IV ... MYCOTAXON 135(2)

MYCOTAXON

VOLUME ONE HUNDRED THIRTY-FIVE (2) — TABLE OF CONTENTS

Nomenclatural novelties-er ty pificatiOnss. . vies eo ab eas eee Ve oes vii

COMA CENAD OY 2) LSA LITE So chee ONE AM ate OM ER GA dM S NPE i viii

TREVIE WETS 8. oe, a that oS ON pi Re eh Pte, cade Shet « Oe wente nto, BX ix

ZO2D SUD MIISSIOTE DIO COC TTE | o. OA cso nity ant OC EE a naa ah MC tice wea hen wee DE as %

PE TATISE TORE CILOPE gee urs ooh wits fines bd ina ttebee abi one guar Wigttin te ghd gee Ak Seared PPh xi

TAXONOMY & NOMENCLATURE

Four new Lepraria species for Iran,

with a key to all Iranian species | SareEH SADAT KAZzEMI, IRAJ MEHREGAN,

YOUNES ASRI, SARAH SAADATMAND, HARRIE J.M. SIPMAN 235

Pluteus dianae and P. punctatus resurrected,

with first records from eastern and northern Europe

Hana SevéiKovA, EKATERINA F. MALYSHEVA,

ALFREDO JUSTO, JACOB HEILMANN-CLAUSEN, MICHAL ToMSoOvSKY 245

Aecidium peristrophes, a new record for Pakistan

N.S. AFSHAN, M. Riaz, A. Saqis 275

Westerdykella aquatica sp. nov., producing phytase

HalI-YAN SONG, ALY FARAG EL SHEIKHA, PING-AN ZHONG,

JIANG-LIN Liao, ZHAO-HalI WANG, YING-JIN HUANG, D1AN-MING Hu 281

Ramiphialis ronuroensis gen. and sp. nov., a hyphomycete

from the Amazonian rainforest FLAVIA RODRIGUES BARBOSA,

Patricia OLIVEIRA FIUZA, RAFAEL F. CASTANEDA-RuIz 293

Phylogenetic placement of Acrospeira

De-WE!I Li, RAFAEL F, CASTANEDA-RutIZz, NEIL P. SCHULTES 299

Golovinomyces asperifolii; first record in China and

Bothriospermum chinense as a new host

Lucuao Bal, HUIYAN XIONG, BAOGUO SHI 309

Cercosporella bundelkhandae comb. nov. from India RAGHVENDRA SINGH,

SANJEET KUMAR VERMA, SANJAY YADAV, SHAMBHU KUMAR 315

Coprinellus ovatus sp. nov. from Pakistan

MUHAMMAD KAMRAN & SANA JABEEN 321

An update of G.K. Merrill’s 1909 “Lichen notes no. 14”

STEVEN B. SELVA & TROY MCMULLIN 333

Elaphomyces citrinus & E. cyanosporus, new for Turkey

YASIN UZUN & ABDULLAH Kaya 339

APRIL-JUNE 2020... V

New records of graphidoid and thelotremoid lichens from India

P. Gupta, P. RANDIVE, S. NAYAKA, R. DAIMARI,

S. JOSEPH, M.K. JANARTHANAM 345

Carbonea assimilis and Rinodina aspersa new to Poland

KATARZYNA SZCZEPANSKA 355

Badhamia versicolor and Trichia subfusca, new records for Belarus

EvGENY Moroz & ANDREI TSURYKAU 365

Vanderbylia cinnamomea sp. nov. from southwestern China

JUN-ZHU CHEN, XIONG YANG, CHANG-LIN ZHAO 371

G.H. Cunningham’ use of te reo Maori in fungal epithets

SHAUN R. PENNYCOOK 383

Re-collection of secotioid Entoloma calongei in Europe

Ivona KAUTMANOVA, VACLAV KAUTMAN, VIKTOR KUCERA, DARINA ARENDT 405

Xerula setulosa, a new Neotropical agaric record for Argentina

Maria M. ALBERTI, NICOLAS NIVEIRO, EDGARDO O. ALBERTO 415

Haematomma rubidum sp. nov. from China

RONG TANG, AN-CHENG YIN, ZUN-TIAN ZHAO 425

Armillaria xiaocaobaensis sp. nov. from China

Jia-Hua PENG & CHANG-LIN ZHAO 431

Phylogeny and morphology of Ellismarsporium parvum and

the new combination E. varium Kar ZHANG, WEIHUA Guo,

DayNET SOSA, FREDDY MAGDAMA, LIZETTE SERRANO,

ELAINE Matosso, De-WEI L1, RAFAEL F. CASTANEDA-Ru1z 443

Acarospora scottii and Sarcogyne paradoxa spp. nov.

from North America Kerry KNUDSEN & JANA KocourkovA 453

MycosioTa (FUNGAE) NEW TO THE MYCOTAXON WEBSITE

A checklist of marine fungi from Australia (SUMMARY)

SALLY C. FryAR, KEVIN D. Hype, Davip E.A. CATCHESIDE 465

A checklist of corticioid fungi (Agaricomycetes, Basidiomycota)

from Brazil (SUMMARY) RENATA S. CHIKOWSKI, CARLA R. S. DE LirA,

Kar. HENRIK LARSSON, TATIANA B. GIBERTONI. 467

Occurrence of Glomeromycota species in aquatic habitats:

a global overview (SUMMARY) MartANA BESSA DE QUEIROZ, KHADIJA JOBIM,

XOCHITL MARGARITO VISTA, JULIANA APARECIDA SOUZA LEROY,

STEPHANIA RUTH BaASiLio SILVA GOMES, BRUNO TOMIO GOTO 469

VI ... MYCOTAXON 135(2)

PUBLICATION DATE FOR VOLUME ONE HUNDRED THIRTY-FIVE (1)

MYCOTAXON for JANUARY-MARCH 2019 (I-xIV + 1-234)

was issued on April 20, 2020

APRIL-JUNE 2020...

NOMENCLATURAL NOVELTIES AND TYPIFICATIONS

PROPOSED IN MYCOTAXON 135(2)

Acarospora scottii K. Knudsen & Kocourk.

[MB 831962], p. 455

Armillaria xiaocaobaensis Jia H. Peng & C.L. Zhao

[MB 832231], p. 436

Cercosporella bundelkhandae (S. Shrivast., N. Verma & A.N. Rai)

Raghv. Singh, S.K. Verma, S. Yadav & Sh. Kumar

[MB 831083], p. 317

Coprinellus ovatus Kamran & Jabeen

[MB 830529], p. 326

Ellismarsporium varium (Alves-Barb., Malosso & R.F. Castafieda)

K. Zhang, Alves-Barb., Malosso & R.F. Castafieda

[MB 832010], p. 448

Haematomma rubidum R. Tang & Z.T. Zhao

[MB 835897], p. 426

Microthecium inferius (Udagawa & Cain) D.W. Li, R.F. Castafieda &

N.P. Schultes

[MB 831435], p. 307

Ramiphialis ER. Barbosa, Fiuza & R.E. Castafieda

[MB 831423], p. 294

Ramiphialis ronuroensis ER. Barbosa, Fiuza & R.E Castaneda

[MB 831424], p. 294

Sarcogyne paradoxa Kocourk. & K. Knudsen

[MB 831963], p. 458

Vanderbylia cinnamomea C.L. Zhao

[MB 831788], p. 375

Westerdykella aquatica H.Y. Song & D.M. Hu

[MB 825645], p. 287

VII

VI ... MYCOTAXON 135(2)

CORRIGENDA

MYCOTAXON 134(2)

p. iv, line 14 For: A. NASEER, S. GHANI, A.R. N1Azi, A.N. KHALID 241

READ: A. NASEER, S. KHANUM, A.R. NriAzi, A.N. KHALID 241

p.241,line6 For: A. NASEER™, S. GHANT’, A.R. Niaz’, A.N. KHALID?

READ: A. NASEER™, S. KHANUM?, A.R. Niazi’, A.N. KHALID?

APRIL-JUNE 2020...

REVIEWERS — VOLUME ONE HUNDRED THIRTY-FIVE (2)

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.

Mohamed A. Abdel-Wahab

Grazina Adamonyté

Vladimir Antonin

Juliano Marcon Baltazar

Flavia Rodrigues Barbosa

Janusz Blaszkowski

Tor Erik Brandrud

Taimy Cantillo-Pérez

Zhimin Cao

Rafael FE Castaneda Ruiz

Yuan- Yuan Chen

Vagner Gularte Cortez

Patricia Oliveira Fiuza

Adam Flakus

Eduardo Furrazola Gomez

Masoomeh Ghobad-Nejhad

Otto Gockman

Shouyu Guo

Luis Fernando Pascholati Gusmao

Alejandro Huereca

Holden Hohaia

Jan Holec

Shah Hussain

Sana Jabeen

Klaus Kalb

Kerry Knudsen

De-Wei Li

Jian Ma

Helmut Mayrhofer

John McCarthy

Eric H.C. McKenzie

Nelson Menolli Jr.

Josiane Santana Monteiro

Gabriel Moreno Horcajada

Olga V. Morozova

Rikke Reese Naesborg

Lorelei L. Norvell

Ka-Lai Pang

Omar Paino Perdomo

Shaun R. Pennycook

Sergio Pérez Gorjon

T.A.M. Jagadeesh Ram

Geoff Ridley

Amy Y. Rossman

Mark R.D. Seaward

Ertugrul Sesli

Danielle Karla Alves da Silva

Harrie Sipman

Susumu Takamatsu

Muhammad Usman

Else C. Vellinga

Allison Walker

Huang Zhang

Changlin Zhao

xX ... MYCOTAXON 135(2)

2020 MYCOTAXON SUBMISSION PROCEDURE

Prospective Mycotaxon authors should download the Mycotaxon 2020 guide,

review & submission forms, and MycoTaxon sample manuscript by clicking the ‘file

download page’ link on our INSTRUCTIONS TO AUTHORS page before preparing their

manuscript. This page briefly summarizes our “4-step’ submission process.

