman) Pennycook & P.M. Kirk,

Index Fungorum no. 418: 1. 2019

= Nectriella flocculenta Henn. & E. Nyman, Monsunia 1: 160. 1899 [“1900”]

= Nectria flocculenta (Henn. & E. Nyman) Hohn., Sitzungsber. Kaiserl.

Akad. Wiss., Wien, Math.-Naturwiss. Cl., Abt. 1, 121: 360. 1912

= Lanatonectria flocculenta (Henn. & E. Nyman) Samuels

& Rossman, Stud. Mycol. 42: 138. 1999

= Actinostilbe flocculenta (Henn. & E. Nyman) Rossman,

Samuels & Seifert, IMA Fungus 4(1): 46. 2013

= Kutilakesopsis macalpinei Agnihothr. & G.C.S. Barua, J.

Indian Bot. Soc. 36: 309. 1957 [as ‘macalpineae’ |

= Sarcopodium macalpinei (Agnihothr. & G.C.S. Barua) B.

Sutton, Trans. Brit. Mycol. Soc. 76: 99. 1981

= Actinostilbe macalpinei (Agnihothr. & G.C.S. Barua) Seifert

& Samuels, Stud. Mycol. 42: 138. 1999

Acknowledgments

We wish to thank Tom May (Royal Botanic Gardens, Melbourne) and Scott

Redhead (Agriculture and Agri-Food Canada, Ottawa) for refereeing the manuscript.

Literature cited

Lombard L, van der Merwe NA, Groenewald JZ, Crous PW. 2015. Generic concepts in

Nectriaceae. Studies in Mycology 80: 189-245. https://doi.org/10.1016/j.simyco.2014.12.002

Pennycook SR, Kirk PM. 2019. Nomenclatural novelties: S$. Pennycook & P.M. Kirk. Index

Fungorum no. 418. 1 p.

http://www.indexfungorum.org/Publications/Index%20Fungorum%20n0.418.pdf

Rossman AY, Samuels GJ, Rogerson CT, Lowen R. 1999. Genera of Bionectriaceae, Hypocreaceae

and Nectriaceae (Hypocreales, Ascomycetes). Studies in Mycology 42. 248 p.

Sarcopodium flocculentum (= S. macalpinei) explained ... 679

Rossman AY, Allen WC, Braun U, Castlebury LA, Chaverri P, Crous PW, Hawksworth

DL & al. 2016. Overlooked competing asexual and sexually typified generic names of

Ascomycota with recommendations for their use or protection. IMA Fungus 7: 289-308.

https://doi.org/10.5598/imafungus.2016.07.02.09

Sutton BC. 1981. Sarcopodium and its synonyms. Transactions of the British Mycological

Society 76: 97-102. https://doi.org/10.1016/s0007-1536(81)80012-5

Turland NJ, Wiersema JH, Barrie FR, Greuter W, Hawksworth DL, Herendeen PS & al.

2018. International Code of Nomenclature for algae, fungi, and plants (Shenzhen Code)

adopted by the Nineteenth International Botanical Congress, Shenzhen, China, July 2017.

Regnum Vegetabile 159. https://doi.org/10.12705/Code.2018

MYCOTAXON

October-December 2019—Volume 134, pp. 681-705

https://doi.org/10.5248/134.681

Pteridicolous ascomycetes from a cloud forest

in eastern Mexico

ROSARIO MEDEL-ORTIZ?, YAJAIRA BAEZA?,

& FRANCISCO G. LOREA-HERNANDEZ?

‘Centro de Investigacion en Micologia Aplicada, Universidad Veracruzana,

Médicos No. 5, Col. Unidad del Bosque, 91010, Xalapa, Veracruz, México.

*Facultad de Agronomia, Universidad Veracruzana,

Zona Universitaria, 91000, Xalapa, Veracruz, México

*Instituto de Ecologia, A.C. Red de Sistemdtica y Biodiversidad,

Carretera antigua a Coatepec 351, 91070, Xalapa, Veracruz, México.

“CORRESPONDENCE TO: romedel@uv.mx

ABSTRACT— Twenty-one ascomycetous taxa collected on ferns in a cloud forest of Veracruz,

Mexico are reported and described. Fourteen represent new records for the Mexican

mycobiota: Bisporella pteridicola, Crocicreas quinqueseptatum, Dasyscyphella dryina, Durella

macrospora, Hamatocanthoscypha helicotricha, Hyaloscypha fuckelii, Lachnum brevipilosum,

L. nudipes, Mollisia ventosa, Orbilia auricolor, Protocreopsis pertusa, Seimatosporium tostum,

Stictis carnea, and S. radiata. Two collections—Cyathicula and cf. Arachnopeziza—that could

not be fully identified may represent new species. The remaining five taxa have previous

Mexican records but are reported here from new fern hosts. This study increases to 25 the

number of ascomycete species known to occur on ferns in the cloud forests of Mexico.

Key worps—Cyatheaceae, Helotiales, Hypocreales, Ostropales, Xylariales

Introduction

The Ascomycota, with around 90,000 species (Cannon & al. 2018), is the

most diverse group in kingdom Fungi. Its diversity significantly complicates

gaining knowledge of individual species, especially those with minute

fruitbodies restricted to specialized substrates. Such is the case of fungi

growing on ferns. Most fungi reported on ferns are saprobes, a group that

682 ... Medel-Ortiz, Baeza, Lorea-Hernandez

receives less attention than parasites (Mehltreter et al. 2010). Currently

69 ascomycete species have been recorded in cloud forest of Veracruz

(Medel 2013), small in comparison to the over 500 fern species known

to inhabit cloud forests (Tejero-Diez & al. 2011). Although the first fern-

related fungal study was devoted to rust fungi (Stevenson 1945), subsequent

studies have been more focused on ascomycetes and their anamorphs (e.g.,

Mycosphaerellaceae and Periconiella; Braun 2004, Kirschner & Liu 2014).

Other studies have centered on endophytic ascomycetes and fern-associated

saprobes (Cannon 1997, Castaheda-Ruiz & Heredia 2000, Del Olmo &

Arnolds 2014, Dingley 1972, Medel & Lorea-Hernandez 2008, Samuels &

Rogerson 1990).

For Mexico there are relatively few studies on the diversity of fern-

inhabiting fungi (Castafieda-Ruiz & Heredia 2000, Haines 1980, Medel &

Lorea-Hernandez 2008, Medel & al. 2010, Samuels & Rogerson 1990). So

far, the seven ascomycete species recorded growing on ferns represent only

three genera: Crocicreas Fr. (1), Dimeriella Speg. (1), and Lachnum Retz. (5).

The few Ascomycota recorded from these forests contrasting with the great

diversity of fern species suggests a high diversity yet to be explored. The

present study was conducted in order to provide more information about

the diversity of fern-inhabiting fungi in Mexican cloud forests.

Materials & methods

Sampling sites in central Veracruz state (eastern Mexico) were selected based

on the high fern diversity within an area. Three sampling sites were chosen: El

Riachuelo, Los Encinales, and Rio Xocoyolapan. Field work was conducted during

August-November 2011. Fungi were collected by opportunistic sampling of

different fern species, with fronds and rachises presenting ascomata or stromata

selected and placed in paper bags (Mueller & al. 2004).

Fruiting bodies were identified and processed following standard mycological

techniques for Ascomycota (Dennis 1978). Specimens were hand _ sectioned,

pretreated with 3% aqueous KOH, and stained with phloxine and Melzer’s reagent;

ascal tip bluing reactions are designated by J- (negative, no blue present) and J+

(positive, blue present). Tissues were examined microscopically using a Zeiss Primo

Star microscope. Averages, ranges, and Q-calculations (Largent & al. 1977) are

based on 30 measurements. Colours are coded according to Kornerup & Wanscher

(1978). Ascomata were photographed digitally with a Sony Cyber-Shot camera

mounted on a Zeiss Stemi DV4 stereomicroscope. The specimens were identified

using specialized literature (Breitenbach & Kranzlin 1984, Carpenter 1981, Dennis

1949, 1956, 1960, 1978). Fungal nomenclature follows Index Fungorum (2019),

and host plants were identified following Mickel & Smith (2004). All specimens are

Pteridicolous ascomycetes (Mexico) ... 683

deposited in the mycological collection of the Herbarium, Instituto de Ecologia A.C.

(XAL). Specimens discovered in XAL and also examined are noted as “additional

specimens examined” in species descriptions where pertinent.

Taxonomy

cf. Arachnopeziza FIGS 1-3

APOTHECIA sessile, cup-shaped, reddish-golden (6C7) to Pompeian

yellow (5C6), <1 mm diam., margin hairy (sparsely pilose), hairs white

(1A1), readily dropping from the margin to give a smooth appearance.

SUBICULUM white to pale yellowish white (1A1, 2A1), not scattered on the

substrate, reticulate or net-like, with hyphae 2-3 um diam., septate, hyaline,

thick-walled (<1 um). Ascr cylindrical, 70-90(-100) x 7-9(-10) um, thin-

walled, hyaline, 1-2-seriate, pore J+. Ascospores cylindrical-ellipsoid,

(13-)14-15 x 5 um (Q = 2.8), smooth, hyaline, 0-1-septate, biguttulate,

with sheath and hyaline appendages observed in Cresyl blue. PARAPHYSES

filiform, 1-2 um diam., hyaline, septate toward the base. EcraL ExcrpuLUM

a textura globulosa to textura angularis, cells 12—25(—40) um diam.

HaBItTat—Gregarious, on rachises of Alsophila firma. Known only from

these collections.

SPECIMENS EXAMINED—MEXICO. VERACRUz: San Andrés Tlalnelhuayocan County,

Rio Xocoyolapan, 19°30.89’N 97°00.39’W, 1590 m, 13 October 2011, Cordova 78.

ADDITIONAL SPECIMENS EXAMINED: MEXICO. Veracruz: San Andrés

Tlalnelhuayocan County, Rancho Agitita Fria, 6 April 2007, Medel 1362; 1363-A.

Arachnopeziza Fuckel is characterized by small apothecia growing on a

densely reticulate, net-like subiculum covered by hairs, and ellipsoid-fusoid

to filiform, 1-7-septate ascospores (Korf 1951). The genus has a worldwide

temperate distribution but is uncommon in the tropics (Gamundi &

Giaiotti 1994). Although the studied specimens suggest an affinity with

Arachnopezizaceae, they do not match any known genus or species covered

in the references consulted (Dennis 1949, Huhtinen 1987, Korf 1951). The

0-1-septate spores covered by a hyaline sheath with a small appendage

resemble those of Arachnopeziza aurelia (Pers.) Fuckel, which differs in its

long hairs, slightly larger spores (17-22 x 3.5-5.5 um, O0-3-septate; Korf

1951). The presence on the apothecial margin of very thick, scarce, and

septate hairs that drop off, leaving a smooth margin (as in our specimen;

Fic. 1) is a feature unknown in Arachnopeziza. The subiculum is restricted

to the apothecial base and not distributed throughout the substrate, unusual

in Arachnopeziza (Seaver 1938), but not uncommon in Parachnopeziza Kort.

684 ... Medel-Ortiz, Baeza, Lorea-Hernandez

In Parachnopeziza, however, apothecia grow directly from a small stipe

immersed in the substrate (Korf 1978) growing directly from a subiculum, as

occurs in Arachnopeziza, and certainly in our specimen.

Bisporella pteridicola F. Ren & W.Y. Zhuang,

Mycosystema 36 (4): 408 (2017). FIGS 4, 5

APOTHECIA superficial, cup-shaped to discoid. Disc concave, yellow

(4A6), pastel yellow (2A4), or chicken yellow (2A6), translucent, waxy in

appearance, 0.2-0.4 mm diam. Stipe short or absent, concolorous with the

disc. Asci claviform (32—)35-40(-45) x 5-6 um, 8-spored, biseriate, apical

pore J-, arising from croziers. AScosPoREs fusiform (5 —)8-9(-10) x 2-3

um (Q = 4.3), hyaline, with two oil drops and one septum visible in KOH.

PARAPHYSES filiform, slightly enlarged at the apex, 1.5-2 um diam. ECTAL

EXCIPULUM a textura angularis, gelatinized, 15-40 um thick, cells 3-6 um

diam., protruding from outer surface <3 um. MEDULLARY EXCIPULUM a

textura intricata, thin, hyaline, not gelatinized, cells 2-3 um diam.

HasitatT—On rachises of unidentified fern as well as on fern rachises

and leaf blades of Alsophila firma, Cyathea bicrenata, Sphaeropteris horrida,

Pecluma alfredii, and Thelypteris sp.

SPECIMENS EXAMINED—MEXICO. VeRAcRUz: San Andrés Tlalnelhuayocan

County, Rio Xocoyolapan, 19°30.89’N 97°00.39’W, 1590 m, 13 October 2011,

Cordova 80c; 4 November 2011, Castillo del Moral 63, 66; 8 December 2011, Medel

& Lorea 2203. El Riachuelo; 19°30.99’N 97°00.39’W, 1626 m, 18 August 2011, Baeza

47a, Cordova 67, 71; 11 November 2011, Medel 2192. Los Encinales, 19°31.10’N

97°00.32’W, 1630 m, 10 August 2011, Baeza 36; 18 August 2011, Baeza 49,55, 57;

Medel & Lorea 2158, Cérdova 73, 74.

Bisporella Sacc. is a genus easily recognized by its small sessile bright yellow

apothecia growing on woody substrata (Carpenter & Dumont 1978a,

Dumont 1981). Bisporella pteridicola was described growing on ferns in Asia

(Zhuang & al. 2017). Ours is the first report of the species from Mexico.

Our collections agree with the description in Zhuang & al. (2017) in color,

size, and shape of the apothecia, ascospore measurements, and ecology

(growth on ferns).

Coenogonium botryosum C. Knight,

Syn. Queensl. Fl. 1(Suppl.): 74 (1886). Fics 6, 7

APOTHECIA superficial, discoid, sessile, flattened, 0.5-1 mm diam., pale

yellow (3A3), waxy in appearance, margin differentiated from the disc,

smooth, yellowish white (3A2), paler than the disc. THaLtus filamentous

or felt-like and pilose, with short, upright algal threads resembling hairs,

Pteridicolous ascomycetes (Mexico) ... 685

greyish yellow to linden green (2B5-2C5). Hymenium J+. Harrs small

and thin, linden green (2C5) to pale yellow (1A3-2A3), filiform, septate.

Ascli cylindric-clavate (48—)50-55(-60) x 4-5 um, thin walled, uniseriate.

Ascosporss ellipsoid (7-)8-9(-12) x 2 um (Q = 4.2), hyaline, smooth,

0-1-septate, with 4 small guttules. ParApHysEs cylindrical, apices 4-5 um

diam., bifurcate, hyaline, septate. ECTAL EXCIPULUM a textura globulosa,

with hyaline cells.

Hasitat—Gregarious, on fronds of Adiantum andicola and

Grammiitis sp.

SPECIMENS EXAMINED—MEXICO. VERACRUz: San Andrés Tlalnelhuayocan County,

El Riachuelo, 19°30.99’N 97°00.39’W, 1626 m, 8 December 2011, Baeza 70, Medel &

Lorea 2211, 2215.

Coenogonium Ehrenb. is a tropical and subtropical ascolichen genus found

on all continents (GBIF 2019, Rivas-Plata & al. 2006, Uyenco 1963). The

filamentous green-pale yellowish thallus attached to the substrate by small

filaments as well as the threadlike algae, absence of septa, uniseriate asci, and

paraphyses with capitate subglobose apices are characteristic of C. botryosum

(Rivas-Plata & al. 2006). Two similar species are C. moniliforme Tuck., which

has moniliform algae without septa, and C. implexum Nyl., which has bright

yellow thalli and a cottony texture.

Known from Neotropics and eastern Paleotropics (GBIF 2019, Rivas-Plata

& al, 2006, Uyenco 1963), Coenogonium botryosum was reported previously

for Veracruz, Mexico, growing on Ficus (Uyenco 1963); this is the first report

of the species on ferns.

Crocicreas quinqueseptatum S.E. Carp.,

Mem. New York Bot. Gard. 33: 160 (1981). Fics 11, 12

APOTHECIA cup-shaped, 0.5-0.8 mm diam. Disc white to yellowish

white (2A2, 2A1). MARGIN denticulate, inrolled over the hymenium when

dry, almost hyaline or translucent with long teeth; teeth fragile, formed by

groups of long, septate, hyaline hyphae, 100-115(-400 um) x 40-48 um,

lighter in colour than the rest of the apothecium, white to yellowish white

(2A1-2A2), 8-12 teeth per apothecium; breaking easily leaving the apothecia

non-denticulate. Stipe cylindrical, thin, concolorous with apothecia.

Ascli cylindrical, tapering toward the base, (60-—)80-105 x 6-8 um, thin-

walled, apical pore J+, biseriate, base without croziers. AscosporEs fusoid

to fusiform with subacute to acute ends, smooth, (20-)26-30 x 3-4 um

(Q = 5.5), 5-septate (rarely 6-7), hyaline. PARAPHYSES filiform, unbranched,

686 ... Medel-Ortiz, Baeza, Lorea-Hernandez

1-2 um diam., hyaline. EcraL ExcipuLum single layered, gelatinous,

composed of long cells in the inner part, extending outward from the disc to

form the teeth. MEDULLARY EXCIPULUM not differentiated.

Hasitat—On fallen stems and rachises of Alsophila firma and Cyathea

bicrenata.

SPECIMEN EXAMINED— MEXICO. VERACRUZ: San Andrés Tlalnelhuayocan County,

Los Encinales, 19°31.10’N 97°00.32’W, 1630 m, 10 August 2011, Medel & Lorea 2168.

Crocicreas quinqueseptatum was described by Carpenter (1981) as having

ascospores with a variable number (4-7) of septa and which are slightly

longer and narrower [(28—)30-33(-35) x (2-)2.5-3 um] than those found

in our specimen. Other features (outer apothecial surface, morphology,

and measurements) are close to C. helios var. parahelios S. E. Carp., which

differs by 0-3-septate ascospores, an absent or poorly developed medullary

excipulum, and broader apothecia (<1.5 mm; Carpenter 1981). Crocicreas

quinqueseptatum has been reported growing on angiosperm stems, especially

fallen herbaceous stems of Asteraceae in South America. This is the first

report of this species on ferns.

Cyathicula sp. Fics 8-10

APOTHECIA cup-shaped, sessile, disc flattened or concave, <1 mm diam.,

pastel yellow (3A4); margin dentate with 6-10 hyaline teeth per ascoma, teeth

composed of groups of long, hyaline, aseptate hyphae that extend beyond the

margin, each tooth separated by c. 420-480 um. Ascr cylindric-clavate, (96-)

110-140 x (7-9)10-11 um, 8-spored, biseriate, apical pore J-. ASCOSPORES

ellipsoidal-fusiform, 13-16 x 5-6 um (Q = 2.6), smooth, hyaline, 1-3-septate,

guttulate. PARAPHYSES filiform, apices simple or branched, 2-3 um diam. at

apex, septate; embedded with asci in a gelatinous matrix and separated only

with difficulty. ECrAL EXCIPULUM a textura prismatica to textura porrecta,

hyphae single-layered, long-celled, subparallel, close together, thick-walled,

hyaline, smooth, lacking crystals. MEDULLAR EXCIPULUM little differentiated,

hyphae 3-5 um diam., not easily observed due to gelatinous consistency.

Hasitat—On rachises of Cyathea bicrenata. Known only from our

collections.

SPECIMENS EXAMINED—MEXICO. VERACRUz: San Andrés Tlalnelhuayocan County;

E] Riachuelo, 19°30.99’N 97°00.39’W, 1626 m, 4 November 2011, Baeza 56; Los

Encinales, 19°31.10’N 97°00.32’W, 1630 m, 13 October 2011, Baeza 56a; 4 November

2011, Baeza 61, Castillo del Moral 64.

We place our material into Cyathicula based on its cupulate, sessile

apothecium with a toothed margin, whitish yellowish to pale brown color,

Pteridicolous ascomycetes (Mexico) ... 687

Lair , $10 pm [ 1mm

Figs 1-15. cf. Arachnopeziza. 1. Apothecia showing scarce subiculum and hairs (top right);

2. Ascospores in Congo red; 3. Excipulum showing hyphae from the subiculum. Bisporella

pteridicola. 4. Apothecia; 5. Asci with ascospores. Coenogonium botryosum. 6. Apothecia;

7. Ascospores. Cyathicula sp.; 8. Apothecia; 9. Cross section of excipulum, 10. One-septate

ascospores. Crocicreas quinqueseptatum. 11. Apothecia showing dentate margin; 12. Ascospores.

Dasyscyphella dryina. 13. Apothecium; 14. Ascospores. Durella macrospora. 15. Apothecia.

688 ... Medel-Ortiz, Baeza, Lorea-Hernandez

septate ascospores, J- asci, and the presence of a gelatinous matrix (Carpenter

& Dumont 1978b). Nonetheless, we must also note that the specimens fit

diagnostic characters of Crocicreas (Carpenter 1981). The controversy over

whether Crocicreas and Cyathicula are congeneric has lasted for decades

(Iturriaga & al. 1999, Zheng & Zhuang 2016). According to one classification

of Helotiales (Baral & al. 2015), Crocicreas is restricted only to the type species

C. gramineum (Fr.) Fr., and Cyathicula is unrelated. Although Wijayawardene

& al. (2018) support this distinction, we agree with the conclusion of Zheng

& Zhuang (2016) that existing data are insufficient for determining the

relationship between Crocicreas and Cyathicula. Molecular analysis is still

needed for us to assign our material definitively to a genus.

Dasyscyphella dryina (P. Karst.) Raitv.,

Akad. Nauk Estonskoi S.S.R., Inst. Zool. Bot., Tartu: 72 (1970). Fics 13, 14

APOTHECIA superficial, cup-shaped, stipitate, covered with white (1A1)

hairs. Disc 0.8-1 mm diam., yellowish white (2A2) to pale yellow (2A3),

smooth. STIPE cylindrical, covered with white (1A1) hairs with small hyaline

crystals at the tips. Harrs cylindrical, septate, slightly granular, hyaline,

easily visible, giving a distinctive villous appearance to the apothecium. AscI

cylindrical, 45-50 x 3-4 um, hyaline, 8-spored, irregularly biseriate, thin

walled, apical pore J+. Ascospores cylindric-ellipsoid to fusoid, (6-—)7-10

x 2-3 um, (Q = 4.2), smooth, hyaline, with rounded ends. PARAPHYSES

filiform, slightly lanceolate, 1 um diam., hyaline, projecting beyond the asci

by 10-20 um.

Hasitat—On rachises of Alsophila firma.

SPECIMENS EXAMINED—MEXICO. VeRAcRuz: San Andrés Tlalnelhuayocan

County, Rio Xocoyolapan, 19°30.89’N 97°00.39’W, 1590 m, 19 August 2011, Medel

2180, Medel 2181.

The studied specimen matches well with the concept of Lachnum

distinguendum sensu Dennis (1949, 1956, 1960), which is a synonym of

Dasyscyphella dryina. Dennis (1956) cited spores 2 um longer than those of

our collections, and filiform paraphyses which agreed with our specimen.

Rick (1906) described L. distinguendum Rick with broader apothecia

(<2 mm diam.), larger asci (50-69 x 7-10 um), larger ascospores (10-15 x

3-4 um), and lanceolate paraphyses (3-4 um diam.). Raitviir (1970) cited

slightly longer ascospores (9-12 x 2-3.2 um) and cylindrical paraphyses with

pointed tips. Nonetheless, our collection fits better Dennis's concept, because

our material has paraphyses that exceed the asci, while Raitviir (2002)

described paraphyses not exceeding the asci.

Pteridicolous ascomycetes (Mexico) ... 689

Dasyscyphella dryina is known from Europe and South America on Betula,

Fagus, and Populus (Dennis 1949, 1956. 1960, Raitviir 1970, Rick 1906).

Durella macrospora Fuckel,

Jahrb. Nassauischen Vereins Naturk. 23-24: 281 (1870). Fics 15-17

APOTHECIA superficial, cup-shaped, stipe short with a wide base. Disc

flat, 0.2-0.5 mm diam., wrinkled, brownish gray (10F2) with dark brown

(7F8) tones, hard consistency. Asci cylindrical (90-)100-130 x 11-12 um,

hyaline, 8-spored, irregularly biseriate, apical pore J-. Ascosporss ellipsoid-

fusiform, (18—)19-22(-23) x 4-5 um (Q = 4.2), smooth, hyaline, 3-septate,

constrained at the septum, ends rounded, with 4 small guttules. PARAPHYSES

cylindrical, slightly wider at the apex (c. 3 um) brownish to reddish brown in

KOH. EcTAL EXCIPULUM Of parallel brown hyphae.

Hasitat—On fronds of Sticherus palmatus.

SPECIMEN EXAMINED—MEXICO. VERACRUz: San Andrés Tlalnelhuayocan County,

El Riachuelo, 19°30.99’N 97°00.39’W, 1626 m, 8 December 2011, Medel 2210.

Spores of Durella macrospora have been cited as 15-21 x 3-5.5 um

(Breitenbach & Kranzlin 1984, Medardi 2004). Although the spores in

our collection are 2 um longer, other morphological characters (black

apothecia, hard consistency, ectal excipulum of parallel brown hyphae)

agree well with the published descriptions (Breitenbach & Kranzlin 1984,

Medardi 2004).

Durella macrospora has been previously recorded growing on wood of

deciduous trees such as Fagus, Malus domestica, and Quercus sp. (Breitenbach

& Kranzlin 1981, Medardi 2004), but not on ferns. Previously known only

from Europe (Breitenbach & Kranzlin 1984, Dennis 1956, Medardi 2004).

Hamatocanthoscypha helicotricha Huhtinen,

Karstenia 29(2): 182 (1990 [“1989”]). Fries 18-20

APOTHECIA cup-shaped, disc concave, pale yellow (3.A3) to pastel yellow

(3A4) shades, 0.5 mm diam., covered with fine, white (1A1) to yellowish

white (2A1-2A2) short hairs. Stipe cylindrical, short, slightly robust,

concolorous with apothecia. Hairs cylindrical, apically coiled to uncinate,

12-32 x 2-4 um, the apex tapering to <1 um diam., hyaline, septate

toward to the base, smooth. Ascr cylindric-clavate, apex conical, base

attenuate, (42—)45-72 x 6-7 um, hyaline, pore J-, 8-spored, biseriate, base

with croziers. Ascospores ellipsoid-fusoid, hyaline, smooth, 7-9(-10)

x 2-3 um (Q = 2.6), 1-septate, biguttulate. PARAPHysEs filiform, 2 um

diam., narrowing toward the base.

690 ... Medel-Ortiz, Baeza, Lorea-Hernandez

Hasitat—On rachises of Cyathea bicrenata and fronds of Sticherus

palmatus.

SPECIMENS EXAMINED—MEXICO. VeERAcRUZz: San Andrés Tlalnelhuayocan

County, El Riachuelo, 19°30.99’N 97°00.39’W, 1626 m, 8 December 2011, Medel &

Lorea 2213. Los Encinales, 19°31.10’N 97°00.32’W, 1630 m, 10 August 2011, Baeza

41,

Huhtinen (1990) and Raitviir (2004) report somewhat larger (9.8-12.8 x

2.8-4 um) spores than found in our specimens, but the presence of strongly

coiled hairs at the apex distinguishes this species. Hamatocanthoscypha

helicotricha has been recorded on fern rachises from Colombia and Cuba

(Huhtinen 1990, Raitviir 2004); this is the first report of the species for

Mexico.

Hyaloscypha fuckelii Nannf.,

Nova Acta Regiae Soc. Sci. Upsal., Ser. 4, 8(2): 273 (1932). Fics 21, 22

APOTHECIUM superficial, sessile, cup-shaped. Disc concave, 0.1-0.3 mm

diam., white (1A1), translucent when fresh or when rehydrated in KOH.

Receptacle with fine, whitish hairs. Harrs filiform, cylindrical, curved in

the apex, thin-walled, slightly granular. Ascr clavate, 50-66(-67) x 4-5

um, hyaline, thin-walled, 8-spored, irregularly biseriate, apical pore J+,

base with croziers. AscosporsEs ellipsoid, (5—)6-8(-10) x 2 um, (Q = 3.2),

0-1-septate, smooth, hyaline, with small oil drops. Parapuyses filiform,

protruding <9 um beyond asci, narrow, smooth, hyaline, apical cell with

small guttules.

Hasirat—On rachises of Alsophila firma.

SPECIMEN EXAMINED—MEXICO. VerRAcRUz: San Andrés Tlalnelhuayocan

County, El Riachuelo, 19°30.99’N 97°00.39’W, 1626 m,10 August 2011, Baeza 38.

Our collection agreed with Hyaloscypha fuckelii as described by Huhtinen

(1990). The similar H. albohyalina (P. Karst.) Boud. differs in its narrow

hairs and smaller spores.

Hyaloscypha fuckelii is widely distributed in Europe and North America

(Huhtinen 1990) and has been reported growing on monocotyledonous

genera such as Juncus, Typha, and Molinia. This is a new record for Mexico

and the first report on ferns.

Lachnum brevipilosum Baral,

Beih. Z. Mykol. 6: 74 (1985). Fics 23, 24

APOTHECIUM superficial, cup-shaped. Disc concave, white (1A1) to

pale (2A2), 0.8-1 mm diam. Receptacle covered with whitish hairs visible

Pteridicolous ascomycetes (Mexico) ... 691

to the naked eye. Stipe cylindrical, 0.6 mm long, covered by hairs generally

concolorous with disc (white, 1A1) except for some with a yellowish gray

(4B2) base. Hairs cylindrical, 130-150 x 4-5 um (the apex tapering to <2-3

um), 5-7-septate, hyaline, covered by granules. Asc clavate-cylindrical,

42-50 x 4-5 um, hyaline, pore J+, 8-spored, biseriate, croziers not observed.

Ascosporss ellipsoid, smooth, with rounded ends, 8-—10(-12) x 2 um (Q 4.2),

hyaline, 1-septate, biguttulate. PaRAPHysES filiform, septate, protruding by

5-10 um above the asci, hyaline.

Hasitat—On rachises of Alsophila firma and Cyathea bicrenata.

SPECIMENS EXAMINED—MEXICO. VERAcRuz: San Andrés Tlalnelhuayocan

County, Rio Xocoyolapan, 19°30.89’N 97°00.39’W, 1590 m, 4 November 2011,

Baeza 58. Paraje El Riachuelo, 19°30.99’N 97°00.39’W, 1626 m, 4 November 2011,

Cordova 86.

The Mexican specimens agree with the morphology of Lachnum

brevipilosum as provided by Dennis (1949, as Dasyscyphus brevipilus Le

Gal) and Ye & al. (2006), except for the length and width of the hair apices.

Lachnum brevipilosum has been reported growing on stems of Clematis,

Fagus, and Rubus from Europe and temperate China (Dennis 1949, Ye &

al. 2006) This is a new record for Mexico and represents the first report on

ferns.

Lachnum nudipes (Fuckel) Nannf.,

Svensk Bot. Tidskr. 22: 124 (1928). Fics 25, 26

APOTHECIUM superficial, cup shaped. Disc concave slightly flattened,

yellowish white (4A2) to pale yellow (4A3), 0.3-0.4 mm in diameter.

Receptacle densely covered with white (1A1) hairs. Stipe cylindrical, with

whitish hairs (1A1), more or less equal to the diameter of the disc. Harrs

cylindrical, ornamented with granules, widening at apex to 3-4 um, longer

than 80 um, hyaline in KOH. Asc clavate, (35-)38-47 x 4-5 um, 8-spored,

biseriate, pore J+, base without croziers. AscosporEs fusiform to elliptical

with rounded ends, (5—)6-8(-9) x 1-2(-3) um (Q = 3.1), smooth, hyaline.

PARAPHYSES lanceolate, septate towards the base, (50-)68-70 x 3-4 um,

clearly extending beyond the asci.

Hasitat—On fronds of Lophosoria quadripinnata.

SPECIMENS EXAMINED—MEXICO. VERACRUz: San Andrés Tlalnelhuayocan County,

Rio Xocoyolapan 19°30.89’N 97°00.39’W, 1500 m, 18 August 2011, Cérdova 72, 13

October 2011, Baeza 51; 4 November 2011, Cordova 65, 84, Castillo del Moral 65.

Our material agrees with Dennis's (1949) descriptions of Dasyscypha nudipes

(Fuckel) Sacc. var. nudipes and D. nudipes var. minor Dennis [both now treated

692 ... Medel-Ortiz, Baeza, Lorea-Hernandez

as synonyms of L. nudipes], but our ascomata and ascospore measurements

are closer to his D. nudipes var. minor.

Lachnum nudipes has been reported on Epilobium, Filipendula, and Spiraea

in Europe (Dennis 1949, as Dasyscypha). This is the first record Lachnum

nudipes for Mexico and the first report of its occurrence on ferns.

Lachnum pteridophyllum (Rodway) Spooner,

Biblioth. Mycol. 116: 470 (1987). Fics 27, 28

Macro and micromorphological characters agree with the description of

Medel & Lorea- Hernandez (2008).

Hasitat—On rachises of Cyathea divergens var. tuerckheimii, Cyathea

sp., Dryopteris sp., and Lophosoria quadripinnata.

SPECIMENS EXAMINED—MEXICO. VERACRUZ, San Andrés Tlalnelhuayocan

County, El Riachuelo, 13 October 2011, Baeza 29, 52, 53; 4 November 2011, Baeza

60, Medel 2196; 8 December 2011, Medel & Lorea 2214. Los Encinales, 19°31.10’N

97°00.32’W, 1630 m, 10 August 2011, Baeza 36. 19 August 2011, Medel 2179, 2182,

2185.

Lachnum pteridophyllum has been reported from Colombia, Dominican

Republic, Jamaica, Mexico, Panama, Peru, Puerto Rico, New Guinea, New

Zealand, Taiwan, Tasmania, and Venezuela (Haines 1980, Medel & Lorea-

Hernandez 2008). Haines (1980) cited this species on Cyathea sp. from

Oaxaca (Mexico), and Medel & Lorea-Hernandez (2008) from Veracruz on

decaying leaves of Dicksonia sellowiana, Cyathea divergens var. tuerckheimii,

and Cyathea spp. We add now two more hosts: Dryopteris sp. and Lophosoria

quadripinnata.