1—PEER REVIEW: Authors first contact peer reviewers (two for journal papers;

three for mycobiota/fungae) before sending them formatted text & illustration

files and the appropriate 2020 MycoTaxon journal or mycota reviewer comment

form. Experts return revisions & comments to BoTH the Editor-in-Chief

<editor@mycotaxon.com> and authors. ALL co-authors Must correct and proof-

read their files before submitting them to the Nomenclature Editor.

2—NOMENCLATURAL REVIEW: Authors email all ERROR-FREE text & illustration

files to the Nomenclature Editor <PennycookS@LandcareResearch.co.nz>.

Place first author surname + genus + ‘“Mycotaxon’ on the subject line, and

(required) attach a completed SUBMISSION FORM. The Nomenclature Editor will

(i) immediately assign the accession number and (ii) after a few weeks return his

notes and suggested revisions to the author(s) and Editor-in-Chief.

3—FINAL SUBMISSION: All coauthors thoroughly revise and proof-read files

to prepare error-free text and images ready for immediate publication. Poorly

formatted copy will be rejected or returned for revision. E-mail the final manuscript

to the Editor-in-Chief <editor@mycotaxon.com>, adding the accession number to

the message and all files, which include a (i) revised 2020 submission form, all (ii)

text files and (iii) jpg images, and (iv) FN, IF, or MB identifier verifications for each

new name or typification. The Editor-in-Chief acknowledges submissions within

two weeks of final submission but requests authors to wait at least 14 days before

sending a follow-up query (without attachments).

4—FINAL EDITORIAL REVIEW & PUBLICATION: The Editor-in-Chief conducts a

final grammatical and scientific review and returns her editorial revisions to all

expert reviewers and coauthors for final author approval. Author-approved files

are placed in the publication queue.

The PDF proof and bibliographic & nomenclatural index entries are sent to all

coauthors for final inspection. After PDF processing, the Editor-in-Chief corrects

ONLY PDF editorial/conversion and index entry errors; corrections of all other

errors are listed in the ERraTA of a subsequent issue for no charge. Authors will pay

fees for mycobiota uploads, optional open access, and correction of major author

errors to the Business Manager <subscriptions@mycotaxon.com> at this time.

MyYcoTAXON LTD— www.mycotaxon.com

The Mycotaxon Webmaster <mycotaxon@gmail.com> posts announcements,

subscription & publication information, and author forms & templates on the official

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of Fungae (regional annotated species lists).

MYCOTAXON ONLINE— www.ingentaconnect.com/content/mtax/mt

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

and subscription articles are offered.

APRIL-JUNE 2020 ... XI

FROM THE EDITOR-IN-CHIEF

COLLABORATION & THE MULTI-AUTHORED PAPER— Mycological taxonomic research

is no longer a simple matter of reaching down, plucking a fungus from its substrate,

seeing something REALLY different, checking the microscopical characters, reading

the one or two papers ever written on aforesaid fungal family, and then writing a

paper. Current fungal taxonomy encompasses a great many disciplines, including

(but not limited to) microscopy, ecology, physiology, mating studies, ultrastructure,

systematics, DNA extractions, sequence analyses, and evolutionary relationships. Yet

we somehow expect students injected into this bewildering maelstrom of disciplines

to be able (eventually) to present their research in the form of a published paper.

As only a limited number of geniuses can master all modern disciplines, there

are now relatively few solo-authored papers. Most research papers are crafted by an

agreeable consortium of individuals, each of whom contributes one or two expert

pieces that flesh out the final manuscript. The individual who bears the greatest

responsibility (and gets—however little deserved—most of the credit) is the first

author. All students must oversee all stages of each manuscript submission to ensure

that they master the scientific publication process. The first named author bears the

responsibility for ensuring that a manuscript meets all journal requirements and

seeing that all facts are coherently written and correct. Generally it is that author

who discusses the research with all whose names will stand on the final publication.

It is also generally the first author who gathers all contributors together to work out

a complication and perfect the final text whenever an error or oversight is detected.

Nonetheless, a multi-authored paper is NoT a work by one author. Journals such

as MyCOTAXON expect ALL contributing authors to read and revise numerous drafts

thoroughly, to check all drawings, tables, and data, and communicate with the first

author throughout the research process and prior to submission. Editors expect to

receive papers that have been—by all authors—thoroughly proofed and that contain

no scientific, grammatical, or visual errors. Only through rigorous oversight by all

collaborators, particularly the more experienced and knowledgeable among them,

is it possible for a beginning scientist to publish in a well-established international

scientific journal.

We heartily urge all of our authors to discuss their research among themselves

and with their lab mates. That way lies progress!

DEAR READERS (PLUS PREMATURE THANKS TO THE MYCOTAXON EDITORIAL BOARD)—

Your editors are currently swamped with submissions—the expected result of

discontinuing required page charges in 2018. For sixteen years, Dr. Pennycook

has tirelessly accessioned and examined manuscripts, gradually adding scientific

and grammatical oversight to his already heavy nomenclatural duties. In the

process, he has nomenclaturally accepted almost 2000 manuscripts, rejected ~100,

and is currently reviewing 105 more. In its search for scientifically rigorous and

nomenclaturally correct papers produced by dedicated authors and their expert

XII ... MYCOTAXON 135(2)

reviewers, Mycotaxon still needs Shaun’s nomenclatural knowledge because so few

mycologists fully understand the rules of the International Code of Nomenclature.

Needless to say, both authors and editors are unhappy with the current lengthy

time between manuscript accession and return of the nomenclatural review.

However, it now appears that all too often, author-selected experts do not devote

the time needed for rigorous review, so that Dr. Pennycook frequently accessions

manuscripts that require much more work. With many problems not detected by

a quick scan of accessioned files, frequently only after Shaun has begun a review

does he discover that the unsatisfactory files must be returned to authors for more

work. Not surprisingly, this further lengthens the time between accession and

nomenclatural approval.

Three months ago we decided that Shaun could work through his backlog more

quickly by focusing only on nomenclature and by returning all other grammatical

and scientific edits to the Editor-in-Chief (me). Unfortunately after encountering far

too many scientific errors or inaccurate citations, I now must also double-check Gen-

Bank, herbarium records, and — yes - even scale bars for accuracy and consistency.

Fortunately, learning in June that your two volunteer editors needed at least one

extra pair of eyes, editorial board member Else Vellinga volunteered to serve as

Mycotaxon’s first ‘official’ PRELIMINARY READER. She offered to glance through still

unreviewed manuscripts to determine which authors properly implemented expert

suggestions, formatted grammatically correct text files, and presented no scientific

contradictions as well as flag those papers needing additional work. Dr. Vellinga

surveyed several manuscripts—happily finding one in excellent shape and less

happily finding several in need of considerable repair or additional research prior to

nomenclatural review.

In our estimation, Else, who delivered thorough reviews forwarded to the

authors, has been a rousing success. We now plan to contact other board members to

help us determine which other accessions are ready for nomenclatural review, which

need additional research, and which should be rejected. Authors able to make any

suggested changes and return them before Shaun begins his review will not lose their

place in the nomenclatural ‘queue:

We sincerely hope this will help shorten our review turn-around time and give

your volunteer editors much needed assistance in their currently overwhelming

editorial duties. We both dearly desire returning to a time when all submissions were

so well prepared that all we need do is to rubber-stamp “approved” after the first

read-through. “Tis a consummation devoutly to be wished.

MycoTAXxON 135(2) provides 22 papers and announces three mycobiota by 88

authors (from 23 countries) reviewed by 54 experts.

The 2020 April-June MycoTaxon sets forth ONE new genus (Ramiphialis from

Brazil) and EIGHT species new to science representing Acarospora & Sarcogyne from

APRIL-JUNE 2020 ... XIII

the US & CANADA; Armillaria, Haematomma, Vanderbylia, and Westerdykella from

Cuina; Coprinellus from Paxistan; and Ramiphialis from Brazit. Authors also

offer three new combinations in Cercosporella, Ellismarsporium, and Microthecium.

Range extensions are reported for [ascomycetes] the powdery mildew

Golovinomyces (CHINA) and truffle Elaphomyces (TuRKEy); [basidiomycetes] the

agarics Pluteus (EUROPE) & Xerula (ARGENTINA), secotioid Entoloma (SLOVAKIA),

and rust Aecidium (PakIsTAN); [lichens] the leprose Lepraria (IRAN), assorted

graphidoid & thelotremoid taxa (INDIA), and crustose Carbonea & Rinodina

(POLAND); and [myxomycetes] the slime molds Badhamia & Trichia (BELARUS).

We also offer phylogenetic explorations of Acrospeira, Armillaria, Coprinellus,

Ellismarsporium, Golovinomyces, Pluteus, Vanderbylia, and Westerdykella; a key to

the Lepraria species of Iran; reevaluation of three Calicium taxa proposed in 1909;

and a comprehensive index of fungal names derived from te reo Maori. Finally, we

announce three new www.mycotaxon.com mycobiota covering Australian marine

fungi, Brazilian corticioids, and a global list of aquatic glomeromycotans.

Best wishes for excellent health and with warm regards,

Lorelei L. Norvell (Editor-in-Chief)

8 July 2020

MYCOTAXON

April-June 2020—Volume 135, pp. 235-244

https://doi.org/10.5248/135.235

Four new Lepraria species for Iran,

with a key to all Iranian species

SAREH SADAT KAZEMI’, [RAJ MEHREGAN”*, YOUNES ASRI’,

SARAH SAADATMAND’, HARRIE J. M. SIPMAN?

' Department of Biology, Science and Research Branch, Islamic Azad University,

Tehran, Iran

? Research Institute of Forests & Rangelands, Agricultural Research,

Education & Extension Organization (AREEO),

P.O. Box 13185-116, Tehran, Iran

° Botanischer Garten und Botanisches Museum, Freie Universitat,

K6nigin-Luise Strafse 6-8, D-14195 Berlin, Germany

* CORRESPONDENCE TO: imehregan@srbiau.ac.ir

ABSTRACT—Lepraria eburnea, L. ecorticata, L. jackii, and L. leuckertiana, found in deciduous

forests in Golestan province, are recorded for the first time from Iran. Descriptions and

illustrations of all four species and a key to all Lepraria species known from Iran are provided.

Key worps—Hyrcanian forest, leprose lichens, sterile lichens

Introduction

Species of Lepraria Ach. (Stereocaulaceae, Lecanoromycetes) are

characterized by thalli completely composed of more or less clusters of soredia

resting on a continuous medulla (Lendemer 2013). They lack ascocarps or

conidangia; therefore with few morphological features available as taxonomic

characters, the presence of secondary metabolites plays a significant role in

distinguishing species (Leuckert & al. 1995, Saag & al. 2009). DNA analyses

by Ekman & Tonsberg (2002) confirmed the taxonomic status of the genus

and its species. ITS analysis by Bayerova & al. (2005) showed that species

may be distinguished through subtle differences in aliphatic acid content,

difficult to trace using the usual TLC methods.

236 ... Kazemi & al.

Lepraria is widely distributed with 61 species currently known worldwide

(Sipman 2004, Saag & al. 2009, Matwiejuk 2017). In Asia it is known from

Armenia, China, India, Mongolia, Pakistan, Singapore, Tajikistan, and

Turkey. Eleven species have been reported from Iran: Sohrabi & Orange (2006)

and Sohrabi & al. (2010) report L. lobificans Nyl. and L. diffusa (J.R. Laundon)

Kukwa from Golestan and Mazandaran provinces, L. vouauxii (Hue) R.C.

Harris from East Azerbaijan and Golestan provinces, L. crassissima (Hue)

Lettau from Mazandaran, and L. alpina (B. de Lesd.) Tretiach & Baruffo and

L. nivalis J.R. Laundon from Zanjan province; Valadbeigi & al. (2009) report

L. isidiata (Llimona) Llimona & A. Crespo from Ilam; Haji Moniri & Kukwa

(2009) report L. caesioalba (B. de Lesd.) J.R. Laundon and L. rigidula (B. de

Lesd.) Tonsberg from Northern Khorasan; Kukwa & Sohrabi (2008) report

L. impossibilis Sipman from Mazandaran province; and Dyanat-Nejad &

Keramedini (1993) report L. membranacea (Dicks.) Vain. from Tehran

province.

The Golestan province is particularly promising for lichens due to

its extensive Hyrcanian forests dominated by Parrotia persica (DC.)

C.A. Mey., Quercus castaneifolia C.A. Mey., Acer velutinum Boiss., Gleditsia

caspia Desf., Pyrus boissieriana Buhse, and Alnus subcordata C.A. Mey.

(Akhani & al. 2010). Only three Lepraria species have previously been

identified from Golestan province (Sohrabi & Orange 2006): L. lobificans,

L. diffusa, and L. vouauxii. We report new to Iran and describe four

additional species from Naharkhoran Forest in Golestan found in wet

and shady habitats, often on thick Quercus bark and on mosses over bark.

We also provide a key to all Lepraria species known from Iran, including

L. finkii (B. de Lesd.) R.C. Harris, which has been confused with

L. lobificans (Lendemer 2013).

Material & methods

The specimens are deposited in the Herbarium, Research Institute of Forests

and Rangelands, Tehran, Iran (TARI) and were compared with herbarium

samples in the Herbarium, Botanischer Garten und Botanisches Museum Berlin-

Dahlem, Berlin, Germany (B). Morphological and anatomical characters were

observed with a Luxeo 4D stereomicroscope and a Zeiss compound microscope.

The chemistry was analyzed with thin layer chromatography (TLC) following

Orange & al. (2010) and Elix (2014) using solvent systems A (toluene, dioxane,

acetic acid), B’ (hexane, diethyl ether, formic acid), and C (toluene, acetic acid).

For identification, the keys of Saag & al. (2009) and Knudsen & Elix (2007) were

also used.

Lepraria spp. new for Iran ... 237

Taxonomy

ae ty Maen x

Fic. 1. Lepraria eburnea (TARI, Kazemi 9023).

Lepraria eburnea J.R. Laundon, Lichenologist 24: 332. 1992. Fig. 1

Type: British Isles, England, London Borough of Hammersmith and Fulham (V.C. 21,

Middlesex), All Saints Fulham churchyard, 51/243759, in slight shade on moss, brick,

and mortar on vertical surface of brick wall on east side of churchyard, 30.vi.1987, on

calcareous rocks, J.R. Laundon 3185 (BM [n.v.], holotype).

Thallus white-grey to white-green, turning pale pink in the herbarium due

to its alectorialic acid content, thick, leprose, placodioid, without wrinkled

margin, diffuse, persistent and clearly two-layered, initially forming small

patches of thallus that are later incorporated into the margin, without lobes;

hyphae unilateral, hyaline, septate; initially with prothallus; hypothallus with

variously developed layer of hyphae around and under the granules, thick and

white, without rhizohyphae, granules spherical, fuzzy, and ecorticate, with

short projecting hyphae, forming multiplex units (consoredia).

CuHEMiIstTRy: thallus K+ yellow, C+ yellow, KC+ pink.

TLC: Alectorialic acid, protocetraric acid.

SPECIMEN EXAMINED: IRAN, GOLESTAN PROVINCE: Gorgan, Alangdareh Forest,

around health road, 36.7928°N 54.4504°E, 341 m, 16.11.2017, Kazemi 9023 (TARI).

COMMENTS: Our material agrees with the most detailed descriptions of

L. eburnea presented by Saag & al. (2009), Kukwa (2006), and Tsurykau &

al. (2016).

238 ... Kazemi & al.

Lepraria eburnea is similar to L. nivalis, in that both species have

protocetraric acid and grow on mosses; however, L. nivalis differs by its

growth on mosses covering calcareous rocks, not bark and its absence of

alectorialic acid.

Lepraria eburnea is morphologically similar to L. finkii in that both have a

thick white medulla; they are separated by chemical differences: L. finkii has

atranorin, stictic acid, and zeorin as secondary metabolites.

Hasitat: Lepraria eburnea grows on sheltered microhabitats in

mountainous regions as well as on partially shaded moss, brick, and mortar

on vertical brick wall surfaces and on acidic substrates, soil, Alnus glutinosa,

and Quercus robur (Laundon 1992, Kukwa 2006, Lendemer 2013). The

specimen reported here was found on mosses on large Quercus castaneifolia

tree in Hyrcanian forest.

GEOGRAPHICAL DISTRIBUTION: Asia (Armenia, Turkey) (Gasparyan &

Sipman 2016, Cobanoglu & Akdemir 2000), Europe, North America (Saag

2009).

Lepraria ecorticata (J.R. Laundon) Kukwa, Mycotaxon 97: 64. 2006 Fic. 2

Tye: British Isles, England, Devon [vice-c 4], Ilfracombe, Torrs Walks, 21/50 47, on

shaded vertical rocks, 1 September 1971, J.R. Laundon 2851 (BM [n.v.], holotype).

Thallus yellowish-green to bluish-grey, leprose; without margin, granules in

to irregular mass, without lobes; medulla thin and pale, rarely differentiated,

often absent; hyphae aggregate, without septa, pale, covered with granular

crystals, soredia abundant, without projecting hyphae, ecorticate, mostly fine

to medium, well separated from one another.

CHEMISTRY: K+ pale yellow, C—, KC-, Pd-.

TLC: Atranorin, usnic acid, zeorin.

SPECIMEN EXAMINED: IRAN, GOLESTAN PROVINCE: Gorgan, Naharkhoran road,

Delbar mountain, 36.7223°N 54.5490°E, 1049 m, 17.11.2017, Kazemi 8784 (TARI).

CoMMENTSs: Our specimen agrees with the detailed descriptions of L. ecorticata

presented by Kukwa (2006), Saag & al. (2009), Laundon (2003), Flakus &

Kukwa. (2007).

In Iran Lepraria ecorticata is similar chemically with L. leuckertiana, as

both have usnic acid and zeorin, but L. ecorticata is distinguished by having

well-separated soredia and lacking a distinct medulla and marginal lobes,

while L. leuckertiana has irregular and greatly coalescing soredia, a thick

medulla, and minute marginal lobes.

Lepraria spp. new for Iran ... 239

Fic. 2. Lepraria ecorticata (TARI, Kazemi 8784).

Hapsirat: Lepraria ecorticata grows on shaded places on acidic rocks, on

siliceous and calcareous rocks and walls, and (rarely) on the bark of Populus,

Tilia, and Fraxinus (Laundon 2003, Tsurykau & al. 2016). The specimen

reported here was found on Parrotia persica in montane Hyrcanian forest.

GEOGRAPHICAL DISTRIBUTION: Asia (China), Europe, North America,

South America (Kukwa 2006).

Lepraria jackii Tonsberg, Sommerfeltia 14: 200. 1992. FIG. 3

Type: Norway, Nord-Trondelag, Grong, W of Abrahammyra, 80-100 m, 16.ix.1981,

on Picea abies, T. Tonsberg 6176 (BG [n.v.], holotype).

Thallus greenish-grey to whitish-green, leprose, without stratified and

alternate margin, at first forming separated granules that later accumulate to

continuous masses, without marginal zone, without lobes; hyphae unilateral,

hyaline; without prothallus; hypothallus absent; without rhizohyphae;

granules spherical, well developed, separate, and forming multiplex units;

medulla present, white, sparse to continuous; soredia abundant, without

projecting hyphae, ecorticate, small, dispersed or forming a thick, continuous

layer, sometimes in larger clusters (consoredia).

CuHeEmistry: Thallus K+ yellow, C-, KC-, P+ yellow.

TLC: Atranorin (fatty acids are also reported but were not clearly visible on

our plates).

240 ... Kazemi & al.

SPECIMEN EXAMINED: IRAN, GOLESTAN PROVINCE: Gorgan, Naharkhoran road,

around environmental station, 36.7911°N 54.4665°E, 387 m, Kazemi 9019 (TARI).

CoMMENTs: The Iranian material does not show significant differences from

the detailed descriptions of L. jackii presented by Kukwa (2006), Czarnota

& Kukwa (2001), Matwiejuk (2017), Saag & al. (2007), and Tsurykau & al.