Lachnum singerianum (Dennis) W.Y. Zhuang & Zheng Wang,

Mycotaxon 67: 27 (1998). Fics 29, 30

Macro and micromorphological characters agree with the description of

Medel & Lorea- Hernandez (2008).

Hasitat—On rachises of Alsophila firma and Dicksonia sellowiana.

SPECIMEN EXAMINED—MEXICO. VERACRUz: San Andrés Tlalnelhuayocan County,

Rio Xocoyolapan, 19°30.89’N 97°00.39’W, 1590 m; 8 December 2011, Medel 2222.

Lachnum singerianum is known from the Andean region in South America

from Bolivia to Venezuela and from China, Jamaica, and Mexico (Haines

1980, Medel & Lorea-Hernandez 2008). Haines (1980) earlier reported

L. singerianum growing on rachises of ferns, while Medel & Lorea- Hernandez

(2008) reported the species on decaying leaves of Alsophila firma and

Dicksonia sellowiana in Veracruz.

Pteridicolous ascomycetes (Mexico) ... 693

20 um

-<¥ um

Fics 16-31. Durella macrospora. 16. Three-septate ascospores; 17. Asci with 1-septate ascospores.

Hamatocanthoscypha helicotricha. 18. Apothecia; 19. Asci and ascospores; 20. Uncinate or

coiled hairs. Hyaloscypha fuckelii. 21. Apothecia; 22. Ascospores. Lachnum brevipilosum.

23. Apothecium; 24. Septate hairs. Lachnum nudipes. 25. Apothecium (left) and ascospores (right);

26. Lanceolate paraphysis. Lachnum pteridophyllum. 27. Apothecium; 28. Ascospores. Lachnum

singerianum. 29. Apothecium; 30. Septate hair in 5% KOH. Lachnum varians. 31. Apothecia.

694 ... Medel-Ortiz, Baeza, Lorea-Hernandez

Lachnum varians (Rehm) M.P. Sharma,

Nova Hedwigia 43: 411 (1986). Fics 31, 32

Macro and micromorphological characters agree with the description of

Medel & Lorea- Hernandez (2008).

Hasitat—On rachises of Cyathea bicrenata and C. divergens var.

tuerckheimii and fronds of Adiantum andicola.

SPECIMENS EXAMINED—MEXICO. VeRAcRuz: San Andrés Tlalnelhuayocan

County, El Riachuelo, 19°30.99’N 97°00.39’W, 1626 m 18 August 2011, Baeza 47b,

Cordova 68, 69; 8 December 2011, Baeza 70.

ADDITIONAL SPECIMENS EXAMINED: MEXICO. VERACRUZ: Xalapa County,

Santuario del Bosque de Niebla, Parque Ecoldgico Francisco Javier Clavijero,

1 September 2013, Medel & Lorea 2259.

Lachnum varians is known from Australia, Hawaii, Mexico, New Guinea,

New Zealand, South America, and the Caribbean (Haines 1980, Medel

& Lorea-Hernandez 2008). The species was previously reported only on

decaying leaves of Alsophila firma (Medel & Lorea-Hernandez 2008);

the present study adds the new hosts Cyathea bicrenata, C. divergens var.

tuerckheimii, and Adiantum andicola.

Lachnum virgineum (Batsch) P. Karst.,

Bidrag Kannedom Finlands Natur Folk 19: 169 (1871). Fics 33, 34

APOTHECIA superficial, cupulate, 1-1.3 mm diam. Disc flat to concave,

white to yellowish white (1A1-1A2). Receptacle densely covered with

short, broad, white (1A1) hairs. Stipe, thin, covered with hairs concolorous

with the apothecia. Hairs cylindrical, 60 x 5-6(-8) um, granulose along

the entire length, 3-4-septate, the apex broadening <8 um diam. AscI

cylindric-clavate, 42-51x 4-5 um, 8-spored, biseriate, pore weakly blue

(J+). AScosPorgs fusiform, ends rounded, with one end sometimes wider

than the other, (6—)7-8 x 1-2 um (Q = 4.6). PaRAPHYSES lanceolate 4-5 um

diam., 2—3-septate, with small refractive guttules in the apex, extending

beyond the asci by 20 um.

Hasrirat—On fronds of Phlebodium aureum.

SPECIMENS EXAMINED—MEXICO. Veracruz: San Andrés Tlalnelhuayocan

County, El Riachuelo, 10 August 2011, Baeza 32; 8 December 2011, Medel 2208.

The Mexican specimens fit well the description of Lachnum virgineum

(Spooner 1987), especially the shape, color, and size of the apothecium

(densely covered by white hairs), spore size, and paraphyses exceeding the

asci by 15-20 um. Lachnum virgineum is similar to L. controversum (Cooke)

Rehm, which differs in obtuse or slightly swollen hair apexes and very slightly

Pteridicolous ascomycetes (Mexico) ... 695

narrower (3.5-4.5 um diam.) paraphyses. Although L. virgineum is generally

regarded as lignicolous, we found it growing on fronds of Phlebodium

aureum. Known from Australia and Europe (Breitenbach & Kranzlin 1984,

Spooner 1987).

Mollisia ventosa P. Karst.,

Bidrag Kannedom Finlands Natur Folk 19: 188 (1871). Fics 35-37

APOTHECIA superficial, sessile, discoid, flat, 0.5-1 mm diam., greenish-

yellow (1A7) with dark grey (1F1) to olive gray (1D2) margin. Receptacle

greenish gray (29F2), smooth. Ascr (72-)85-115 x 7-8(-10) um, apical

pore J+. Ascospores ellipsoid, (9-)12-15(-16) x 1.5-2 um (Q = 7.5),

1-3-septate, guttulate. PARAPHYSES filiform, slender, septate, branched or

not. ECTAL EXCIPULUM a textura angularis to textura globulosa, cells thick-

walled, brownish, 6-10(-12) x 6-10 um.

Hasrirat—On rachises of Cyathea bicrenata.

SPECIMENS EXAMINED—MEXICO. VeERaAcRuUz: San Andrés Tlalnelhuayocan

County, El Riachuelo, 19°30.99’N 97°00.39’W, 1626 m, 4 November 2011, Medel

2196; 8 December 2011, Baeza 71, Baeza 72.

Mollisia ventosa has been reported from Europe, and America (Breitenbach

& Kranzlin 1984, Dennis 1950, 1978, Medel & Chac6én 1997). Dennis (1950,

1978) described M. ventosa with spores larger (10-20 x 2-3.5 um) than

those in our Mexican collections, which instead match the measurements

cited by Breitenbach & Kranzlin (1984). They cited M. ventosa as growing on

Alnus and decorticated wood, but not on ferns. Mollisia undulatodepressula

(Feltgen) Le Gal & EF Mangenot, the only other Mollisia species known to

inhabit cloud forests in Mexico, grows on rotten wood (Medel & Chacén

1997).

Orbilia auricolor (A. Bloxam) Sacc.,

Syll. Fung. 8: 625 (1889). Figs 38, 39

APOTHECIA Superficial, scattered, sessile, cup-shaped. Discconcave,0.8-1mm

diam., yellow brownish to yellow gray (4B3-4B5) translucent when

rehydrated, margin crenulate. RECEPTACLE concolorous with disc, smooth.

Asct cylindric-clavate, 30-40(-42) x 2-4 um, hyaline, thin-walled, 8-spored,

biseriate, pore J—, base forked. Ascosporgs curved-filiform, (8—)9-10 x 1 um

(Q = 8.8), smooth, hyaline. PARAPHysEs cylindrical, apex capitate, with

incrustations, 3-4 um diam., septate, branched at base. ECTAL EXCIPULUM

textura globulosa with hyaline cells.

696 ... Medel-Ortiz, Baeza, Lorea-Hernandez

Hasitat—On rachises of Cyathea bicrenata and Marattia laxa.

SPECIMENS EXAMINED—MEXICO. VERACRUZ: San Andrés Tlalnelhuayocan

County, Rio Xocoyolapan 19°30.89’N 97°00.39’W, 1590 m, 8 December 2011, Baeza

67, Medel 2202.

The morphology of the apothecia, ascospores, asci, and paraphyses in our

Mexican specimens agree with Orbilia auricolor as described by Spooner

(1987) and Mo & al. (2005). The asexual stage of O. auricolor has been

recorded (as Arthrobotrys oligospora Fresen.) from Xochimilco, Mexico

(Chavarria & al. 2010).

Orbilia auricolor is known from Asia, Australia, Europe (Breitenbach

& Kranzlin 1984, Dennis 1981, GBIF 2019, Mo & al. 2005, Spooner 1987).

The species has previously been recorded growing on dead wood and

woody stems and other plant matter (Spooner 1987, Mo & al. 2005), but

not on ferns.

Protocreopsis pertusa (Pat.) Samuels & Rossman,

Stud. Mycol. 42: 66 (1999). FIGs 40, 41

PERITHECIA aggregated, globose, sessile, 0.8-1 mm diam., covered by

whitish (2A1) to yellowish white (3A2) hairs. Hairs cylindrical, curved

to wavy, smooth, thick-walled, hyaline, apices simple or forked, septate.

SuBICcULUM of thin whitish (2A1) hyphae, attached to the perithecia.

Asci clavate-cylindrical, 75-95(-100) x 10-12 um, thin-walled, hyaline,

ascospores irregularly biseriate, 8-spored, base with croziers. ASCOSPORES

ellipsoid, (13—)14-16 x 5 um (Q = 2.7), smooth, hyaline, 1-septate, with two

large guttules in the ends. PARAPHYSES filiform and hyaline.

Hasrrat—On rachises of Alsophila firma and Marattia laxa.

SPECIMENS EXAMINED—MEXICO. VeERAcRuz: San Andrés Tlalnelhuayocan

County, Rio Xocoyolapan, 19°30.89’N 97°00.39’W, 1590 m, 13 October 2011,

Cordova 77; 8 December 2011, Baeza 68, Medel & Lorea 2205.

ADDITIONAL SPECIMENS EXAMINED: MEXICO. VERAcRUz: San Andrés

Tlalnelhuayocan County, Rancho Agiiita Fria, 6 April 2007, Medel 1362.

Rossman & al. (1999) and Chaverri & al. (2010) described Protocreopsis

pertusa as having thin white mycelium beneath each perithecium,

thick-walled hairs densely united around the perithecial opening, and

1-3-septate spores measuring 13-17 x 4-5 um, all characters agreeing

with our Mexican specimens. This species has been reported from tropical

to subtropical regions on leaves of Musa, Heliconia, palms, bamboos, and

fern rachises. It is a new record for Mexico.

Pteridicolous ascomycetes (Mexico) ... 697

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Figs 32-48. Lachnum varians. 32. Asci with ascospores. Lachnum virgineum. 33. Apothecia;

34. Hair with enlarged apex. Mollisia ventosa. 35. Apothecia; 36. Excipulum at margin and lower

flank; 37. Asci with ascospores. Orbilia auricolor. 38. Apothecia 39. Paraphysis. Protocreopsis

pertusa. 40. Apothecia 41. Ascospores. Seimatosporium tostum. 42. Stromata 43. Asci with

ascospores, 44. Ascospores. Stictis carnea. 45. Apothecia. 46. Ascospores. Stictis radiata.

47. Apothecia; 48. Asci and ascospores.

698 ... Medel-Ortiz, Baeza, Lorea-Hernandez

Seimatosporium tostum (Berk. & Broome) Rossman & W.C. Allen, IMA Fungus

7(1): 5 (2016). Figs 42-44

StromartaA shield-like, growing within host tissue, black, grey (30F1) to

greenish grey (30F2) pigments are observed in KOH. Two perithecia per

stroma. Asctr cylindrical, 62-75 x 5-7(-8) um, thin-walled, apical pore

J+, spores 1-2-seriate, without croziers. Ascosporegs ellipsoidal-fusiform,

1-septate, constricted at the septum, smooth, poles subacute to acute,

(9-)10-11(-12) x 3-4 um (Q = 2.3). PARAPHYSES filiform, <1 um diam.,

extending beyond the ascus by 3-5 um, with tiny hyaline guttules at the

apex.

Hasitat—On rachises of Alsophila firma and Cyathea bicrenata.

SPECIMENS EXAMINED—MEXICO. VERACRUZ: San Andrés Tlalnelhuayocan County,

Rio Xocoyolapan, 19°30.89’N 97°00.39’W, 1590 m, 8 December 2011, Medel & Lorea

2108; 26 February 2001, Medel & Lorea 2201; Los Encinales, 19°31.10’N 97°00.32’W,

1630 m, 30 March 2014, Medel & Lorea 2379.

ADDITIONAL SPECIMENS EXAMINED: MEXICO. Veracruz: San Andrés

Tlalnelhuayocan, Rancho Agiiita Fria, 7 February 2007, Medel 1342.

Seimatosporium tostum is described by Paulus & al. (2006) as having larger

(>14 x 5 um) hyaline ascospores with only one septum, while Brockmann

(1976) described North American material (as Discostroma tostum) with

1-6-septate ascospores. The species has been reported growing on stems

of Epilobium. Known from North America (Brockmann 1976, Paulus & al.

2006), S. tostum establishes a new record for Mexico and the first report on

ferns.

Stictis carnea Seaver & Waterston,

Mycologia 33: 311 (1941). Fics 45, 46

APOTHECIA immersed in host tissue, hymenium greyish yellow to light

yellow (1A5-1B6), 05-0.7 mm diam., surrounded by an elliptical white

(1A1) ring. Ascr cylindric-clavate, 110-150 x 6-7 um, 8-spored, hyaline, J-.

Ascosporss long filiform, (80-)85-130(-150) x 3 um, tapering to one end,

multi-septate, twisted within the ascus. PARAPHYSES filiform, <3 um wide,

longer, hyaline, septate.

Hasitat—On rachises of Cyathea bicrenata.

SPECIMEN EXAMINED—MEXICO. VERACRUz: San Andrés Tlalnelhuayocan County;

Rio Xocoyolapan; 19°30.89’N 97°00.39’W, 1590 m, 4 November 2011, Medel 2112.

There are two species of Stictis growing on ferns in our study site, S. carnea

and S. radiata. ‘The differences in spore length and apothecial morphology

of the apothecia are sufficient to separate these two species (Sherwood 1977,

Pteridicolous ascomycetes (Mexico) ... 699

Johnston 1983). Stictis carnea has been reported from Asia, North America

(including Mexico), and New Zealand (Johnston 1983, Sherwood 1977),

growing on dead tissues of many herbaceous and woody plants and the

fern Cyathea medullaris (Johnston 1983). This species is a new report from

Mexico.

Stictis radiata (L.) Pers., Observ. Mycol. 2: 73 (1800 [“1799”]). Fics 47, 48

APOTHECIA deeply sunken in host tissue, hymenium vivid yellow to yellow

(3A7-3A8), <1 mm diam., surrounded by an irregular white (1A1) ring. Asc1

cylindric-clavate, 240-290 x (7-)10-12 um, 8-spored, hyaline, narrowed

toward the apex, pore J-. Ascospores long filiform, (219-)245-280(-283)

x 2-3 um, multiseptate, spirally arranged within the ascus. PARAPHYSES

filiform, 1 um diam., slightly widening (<2 1m) at the apex.

Hasitat—On _ rachises of Alsophila firma, Cyathea _ bicrenata,

Sphaeropteris horrida, and fronds of Polypodium sp.

SPECIMENS EXAMINED—MEXICO. VeERAcRuz: San Andrés Tlalnelhuayocan

County; Rio Xocoyolapan; 19°30.89’N 97°00.39’W, 1590 m, 4 November 2011,

Medel 2199; 8 December 2011, Baeza 69, Medel & Lorea 2200. Paraje El Riachuelo, 8

December 2011; Medel & Lorea 2207. Los Encinales, 19°31.10’N 97°00.32’W, 1630 m,

10 August 2011, Baeza 38b, Medel & Lorea 2169.

Diagnostic characters of Stictis radiata include ascomata with a deeply sunken

orange yellow hymenium and filiform multiseptate spores with interlaced

arrangement within the ascus and variably sized (180-325 x 1.9-2.8(-3) um;

Breitenbach & Kranzlin 1984, Johnston 1983, Sherwood 1977). Stictis radiata

resembles S. carnea but is distinguished by its shorter and wider ascospores.

Known from Mexico, New Zealand, and Australia (Johnston 1983, Sherwood

1977). In New Zealand, S. radiata has been reported on the ferns Cyathea

dealbata and Dicksonia squarrosa (Johnston 1983). We found S. radiata also

growing on Sphaeropteris horrida and Polypodium sp.

Discussion

Ferns, which comprise many genera and families, serve as habitat for a large

number of fungi. Our study indicates that most of the fungi on ferns belong

to Lachnaceae [previously included in Hyaloscyphaceae s.l.], represented by

eight species of Lachnum. Haines (1980), Samuels & Rogerson (1980), and

Medel & Lorea-Hernandez (2008) previously reported Hyaloscyphaceae s.l. on

ferns. Medel (2013), who conducted earlier research on Mexican cloud forest

ascomycetes, pointed out that Lachnaceae is “recurrent” on ferns in Cyatheaceae,

indicating that some of these fungi may live as endophytes in this fern family.