(2016).

Lepraria jackii can be confused with L. caesioalba as both have atranorin

and similar reactions with K and P, but L. caesioalba can be distinguished

morphologically by its obscure sublobes while L. jackii completely lacks lobes.

Fic. 3. Lepraria jackii (TARI, Kazemi 9019).

a. Thallus; b. Photobiont cells.

Hasitat: Lepraria jackii grows on conifers (Picea abies) and rarely on

sheltered rocks or humus, on mosses and soil, and sometimes on other lichens

(Tonsberg 1992, Saag & al. 2007, Lendemer 2013). The specimen reported

here was found on Zelkova carpinifolia bark in a Hyrcanian forest.

GEOGRAPHICAL DISTRIBUTION: Asia (Armenia) (Gasparyan & Sipman

2016), Europe, North America, Australia (Saag 2009).

Lepraria leuckertiana (Zedda) L. Saag, Lichenologist 41: 41. 2009. Fic. 4

TyPE: Italy: Sardinia: Prov. Sassari: Illorai, Monte Artu, 1494647E, 4472420N (Gauss-

Boaga), 840 m, on bark and mosses on old Quercus pubescens Willd. trees in a mixed

wood of Q. pubescens and Q. ilex L., in open conditions, 9.4.1997, L. Zedda 1800(2) (B

[n.v.], holotype; herb Zedda and SS [n.v.], isotypes).

Thallus greenish-white to greenish-grey, leprose, thick and powdery

to granular; without margin, often sublobate, hyphae condensed, pale;

prothallus absent; medulla white, exposed in some parts, forming a thick

layer; soredia abundant, granular, fine to coarse, gathered into consoredia.

Soralia ecorticate, with short projecting hyphae, hyphae to 4-5 um diam., pale

and branched, unorganized.

CHEMIstTRY: Thallus K-, C-, Pd-, KC-.

Lepraria spp. new for Iran... 241

TLC: Usnic acid, zeorin; isousnic acid is also reported but was not clearly

visible on our plates.

SPECIMEN EXAMINED: IRAN, GOLESTAN PROVINCE: Gorgan, Naharkhoran road,

Delbar mountain, 36.7467°N 54.5001°E, 1092 m, 17.11.2017, Kazemi 8784 (TARI).

COMMENTS: Our specimen agrees fully with the detailed descriptions of

L. leuckertiana presented by Saag & al. 2009, Zedda 2000.

Morphologically L. leuckertiana resembles L. vouauxii but differs in spot

test and chemistry; chemically L. leuckertiana resembles L. ecorticata and

differs by the strongly agglutinating, not well separated soredia.

Hasitat: Lepraria leuckertiana grows on old trees (especially on

Quercus), on palm and olive trees, and on sandy soil and roots (Zedda 2000).

The specimen reported here was found on Quercus castaneifolia bark in a

montane Hyrcanian forest.

GEOGRAPHICAL DISTRIBUTION: Asia (Gasparyan & Sipman 2016, Paukov

& al. 2017), Africa, Europe, South America (Saag 2009).

Key to the Lepraria species in Iran

1 Thallus surface granular; soredia coarse, >100 um in diam.,

somefinres-clongatecand isidiuinr= like) FF 8 cee Be ans Sea eh ane rates eee «een eae SS 2

Thallus surface powdery; soredia mostly <80 um in diam. .................0.. 5

2 On mineral soil; fumarprotocetraric acid present;

granules indistinct, partly elongate and isidioid ................... L. isidiata

On rock or moss over rock; fumarprotocetraric acid absent;

STAniMles-CistiGt-AVOtISTAIOLAS 0. Fs Pee nak s espeuk Sone .a/ fee ccaiolt ee cnet k Specs eeu rae 3

242 ... Kazemi & al.

3 Stictic acid present; thallus K+ yellow to orange .................00. L. lobificans

Stictic acid absent; thallus K- or + pale yellow to pale brown .................. =

4 Thallus K+ brown, Pd + red, with protocetraric acid................2--- L. nivalis

Thallus K+ yellow, Pd+ yellow, with psoromic acid and atranorin L. caesioalba

5 Alectorialic and protocetraric acids present (staining paper brown in herbarium);

Mme aulladthickeaiid (GaStiiet= is Hcs ek whee ede Lod ace fe deer huge age L. eburnea

Alectorialic acid absent (not staining paper in herbarium) .................... 6

6 Divaricatic acid with nordivaricatic acid and zeorin present,

Sore MAT TEM: sos seguir dG Meee eee omen agendas oe dine Bad hs L. crassissima

IV ARIC ALICIA CIC ADSONE. o's omer naslat longo Ale arcs gyrtetat eitngs seeks china Gedo wie ae ep gn ve

7 Stictic acid present; soredia K+ yellow to orange .................. 00000 L. finkii

SUCH GACICRADS OE on Pees Baba aes Waele eats Sea's & ae Ree eed TRE eA ot 8

8 Pannaric acid (or pannaric acid derivatives) as primary substance ............... 9

Pannaric acid and its derivatives absent or present only in trace

to minor amounts; other major compound(s) present ................2--- 12

9 4-Oxypannaric acid 2-methylester as primary substance ................ L. diffusa

4-Oxypannaric acid 2-methylester absent or present in only trace to minor

amounts; other major dibenzofurans present ............. 0... c cece eee eee 10

10 Thallus with distinct lobes; pannaric acid as primary substance,

other minor dibenzofurans also present...............----- L. membranacea

Thallus indistinctly or not lobed; pannaric acid absent or present only in

trace to minor amounts, pannaric acid-6-methylester present ............. 11

11 Thallus with distinct lobes with raised marginal rim; lecanoric acid and

pannaric acid-6-methylester present; C+ red .................. L. impossibilis

Thallus with indistinct lobes without marginal rim;

pannaric acid 6-methylester alone present; C- .................. L. vouauxii

2°Porphiyrilic acid present: A wa tyra oe wi wae rat ae debe bak ONY Sig dt Sten L. alpina

Porphyrilieacid ‘absent thallus powdery .n.cs - 1 28 «os Se et hace geediece ce ole enaltnen 13

LS Vshic ACM ANCEZeOrIN resem tre ete AE A YS we Pe Bes oe tet oe 14

Usnic acid‘absent;-zeorin-absent.or present. 2.4 fP.. 64 74¢ang Cad odthkpe ee mes 15

PA viedullathyele and cOteony sack 5. bez dri: sehen web stan ve evae esp Puede A dbp ape ddd L. leuckertiana

WMediilla thin GADSCIt wt. gc lt tuts set Pete ee ese ee .s eulee 2 Sail eae 2 L. ecorticata

15 Soredia small, without projecting hyphae ......................0 000 eee L. jackii

Soredia, large, with projecting Nyphae wuss cee ela vee la to dea Maley pele one L. rigidula

Acknowledgments

The authors appreciate the cooperation of the Research Institute of Forests and

Rangelands and the Herbarium of Berlin Botanical Museum for the facilities provided.

The authors are grateful to Prof. Mark R. D. Seaward (University of Bradford, UK) and

Prof. Helmut Mayrhofer (University of Graz, Austria) for reviewing the manuscript.

Lepraria spp. new for Iran ... 243

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MYCOTAXON

April-June 2020—Volume 135, pp. 245-274

https://doi.org/10.5248/135.245

Pluteus dianae and P. punctatus resurrected,

with first records from eastern and northern Europe

HANA SEVCfKOVA"’, EKATERINA F. MALYSHEVA?,

ALFREDO JUSTO’, JACOB HEILMANN-CLAUSEN‘*, MICHAL TOMSOVSKY°

" Department of Botany, Moravian Museum, Zelny trh 6, CZ-659 37 Brno, Czech Republic

? Komarov Botanical Institute of the Russian Academy of Sciences,

RUS-197376, Saint Petersburg, Russia

> New Brunswick Museum, 277 Douglas Ave, Saint John, E2K 1E5-NB, Canada

* Centre for Macroecology, Evolution & Climate, Natural History Museum of Denmark,

University of Copenhagen, Universitetsparken 15, DK-2100 Copenhagen, Denmark

° Faculty of Forestry and Wood Technology, Mendel University in Brno,

Zemédélska 1, CZ-613 00, Brno, Czech Republic

* CORRESPONDENCE TO: hsevcikova@mzm.cz

ABSTRACT— The type specimens of Pluteus dianae and P. punctatus from the Czech Republic

were studied morphologically and molecularly. New collections identified by nrITS sequence

analyses extend the distribution of P dianae to Denmark, European Russia, and the Asian

part of Turkey and of P. punctatus to Sweden. The application of these names is discussed;

both belong in the P. plautus complex, and data on European and North American taxa in

this complex are summarised and compared with P. dianae and P. punctatus. Pluteus aestivus

is considered a nomen dubium.

Key worps—Agaricales, Czechia, Pluteaceae, Pluteus sect. Hispidoderma, taxonomy

Introduction

Pluteus Fr. (Pluteaceae, Agaricales) is an agaricoid genus forming

basidiomata characterized by free lamellae, a pinkish spore print, smooth

globose to ellipsoid (rarely oblong) basidiospores, an inverse hymenophoral

trama, and the presence of cheilocystidia and (often) pleurocystidia (Singer

1986, Vellinga 1990). Singer (1986) distinguished three sections in the genus:

246 ... Sevéikova & al.

Pluteus sect. Pluteus, P. sect. Hispidoderma Fayod, and P. sect. Celluloderma

Fayod (the last comprising P. subsect. Eucellulodermini Singer and P. subsect.