700 ... Medel-Ortiz, Baeza, Lorea-Hernandez

The dominant plant associates in this study belong to the “tree ferns” —

Alsophila firma, Cyathea bicrenata, C. divergens var. tuerckheimii, and

Sphaeropteris horrida. Herbaceous fern species with short stems at or

barely above ground level include Dryopteris sp., Lophosoria quadripinnata,

Marattia laxa, and ‘Thelypteris spp. Adiantum andicola and Sticherus

palmatus are classed as herbs with underground stems, while Grammitis

sp., Phlebodium aureum, Pecluma alfredii, and Polypodium sp., represent a

diverse epiphytic fern group. Two fern species hosted the highest number

of fungal species: Alsophila firma (11 species) and Cyathea bicrenata (12

species). The ascomycetes identified in this study grew more frequently on

petioles and rachises of tree ferns and some herbaceous ferns.

Our work stresses the importance of studying the highly diverse small

ascomycetes given that new records have been established by every study

(Braun 2004, Braun & al. 2013, Cannon 1997, Castaneda-Ruiz & Heredia

2000, Del Olmo & Arnolds 2014, Dingley 1972, Haines 1980, Kirschner

& Liu 2014, Medel & Lorea-Hernandez 2008, Samuels & Rogerson 1990,

Stevenson 1945); unfortunately, few have studied species from the Western

Hemisphere. This paper contributes new fern families and genera to the

list of ascomycetes hosts: Dicksoniaceae (genus Lophosoria), Marattiaceae

(Marattia), and Polypodiaceae (Grammitis and Pecluma). This is important

since there are few studies on the diversity of fungi on ferns, and most have

dealt only with genera of tree ferns in Cyatheaceae and Dicksoniaceae.

According to the literature surveyed, only seven ascomycete species

had previously been registered on ferns from Veracruz. This study adds 18

more records associated with ferns in Mexican cloud forests, bringing the

TABLE 1. Ascomycete fungi and fern hosts recorded from Mexico

SPECIES Host REFERENCE

HELOTIALES

Arachnopezizaceae

cf. Arachnopeziza Alsophila firma This paper

Dermataceae

Mollisia ventosa* Cyathea bicrenata This paper

Helotiaceae

Bisporella pteridicola* Alsophila firma, Cyathea bicrenata, This paper

Pecluma alfredii, Sphaeropteris horrida,

Thelypteris sp.

Crocicreas Cyathea bicrenata This paper

quinqueseptatum*

Crocicreas sessilis

Cyathicula sp.

Durella macrospora*

Hyaloscyphaceae

Dasyscyphella dryina*

Hamatocanthoscypha

helicotricha*

Hyaloscypha fuckelii*

Lachnaceae

Lachnum brevipilosum*

Lachnum fimbriiferum

Lachnum nudipes *

Lachnum oncospermatum

Lachnum pteridophyllum

Lachnum singerianum

Lachnum varians

Lachnum virgineum

Paradiopsidaceae

Dimeriella polypodii

HyPocrEALESs (Bionectriaceae)

Protocreopsis pertusa*

ORBILIALES (Orbiliaceae)

Orbilia auricolor*

OSTROPALES

Coenogoniaceae

Coenogonium botryosum

Stictidaceae

Stictis carnea*

Stictis radiata*

XYLARIALES (Sporocadaceae)

Seimatosporium tostum

Pteridicolous ascomycetes (Mexico) ... 701

Cyathea divergens var. tuerckheimii

Cyathea bicrenata

Sticherus palmatus

Alsophila firma

Cyathea bicrenata, Sticherus palmatus

Alsophila firma

Alsophila firma, Cyathea bicrenata

Alsophila firma, Cyathea bicrenata,

Cyatheaceae, Dicksonia sellowiana

Lophosoria quadripinnata

Dicksonia sellowiana

Cyathea divergens var. tuerckheimii,

Cyathea sp., Dicksonia sellowiana,

Dryopteris sp., Lophosoria

quadripinnata

Alsophila firma, Dicksonia sellowiana

Alsophila firma, Cyathea bicrenata,

C. divergens var. tuerckheimii,

Adiantum andicola

Phlebodium aureum

Polypodium montigenum, P. madrense

Alsophila firma, Marattia laxa

Cyathea bicrenata, Marattia laxa

Adiantum andicola, Grammitis sp.

Cyathea bicrenata

Alsophila firma, Cyathea bicrenata,

Sphaeropteris horrida, Polypodium sp.

Alsophila firma, Cyathea bicrenata.

Bold face indicates new hosts for the corresponding fungus

* indicates new fungal records for Mexico

Samuels & Rogerson

1990

This paper

Medel & Lorea-

Hernandez 2008

This paper

Medel & Lorea-

Hernandez 2008

Haines 1980; Medel &

Lorea-Hernandez 2008

Medel & Lorea-

Hernandez 2008

Haines 1980; Medel &

Lorea-Hernandez 2008

This paper

Samuels & Rogerson

1990

This paper

702 ... Medel-Ortiz, Baeza, Lorea-Hernandez

total to 25 ascomycete species distributed among Helotiales (20 species),

Hypocreales (1), Ostropales (3), and Xylariales (1). Taxonomic diversity of

Ascomycota reported in similar studies from other regions of the world show

a wide diversity of ascomycetes involved in biological interaction with ferns.

For example, Del Olmo & Arnolds (2014) reported the primary endophytes

to inhabit ferns in Costa Rica belonged to Dothideomycetes, Eurotiomycetes,

and Sordariomycetes, while mycosphaerellaceous fungi were more abundant

in Taiwan (Kirschner & Liu 2014) and other countries in Asia and Africa

(Braun & al. 2013). Only Haines (1980) cited Helotiales (Hyaloscyphaceae

s.l.) as the predominant fungi on tropical ferns, further supported by this

study where Lachnaceae was the most diverse family represented. To find

such diversity of ascomycetes associated with ferns, almost quadrupling the

number of species previously known for Mexico, suggests there are several

taxa yet to discover. We agree with Kirschner & Liu (2014) that research

on ascomycetes growing on ferns is still in its infancy. Additional work is

needed to establish the ecology of fern-associated fungi, particularly the

recurrence of some taxa on fern hosts.

Acknowledgments

The first author expresses her gratitude to Gast6n Guzman (+), who was a

constant guide during her studies. Thanks to DGI-Universidad Veracruzana for the

research assistant fellowship granted to Yajaira Baeza and to Juan Lara Carmona,

XAL mycological collection assistant, who processed loans during the our research.

Special thanks to Sharon Cantrell, Teresa Iturriaga, and Andrea I. Romero for

comments that greatly improved an early version of the manuscript. Luis Quijada

(Department of Organismic and Evolutionary Biology, Harvard Herbarium,

Cambridge MA, U.S.A.) and José Marmolejo (Facultad de Ciencias Forestales,

Universidad Autonoma de Nuevo Leon, México) made valuable suggestions during

their expert peer reviews. Special thanks to Shaun Pennycook for his exhaustive

revision.

Literature cited

Baral HO, Haelewaters D, Partel K. 2015. A new attempt to classify the families of the Helotiales.

Poster presented at the Second International Workshop on Ascomycete Systematics. CBS

symposium, 22-24 April 2015, Amsterdam.

Braun U. 2004. Periconiella species occurring on ferns. Feddes Repertorium 115(1-2): 50-55.

https://doi.org/10.1002/fedr.200311025

Braun U, Nakashima Ch, Crous PW. 2013. Cercosporoid fungi (Mycosphaerellaceae) 1.

species on other fungi, Pteridophyta and Gymnospermae. IMA Fungus 4: 265-345.

https://doi.org/10.5598/imafungus.2013.04.02.12

Breitenbach J, Kranzlin F. 1984. Fungi of Switzerland. Volume 1: Ascomycetes. Luzern,

Switzerland: Verlag Mykologia. 310 p.

Pteridicolous ascomycetes (Mexico) ... 703

Brockmann I. 1976. Untersuchungen tber die Gattung Discostroma Clements (Ascomycetes).

Sydowia 28: 275-38.

Cannon PF. 1997. Two new genera of Ascomycota, and other new or interesting fungi from

Slapton Ley National Nature Reserve and its environs. Systema Ascomycetum 15: 121-138.

Cannon PFE, Aguirre-Hudson B, Aime MC, Ainsworth AM, Bidartondo MI, Gaya E,

Hawksworth DL, Kirk PM, Leitch IJ, Liicking R. 2018. Definition and diversity. 5-11, in:

K Willis, R Smith (eds). State of the World’s Fungi. Royal Botanic Gardens, Kew.

Carpenter SE. 1981. Monograph of Crocicreas (Ascomycetes, Helotiales, Leotiaceae). Memoirs of

the New York Botanical Garden. 33. 290 p.

Carpenter SE, Dumont KP. 1978a. Los Hongos de Colombia - IV. Bisporella triseptata and its

allies in Colombia. Caldasia 12(58): 339-348.

Carpenter SE, Dumont KP. 1978b. Leotiaceae I. Nannfeldt’s Phialeoideae: the genera

Belonioscypha, Cyathicula and Phialea. Mycologia 70: 1223-1238.

https://doi.org/10.1080/00275514.1978.12020340

Castaneda-Ruiz RF, Heredia G. 2000 Two new dematiaceous hyphomycetes on Cyathea from

Mexico. Cryptogamie, Mycologie 21: 221-228.

https://doi.org/10.1016/S0181-1584(00)01047-2

Chavarria A, Gonzalez MC, Dantan E, Cifuentes J. 2010. Evaluacion espacial y temporal de la

diversidad de los ascomicetes dulceacuicolas del canal turistico Santa Cruz, Xochimilco,

México. Revista Mexicana de Biodiversidad 81: 733-744.

Chaverri P, Huhndorf SM, Rogers JD, Samuels G. 2010. Microhongos comunes de Costa Rica

y otras regiones tropicales (Ascomycota, Pezizomycotina, Sordariomycetes). INBio. 241 p.

Del Olmo-Ruiz A, Arnolds E. 2014. Interannual variation and host affiliations of

endophytic fungi associated with ferns at La Selva, Costa Rica. Mycologia 106: 8-21.

https://doi.org/10.3852/13-098

Dennis RWG. 1949. A revision of the British Hyaloscyphaceae with notes on related European

species. Mycological Papers 32. 97 p.

Dennis RWG. 1950. Karsten’s species of Mollisia. Kew Bulletin 5(2): 171-187.

https://doi.org/10.2307/4117221

Dennis RWG. 1956. A revision of the British Helotiaceae in the Herbarium of the Royal Botanic

Garden Kew, with notes on related European species. Mycological Paper 62. 216 p.

Dennis RWG. 1960. Discomycetes described by Rick from South Brasil. Kew Bulletin 14(1):

114-125.

Dennis RWG. 1978. British Ascomycetes. Revised edition. J. Cramer, Vaduz. 585 p.

Dennis RWG. 1981. British Ascomycetes. Supplement to 1978 2nd revised edition. J. Cramer,

Vaduz. 4 p.

Dingley JM. 1972. Some foliicolous ascomycetes on ferns in Australia and New Zealand. The

genera Rhagadolobium P. Henn. & Lind. and Lauterbachiella Theiss. & Syd. New Zealand

Journal of Botany 10: 74-86. https://doi.org/10.1080/0028825X.1972.10430213

Dumont K. 1981. Leotiaceae II. A preliminary survey of the neotropical species referred to

Helotium and Hymenoscyphus. Mycotaxon 12(2): 313-371.

Gamundi JJ, Giaiotti AL. 1994. Notas sobre discomycetes andino-patagonicos I. Arachnopeziza

Fuckel y Parachnopeziza Korf. Sydowia 46(1): 12-22.

GBIF. 2019. GBIF home page. https://www.gbif.org (accessed March 14, 2019).

Haines JH. 1980. Studies in the Hyaloscyphaceae I: some species of Dasyscyphus on tropical

ferns. Mycotaxon 11(1): 189-216.

704 ... Medel-Ortiz, Baeza, Lorea-Hernandez

Huhtinen S. 1987. Taxonomic studies in the genera Protounguicularia, Arachnopeziza and

Dematioscypha. Mycotaxon 30: 9-28.

Huhtinen S. 1990 [“1989”]. A monograph of Hyaloscypha and allied genera. Karstenia 29(2):

45-252

Index Fungorum. 2019. http://www.indexfungorum.org/names/names.asp (accessed:15 January

2019).

Iturriaga T, Korf RP, Babcock JF. 1999. Fungi on Epifagus Crocicreas epifagicola sp. nov., with

comments on the generic names Crocicreas and Cyathicula. Mycological Research 103:

28-30. https://doi.org/10.1017/S0953756298007060

Johnston PR. 1983. Stictis and its anamorphs in New Zealand. New Zealand Journal of Botany

21: 249-279. https://doi.org/10.1080/0028825X.1983.10428557

Kirschner R, Liu LC. 2014. Mycosphaerellaceous fungi and new species of Venustosynnema

and Zasmidium on ferns and fern allies in Taiwan. Phytotaxa 176: 309-326.

https://doi.org/10.11646/phytotaxa.176.1.29

Korf R. 1951. A monograph of Arachnopezizaceae. Lloydia 14(3): 129-180.

Korf R. 1978. Revisionary studies in the Arachnopezizoideae: a monograph of the Polidesmieae.

Mycotaxon 7(3): 457-492.

Kornerup A, Wanscher JH. 1978. Methuen handbook of color. Polotokens, Forlag, Copenhagen.

258 p.

Largent D, Johnston D, Watling D. 1977. How to identify mushrooms to genus II: microscopic

features. Eureka, CA, Mad River Press Inc. 146 p.

Medardi G. 2004. Etudes sur le genre Durella. Documents Mycologiques 33(131) : 29-35.

Medel R. 2013. Hongos Ascomicetos del bosque mesofilo de montafa en México. Acta Botanica

Mexicana. 105: 87-106. https://doi.org/10.21829/abm105.2013.224

Medel R, Chacon S. 1997. Ascomycetes poco conocidos de México VII. Algunas especies del

bosque mesofilo de Veracruz. Acta Botanica Mexicana 39: 43-52.

https://doi.org/10.21829/abm39.1997.775

Medel R, Lorea-Hernandez F. 2008. Hyaloscyphaceae (Ascomycota) growing on tree ferns in

Mexico. Mycotaxon 106: 209-217.

Medel R., Lorea- Hernandez F, Guzman G. 2010. Fungi growing on Mexican tree ferns II. First

record of Favolaschia singeriana (Agaricales, Marasmiaceae). Sydowia 62(2): 277-281.

Mehltreter K. 2010. Interactions of ferns with fungi and animals. 220-254, in: K Mehltreter &

al. (eds). Fern Ecology. Cambridge University Press, Cambridge, UK.

Mickel JT, Smith AR. 2004. The pteridophytes of Mexico. Memoirs of the New York Botanical

Garden. 88. 1054 p.

Mo MH, Huang XW, Zhou W, Huang Y, Hao YE, Zhang KQ. 2005. Arthrobotrys yunnanensis sp.

nov., the fourth anamorph of Orbilia auricolor. Fungal Diversity 18: 107-115.

Mueller GM, Schmit JP, Huhndorf SM, Ryvarden L, O’Dell TE, Lodge DJ, Leacock PR, Mata

M, Umana L, Wu QX, Czederpiltz DL. 2004. Recommended protocols for sampling

macrofungi. 168-172, in: GM Mueller & al. (eds). Biodiversity of Fungi: Inventory and

Monitoring Methods. Amsterdam, Elsevier.

https://www.fpl.fs.fed.us/documnts/pdf2004/fpl_2004_mueller001.pdf

Paulus BC, Gadek PA, Hyde KD. 2006. Discostroma ficicola sp. nov. (Amphisphaeriaceae) and a

key to species of Discostroma. Sydowia 58(1): 76-90.

Raitviir A. 1970. Synopsis of the Hyaloscyphaceae. Scripta Mycologica 1. 115 p.

Raitviir A. 2002. A revision of the genus Dasyscyphella (Hyaloscyphaceae, Helotiales). Polish

Botanical Journal 47: 227-241

Pteridicolous ascomycetes (Mexico) ... 705

Raitviir A. 2004. Revised synopsis of the Hyaloscyphaceae. Estonian Agricultural University

Institute of Zoology and Botany. Scripta Mycologica 20. 133 p.

Rick J. 1906. Pilze aus Rio Grande do Sul. Brotéria 5: 5-53.

Rivas-Plata E, Licking R, Aptroot A, Sipman HJM, Chaves JL, Umafia L, Lizano D. 2006. A first

assessment of the Ticolichen biodiversity inventory in Costa Rica: the genus Coenogonium

(Ostropales: Coenogoniaceae) with a world-wide key and checklist and phenotype-based

cladistic analysis. Fungal Diversity 23: 255-321.

Rossman AY, Samuels GJ, Rogerson CT, Lowen R. 1999. Genera of Bionectriaceae, Hypocreaceae

and Nectriaceae (Hypocreales, Ascomycetes). Studies in Mycology 42. 248 p.

Samuels G.J, Rogerson C. 1990. Some ascomycetes (Fungi) occurring on tropical ferns. Brittonia

42(2): 105-115. https://doi.org/10.2307/2807623

Seaver FJ. 1938. Photographs and descriptions of cup fungi: XXX. Arachnopeziza. Mycologia

30(6): 659-663. https://doi.org/10.2307/3754363

Sherwood MA. 1977. The ostropalean fungi. Mycotaxon 5(1): 1-277.