Mixtini Singer). Pluteus sect. Hispidoderma is characterized by non-metuloid

cystidia and a pileipellis arranged as a trichoderm or hymeniderm (Vellinga &

Schreurs 1985, Justo & al. 2011a); P dianae Pilat and P. punctatus Wichansky

belong in this section. Within the section, according to Pilat (1968), Vellinga

(1990), and in concordance with our morphological and phylogenetical

studies, both species can be broadly ascribed to the species complex around

P. plautus (Weinm.) Gillet. Although many species have been described in

the P plautus complex, there still exists taxonomic uncertainty regarding the

identity of many of its species. Vellinga & Schreurs (1985) placed 13 names

in synonymy with P plautus, while other authors who maintained a narrower

taxonomic concept (e.g., Orton 1986) accepted several species in this group

(e.g., PR. dryophiloides P.D. Orton, P. punctipes P.D. Orton, P granulatus Bres.,

P. depauperatus Romagn.). Recent molecular phylogenies challenge the

broad taxonomic concept of Vellinga & Schreurs and emphasize the need for

reevaluating names in this complex based on a combination of morphological,

ecological, and molecular data (Justo & al. 2011b). In this article we take a first

step toward clarifying the taxonomy of the P plautus complex by focusing on

type studies of P dianae and P. punctatus.

Pluteus dianae was described by Pilat (1968) and P. punctatus by Wichansky

(1972) based on collections from Czechia (formerly Czechoslovakia). However,

no subsequent collections have been reported and no further information

on these species has ever been published. Pilat (1968) described P dianae as

macroscopically similar to P. pellitus (Pers.) P. Kumm., belonging to P. subsect.

Depauperati Lange, a taxon that almost corresponds to P. sect. Hispidoderma

(Lange 1917, Lange 1936, Vellinga & Schreurs 1985). Wichansky (1972)

characterized P punctatus by a greyish-brown pileus with dark brown floccose

squamules, a whitish stipe densely covered with brown felted squamules, and a

pileipellis consisting of very long cylindrical elements.

Materials & methods

Morphology

Macroscopic descriptions of the collected specimens are based on fresh basidiomata.

Colour abbreviations follow Kornerup & Wanscher (1983). Morphological terminology

follows Vellinga (1988). Microscopic features were studied by H. Sevéikova (holotypes

of P. dianae, PB. depauperatus, P. hiatulus, P. inflatus, P puberulus, P. punctatus, and

Czech collections of P granulatus, P. plautus sensu lato, and P. semibulbosus sensu lato)

on dried material mounted in Congo red using an Olympus BX-50 light microscope

Pluteus dianae & P. punctatus in eastern & northern Europe ... 247

with magnifications of 400x and 1000x. All Russian collections were studied by E.F.

Malysheva using an ammonia Congo red solution under a Zeiss Axio Scope.Al

microscope. Collections studied by A. Justo include material from Denmark, Sweden,

and all North American holotypes; microscopic features were studied on dried material

mounted in 5% KOH or an ammonia Congo red solution under a Leica DFC2500

microscope. Microscopic descriptions were based on at least 30 measurements each of

basidiospores, cheilocystidia, pleurocystidia, and caulocystidia and 25 measurements

of basidia. The expression 1/1/30 for basidiospores means 1 collection / 1 basidioma /

30 basidiospores measured in total. Abbreviations: avl = mean of basidiospore length;

avw = mean of basidiospore width; avg. = average; Q = quotient of length and width in

any one basidiospore; avQ = mean of basidiospore Q-values. Herbarium abbreviations

follow Thiers (2018). Authors of fungal names are cited according to the International

Plant Names Index website (http://www.ipni.org).

Molecular phylogeny

DNA EXTRACTION—The type collections of P dianae and P. punctatus were

sequenced to establish their position in P. sect. Hispidoderma. Only dried herbarium

specimens were used for DNA extraction. M. TomSovsky extracted the DNA from

both holotypes following the protocol of the Qiagen PowerSoil DNA Isolation Kit to

step no. 13 (purification of DNA with C2 and C3 buffers) and then continued with

MagNA Pure Compact Nucleic Acid Isolation Kit I performed automatically in

MagNA Pure Compact Instrument; M. Sochor (Crop Research Institute, Olomouc,

Czechia) extracted DNA from the Czech specimens of P. granulatus using the CTAB

method (Doyle & Doyle 1987); and E.E Malysheva extracted DNA from the Russian

specimens following the manufacturing protocol of the Macherey-Nagel Nucleo-Spin

Plant II Kit.

PCR AMPLIFICATION AND SEQUENCING—The ITS1-5.8S-ITS2 region from the

ribosomal DNA was amplified and sequenced with primer pairs ITS1f-ITS4b or

ITS1f-ITS4 (White & al. 1990; Gardes & Bruns 1993) and ITS5-ITS4 (White & al.

1990, Nikolcheva & Barlocher 2004). PCR products were purified using the Thermo

Scientific GeneJET Gel Extraction Kit or by precipitation with polyethylene glycol

(10% PEG 6000 and 1.25 M NaCl in the precipitation mixture) and then sequenced

with the Sanger method. Raw data were edited and assembled in Sequencher 4.7 and

MEGA 6 (Tamura & al. 2013). The sequences generated were deposited in GenBank

with corresponding accession numbers (TABLE 1).

PHYLOGENETIC RECONSTRUCTION—In addition to 12 ITS sequences newly

generated for the study, 77 sequences including the outgroup Pluteus chrysaegis

(Berk. & Broome) Petch were retrieved for phylogenetic reconstruction from

GenBank (http://www.ncbi.nlm.nih.gov/genbank/). The taxonomic identities of these

sequences are given as they appear in GenBank (TABLE 1). The sequences were first

aligned with the Muscle method/procedure (Edgar 2004) embedded in MEGA 6 and

then corrected manually using the same program. The alignment was deposited in

TreeBASE (S23054). Phylogenetic reconstructions for the dataset were performed with

maximum likelihood (ML) and Bayesian (BA) analyses. Before the analyses, the best-fit

ikova & al.

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Pluteus dianae & P. punctatus in eastern & northern Europe... 251

substitution model for the alignment was estimated based on the Akaike Information

Criterion (AIC) using the FindModel web server (http://www.hiv.lanl.gov/content/

sequence/findmodel/findmodel.html). Both ML and BA analyses employed the GTR

model. ML analysis was run on the PhyML server, v. 3.0 (http://www.atgc-montpellier.

fr/phyml/), with one hundred rapid bootstrap replicates. BA was performed using the

MrBayes 3.2.5 software (Ronquist & al. 2012) for two independent runs, each with

7 million generations with sampling every 100 generations, under described model

and four chains. To quickly diagnose convergence of MCMC analyses and to obtain

estimates of the posterior distribution of parameter values, Tracer v1.6 was used

(Rambaut & al. 2014). Bootstrap (BS) values =70% and posterior probability (PP)

values =0.95 are considered significant.

Results

Molecular phylogeny

The dataset comprises 88 sequences of Pluteus sect. Hispidoderma and one

outgroup sequence (P. chrysaegis) and consists of 693 characters (with gaps).

The overall topologies of the ML and BA trees are substantially the same.

The P. dianae and P. punctatus clades received high statistical support in both

analyses. All sequenced collections of P. dianae appeared in a single clade in the

phylogenetic tree (Fic. 1), including the sequence generated from the holotype

collection. Genetic distance between these collections is less than 0.5%. One

collection from Spain (AJ209) that appeared extremely close to the P dianae

clade but differed in seven nucleotide positions (and with a genetic distance

from other collections ca. 1%) is provisionally named Pluteus aff. dianae.

Collections assigned to the P. plautus complex in the sense of Vellinga &

Schreurs (1985) did not form a monophyletic clade (Fic. 1). For the time being,

we have chosen to assign the published names only to the clades corresponding

to PB granulatus, P. longistriatus, P. semibulbosus, and P. velutinus C.K. Pradeep

& al. The other six species sampled here and belonging to the P plautus

complex are labelled simply Pluteus sp. I, II, HI, IV, V, and VI. Work is currently

underway to determine which clade should bear the name P. plautus and to

designate a type that would stabilize the usage of the name.

Taxonomy

Pluteus dianae Pilat, Ceska Mykol. 22(3): 171 (1968) Fics 2, 3, 6a,b,d-f

ORIGINAL DESCRIPTION. “Pileus 50-70 mm diam., regulariter orbicularis, adultus

plane convexus vel planus, cacumine minime rugoso-reticulatus et ibi tinctu debili

luteo-brunneolo, ceterum albus, margine ad dimidium radii rimosus, ceterum laevis

glaberque, cuticula marginem in forma membranae tenuissimae superanti, exsiccatus

albus tinctu debili sordide isabellino. Lamellae liberae, sat latae et ventricosae, novae

252 ... Sevéikova & al.

97/4

Pluteus sp. |

89/1

20/0.97| |

99/1

Pluteus aff, plau ‘us KX21 2

Plut ff. dianae HM562055

P. granulatus

92/0.97

Pluteus dianae

I peutas MG544918

lute fautus MG544917

Pluteus sp. Il

P. velutinus

Pluteus sp. III

—__— Pluteus fibrillosus KR022018

|+—Pluteus heteromarginatus HM562058

Pluteus sp. KRO22016

Pluteus sp. KR022014

99/4} luteus plautus KF306033

Pluteus sp. KR022015 Pluteus sp. IV

Pluteus sp. KC147677

Pluteus romellii KF306034

Pluteus sp. KRO22017

Pluteus cf. fernandezianus JQ065028

Uncultured fungus clone HQ257439

Pluteus sp. KRO22019

Pluteus semibulbosus MG544922

Pluteus sp. KU131672

Pluteus sp. KU131675

Pluteus sp. KU131674

Pluteus sp. KU131673

Pluteus semibulbosus KF668315

Pluteus semibulbosus MF437007

Bus ci enn posus SS rentaee

luteus aff. semibulbosus j

Pluteus semibulbosus MG544919 ease NeuWOSUS group

Pluteus semibulbosus KR022023

Pluteus semibulbosus KRO22024

Pluteus semibulbosus KR022021

Pluteus semibulbosus KX216353

Pluteus semibulbosus FJ774080

Pluteus semibulbosus MG544921

Pluteus semibulbosus KRO22022

Pluteus semibulbosus MG544920

Pluteus longistriatus KU953374

Pluteus longistriatus HM562149

Pluteus longistriatus HM562158

Pluteus longistriatus HM562172

Pluteus longistriatus MH211798

Pluteus longistriatus MF161220

Pluteus longistriatus HM562082

Pluteus longistriatus KT695338

Pluteus longistriatus KY777367

Pluteus longistriatus MH211936

Pluteus longistriatus KM052568

Pluteus longistriatus KX216355

93/0.98 __ Piyteus sp. KRO22028

Pluteus sp. KU131676

Pluteus sp. KF306015

Pluteus sp. KRO22026

Pluteus sp. uss 220255

74/0.93)

92/0.98

P. longistriatus group

100/1

Pluteus sp. V

Pluteus sp. VI

status (holotype

atus 7 Vou Pluteus punctatus

Pluteus chrysaegis JN603206

0.2

Pluteus dianae & P. punctatus in eastern & northern Europe ... 253

albae, adultae albido-salmoneae. Stipes 50-60 x 6-7 mm, albus, minime longitudinaliter

rugosus, subcylindraceus, deorsum paulum incrassatus basique truncata ad lignum

putridum insidens, parte tertia basali haud raro minime disperse obscurius fibrilloso-

subsquamulosus.