Spooner BM. 1987. Helotiales of Australasia: Geoglossaceae, Orbiliaceae, Sclerotiniaceae,

Hyaloscyphaceae. Bibliotheca Mycologica 116. 711 p.

Stevenson JA. 1945. Ferns and fungi. American Fern Journal 35: 97-104.

https://doi.org/10.2307/1545645

Tejero-Diez D, Torres Diaz A, Mickel J, Mehltreter K, Krémer T. 2011. Helechos y licopodios.

97-115, in: AC Angon & al. (eds). La Biodiversidad en Veracruz estudio del Estado,

volumen II, Diversidad de Especies: Conocimiento Actual. Gobierno del Estado de

Veracruz. CONABIO- Universidad Veracruzana, INECOL.

Uyenco FR. 1963. The species of Coenogonium in the United States. Bryologist 66(4): 217-224.

https://doi.org/10.1639/0007-2745

Wijayawardene, N. Hyde KD, Lumbsch T, Liu JK, Maharachchikumbura SN, Ekanayaka AH

Qing T, Phookamsak R. 2018. Outline of Ascomycota: 2017. Fungal Diversity 88: 167-263.

https://doi.org/10.1007/s13225-018-0394-8

Ye M., Cao SQ, Jiang ST, Pan LJ, Luo SZ, Li XJ. 2006. Species diversity of Lachnum (Helotiales,

Hyaloscyphaceae) from temperate China. Journal of Zhejiang University Science B 7: 20-27.

https://doi.org/10.1631/jzus.2006.B0020

Zheng HD, Zhuang WY. 2016. Two new species of Crocicreas (Helotiaceae, Ascomycota)

revealed by morphological and molecular data. Phytotaxa 272: 149-152.

https://doi.org/10.11646/phytotaxa.272.2.6

Zhuang WY, Zheng HD, Ren FE. 2017. Taxonomy of the genus Bisporella (Helotiales) in

China with seven new species and four new records. Mycosystema 36(4): 401-420.

https://doi.org/10.13346/j.mycosystema.160193

MY COTAXON

October-December 2019—Volume 134, pp. 707-717

https://doi.org/10.5248/134.707

First sexual morph record of Sarcopodium vanillae

NAPALAI CHAIWAN?”, SAJEEWA S.N. MAHARACHCHIKUMBURA},

DHANUSHKA N. WANASINGHE’, MINGKWAN DOILOM?’,

RUVISHIKA JAYAWARDENA’, KEVIN D. HyDE*””

"Center of Excellence in Fungal Research, Mae Fah Luang University,

Chiang Rai 57100, Thailand

? Key Laboratory for Plant Diversity and Biogeography of East Asia, Kunming Institute of Botany,

Chinese Academy of Science, Kunming 650201, Yunnan, Peoples Republic of China

° School of Life Science and Technology, University of Electronic Science and Technology of China,

Chengdu 611731, Peoples Republic of China

* CORRESPONDENCE TO: kdhyde3@gmail.com

ABSTRACT—Sarcopodium vanillae was isolated from a dead leaf of Dracaena in Chiang

Rai Province, Thailand. Combined analyses of ACT, ITS, LSU, and TuB2 sequence data

obtained from the cultures derived from single spore isolates confirm that our collections

represent S. vanillae. This is the first record of the sexual morph, and the first record of S.

vanillae from Dracaena. A description and illustrations of both sexual and asexual stages of

S. vanillae are provided.

KEY worps—multigene, new host record, Nectriaceae, saprobe, taxonomy

Introduction

During our research on the diversity of microfungi in Thailand, we

recovered isolates from Dracaena in Chiang Rai Province that were typical

of Nectriaceae based on our preliminary morphological studies. Further

morphological and molecular characterization (multigene-based phylogeny

of nuclear ribosomal and protein-coding loci ACT, ITS, LSU and Tus2

sequence data) revealed the taxon as Sarcopodium vanillae. Nectriaceae

(Hypocreales; Rossman & al. 1999, Maharachchikumbura & al. 2015) is

typified by a characteristic perithecial wall structure and specific anamorphic

708 ... Chaiwan & al.

states. Its uniloculate perithecioid ascomata are yellow, orange, red, purple or

white, and the asexual morph is phialidic (Rogerson 1970, Rehner & Samuels

1995). The genus Sarcopodium Ehrenb. has had numerous nomenclatural

changes since it was first described in 1818 that have been synonymised

under Sarcopodium.

Cyphina Sacc., had already been synonymised under Sarcopodium by

Sutton (1977) and under “one fungus: one name” by Rossman & al. (2016)

supported the placement of the (asexual) Actinostilbe and the (sexual)

Lanatonectria under the much earlier (asexual) Sarcopodium. Sarcopodium

Ehrenb. is closely related to Pseudonectria Seaver. (Rossman & al. 1999).

Sutton (1981) synonymized Actinostilbe Petch. under Sarcopodium; and all

five species of Lanatonectria Samuels & Rossman (a genus described from a

sexual morph) have been transferred to Sarcopodium following phylogenetic

analyses (Lombard & al. 2015, Pennycook & Kirk 2019a). The new epithet for

the type species (“flocculentum”) has priority over the previous heterotypic

synonym, Sarcopodium macalpinei (Pennycook & Kirk 2019b).

Some Sarcopodium species have both sexual and asexual morphs, but S.

vanillae has been known only from its asexual morph (Sutton 1981; Rossman

1983, 1996, 2000; Rossman & al. 1993, 2013, 2016). This paper describes the

first record of a sexual morph for S. vanillae and reports Dracaena as a new

host record for this fungus.

Materials & methods

Sample collection, morphological study and isolation

Dead leaves of Dracaena were collected from Chiang Rai Province in Thailand

during November 2017. The samples were taken to the laboratory in Zip-lock bags.

Micro-morphological structures were observed and photographed with a Canon

EOS 600D digital camera fitted to a Nikon Eclipse Ni compound microscope. Single

spore isolates were obtained using the method described in Chomnunti & al. (2014).

Germinated spores were transferred to potato dextrose agar (PDA) plates and

incubated at 25-30 °C. Growth rates and culture characteristics were recorded after

three weeks. The morphological characters were measured using Tarosoft (R) Image

Frame Work v. 0.9.7 software. Figures were processed with an Adobe Photoshop CS6

Extended v. 10.0 software. The cultures are deposited in Mae Fah Luang University

Culture Collection, Chang Rai, Thailand (MFLUCC). Specimens are deposited in

the herbarium of Mae Fah Luang University, Chang Rai, Thailand (MFLU).

DNA extraction, PCR amplification and sequencing

Fungal isolates were grown on PDA for 3-4 weeks at 30 °C. The genomic DNA

was extracted from mycelium using the E.Z.N.A. Forensic DNA Kit. Amplification

Sexual morph of Sarcopodium vanillae (Thailand) ... 709

TABLE 1. Sequences from Pseudonectria and Sarcopodium species

(and Stachybotrys chartarum outgroup) used in the phylogenetic analysis

GENBANK ACCESSION NO.

TAXON CULTURE NO.

ACT ITS LSU TUB2

P. buxi CBS 324.53 KM231171 KM231778 KM231644 KM232037

P foliicola CBS 122566 KM231170 KM231777 KM231643 KM232036

S. circinatum CBS 587.92 KM231180 KM231787 KM231651 KM232046

CBS 100998 KM231179 KM231786 KM231650 KM232045

S. circinosetiferum CBS 100251 KM231175 KM231782 KM231646 KM232041

S. flavolanatum CBS 112283 KM231178 KM231785 KM231649 KM232044

CBS 128370 KM231177 KM231784 KM231648 KM232043

S. macalpinei CBS 115296 KM231176 KM231783 KM231647 KM232042

S. vanillae CBS 100582 KM231173 KM231780 HQ232174 KM232039

MEFLU 17-2595 MK692541 MK608516 MK691503 MK962543

MFLU 17-2597 MK692542 MK685870 MK691502 MK692544

Stachybotrys CBS 129.13 KM231268 KM231858 KM231738 KM232127

chartarum

New sequences are set in bold font.

primers used were: internal transcribed spacer (ITS)—ITS5 and ITS4 (White & al.

1990); 28S large subunit ribosomal RNA (LSU)—LROR and LRS5 (Rehner & Samuels

1994, Vilgalys & Hester 1990); partial actin gene (ACT)—ACT-512F and ACT-783R

(Carbone & Kohn 1999); partial beta-tubulin gene (ruB2)—Bt-2a and Bt-2b (Glass

& Donaldson 1995). The PCR mixer comprised 1 ul forward primer, 1 pul reverse

primer, 9.5 ul distilled deionized (DD) water, and 12.5 ul mixer. The PCR conditions

for ITS and LSU were 3 min at 94 °C; followed by 35 cycles of 30 s at 94 °C, 50 s at 55

°C, and 90 s at 72 °C; and a final elongation step at 72 °C for 10 min. Conditions for

ACT were an initial elongation step of 2 min at 95 °C; followed by 35 cycles of 45 s

at 95 °C, 45 s at 55 °C, and 1 min at 72 °C; and a final elongation step of 10 min at 72

°C. Conditions for TUB2 were an initial 8 min of 95 °C; followed by 35 cycles of 30 s

at 95 °C, 30 s at 55 °C, and 1 min at 72 °C; and a final elongation step at 72 °C for 5

min. The PCR products were purified and sequenced at Shanghai Sangon Biological

Engineering Technology and Service Co. Isolates including accession numbers of

gene sequences are listed in TABLE 1.

Phylogenetic analysis

Sequence data of Sarcopodium vanillae and related taxa (TABLE 1) were

downloaded from GenBank following Zeng & al. (2018) and Yang & al. (2018). The

multiple sequence alignments were produced with MAFFT v. 7 (http://maftft.cbre.jp/

710... Chaiwan & al.

alignment/server/index.html) and Biokdit v. 7.0.5.2 (Hall 1999). The phylogenetic

analyses were performed using maximum likelihood (ML) trees and generated using

the RAxML-HPC2 on XSEDE (8.2.8) (Stamatakis & al. 2008, Stamatakis 2014) in

the CIPRES Science Gateway platform (Miller & al. 2010) using GTR+I+G model

of evolution. Parsimony analysis was carried out with the heuristic search option

in PAUP v. 4.0b10 (Swofford 2002) using the following parameters: characters

unordered with equal weight, random taxon addition, branch swapping with tree

bisection-reconnection (TBR) algorithm, branches collapsing if the maximum

branch length was zero. Alignment gaps were treated as missing characters in the

combined data set, where they occurred in relatively conserved regions. Trees were

inferred using the heuristic search option with 1000 random sequence additions,

with maxtrees set at 1000. Descriptive tree statistics for parsimony; tree length (TL),

consistency index (CI), retention index (RI), relative consistency index (RC) and

homoplasy index (HI) were calculated for trees generated. The Kishino-Hasegawa

tests (Kishino & Hasegawa 1989) were performed to determine whether trees

were significantly different. Bayesian analysis was conducted with MrBayes v. 3.1.2

(Huelsenbeck & Ronquist 2001) to evaluate posterior probability (PP) (Rannala &

Yang 1996) by Markov chain Monte Carlo sampling. GTR+I+G was used in the

command. Six simultaneous Markov chains were run for 2,000,000 generations with

trees sampled every 200th generation. The distribution of log-likelihood scores was

examined to determine stationary phase for each search and to decide if extra runs

were required to achieve convergence, using the program Tracer 1.4 (Rambaut &

Drummond 2007). First 10% of generated trees were discarded and remaining 90%

of trees were used to calculate posterior probabilities of the majority rule consensus

tree. The phylogenetic tree was figured in FigTree v. 1.4 (Rambaut 2014) and edited

using Microsoft Office Power Point 2007. Sequences derived in this study were

deposited in GenBank (TABLE 1).

Phylogenetic results

The combined sequence alignments comprised 12 isolates, with

Stachybotrys chartarum (CBS 129.13) as the outgroup taxon. The combined

dataset comprised 3065 characters including alignment gaps, of which 844

were derived from ITS, 624 from LSU, 812 from ACT, and 785 from TuB2.

The MP analysis for the combined dataset had 415 parsimony informative,

2400 constant, 250 parsimony uninformative characters and yielded a single

most parsimonious tree (TL = 1109, CI = 0.812, RI = 0.761, HI = 0.188,

RC=0.617; Fic 1). The RAxML analysis of the combined dataset yielded a

best scoring tree (Fic. 1) with a final ML optimization likelihood value of

-~9266.358949. The matrix had 656 distinct alignment patterns, with 17.28% of

undetermined characters or gaps. Estimated base frequencies were as follows:

A = 0.220970, C = 0.279441, G = 0.267083, T = 0.232506; substitution rates

Sexual morph of Sarcopodium vanillae (Thailand) ... 711

Pseudonectria follicola CBS122566

100/400/1

Pseudonectria buxi CBS324.53

Sarcopodium flavolanatum CBS 112283

100/101

Sarcopodium flavolanatum CBS128370

Sarcopodium circinatum CBS 100998

100/401

Sarcopodium circinatum CBS587.92

99/82/0.99

Sarcopodium circinosetiferum CBS100251

100/100/1

Sarcopodium macalpinel CBS115296

Sarcopodium vanilla CBS100582

100/101

64/100/0.95

Fic. 1. Phylogram generated from RAxML based on combined ITS, LSU, ACT, and Tus2 sequence

data. Bootstrap support values for maximum likelihood (ML) 260%, maximum parsimony (MP)

260%, and Bayesian posterior probabilities (PP) 20.95 are indicated as ML/MP/PP at the nodes.

Ex-type strains are in bold and the strains from this study are indicated in green.

712 ... Chaiwan & al.

AC = 0.730175, AG = 1.879094, AT = 1.313827, CG = 0.798328, CT = 4.086170,

GT = 1.000000; gamma distribution shape parameter a = 0.147281.

Bayesian posterior probabilities from Bayesian inference analysis were

assessed with a final average standard deviation of split frequencies = 0.0056.

The phylogenetic tree in this study showed that our strains (Sarcopodium

vanillae MFLUCC 17-2595 and MFLUCC 17-2597) grouped in the

Sarcopodium clade, and formed a well-supported cluster with S. vanillae

(CBS 100582) with 64% ML, 100% MP, and 0.95 PP.

Taxonomy

Sarcopodium vanillae (Petch) B. Sutton,

Trans. Brit. Mycol. Soc. 76: 99 (1981) PLATES 1, 2

= Actinostilbe vanillae Petch, Ann. Roy. Bot. Gard. (Peradeniya) 9(3):327 (1925)

SAPROBIC on dead leaves of Dracaena (Asparagaceae).

SEXUAL MORPH: ASCOMATA 150-200 um high, 160-240 um wide (x = 173

x 189 um, n = 5), perithecial, subglobose, solitary or in groups, soft-textured,

pale yellow or rarely orange, superficial on a leaf or erumpent, with a papillate

ostiole. PERIDIUM 15-25 um wide (x = 47 um, n = 5), composed of several

layers of white to light orange cells of textura angularis. Asci 36-52 x 3-5 um

(x = 44 x 4.5 um, n = 20), 4-spored, unitunicate, cylindrical, rounded at apex,

clavate to fusiform, short pedicellate. Ascosporss 8-12 x 3-4.5 um (x = 11

x 3.9 um, n = 20), fasciculate, broadly elongate, 1-septate.

ASEXUAL MORPH: Myce.Lium, branched, septate, hyaline, smooth.

CONIDIOMATA 200-210 x 220-240 um (x= 205 x 230 um, n= 5), sporodochial,

solitary or gregarious, setiferous, yellow to bright yellow or rarely orangish

brown, soft-textured, superficial, separate, gregarious or confluent, sessile,

attached to the substratum by a small stroma concentrated in the epidermis

or outer layers of peridermal tissue, pulvinate, setose. SETAE 100-110 x 5-10

um (x = 105 x 7.5 um, n = 5), septate, unbranched, cylindrical, incurved,

erect, very thick-walled, medium to pale golden brown, more or less straight

(occasionally slightly bent either at the base or nearer the apex), erect,

hyaline, pointed or rounded at the apex. CONIDIOPHORES mononematous,

verticillately or penicillately branched, straight or flexuous, smooth, hyaline,

short, septate, with 1-4 monochasial branching, compactly arranged,

cylindrical, intermixed with long setae. CONIDIOGENOUS cells enteroblastic,

monophialidic, integrated, cylindrical or more frequently tapered towards

the apices, subulate, widest from middle to base, 10-20 x 1.2-2.7 um

(x = 15 x 1.95 um, n = 5), with inconspicuous collarette, hyaline, smooth,

Sexual morph of Sarcopodium vanillae (Thailand) ... 713

PLATE 1. Sarcopodium vanillae (MFLU 19-0567, herbarium specimen): A, B. Perithecia on

host surface; C-E. Perithecial cross sections; F Crush mount of perithecium; G. Conidioma on

perithecium; H, I. Asci, J. Ascospores; K, L. PDA cultures. Scale bars: A, B = 200 um; C-G = 50 um;

H, I= 20 um; J = 10 um

formed as the ultimate branches of conidiophores and completely covering the

external face of the conidiomata. Conrip1a 5-9 x 2.1-2.6 um (x = 6 x 2.5 um,

n = 20), cylindrical, 0-1-septate, smooth, hyaline, rounded at both ends, held

together in a slimy mass, ellipsoid to oval, straight.

CULTURE CHARACTERISTICS: Colony on PDA reaching 30-40 mm diam.

after 3 weeks at 25-30 °C; from above, white to yellow at margin, white to

714 ... Chaiwan & al.

PLATE 2. Sarcopodium vanillae (MFLU 19-0566, herbarium specimen): A-C. Conidiomata on host

surface; D. Conidioma; E-I. Setae; J, K. Setal bases; L. Setal apex; M. Conidia; N, O. PDA cultures.

Scale bars: A-C = 200 um; D, J, K = 100 um; E-I, L, M = 50 um.

orange in the middle, white at centre; from below, yellow, medium dense,

irregular, slightly raised to umbonate, surface slightly rough, dull with

umbonate edge, concave at centre, fluffy to floccose, with white tufts at centre.

SPECIMENS EXAMINED: THAILAND, CHIANG RAI PROVINCE, Mae Lao District, on

dead leaf of Dracaena, 17 November 2017, Napalai Chaiwan NCCR003 (MFLU 19-

0566 [asexual morph]; living culture MFLUCC 17-2595); Napalai Chaiwan NCCR004

(MFLU 19-0567 [sexual + asexual morph]; living culture MFLUCC 17-2597).

Discussion

Our strains share similar morphological characters with S. vanillae strain

CBS 100852, which was isolated from Anthurium sp. in Ecuador (Lombard &

al. 2015). The phylogenetic analysis also supports the close relationship. The

genus Sarcopodium has both sexual and asexual morphs (Wijayawardene

& al. 2017a,b, 2018). The conidial morphology of our strain is similar to

S. circinatum (the type species of the genus); however our conidiomata

and conidiophores more closely resemble Volutella ciliata (CBS 483.61)

Sexual morph of Sarcopodium vanillae (Thailand) ...715

(Lombard & al. 2015). Previously, only the asexual morph has been observed

for S. vanillae (Sutton 1981), and this study is the first report of the sexual

morph. Sarcopodium vanillae has been reported from Abelmoschus manihot

in Papua New Guinea, Citrus nobilis in Brunei, Vanilla planifolia in Sri Lanka,

and V. tahitensis in Papua New Guinea (Farr & Rossman 2019). This study

provides the first report of S. vanillae from Dracaena, and its first report from

Thailand.

Acknowledgments

N. Chaiwan thanks the Thailand Research Fund (PHD60K0147) and Kunming

Institute of Botany for financial support and the molecular laboratory for support.

K.D. Hyde thanks the grants entitled: 1 the future of specialist fungi in a changing

climate: baseline data for generalist and specialist fungi associated with ants,

Rhododendron species and Dracaena species (Grant number: DBG6080013) and

2. The climate changes grant: Impact of climate change on fungal diversity and

biogeography in the Greater Mekong Subregion (Grant number: RDG613001).

M. Doilom thanks the 5th batch of Postdoctoral Orientation Training Personnel in

Yunnan Province and the 64th batch of China Postdoctoral Science Foundation.

D.N. Wanasinghe thanks the CAS President’s International Fellowship Initiative

(PIFI) for funding his postdoctoral research (number 2019PC0008), the National

Science Foundation of China and the Chinese Academy of Sciences for financial

support under the following grants:41761144055, 41771063 and Y4ZK111B01.

S.C. Karunarathna thanks the CAS President's International Fellowship Initiative

(PIFI) for funding his postdoctoral research (number 2018PC0006). The National

Science Foundation of China (NSFC) for funding this work under the project code

31750110478. Jianchu Xu thanks the Key Research Program of Frontier Sciences

“Response of Asian mountain ecosystems to global change’, CAS, Grant No.

QYZDY-SSW-SMC014. All authors are grateful to peer experts D. Jayarama Bhat

(Emeritus Professor of Botany. Goa University, India) and Eric H.C. McKenzie

(Manaaki Whenua-Landcare Research, Auckland, New Zealand) for their assistance

and presubmission review.

Literature cited

Carbone I, Kohn LM. 1999. A method for designing primer sets for speciation studies in

filamentous ascomycetes. Mycologia 91: 553-556. https://doi.org/10. 2307/3761358

Chomnunti P, Hongsanan S, Hudson BA, Tian Q, Persdh D, Dhami MK, Alias AS, Xu J,

Liu X, Stadler M, Hyde KD. 2014. The sooty moulds. Fungal Diversity 66: 1-36.

https://doi.org/10.1007/s13225-014-0278-5

Farr DF, Rossman AY. 2019. Fungal databases, U.S. National Fungus Collections, ARS, USDA.

https://nt.ars-grin.gov/fungaldatabases/

Glass NL, Donaldson GC. 1995. Development of primer sets designed for use with the PCR

to amplify conserved genes from filamentous ascomycetes. Applied and Environmental

Microbiology 61: 1323-1330.

716... Chaiwan & al.

Hall TA. 1999. BioEdit: a user-friendly biological sequence alignment editor and analysis

program for Windows 95/98/NT. Nucleic Acid Symposium Series 41: 95-98.

Huelsenbeck JP, Ronquist F. 2001. MrBayes: Bayesian inference of phylogeny. Bioinformatics 17:

754-755. doi.org/10.1093/bioinformatics/17.8.754

Kishino H, Asegawa M. 1989. Evaluation of the maximum likelihood estimate of the evolutionary

tree topologies from DNA sequence data, and the branching order in Hominoidea. Journal of

Molecular Evolution 29: 170-179. https://doi.org/10.1007/BF02100115

Lombard L, van der Merwe NA, Groenewald JZ, Crous PW. 2015 Generic concepts in

Nectriaceae. Studies in Mycology 80: 189-245. https://doi.org/10.1016/j.simyco.2014. 12.002.

Maharachchikumbura SSN, Hyde KD, Gareth JEB, McKenzie EHC, Huang SK & al. 2015.

Towards a natural classification and backbone tree for Sordariomycetes. Fungal Diversity 72:

199-301. https://doi.org/10.1007/s13225-015-0331-z

Miller MA, Pfeiffer W, Schwartz T. 2010. Creating the CIPRES science gateway for

inference of large phylogenetic trees. 1-8, in: Proceedings of the Gateway Computing

Environments Workshop (GCE), November 14, 2010, New Orleans, Louisiana.

https://doi.org/10.1109/GCE.2010.5676129

Pennycook SR, Kirk PM. 2019a. Nomenclatural novelties: S. Pennycook & P.M. Kirk. Index

Pungorum No. 418.

http://www.indexfungorum.org/Publications/Index%20Fungorum%20no0.418.pdf

Pennycook SR, Kirk PM. 2019b. Sarcopodium flocculentum, the correct name for S. macalpinei.

Mycotaxon 134: 677-679. https://doi.org/10.5248/134.677

Rambaut A. 2014. FigTree v1.4: Tree figure drawing tool. http://treebio.ed.ac.uk/software/figtree/.

Rambaut A, Drummond AJ. 2007. Tracer v1.4, Available at: http://beast.bio.ed.ac.uk/Tracer

Rannala B, Yang, Z. 1996. Probability distribution of molecular evolutionary trees: a

new method of phylogenetic inference. Journal of Molecular Evolution 43: 304-311.

https://doi.org/10.1007/BF02338839

Rehner SA, Samuels GJ. 1994. Taxonomy and phylogeny of Gliocladium analyzed by large

subunit rDNA sequences. Mycological Research 98: 625-634.

https://doi.org/10.1016/s0953-7562(09)80409-7

Rehner SA, Samuels GJ. 1995. Molecular systematics of the Hypocreales: a teleomorph gene

phylogeny and the status of their anamorphs. Canadian Journal of Botany 73: S$816-S823.

https://doi.org/10.1139/b95-327

Rogerson CT. 1970. The hypocrealean fungi (Ascomycetes, Hypocreales). Mycologia 62: 865-910.

https://doi.org/10.1080/00275514.1970.12019033

Rossman AY. 1983. The phragmosporous species of Nectria and related genera. Mycological

Papers 150: 1-164.

Rossman AY. 1996. Morphological and molecular perspectives on systematics of the Hypocreales.

Mycologia 88: 1-19. https://doi.org/ 10.2307/3760780

Rossman AY. 2000. Towards monophyletic genera in the holomorphic Hypocreales. Studies in

Mycology 45: 27-34.

Rossman AY, Samuels GJ, Lowen R. 1993. Leuconectria clusiae gen. nov. and its anamorph

Gliocephalotrichum bulbilium with notes on Pseudonectria. Mycologia 85: 685-704.

https://doi.org/10.2307/3760514

Rossman AY, Samuels GJ, Rogerson CT, Lowen R. 1999. Genera of Bionectriaceae, Hypocreaceae

and Nectriaceae (Hypocreales, Ascomycetes). Studies in Mycology 42: 1-248.

Rossman AY, Seifert KA, Samuels GJ, Minnis AM, Schroers HJ, Lombard L & al. 2013. Genera of

Bionectriaceae, Hypocreaceae and Nectriaceae (Hypocreales) proposed for acceptance and rejection.

IMA Fungus 4: 41-51. https://doi.org/10.5598/imafungus.2013.04.01.05. Epub 2013 Apr 4.

Sexual morph of Sarcopodium vanillae (Thailand) ... 717

Rossman AY, Allen WC, Braun U, Castlebury LA, Chaverri P, Crous PW & al. 2016.

Overlooked competing asexual and sexually typified generic names of Ascomycota

with recommendations for their use or protection. IMA Fungus 7: 289-308.

https://doi.org/10.5598/imafungus.2016.07.02.09

Stamatakis A. 2014. RAxML version 8: a tool for phylogenetic analysis and post-analysis of

large phylogenies. Bioinformatics 30(9): 1312-1313.

https://doi.org/10.1093/bioinformatics/btu033

Stamatakis A, Hoover P, Rougemont J. 2008. A rapid bootstrap algorithm for the RAxML web

servers. Systematic Biology 57(5): 758-771. https://doi.org/10.1080/10635150802429642.

Sutton BC. 1981. Sarcopodium and its synonyms. Transactions of the British Mycological

Society 76: 97-102. https://doi.org/10.1016/s0007-1536(81)80012-5

Swofford DL. 2002. PAUP: phylogenetic analysis using parsimony, version 4.0 b10. Sinauer

Associates, Sunderland.

Vilgalys R, Hester M. 1990. Rapid genetic identification and mapping of enzymatically

amplified ribosomal DNA from several Cryptococcus species. Journal of Bacteriology 172:

4238-4246. https://doi.org/10.1128/jb.172.8.4238-4246.1990

White TJ, Burns T, Lee S, Taylor JW. 1990. Amplification and direct sequencing of fungal

ribosomal RNA genes for phylogenetics. PCR protocols: A Guide to Methods and

Applications (Innis MA, Gelfand DH, Sninsky JJ, White TJ, eds). Academic Press, San

Diego, California, USA, 315-322.

Wijayawardene NN, Hyde KD, Lumbsch HT, Liu JK, Maharachchikumbura SSN, Ekanayaka

AH, Tian Q, Phookamsak R. 2017a. Outline of Ascomycota: 2017. Fungal Diversity 9(1):

115-140. https://doi.org/10.1007/s13225-018-0394-8

Wijayawardene NN, Hyde KD, Tibpromma S, Wanasinghe DN, Thambugala KM, Tian Q, Wang

Y, Fu L. 2017b. Towards incorporating asexual fungi in a natural classification: checklist and

notes 2012-2016. Mycosphere 8(9): 1457-1555. https://doi.org/10.5943/mycosphere/8/9/10

Wijayawardene NN, Hyde KD, Divakar PK, Rajeshkumar KC, Weerahewa D, Delgado G,

Wang Y, Fu L. 2018. Notes for genera update — Ascomycota: 6616-6821. Mycosphere 9(1):

115-140. https://doi.org/10.5943/mycosphere/9/1/2

Yang Q, Du Z, Liang YM, Tian CM. 2018. Molecular phylogeny of Nectria species associated

with dieback and canker diseases in China, with a new species described. Phytotaxa

356(3): 199-214. https://doi.org/10.11646/phytotaxa.356.3.2

Zeng ZQ, Zhuang WY, Yu ZH. 2018. New species and new Chinese records of Nectriaceae

from Tibet, China. Nova Hedwigia 106: 283-294.

https://doi.org/10.1127/nova_hedwigia/2017/0435

MY COTAXON

October-December 2019—Volume 134, pp. 719-730

https://doi.org/10.5248/134.719

Notes on rust fungi in China 7. Aecidium caulophylli

life cycle inferred from phylogenetic evidence and

renamed as Puccinia caulophylli comb. nov.

JING-XIN Jr'4, ZHUANG LY, Yu L1’, MAKOTO KAKISHIMA”?4®

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

Jilin Agricultural University, Changchun, Jilin 130118, China

College of Plant Pathology, Shandong Agricultural University, Taian 271000, China

> College of Forestry, Beijing Forestry University, Beijing 100083, China

‘University of Tsukuba, Tsukuba, Ibaraki 305-8572, Japan

* CORRESPONDENCE TO: *1096314395@qq.com * kakishima.makoto.ga@u.tsukuba.ac.jp

ABSTRACT—Spermogonial and aecial stages of Aecidium caulophylli on Caulophyllum

robustum (Berberidaceae) were shown by phylogenetic analyses of ITS and 28S sequence data

to be identical to a uredinial and telial rust on Milium effusum (Poaceae). A new combination,

Puccinia caulophylli is proposed for this species, and an epitype is designated.

Key worps—Pucciniaceae, Pucciniales, taxonomy

Introduction

Aecidium caulophylli was described in 1898 based on spermogonial and

aecial stages of a specimen on Caulophyllum robustum (Berberidaceae)

collected by V. Komarov in Amur, Siberia, Russian Far East, during June

1895 (Saccardo & Sydow 1902). This rust has been reported from China,

Russian Far East, and Japan (Miura 1928, Ito 1950, Tai 1979, Harada 1984,

Hiratsuka & al. 1992, Azbukina 2005). However, its uredinial and telial stages

have not been clearly demonstrated, although Azbukina (1984, 2005) listed

this species as spermogonial and aecial stages of Puccinia brachypodii var.

poae-nemoralis (G.H. Otth) Cummins & H.C. Greene [= P. poae-nemoralis

G.H. Otth], without any evidence.

ji & al.

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Puccinia caulophylli comb. nov. (China) ... 721

During an investigation of rust fungi in Jilin Province, China, during

2015-2018, A. caulophylli was observed on C. robustum. Near the infected

C. robustum, uredinial and telial stages of Puccinia species were also

observed on plants of Poaceae and Cyperaceae. We suspected that this rust

may have an heteromacrocyclic life cycle, producing uredinia and telia

on one of these plants. Inoculation of plants with spores is an appropriate

method to clarify rust fungus life cycles (Ji & al. 2017a,b), but often plant

collection may be restricted in conservation areas. As it may be difficult

to supply appropriate growth conditions after transplantation, molecular

analyses have been applied to clarify rust life cycles (Liu & Hambleton

2013, Ji & al. 2016, Padamsee & McKenzie 2017, Scholler & al. 2019). We

report here the results of phylogenetic and morphological analyses using

spermogonial/aecial specimens on A. caulophylli and potentially related

uredinial/telial specimens on Poaceae and Cyperaceae.

Materials & methods

Molecular analyses

Spermogonial (0) and aecial (I) stages of A. caulophylli on C. robustum were

collected in Jiaohe, Jilin Province, China, and used for molecular analyses. Rust

specimens on Milium effusum (Poaceae) were collected in several areas in Jilin and

Heilongjiang Provinces, China, as preliminary phylogenetic analyses suggested they

represented the uredinial (II) and telial (III) stages of A. caulophylli. The specimens

were collected during surveys of rust fungi in Jilin Province from 2015 to 2018. For

comparative analyses, sequences were included from two life cycles that previously

clarified by inoculations (Ji & al. 2017a,b): (1) Puccinia klugkistiana (Dietel) Jing

X. Ji & Kakish. on Ligustrum obtusifolium Siebold & Zucc. (0, I) and Cleistogenes

hackelii (Honda) Honda (II, II), collected in Changchun, Jilin Province; and

(2) PB adenocauli (Syd. & P. Syd.) Jing X. Ji & Kakish. on Adenocaulon himalaicum

Edgew. (0, I) and Carex onoei Franch. & Sav. (II, IID).

Total genomic DNA was directly extracted from about 200 spores obtained from

single sori on the leaves of each specimen, using similar methods reported by Ji & al.

(2016, 2019). Specimens used in the experiments were deposited in the Herbarium

of Mycology, Engineering Research Center of Chinese Ministry of Education for

Edible and Medicinal Fungi, Jilin Agricultural University, China (HMJAU) and all

data sequenced in this experiment were deposited at GenBank (TABLE 1).

Sequences from the specimens were aligned following Ji & al. (2019). ITS and

28S sequence data retrieved from GenBank were added to phylogenetic analyses.

Accession numbers for these data are shown in the phylogenetic trees (Fics 1,

2). Phylogenetic trees were constructed with the sequences of Gymnosporangium

yamadae Miyabe ex G. Yamada and G. asiaticum Miyabe ex G. Yamada as outgroup,

722... Ji &al.