“Carposoma, praecipue trama lamellarum et cutis pilei ex hyphis dimiticis,

tenuiter tunicatis hyalinis, 4-8 ut crassis et hyphis crassius vel crasse tunicatis, 5-8 u

crassis, conspecte curvatis et haud raro tinctu debili luteolo conspectis, irregulariter

et in angulo recto curvatis et ramosis, parum septatis. Cutis pilei ex hyphis similibus

et ex finibus hypharum clavato-saccatis (pileocystidiis) tenuiter tunicatis, hyalinis,

decumbentibus, 60-100 x 14-25 up magnis. Metuloideis crasse tunicatis corniculatisque

in hymenio absentibus. Cheilocystidia in acie lamellarum copiosa, ovoidea, saccata vel

sublageniformia, tenuiter tunicata, hyalina, 25-60 x 12-25 u. Sporae globoso-ovoideae,

subsalmoneae, laeves, apiculo parvo deflexo praeditae, 7,5-9 x 5-6,5 [?

Hototypus—Czechoslovakia (currently Czechia), Rozvadov, Diana Nature reserve, in

primeval forest, lying rotten trunk of Fagus sylvatica L. infected by Fomes fomentarius

(L.) Fr., 18. VII. 1967 leg. et det. A. Pilat (PRM 629413).

TYPE REVISION. Basidiospores [1/1/30] (6.0-)6.5-8.5(-9.5) x 5.0-

6.5(-7.0) um, avl x avw = 7.2 x 6.3 um, Q = 1.00-1.40(-1.60), avQ = 1.16,

globose, subglobose or broadly ellipsoid, rarely ellipsoid. Basidia (18-)

21-29(-32) x 7-9(-11) um, tetrasterigmate, narrowly clavate to

subcylindrical or subutriform, colourless. Pleurocystidia moderately

abundant, (32-)35-82(-90) x (13-)15-26(-32) um, fusiform, lageniform,

less frequently utriform, usually with a <15 um (very rarely <35 um) long

narrowed apex and 3.0-15.0 um long pedicel, thin-walled, colourless.

Lamellar edges sterile. Cheilocystidia (25-)30-48(-62) x (12-)14.5-26 um,

narrowly to broadly clavate, narrowly ventricose, scarcely sublageniform

or cylindrical, thin-walled, colourless. Pileipellis a trichoderm, terminal

elements 55-180(-280) x 14-24 um, very variable in size, sometimes

in similarly sized tufts (more often smaller than larger-sized elements),

narrowly clavate to clavate, sometimes with a narrowed or strangulated

apex, sometimes with pale ochre intracellular pigment. Branched hyphae

present in the subpileipellis. Stipitipellis a cutis of 4.0-12.0(-18.0) um diam.

cylindrical colourless hyphae. Caulocystidia in tufts, (30-)45-75(-82)

x (9-)12-17.5(-20) um, cylindrical or narrowly clavate, rarely with short

obtuse rostrum, thin-walled, colourless or with very pale intracellular

pigment. Clamp connections absent in all studied tissues.

Fic. 1. Best tree from Maximum likelihood analysis for the nrITS dataset showing some lineages

within Pluteus sect. Hispidoderma. For sequences retrieved from GenBank the corresponding

accession numbers are given. The newly generated sequences are indicated with the collection

numbers (in parentheses). Support values given above the branches are BS 270% /PP20.95. Scale

bar shows expected changes per site.

254 ... Sevéikova & al.

Characteristics of the type locality and type specimen

Diana Nature Reserve is a protected area in western Czechia close to the

German border and situated in the Cesky les mountains near the village of

Rozvadov at 500-532 m a.s.l. (central point: 49°37’57”N 12°34’43”E). The

Cesky les mountain range belongs to the Bohemian massif Mesophyticum

(Skalicky 1988) with a ca. 7-8 °C mean annual temperature and 700-800

mm mean annual precipitation. Climatically, this region is classified

as mildly warm zone (MT3 on the Quitt scale, Vrska & al. 2012). The

reserve comprises predominantly near-natural herb-rich beech forests, in

particular the Dentario enneaphylli-Fagetum association in an advanced

stage of degradation (Zahradnicky & Mackovcin 2004). Based on their

mensurational surveys, Vrska & al. (2012) detected an extensive disturbance

in a part of the reserve, thus establishing that Diana is not now a natural

forest. The protected area comprises a 300-year old forest of Tilia cordata,

Quercus robur, and Picea abies. Pilat (1968), who collected the holotype

collection from a rotting trunk of Fagus sylvatica lying on the ground, stated

that he collected and photographed two basidiomata. However, only one

basidiocarp is shown in the photograph (Pilat 1968: 172). The original

material is preserved in dried form and in good condition in the herbarium

of the Mycological Department of the National Museum in Prague (PRM

629413!).

Additional collections studied

Pileus 20-120 mm in diam., broadly conical, convex to applanate,

sometimes with broad low umbo; creamy-grey or cinnamon buff, greyish-

brown or brown; drying + whitish-grey, surface felty or covered with white

appressed hairs; in some specimens irregularly wrinkled in centre, in some

specimens striate at margin. Lamellae pink with concolorous edges. Stipe

30-70(-100) x 2.5-10 mm, broadened towards base or bulbous; pure white,

whitish, or yellowish; longitudinally striate, whitish floccose to granulose-

pruinose. Smell indistinct or pleasant, sweetish-spicy.

Basidiospores [5/5/150] 5.5-8.0(-9.0) x 4.5-6.5(-6.8) um, avl x avw =

7.2 x 5.5 um, Q = (1.10-)1.17-1.46, avQ = 1.30, broadly ellipsoid to ellipsoid

or ovoid, rarely subglobose. Basidia 25-37 x 8-12 um, tetrasterigmate.

Pleurocystidia abundant, 54-86(-103) x (14-)18-27(-34) um, mostly

(broadly) lageniform, (narrowly) utriform, or fusiform, commonly with

a small apical excrescence, more rarely with more than one on the same

cystidium, colourless. Cheilocystidia abundant, forming a sterile layer,

very variable in size, (41-)52-95(-115) x (14-)17-27(-32) um, narrowly

Pluteus dianae & P. punctatus in eastern & northern Europe ... 255

Fic. 2. Pluteus dianae (holotype, PRM 629413): a. pileipellis elements; b. basidiospores; c. basidia;

d. cheilocystidia; e. pleurocystidia; f. caulocystidia. (del. H. Sevéikova).

256 ... Sevéikova & al.

20 um

Hoth

HN) 0

Fic. 3. Pluteus dianae (LE312950): a. pileipellis elements; b. caulocystidia; c. cheilocystidia;

d. basidiospores; e. pleurocystidia. (del. E.E. Malysheva); (JHC 99_30): f. cheilocystidia;

g. pleurocystidia. (del. A Justo).

Pluteus dianae & P. punctatus in eastern & northern Europe ... 257

to broadly clavate, narrowly utriform to utriform, ovoid or cylindrical,

lageniform, pedicellate, some with single apical papilla, thin- or slightly

thick-walled. Pileipellis a trichoderm, terminal elements (50-)79-125(-220)

x 14-36 um, clavate, utriform or cylindrical, some with tapering apex.

Caulocystidia in clusters, 43-140 x 14-27 um; narrowly clavate, narrowly

utriform, narrowly fusiform, or narrowly lageniform; thin-walled,

colourless. Clamp connections absent.

COLLECTIONS STUDIED—DENMARK, Sja&LLAND, Nesbyholm Storskov [SW of

Ringsted], 55°22’26”N, 11°35°50”E, on fallen log of Fagus sylvatica, 24 VII 1999, leg.

J. Heilmann-Clausen (JHC99-030, as P. cf. depauperatus). RUSSIA: SAMARA REGION,

Zhiguli State Nature Reserve, broadleaved forest (Tilia cordata, Acer platanoides), on

fallen trunk of deciduous tree, 9 VII 2003, leg. E. Malysheva (LE 312950, as Pluteus

plautus); broadleaved forest, on fallen trunk of deciduous tree, 27 VIII 2001, leg.

E. Malysheva (LE 312978, as Pluteus plautus); Moscow REGION, Zvenigorod Biological

Station of Moscow State University, 55°41’00”N 36°43’00”E, mixed forest, on fallen

trunk of Populus tremula, 19 V1 2013, leg. E. Voronina (LE 296318, as Pluteus plautus);

Picea forest (with Pinus and Betula), on decayed deciduous tree, 10 VI 2014, leg.

E. Voronina (LE 303485, as Pluteus plautus).

CoMMENTS—The Danish collection was from a somewhat open,

seminatural, deciduous woodland dominated by Fagus sylvatica and on

a north-facing slope facing the river Susaen. The annual temperature of

the site averages 8.6-8.8 °C, and the annual precipitation averages 600-

700 mm (Wang 2013). The Russian collection sites lie in the submontane

belt and mountain territory (Zhiguli State Nature Reserve) and plains

(Moscow State University Zvenigorod Biological Station) with the annual

temperature averaging 10.3 °C and the total annual precipitation averaging

904.4 mm _ (http://www.earthonlinemedia.com/ipg/outlines/climates_

humid_continental.html).