Puccinia caulophylli HMJAU8620 [Caulophyllum robustum] (SA)

Puccinia caulophylli HMJAU8619 [Milium effusum)] (UT)

Puccinia caulophylli HMJAU8623 [Milium effusum] (UT)

Puccinia caulophylli HMJAU8618 [Milium effusum] (UT)

Puccinia caulophylli HMJAU8625 [Milium effusum] (UT)

Puccinia caulophylli HMJAU8626 [Milium effusum] (UT)

059/50" Puccinia caulophylli HMJAU8621 [Milium effusum)] (UT)

Puccinia caulophylli HMJAU8627 [Milium effusum] (01)

Puccinia caulophylli HMJAU8534 [Caulophyllum robustum] (SA)

Puccinia caulophylli HMJAU8622 [Milium effusum] (U1)

Puccinia caulophylli HMJAU8624 [Caulophyllum robustum] (SA)

Puccinia caulophylli HMJAU8531 [Caulophyllum robustum] (SA)

osoisyg) -Puecinia caulophylli HMJAU8532 [Caulophyllum robustum) (SA)

Puccinia sessilis (AY 217134)

Puccinia sessilis (AY 217135)

Puccinia hordei (KY764128) [Ornithogahim arabicum]

Puccinia recondita (KY798399) [Elymus spicatus]

wioonooPuecinia magnusiana (GU058000) [Phragmites sp.]

Puccinia magnusiana (KY 764139) [Phragmites sp.]

Puccinia triticina (DQ664194) [Triticum aestivum]

Puccinia triticina (KY764169) [Triticum aestivum]

Puccinia recondita (DQ417424) [Aegilops ovata]

‘Puccinia recondita (AF511082)

Puccinia recondita f. sp. secalis (DQ417423) [Secale cereale]

Puccinia clavata (KX985761) [Clematis foetida]

Puecinia hordei (KX985762) [Holcus lanatius]

osgousgPuccinia sorghi (KY764162) [Zea mays]

Puccinia sorghi (GU057994) [Zea sp.]

Puccina polysora (GU058024) [Zea mays]

'Puccina polysora (MF033480) [Zea sp.]

‘Puccinia brachypodii (KX999868) [Poa annua]

‘Puccinia poae-nemoralis (KY 798384) [Calamagrostis sp.]

oossiggjPuccinia coronata (AF426207) [Rhamnus cathartica]

'Puccinia coronata (EU851141) [Holcus lanatus]

4 Puccinia coronata (DQ355448) [Bromus sp.]

Puccinia coronata (AB693935)

0.95/°/73 Puccinia graminis (KM249852) [Glyceria maxima]

oseomeyPuccinia graminis (KY798389) [Poa annua]

'Puccinia graminis (AF522177)

Puccinia adenocauli HMJAU 8628 [Adenocaulon himalaicum] (SA)

nooo Puccinia adenocauli HMJAU 8630 [Carex onoei] (UT)

Puccinia adenocauli HMJAU 8274 [Carex onoei] (UT)

Puccinia adenocauli HMJAU 8629 [Adenocaulon himalaicum] (SA)

Puccinia klugkistiana HMJAU 8280 [Cleistogenes hackelii] (U1)

Puccinia klugkistiana HMJAU 8633 [Ligustrum obtusifolium] (SA)

soon oo| Puccinia klugkistiana HMJAU 8632 [Cleistogenes hackelii] (UT)

Puccinia klugkistiana HMJAU 8198 [Ligustrum obtusifolium] (SA)

Puccinia klugkistiana HMJAU 8282 [Cleistogenes hackelii] (UT)

Puccinia klugkistiana HMJAU 8631 [Ligustrum obtusifolium] (SA)

/100/100) Gymnosporangium asiaticum HMJAU 8324 [Pyrus sp.]

Gymnosporangium yamadae HMJAU 8096 [Malus baccata]

1/100/100)

Fic. 1. Phylogenetic tree constructed by MP method based on sequences of 28S regions of rDNA.

Bootstrap values of MP and ML are followed by the Bayesian posterior probabilities (BPP) on

the nodes in the topology. Asterisk (*) represents bootstrap values <50% or BPP <0.5 in the

topology. Sample data are shown with species name, voucher specimen number or GenBank

accession number (in parentheses), and host plant. Sequence data determined in this study are

shown in bold face. SA: Spermogonial and aecial stages, UT: Uredinial and telial stages.

according to Ji & al. (2019). The alignment and trees were deposited in TreeBase

under http://purl.org/phylo/treebase/phylows/study/TB2:S24307 (Fic. 1) and

TB2:824308 (Fic. 2).

Morphological observations

Light (LM) and scanning electron (SEM) microscopy were used to examine

morphological characters of rust specimens including the size and shape of sori and

spores following Ji & al. (2019).

Puccinia caulophylli comb. nov. (China) ... 723

Puccinia caulophylli HMJAU8620 [Caulophyllum robustum] (SA)

Puccinia caulophylli HMJAU8619 [Milium effusum] (U1)

Puccinia caulophylli HMJAU8618 [Milium effusum] (UT)

Puccinia caulophylli HMJAU8532 [Caulophyllum robustum] (SA)

Puccinia caulophylli HMJAU8534 [Caulophyllum robustum] (SA)

Puccinia caulophylli HMJAU8625 [Milium effusum] (U1)

Puccinia caulophylli HMJAU8627 [Milium effusum] (UT)

Puccinia caulophylli HMJAU8623 [Milium effusum] (UT)

Puccinia caulophylli HMJAU8626 [Milium effusum) (U1)

vosioo| Puccinia caulophylli HMJAU8531 [Caulophyllum robustum] (SA)

Puccinia caulophylli HMJAU8621 [Milium effusum] (UT)

99/55/62] Puccinia caulophylli HMJAU8624 [Caulophyllum robustum] (SA)

Puccinia caulophylli HMJAU8622 [Milium effusum] (UT)

Puccinia sessilis (AY 217134)

1100/1001 Pyecinia sessilis (AY 217135)

joanoor Puccinia triticina (DQ417417) [Triticum turgidum L. var. durum |

o.p/ST/EM Puccinia triticina (DQ417417) [Triticum sp.]

Puccinia triticina (KT982695) [Hordeum vulgare]

1007108 Puccinia triticina (DQ460717) [Hordeum vulgare]

Puccinia recondita f. sp. secalis (DQ417426) [Secale cereale]

M0000 Puccinia recondita (AY956562) [Cerinthe minor]

0.87/#/4 Puccinia coronata (DQ355444) [Holcus lanatus|

0.99/88 Puccinia coronata f. sp. graminicola (HM131240) [Arrhenatherum elatius]

Puccinia coronata var. coronata (HM057141) [Calamagrostis epigejos]

Puccinia coronata f. sp. avenae (EU014044) [Lolium perenne]

Puccinia coronati-hordei (HM131229) [Elymus repens]

Puccinia coronati-japonica (HM131317) [Calamagrostis arundinacea]

Puccinia coronati-agrostidis (HM131319) [Agrostis stolonifera]

Puccinia coronati-calamagrostidis (HM131350) [Elymus sp.]

venoqPuccinia sorghi (AY 114291)

Puccinia sorghi (HQ154038) [Zea mays]

vioggg Puccinia brachypodii (KM 391664)

i600} | Puccinia brachypodii (KM 391669)

Puccinia brachypodii (GQ457303)

Puccinia graminis (AY874140)

oom Puccinia graminis f. sp. tritici (DQ 417379)

Puccinia graminis (HM131357) [Elymus repens]

Puccinia klugkistiana HMJAU 8633 [Ligustrum obtusifolium] (SA)

Puccinia klugkistiana HMJAU 8282 [Cleistogenes hackelii] (UT)

Puccinia klugkistiana HMJAU 8198 [Ligustrum obtusifolium] (SA)

Puccinia klugkistiana HMJAU 8631 [Ligustrum obtusifolium] (SA)

sno Puccinia klugkistiana HMJAU 8632 [Cleistogenes hackelii| (UT)

0.98/52/8 Puccinia klugkistiana HMJAU 8280 [Cleistogenes hackelii| (UT)

oonoor Puccinia polysora (HQ189433) [Zea mays]

Puccinia polysora (HM467909)

Puccinia adenocauli HMJAU 8274 [Carex onoei] (UT)

Puccinia adenocauli HMJAU 8628 [Adenocaulon himalaicum] (SA)

1100/1001 '— Puccinia adenocauli HMJAU 8629 [Adenocaulon himalaicum] (SA)

Puccinia adenocauli HMJAU 8630 [Carex onoei] (UT)

1/100/100 Gymnosporangium asiaticum HMJAU 8324 [Pyrus sp.]

Gymnosporangium yamadae HMJAU 8096 [Malus baccata]

/54/84)

99}

0.54/52/56

0,98/*/67

/97/100

0.97738763)

0.98/*/86)

+/#/8;

/1.00/100

Fic. 2. Phylogenetic tree constructed by MP method based on ITS regions of rDNA. Bootstrap

values of MP and ML are followed by the Bayesian posterior probabilities (BPP) on the nodes in

the topology. Asterisk (*) represents bootstrap values <50% or BPP <0.5 in the topology. Sample

data are shown with species name, voucher specimen number or GenBank accession number

(in parentheses), and host plant. Sequence data determined in this study are shown in bold face.

SA: Spermogonial and aecial stages, UT: Uredinial and telial stages

Results & discussion

Phylogeny and life cycle

The 28S dataset comprised 51 sequences of 50 taxa with 525 total

characters, including 430 constant characters, 25 parsimony-uninformative

variable characters, and 70 parsimony-informative characters. Parsimony

analysis yielded one parsimonious tree with TL = 159, CI = 0.679, RI = 0.864

724 ... Ji & al.

and RC = 0.587. Bayesian analysis resulted in average standard deviation of

split frequencies of 0.005869. The final ITS dataset comprised 51 sequences

of 50 taxa with 820 total characters, of which 336 were parsimony-

informative. Parsimony analysis yielded one parsimonious tree with

TL=995, CI = 0.623, RI = 0.840 and RC = 0.523. Bayesian analysis resulted in

average standard deviation of split frequencies of 0.004251. Tree topologies

formed by MP, ML, and MCMC methods were identical among trees.

The phylogenetic trees generated through Bayesian analysis are shown in

Fiac..1 (28S) and! Fie. 2(ITS):

Both 28S and ITS phylogenetic trees placed spermogonial and aecial

stages on C. robustum (HMJAU 8531, 8532, 8534, 8620, 8624) and uredinial

and telial stages on M. effusum (HMJAU 8618, 8619, 8621, 8622, 8623,

8625, 8626, 8627) within a monophyletic clade (Fries 1, 2). All stages of

P. klugkistiana and P adenocauli were placed in separate monophyletic

clades, confirming that heteroecious life cycles of rust fungi can be revealed

by phylogenetic analyses as demonstrated by Liu & Hambleton (2013),

Padamsee & McKenzie (2017), and Scholler & al. (2019). The phylogenetic

analyses also supported the life cycle connection between the rust on

C. robustum and the rust on M. effusum.

Although the Caulophyllum/Milium rust was shown to be phylogenetically

close to P. sessilis W.G. Schneid. ex J. Schrot., P. triticina Erickss., and

P. recondita Roberge ex Desm., these three rusts inhabit different host

plants. This rust is also genetically distant from P brachypodii G.H. Otth and

P. poae-nemoralis, two other telial rusts reported on Milium spp. (Cummins

1971, Zhuang & al. 1998, Azbukina 2005). Therefore, we conclude that this

rust is distinct from other species.

Morphology & taxonomy

From the phylogenetic analyses, the spermogonia and aecia on

A. caulophylli and the uredinia and telia on M. effusum are produced

by one and the same heteromacrocyclic rust species. LM and SEM

observations showed that overall morphology of the rust on C. robustum

is identical with that of A. caulophylli described by Saccardo & Sydow

(1902), Ito (1950), Harada (1984), and Hiratsuka & al. (1992) (Fics 3,

54,B). The morphologies of uredinial and telial specimens on M. effusum

were similar to each other (Fics 4, 5c-£), and the two-celled teliospores

refer the rust to the genus Puccinia (Cummins & Hiratsuka 2003).

Specimens on Milium effusum are phylogenetically close to P. sessilis,

but urediniospores on M. effusum are bigger than those of P. sessilis (20-32

Puccinia caulophylli comb. nov. (China) ... 725

ye ey

S ay, 4 io

FAM

Fic. 3. Puccinia caulophylli on Caulophyllum robustum: spermogonial and aecial stages.

A. Yellow lesions on the leaves producing spermogonia and aecia; B. Aecia produced

around spermogonia on lower surface of the plant; C. Catenulate aeciospores surrounded

by peridium in a vertical section of an aecium; D. Vertical section of a spermogonium;

E. Aeciospores. Scale bars: C, D = 30 um; E = 15 um.

x 19-25 um), and teliospores are smaller than those of P. sessilis (32-58 x

13-20 um). Additionally, the rust on M. effusum has uredinial paraphyses

that have not been reported in P. sessilis (Hiratsuka & al. 1992, Zhuang & al.

1998). PB. brachypodii var. poae-nemoralis occurs on M. effusum (Cummins

1971, Zhuang & al. 1998) and its telial structures and teliospores are

726 ... Ji & al.

morphologically similar to the present rust fungus. However, urediniospores

of the current rust are bigger than those of P. brachypodii var. poae-nemoralis

(18-25 x 15-23 um). The aecial stage of P brachypodii var. poae-nemoralis

occurs on Berberis spp. (Cummins 1971).

Aecidium caulophylli is a legitimate name under the INTERNATIONAL CODE

OF NOMENCLATURE FOR ALGAE, FUNGI, AND PLANTS (Shenzhen Code, Art.

F.8, 2018). However, the application of an asexual name, such as Aecidium,

to a sexual species can cause confusion (Ono 2016). Therefore, we propose

a new combination in Puccinia for A. caulophylli. The holotype specimen

on C. robustum has only spermogonial and aecial stages of the rust; because

the uredinial and telial stages define the rust genus, we designate an epitype

specimen on M. effusum.

Puccinia caulophylli (Kom.) Jing X. Ji & Kakish., comb. nov. Figs 3-5

MB 830631

= Aecidium caulophylli Kom., in Jaczewski & al., Fungi Rossiae Exsicc. 4: no 176, 1898.

Types: Russian Federation, Russian Far East, Siberia, Amur, Mt. Burejenses, stages

0, I on Caulophyllum robustum Maxim. [= C. thalictroides subsp. robustum (Maxim.)

Kitam.], June 1895, leg. V. Komarov (holotype, LE; isotype, NY 00610976). China, Jilin

Province, Yanbian, Erdaobaihe, stages II, III on Milium effusum L., 2 September 2018,

leg. J.X. Ji & M. Kakishima (epitype designated here, HMJAU 8627; MBT 386779).

SPERMOGONIA amphigenous, pale yellow to yellowish brown, subepidermal,

type 4 of Cummins & Hiratsuka (2003). Azcta hypophyllous, yellow,

subepidermal, erumpent, Aecidium-type with firmly connected peridia.

AECIOSPORES Catenulate, subglobose, ovate to ellipsoid, 14-24 x 13.5-20

um (av. 18.5 x 15.5 um), walls hyaline, 0.5-2 um thick (av. 1 um), densely

verrucose.

UreEDINIA mostly hypophyllous, pale yellow to cinnamon-brown,

subepidermal, erumpent, with abundant peripheral and intermixed paraphyses.

PARAPHYSES cylindric to capitate, 16.5-41 x 3-9.5 um (av. 30 x 5 um), walls

hyaline, 0.5-1.5 um thick (av. 1 um). UREDINIOSPORES pedicellate, globose

to subglobose, 22-35 x 21-30.5 um (av. 30 x 27 um), walls hyaline or pale

yellow, 1-3.5 um thick (av. 1.5 um), echinulate, germ pores obscure. TELIA

mostly hypophyllous, dark brown to black, subepidermal, covered by

epidermis, without paraphyses. TELIOSPORES 2-celled by transverse septum,

borne singly on pedicels, clavate to oblong, with round to obtuse apex and

attenuate towards base, 28.5-40.5 x 9.5-17 um (av. 35.5 x 13 um), walls pale

brown to dark brown, 0.5-1.5 um thick at sides (av. 1 um), 1-5 um thick at

apex (av. 2.5 um), smooth; pedicels short, hyaline.

Puccinia caulophylli comb. nov. (China) ... 727

Fic. 4. Puccinia caulophylli on Milium effusum: uredinial and telial stages. A. Uredinia and

telia produced on the leaves; B. Pale yellow uredinia (U) on lower leaf surface; C. Dark brown

telia on lower leaf surface; D. Echinulate urediniospores; E. Vertical section of uredinium with

urediniospores and paraphyses (P); F. Teliospores; G. Vertical section of telia covered by host

epidermis. Scale bars: D, E = 30 um; F, G = 20 um.

728 ... Ji & al.

Fic. 5. Puccinia caulophylli observed by SEM. A. Aecium with catenulate aeciospores and

peridium; B. Aeciospore with densely verrucose surface; C. Uredinium with urediniospores and

paraphyses (P); D. Echinulate urediniospore; E. Vertical section of a telium covered by host

epidermis. Scale bars: A = 30 um; B, D = 5 um; C, E= 20 um.