Pluteus punctatus Wichansky, Mykol. Sborn. 49(1-2): 1 (1972) FIGS 4, 5, 6C

ORIGINAL DESCRIPTION. “Pileus tenuiter carnosus, 5.5 cm diam., pallide griseo-

brunneolus, late convexus, dimidio squamulis brunneis minutis flocculosis

punctiformibus ad marginem absentibus obtextus. Lamellae confertae, ventricosae

usque 10 mm latae, remotae, albidae, dein incarnatae. Stipes 4 cm longus atque 8

mm crassus, solidus, cylindraceus, ad basim paulum incrassatus, albus, nudo apice

excepto aquamulis brunneis, minutis, flocculosis dense punctatus. Cystidia varie

formata, ellipsoideo- vel ovato-clavata, nonnullae ventricoso fusiformia. Sporae late

ellipsoideae, 7-8.5 x 5.5-6.5 um. Cellulae cuticulae pilei cylindricae, longissimae,

13-26 um crassae. Caro pilei et stipitis alba. Odor et sapor nullae.”

HoLotypus—Carposoma solitarium do. Frant. Fuchs 4. X. 1969 ad codicem Populi

albae in Revnice (Czechoslovakia [currently Czechia], Bohemia centralis) legit et mihi

donavit. Typus in herbario Musei nationalis Pragae asservatur (PRM 682743).

258 ... Sevéikova & al.

TYPE REVISION. Basidiospores [1/1/30] (5.0-)7.0-8.5(-9.5) x 5.0-7.0(-7.5)

um, avl x avw = 7.6 x 6.5 um, Q = (1.00-)1.05-1.33(-1.40), avQ = 1.2,

subglobose or broadly ellipsoid, rarely globose or ellipsoid. Basidia (18-)22-28

(-30) x 8-10(-12) um, tetrasterigmate, rarely bisterigmate, narrowly clavate

to subcylindrical or subutriform, colourless. Pleurocystidia scattered, 28-65

x (13-)15-26(-32) um, fusiform, lageniform, less frequently utriform, often

with <9 um long obtuse apex, sometimes with <13 um long pedicel, thin-

walled, colourless. Lamellar edges sterile. Cheilocystidia abundant (28-)44-65

(-86) x (14-)16-33 um, broadly clavate, narrowly ventricose, fusiform to

broadly fusiform, obovoid, scarcely sublageniform, sometimes with <12 um

long pedicel, rarely with a short rounded apex, thin-walled, hyaline. Pileipellis

a trichoderm, terminal elements 55-280(-380) x 13-26(-38) um, variable

in size, narrowly clavate to clavate, sometimes with a narrowed or slightly

strangulated apex, with or without colourless or with pale brown intracellular

pigment. Stipitipellis a cutis of 5.0-11.0(-16.0) um diam. cylindrical hyaline

hyphae. Caulocystidia solitary or in small tufts, scattered, (24-)28-60 x

10-25 um, cylindrical or narrowly clavate, obovoid with short pedicel or

obfusiform, thin-walled, with or without very pale brown intracellular

pigment. Clamp connections absent in all studied tissues.

Characteristics of the type locality and type specimen

Revnice is a town situated about 10 km southwest of the Prague city limits

on the Berounka river, lying at 200 m a.s.l. (surrounded by ca. 500 m hills

(central point: 49°54’50”N 14°14’09”E). The area belongs to the Bohemian

Massif, Thermophyticum (Skalicky 1988) with annual averages of 8.5-9 °C

and 504-527 mm precipitation (Merkel 2018, Tolasz 2007). Wichansky (1972)

did not describe the ecology in detail, citing only the growth on a Populus sp.

stump. Therefore it is unclear whether the type specimen was found in a semi-

natural alluvial woodland, a disturbed stand, or an ornamental tree in the town.

Wichansky (1972) mentions only one basidiocarp, and no other original

material exists. This basidiocarp is preserved as a holotype in the herbarium of

the Mycological Department of the National Museum in Prague (PRM 682743

contains one dried basidiocarp in good condition).

Additional collection studied

Pileus scales greyish-brown in centre, outwards cream to ochraceous, < 25

mm diam. (dried specimen). Lamellae free, 1-2 tiers of lamellulae, crowded,

lamellar edge concolorous. Stipe 32 x 2 mm (dried specimen), cylindrical,

broadened towards base, finely pruinose. Smell and taste not recorded.

Pluteus dianae & P. punctatus in eastern & northern Europe ... 259

Fic. 4. Pluteus punctatus (holotype, PRM 682743): a. pileipellis elements; b. basidiospores;

c. basidia; d. cheilocystidia; e. pleurocystidia; f. caulocystidia.

260 ... Sevéikova & al.

Warinine

Mpls

HOY ss

Fic. 5. Pluteus punctatus (JHC 04_298): a. pileipellis elements; b. cheilocystidia; c. pleurocystidia;

d. caulocystidia. (del. A. Justo). e. spores. (del. H. Sevcikova). Scale bars = 10 yum.

Pluteus dianae & P. punctatus in eastern & northern Europe ... 261

Basidiospores [1/1/30] 6.5-8.5 x 4.5-7.0 um, avl x avw = 7.3 x 5.8 um,

Q = (1.10-)1.15-1.39, avQ = 1.27, broadly ellipsoid to ellipsoid, rarely

subglobose. Basidia 24-32 x 7-11 Um, tetrasterigmate. Pleurocystidia

abundant, 50-80(-100) x 17-32(-44) um, mostly lageniform, narrowly utriform

or fusiform; hyaline. Cheilocystidia abundant, 58-106 x 17-46 um, mostly

clavate, narrowly utriform or fusiform; hyaline. Pileipellis a trichoderm,

terminal elements 85-250 x 15-27(-35) um, (narrowly) clavate, narrowly

utriform, fusiform, or cylindrical. Caulocystidia abundant, isolated or in

loosely arranged clusters; 61-100 x 17-31 um, narrowly clavate, narrowly

utriform, lageniform, or fusiform; hyaline. Clamp connections absent in all

studied tissues.

COLLECTION STUDIED: SWEDEN, HALLAND, Domestorp, Musikedalen, Vallasen,

56°22/17”'N 13°05’45”E, on Fagus log, 5 X 2004, leg. Jacob Heilmann-Clausen (JHC 04-

298, originally as P. cf. depauperatus).

CoMMENTS— The Swedish specimen was collected from a fallen beech log in

a small (4.4 ha) almost pure beech stand containing a considerable amount

of dead wood, often in sunlit gaps. The ~210-year-old stand is rather even-

aged and part of the larger (110 ha) Musikedalen nature reserve, protected

since 2010. The stand has been without active management of the living trees

since 1946 and under strict protection since 1996 (Lanstyrelsen Halland

2010). The forest lies at 180 m a.s.l. adjacent to a larger bog area and elsewhere

surrounded by spruce plantations. The climate is temperate with an Atlantic

influence with an annual average precipitation of 1000-1100 mm (SMHI

2018a) and ca. 7 °C temperature (SMHI 2018b).

Distribution & ecology

Pluteus dianae was previously known only from its type locality in Czechia.

Our study extends the known distribution to Denmark and Russia. In addition,

phylogenetically identical collections are reported from Turkey (GenBank

Acc. No. MG544918.1 and MG544917.1, as Pluteus plautus (Kaygusuz & al.

2019). Pluteus dianae can now be said to occur through northern, central,

and eastern Europe into Turkish Asia. Climatically, the European collection

sites lie in the temperate continental zone (SMHI 2018b, Tolasz 2007) with

precipitation averaging 600-900 mm annually (Merkel 2018, Wang 2013). The

sites are located from the lowlands to the mountains. The Turkish collections

were found near mountains at 250-850 m elevations in northwest Turkey

in areas of regular and high rainfall (Kaygusuz & al. 2019). Pluteus dianae,

which grows on decaying trunks of deciduous trees, is known from Fagus spp.,

Populus tremula, and a fallen trunk of an unidentified deciduous tree.

262 ... Sevéikova & al.

Pluteus punctatus was also known only from its type locality in Czechia.

The Swedish collection adds important ecological and distributional data. Its

collection sites lie in the European temperate area (SMHI 2018b, Tolasz 2007)

with a mean annual precipitation of 504-1100 mm (Merkel 2018, SMHI

2018a). Both sites are located in the lowlands. Pluteus punctatus, which grows

on dead wood of deciduous trees, has been collected from a Populus sp. stump

and fallen Fagus sylvatica trunk.

OTHER STUDIED COLLECTIONS OF SIMILAR TAXA:

Pluteus atriavellaneus: USA: TENNESSEE, Unicoi Co., Unaka Springs, dead wood,

18-24 Aug. 1904, W. A. Murrill 673 (NY, type).

Pluteus avellaneus: USA: NEw York, Essex Co., Lake Placid, Adirondacks, dead

wood, 17-29 Jul. 1912, W. A. Murrill 91 (NY, type).

Pluteus compressipes: JAMAICA: Castleton Gardens, dead wood, wet and shaded, 15

Dec. 1908, W. A. Murrill 118 (NY, type).

Pluteus depauperatus: FRANCE: Chateau de La Grange, Yerres, stumps and trunks of

Fagus, Aug. 1936 leg. & det. H. Romagnesi (PC, type).

Pluteus fibrillosus: USA: Loutstana, New Orleans, Chalmette, New Orleans, in soil

in a wet thicket, 8 Sept. 1908, F. S. Earle 129 (NY, Type).

Pluteus fuliginosus: USA: NEw YorRK, Essex Co., Lake Placid, Adirondacks, 17-29 Jul.

1912, white pine stump decayed, coniferous or mixed forest, W. A. Murrill 118 (NY,

type).

Pluteus granulatus: CZECHIA: Kladeruby nad Oslavou, VIci kopec, cultivated

spruce forest, on a rotten stump of Picea abies, 3 Oct. 2013 leg. & det. H. Sevcéikova

(BRNM 761707); Ceska Trebova, Kiivolik stream, on rotten stump of Picea abies lying

in a stream, 15 Aug. 2017 leg. M. Micka, det. H. SevZikova (BRNM 807610); Psary,

cultivated spruce forest, near a road, cavity of rotting stump of Picea abies, 20 Sept.