ADDITIONAL SPECIMENS EXAMINED—Stages 0, I on Caulophyllum robustum:

CHINA: JILIN PROVINCE, Jilin, 23 June 2015 (HMJAU 853); 24 June 2015 (HMJAU

8531, HMJAU 8534); 29 June 2017 (HMJAU 8620); 1 July 2018 (HMJAU 8624).

Stages II, III on Milium effusum: CHINA: JILIN PROVINCE, Jilin, 12 September

2017 (HMJAU 8622); 1 July 2018 (HMJAU 86232). Yanbian, 28 July 2015 (HMJAU

8618).; Baishan, 2 September 2018 (HMJAU 8625); 3 September 2018 (HMJAU 8626).

HEILONGJIANG PROVINCE: Wuchang, 5 July 2016 (HMJAU 8619); 9 September 2017

(HMJAU 8621).

Puccinia caulophylli comb. nov. (China) ... 729

HostTs & DISTRIBUTION—Stages 0, I on Caulophyllum robustum: China (Miura

1928, Tai 1979), Japan (Ito 1950, Harada 1984, Hiratsuka & al. 1992). Stages II,

UI on Milium effusum: China.

Acknowledgments

This work was financed by the Fungal Flora in Jilin Province (20130206073NY).

We thank Dr E.H.C. McKenzie (Manaaki Whenua Landcare Research, Auckland,

New Zealand) and Dr C.M. Denchev (Bulgarian Academy of Sciences, Sofia,

Bulgaria) for critical reading of the manuscript and suggestions. We also thank Dr

H. Koba (College of Arts and Sciences, J. F Oberlin University, Tokyo, Japan) for

identification of M. effusum.

Literature cited

Azbukina ZM. 1984. The manual of rust fungi in Soviet Far East. Nauka, Moscow. (In Russian)

Azbukina ZM. 2005. Rust fungi. Cryptogamic plants, fungi and mosses of the Russian Far East,

vol. 5. Dalnauka, Vladivostok. (In Russian)

Cummins GB. 1971. The rust fungi of cereals, grasses and bamboos. Springer-Verlag, New York.

Cummins GB, Hiratsuka Y. 2003. Illustrated genera of rust fungi, 3 ed. American

Phytopathological Society, St. Paul, Minnesota.

Harada Y. 1984. Materials for the rust flora of Japan IV. Transactions of Mycological Society of

Japan 25: 287-294.

Hiratsuka N, Sato S, Katsuya K, Kakishima M, Hiratsuka Y, Kaneko S, Ono Y, Sato T, Harada

Y, Hiratsuka T, Nakayama K. 1992. The rust flora of Japan. Tsukuba-shuppankai, Tsukuba.

Ito S. 1950. Mycological flora of Japan, vol. 2, no 3. Yokendo, Tokyo.

Ji JX, Li Z, Wan Q, Li Y, Kakishima M. 2016. Notes on rust fungi in China 1. Autoecious

life cycle of Puccinia tatarinovii on Prenanthes. Mycotaxon 131: 653-661.

https://doi.org/10.5248/131.653

Ji JX, Li Z, Wan Q, Li Y, Kakishima M. 2017a. Life cycle of Aecidium klugkistianum on

Ligustrum and its new combination, Puccinia klugkistiana. Mycoscience 58: 307-311.

https://doi.org/10.1016/j.myc.2017.01.004

Ji JX, Li Z, Wan Q, Li Y, Kakishima M. 2017b. Notes on rust fungi in China 3. Puccinia

adenocauli comb. nov. and its life cycle and new host. Mycotaxon 132: 141-148.

https://doi.org/10.5248/132.141

Ji JX, Li Z, Li Y, Kakishima M. 2019. Two new species of Pucciniastrum producing

dimorphic sori and spores from northeast of China. Mycological Progress 18: 529-540.

https://doi.org/10.1007/s11557-018-1460-z

Liu M, Hambleton S. 2013. Laying the formation for a taxonomic review of

Puccinia coronata s.l. in a phylogenetic context. Mycological Progress 12: 63-89.

https://doi.org/10.1007/s11557-012-0814-1

Miura M. 1928. Flora of Manchuria and east Mongolia 3. Cryptogams, Fungi.

Minamimanshutetsudo, Dalian.

Ono Y. 2016. Phakopsora hornotina, an additional autoecious rust species on Meliosma

in the Philippines and the Ryukyu Islands, Japan. Mycoscience 57: 71-78.

https://doi.org/10.1016/j.myc.2015.09.003.

Padamsee M, McKenzie EHC. 2017. The intriguing and convoluted life of a heteroecious rust

fungus in New Zealand. Plant Pathology 66: 1248-1257. https://doi.org/10.1111/ppa.12672

730 ... Ji & al.

Saccardo PA, Sydow P. 1902. Supplementum universale, pars 5. Sylloge fungorum 16. 1291 p.

Scholler M, Lutz M, Aime MC. 2019. Repeated formation of correlated species in Tranzschelia

(Pucciniales). Mycological Progress 18: 295-303. https://doi.org/10.1007/s11557-018-1417-2

Tai FL. 1979. Sylloge fungorum sinicorum. Science Press, Beijing.

Zhuang JY, Wei SX, Wang YC. 1998. Flora fungorum sinicorum, vol.10, Uredinales (1). Science

Press, Beijing.

MY COTAXON

October-December 2019—Volume 134, pp. 731-735

https://doi.org/10.5248/134.731

Exserticlava aquatica sp. nov.,

a microfungus from the Brazilian Amazon

LUANA TEIXEIRA DO CARMO’, DIOGO CARELI DOS SANTOS?,

CAROLINA RIBEIRO SILVA”, SHEILA MIRANDA LEAO FERREIRA’,

THAMARA ARAO FELETTI’, Luis FERNANDO PASCHOLATI GUSMAO”?

' Universidade Estadual de Feira de Santana, Programa de Pés-graduagao em Botanica,

Av. Transnordestina s/n, Novo Horizonte, 44036-900, Feira de Santana, Brazil

? Universidade Federal de Pernambuco, Centro de Biociéncias, Depto de Micologia,

Av. Prof. Nelson Chaves, s/n, Cidade Universitaria, 50670, Recife, Pernambuco, Brazil

* Correspondence to: lgusmao@uefs.br

ABSTRACT—A new species, Exserticlava aquatica, collected on submerged decaying twigs

in Para State in the Brazilian Amazon, is described and illustrated. The microfungus is

characterized by monoblastic conidiogenesis with repeated percurrent conidiophore

extensions and conidiogenous cells with a slightly swollen apex that does not protrude

beyond the ruptured outer wall.

Key worps—Ascomycota, Chaetosphaeriaceae, conidial fungi, freshwater fungi, taxonomy

Introduction

Exserticlava S. Hughes is characterized by distoseptate conidia and a hyaline

apical swelling of the conidiogenous cell formed by the inner wall disrupting a

pigmented outer wall (Hughes 1978). Tsui & al. (2001) reviewed the morphology

of the genus and provided a key to five species; subsequently two more species

were described: E. yunnanensis and E. manglietiae (Cai & Hyde 2007, Ren & al.

2012). Exserticlava is associated with the perithecial genus Chaetosphaeria Tul.

& C. Tul. (Reéblova & Seifert 2003, Seifert & al. 2011: 204). During a survey of

microfungi associated with twigs in estuary areas, an interesting specimen was

collected and is described here as a new Exserticlava species.

732 ... Carmo & al.

Material & methods

Samples of submerged decaying twigs were washed in river water to remove

sand and placed in plastic bags. In the laboratory, the samples were processed

according Castafeda-Ruiz & al. (2016) and regularly examined for 40 days under

stereomicroscope for observation of the reproductive structures. Slide mounts

were prepared in PVL (polyvinyl alcohol, lactic acid, and phenol) and/or in

lactic acid. Microphotographs were obtained using a Olympus BX51 microscope

with Nomarski interference optics. The type specimen has been deposited in the

Herbarium of Universidade Estadual de Feira de Santana, Brazil (HUEFS).

Taxonomy

Exserticlava aquatica L.T. Carmo, C.R. Silva, Careli, S.M. Ledo, Feletti & Gusmao,

sp. nov. PLATE 1

MB 831391

Differs from Exserticlava triseptata by its monoblastic conidiogenesis, its conidiogenous

cells with a slightly swollen apex that does not protrude beyond the ruptured outer wall,

and its smaller oblong-obtuse conidia.

Type: Brazil, Parad State: Belém, Mosqueiro island, 1°03’45”S 48°20'14”W, on

submerged decaying twigs of unidentified plant, 2.V1.2018, coll. L.T. Carmo (Holotype,

HUEFS249946).

EryMo.oey: the specific epithet refers to the aquatic habitat from which the species

was collected.

CoLonigs effuse, brown. MyceLium superficial and immersed, composed

of septate, branched, smooth, pale brown hyphae. CoNnrIDIOPHORES

macronematous, mononematous, unbranched, erect, straight or slightly

flexuous, smooth, thick-walled, dark brown to paler toward the apex, slightly

swollen at the base, surrounded by pseudo-parenchymatous cells forming a

brown stroma 22.5-27 um wide, (6-)8-13-septate, 330-455 x 6.5-7.5 um.

CONIDIOGENOUS CELLS monoblastic, enteroblastic, percurrent and progressive

leading to the production of up to 7 conidiogenous cell extensions, integrated,

terminal, clavate, slightly swollen at the apex, without conspicuous expansion

of the inner wall and not protruding from the fragmented and pigmented outer

wall of conidiogenous cells, smooth, pale brown, 18.5-46.5 x 5-7.5 um. Conidial

secession schizolytic. Conip1a acrogenous, holoblastic, solitary, smooth,

oblong-obtuse, thick-walled, 3-distoseptate, sometimes with conspicuous

PuaTE 1. Exserticlava aquatica (holotype, HUEFS 249946): A. General aspect of conidiophore

with attached conidium; B. Conidiogenous cell development; C. Conidium attached to

conidiogenous cell; D. Conidiogenous cell; E. Proliferating conidiogenous cells; F. Conidiogenous

cell producing a conidium after extension; G-I. Conidia. Scale bars: A = 50 um; B-I = 10um.

Exserticlava aquatica sp. nov. (Brazil) ... 733

734 ... Carmo & al.

aperture on distosepta, pale brown, 21-26.5 x 10-11.5 um; conidia sometimes

produced within intercalary old conidiogenous cells. SEXUAL MORPH: not

observed.

Note: Exserticlava triseptata, E. keniensis, and E. vasiformis most closely

resemble E. aquatica by producing 3-distoseptate conidia; the four other

accepted species are 1-distoseptate (Tsui & al. 2001, Rao & de Hoog 1986).

However, E. triseptata and E. keniensis have brown elliptic-obovoid conidia

and the conidiogenous cells extend forming a hyaline swelling that produces

conidia; and E. vasiformis is distinguished by vigorous inner wall extension of

funnel-shaped conidiogenous loci (Hughes 1978, Matsushima 1975, Tsui &

al. 2001). Exserticlava aquatica is easily separated from the other Exserticlava

species by its pale brown conidia, its several percurrent conidiogenous cell

extensions, and the distinctive crown-like appearance of the outer wall of the

conidiogenous cell; the new species is further characterized by monoblastic

conidiogenous cells and an absence of conspicuous inner wall expansion to

fragment the pigmented outer conidiogenous cell wall.

Exserticlava triseptata and E. vasiformis have been widely recorded in

Brazil, including in the Amazon region (Cruz & al. 2008).

Acknowledgments

We are indebted to Dr. De-Wei Li and Dr. Rafael F. Castafieda Ruiz for critical

review of the manuscript. LTC and TAF thank the Programa de Pds-graduacao em

Botanica (PPGBot/UEFS), the Nacional Council for Scientific and Technological

Development (CNPq), and the “Coordenacgao de Aperfeigoamento de Pessoal de

Nivel Superior (CAPES). DCS and CRS thank the Programa de Pés-graduacao

em Biologia de Fungos (PPGBF/UFPE). LFPG is grateful to the CNPq (Proc.

312984/2018-9).

Literature cited

Cai L, Hyde KD. 2007. Anamorphic fungi from freshwater habitats in China: Dictyosporium

tetrasporum and Exserticlava yunnanensis spp. nov. and two new records for

Pseudofuscophialis lignicola and Pseudobotrytis terrestris. Mycoscience 48(5): 290-296.

https://doi.org/10.1007/s10267-007-0369-1

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... New York, Springer

International Publishing. https://doi.org/10.1007/978-3-319-29137-6_9

Cruz ACR, Hernandez-Gutiérrez A, Gusmao LFP. 2008. O género Exserticlava (fungo

anamorfo—Hyphomycetes) no Brasil. Revista Brasileira de Botanica 31(2): 357-361.

https://doi.org/10.1590/S0100-84042008000200018.

Hughes SJ. 1978. New Zealand fungi 25. Miscellaneous species. New Zealand Journal of

Botany16(3): 311-370. https://doi.org/10.1080/0028825X.1978.10425143

Exserticlava aquatica sp. nov. (Brazil) ... 735

Matsushima T. 1975. Icones microfungorum a Matsushima lectorum. Kobe, Published by the

author. 209 p.

Rao V, de Hoog GS. 1986. New or critical hyphomycetes from India. Studies in Mycology 28.

84 p.

Réblova M, Seifert KA. 2003. Six new species of Chaetosphaeria from tropical rain forests in

Thailand and redescription of Chaetosphaeria hiugensis. Sydowia 55: 313-347.

Ren SC, MaJ, Zhang XG. 2012 [“2011”]. Two new species of Exserticlava and Spiropes on decaying

wood from Guangdong, China. Mycotaxon118: 349-353. https://doi.org/10.5248/118.349

Seifert K, Morgan-Jones G, Gams W, Kendrick B. 2011. The genera of hyphomycetes. CBS

Biodiversity Series 9. 997 p.

Tsui CK, Goh TK, Hyde KD. 2001. A revision of the genus Exserticlava, with a new species.

Fungal Diversity 7: 135-143.

MYCOTAXON

October-December 2019—Volume 134, pp. 737

https://doi.org/10.5248/134.737

Regional annotated mycobiotas new to the Mycotaxon website

ABSTRACT—Mycotaxon is pleased to add to our ‘web-list’ page the following new annotated

species distribution list under South America (Brazil): “Ascomycota (lichenized and non-

lichenized) on Syagrus coronata in the Caatinga biome: new and interesting records for

Brazil and South America” by Maiara A.L. dos Santos, Nilo G.S. Fortes, Tassio E.F. Silva,

Nadja S. Vitoria. This brings to 133 the number of free access Fungae now available on our

website: http://www.mycotaxon.com/mycobiota/index.html

SOUTH AMERICA

Brazil

Matra A.L. DOs SANTOS, NILO G.S. FoRTES, TAssio E.F. Sitva, NADJA S. VITORIA.

Ascomycota (lichenized and non-lichenized) on Syagrus coronata in the

Caatinga biome: new and interesting records for Brazil and South America.

10 p.

ABSTRACT—The Caatinga biome occupies most of the semiarid region of

northeastern Brazil, with varied landscapes and notable endemism. Among the

plants having significant importance in the Caatinga environment is the palm tree

Syagrus coronata, which is known as the “life-saving plant” due to its high socio-

biological and economic value. To better understand the mycota of the Arecaceae,

collections were undertaken in the municipalities of Paulo Afonso and Nova Gloria

within the Raso da Catarina eco-region in the drylands (“sertao”) of Bahia State,

Brazil. Twenty species of Ascomycota were identified during the present work:

three are new records for South America (Diplodia galiicola, Seimatosporium

corni, and Wojnowiciella viburni); eleven are new records for Brazil (Anthostomella

caricis, Caryospora callicarpa, C. putaminum, Chaetomium subaffine, Diatrype

bermudensis, Diatrypella persicae, Didymosphaeria massarioides, Eutypella

fraxinicola, Munkovalsaria donacina, Oedohysterium sinense, and Pleospora

calvescens); while six are new records for Bahia State (Dirinaria confusa, Lecanora

achroa, Phaeosphaeria sp., Pleospora herbarum, Polymeridium julelloides, and

Saccardoella macrasca). Syagrus coronata represents a new botanical host for all

taxa identified here.

Key worps—Pezizomycotina, semiarid, taxonomy

MYCOTAXON

October-December 2019—Volume 134, pp. 739

https://doi.org/10.5248/134.739

Regional annotated mycobiotas new to the Mycotaxon website

ABSTRACT—MycotTaxon is pleased to add a new annotated species distribution list

to our 134 previously posted free access fungae. The 22-page “Checklist of Bolivian

Agaricales. 1: Species with dark and pink spore prints.” by E. Melgarejo-Estrada, M.E.

Suarez, D. Rocabado, O. Maillard, and B.E. Lechner may be downloaded from our

website via http://www.mycotaxon.com/mycobiota/index.html

SOUTH AMERICA

Bolivia

E. MELGAREJO-EsTRADA, M.E. SUAREZ, D. ROCABADO, O. MAILLARD, B.E. LECHNER.

Checklist of Bolivian Agaricales. 1: Species with dark and pink spore prints.

22 p.

ABSTRACT—We provide a literature-based checklist of Agaricales reported from

Bolivia. In this first contribution, 101 species belonging to 28 genera and 9 families

are listed. Pluteaceae, Agaricaceae and Hymenogastraceae are the most species-

abundant families.

Key worps—Basidiomycota, distribution, diversity, Gasteromycetes, Neotropics,

macromycetes, South America