2015 leg. J. Her¢ik, det. H. Sevcikové (BRNM 807611). ITALY, probably Trentino, Val

di Sole, rotten ? of Abies near meadow, Aug. 1882 leg. & det. G. Bresadola (S F14380

lectotype).

Pluteus hiatulus: FRANCE: Oise, Lamorlaye, rotten deciduous stump (Populus sp.?)

10 Sept. 1946 leg. Romagnesi H.? (PC, type); Chateau de La Grange, Yerres, stumps of

Fagus, Aug. 1936 leg. & det. H. Romagnesi (PC, herbarium of Romagnesi).

Pluteus inflatus: CZECHIA: Mnichovice, trunk of Alnus (A. glutinosa according to

observations by H.S.), Aug. 1944, leg. & det. J. Velenovsky (PRM 154569, type).

Pluteus latifolius: USA: WASHINGTON, Seattle, on dead alder, 20 Oct. - 1 Nov. 1911,

W. A. Murrill 510 (NY, type).

Fic. 6. Pluteus dianae: a. JHC 99_30) basidiomata (photo J. Heilmann-Clausen); b. (holotype,

PRM 629413) dried basidioma (photo H. Sevéikova). Pluteus punctatus c. (holotype, PRM

682743): dried basidioma (photo H. Sevéikova). Pluteus dianae: (LE312950): d. pleurocystidia;

e. basidium with spores and pleurocystidia (photos E.F. Malysheva). f. (JHC 99_30). caulocystidium

(photo H. Sevéikova). Scale bars: b-c = 1 cm, d-f = 20 um.

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Pluteus plautus sensu lato: CZECHIA: Babice, PR Cihadlo, Fagetum, fallen stem of

Fagus sylvatica, 17 May 2017 leg. & det. H. Sevcikova (BRNM 807612, HS9); Valtice,

Rendezvous National Nature Monument, fallen deciduous stem, 2017 leg. P. Véelicka,

det. H. Sevéikova (BRNM 807613, HS18_6).

Pluteus puberulus: CZECHIA: Chotec¢, Radotin valley, rotten stump of Alnus sp.

(probably A. glutinosa), Aug. 1918, leg. & det. J. Velenovsky (PRC, Velenovsky collection,

bottle 78 f, type).

General discussion

Taxonomy

Pluteus dianae was originally characterized by its large whitish pileus tinged

slightly yellowish-brown at the centre and a whitish striate stipe subpruinose

only in the lower part (Pilat 1968). However, all studied Russian collections

with similar ITS sequence data are described has having a brown or greyish-

brown pileus with a velvety surface or covered with white appressed hairs and

a whitish or yellowish striate often granulose-pruinose stipe. Microscopically,

Pilat (1968) emphasized the absence of metuloid cystidia with hooks but did

not explicitly describe any other kinds of pleurocystidia. Our examination

of the holotype (PRM 629413!) revealed (narrowly) lageniform or fusiform

pleurocystidia with a narrowed apex and a pedicel (Fic. 2e), also found in all

other studied collections (Fics 3e, g; 6d—e). Pilat (1968) cited basidiospores

measuring 7.5-9.0 x 5.0-6.5 um, while our type revision showed a slightly

wider range [(6.0-)6.5-8.5(-9.5) x 5.0-6.5(-7.0) um] in concordance with the

Russian collections. The Danish collection is morphologically similar to the

type and Russian collections except for the scattered caulocystidia, apparently

reflecting the rather poor condition of the dried material, as the macroscopical

notes mention the stipe as being whitish floccose.

Although the Spanish collection AJ209 (Justo & al. 2011b) is phylo-

genetically close to Pluteus dianae, we maintain its identification as Pluteus aff.

dianae based on its different ITS sequence and the following morphological

differences from P. dianae: (i) smaller pilei (3.5-5 cm diam.); (ii) broadly

ellipsoid to ellipsoid (rarely subglobose) and smaller basidiospores [5.5-7.5 x

4.4—6.2 um, avl x avw = 6.6 x 5.3 um, Q = (1.08) 1.17-1.40, avQ = 1.24]; and

(iii) pleurocystidia without apical excrescences.

Pluteus punctatus was described with a pale grey-brown pileus with dark-

brown floccose squamules, a whitish stipe with brown tomentose squamules,

and variably shaped cystidia (Wichansky 1972: 1). The pileus scales of the

Swedish collection are grey-brown in centre, outwards cream to ochraceous.

Our microscopical examination of the holotype (PRM 682743!) and

Pluteus dianae & P. punctatus in eastern & northern Europe ... 265

in concordance with the Swedish collection reveals striking fusiform,

lageniform, or utriform pleurocystidia <80 um long (Fics 4e, 5c) and

caulocystidia not arranged in large tufts but isolated or in loosely arranged

clusters (Fics 4f, 5d).

Similar taxa

According to Pilat (1968), Pluteus dianae resembles Pluteus petasatus

(Fr.) Gillet and Pluteus pellitus in having a large white pileus. However,

these species belong to Pluteus sect. Pluteus, characterized by metuloid

pleurocystidia and a cutis pileipellis.

Both Pluteus dianae and P. punctatus belong to the taxonomically

complicated P plautus complex. Different authors have interpreted the

species in this complex in noticeably diverse ways. Whereas Citérin &

Eyssartier (1998) mentioned about six species in Europe, Vellinga (1990)

recognized only one broad species—Pluteus plautus—incorporating

nine taxa described by Lasch (Fries 1838), Bresadola (1881, Schulzer

1885), Romagnesi (Ktihner & Romagnesi 1953), Orton (1960, 1969), and

Wichansky (1972). Other poorly known species of this complex were

described by Velenovsky (1921) and Pilat (1968). Several additional species

in the complex described from North America (Murrill 1917) may well also

occur in Europe. Below we provide an overview of names published in this

complex in Europe and North America, discussing their similarity to Pluteus

dianae and P. punctatus so as to establish a basis for future taxonomic work

in this species complex.

Pluteus plautus (Weinmann 1836: 394), was originally characterized by

an alutaceous to fuligineous pileus with a velvety brown to blackish brown

stipe and stipe context. Gillet (1876: 394) interpreted it as a small species

without specifying basidioma dimensions. Pluteus punctatus was originally

described by Wichansky (1972) as macroscopically similar to P plautus,

but it differs from P punctatus (and P. dianae) in having a darker stipe and

brown stipe context. Based on Weinmann’s type description and our own

field experience, we consider the dark colour of the context (especially in

the stipe) an important diagnostic character of P. plautus sensu stricto. As

P. plautus is interpreted differently by several agaricologists (e.g. Orton

1986, Vellinga 1990, Citérin & Eyssartier 1998), a neotypification is

needed.

Following the tradition of Kihner & Romagnesi (1953), Pluteus

semibulbosus (Lasch) Quél. also belongs to the P. plautus complex. It was

originally characterized as a small whitish species with a softly atomate,

266 ... Sevéikova & al.

sulcate pileus and pubescent stipe with a distinctly bulbous base (Fries

1838). Lasch (in Fries 1838) did not mention basidioma dimensions, but

Saccardo (1887: 674) mentioned a 13 mm broad pileus and 25 mm long

stipe. Orton (1960: 349, 1986: 55) interpreted P semibulbosus as belonging

to P. subsect. Eucellulodermini, and published the name Pluteus boudieri

P.D. Orton for P semibulbosus sensu Boudier, Kihner & Romagnesi.

However, for us, the original description of the stipe as pubescent strongly

supports P. semibulbosus in P. sect. Hispidoderma and within the P. plautus

complex. This interpretation would place the name P boudieri in synonymy

with P semibulbosus. Recent collections referred to P. semibulbosus by us

demonstrate that this broad morphological species concept corresponds to

a polyphyletic group (unpublished data) and that further studies, including

the selection of a type for P semibulbosus, are needed. Even in its broad

concept Pluteus semibulbosus can be distinguished from P. punctatus and

P. dianae at least by its smaller basidiomata and bulbous stipe bases, while

pale colouration is more diagnostic of P punctatus.

Pluteus aestivus Velenovsky (1921: 607) was described as similar to

P. semibulbosus but having cuneiform-ellipsoid, non-translucent brown

basidiospores, suggesting it belongs in a different genus than Pluteus. As

original material is no longer extant, the name is impossible to interpret

exactly, and we consider it a nomen dubium.

Pluteus candidus Patouillard (1887: 156) was described as a small

species with a white, non-striate, silky pileus up to 10 mm broad, a

pale thin stipe, and fusiform cystidia. It differs from both P dianae and

P. punctatus by its smaller basidiomata and silky pileus, and P punctatus

is further distinguished by the darker pileus colour. Pluteus candidus

probably belongs to the P semibulbosus complex. A smooth non-striate

pileus is an important diagnostic feature for P candidus. Nonetheless,

finding a holotype of P candidus is important for future research.

Pluteus stylobates Velenovsky (1921: 608) was described as a small

whitish agaric with a <10-15 mm broad white pileus with dark squamules

and a discoid stipe base resembling Mycena stylobates (Pers.) P. Kumm.

The species therefore seems to belong to the P semibulbosus complex.

Unfortunately the holotype (PRC, Velenovsky collection, bottle no. 396),

no longer exists (it dried up and was destroyed years ago). This taxon may

be a good species, but neotypification is necessary. Both P dianae and

P. punctatus have larger basidiomata and lack a discoid stipe base, and

P, punctatus differs also by the darker colours.

Pluteus dianae & P. punctatus in eastern & northern Europe ... 267

Pluteus gracilis (Bres.) J.E. Lange was originally described by Bresadola

(Schulzer 1885: 134) as P. pellitus var. gracilis Bres. Lange (1936: 84)

assigned this taxon to P. subsect. Depauperati. His description and painting

(Lange 1936: tab. 71 E) showed a whitish to very pale ochre Pluteus with

dirty brown squamules on the pileus and stipe, presumably belonging to

the P. plautus complex. However, it is unclear based on which material

Bresadola described this taxon as a variety of Pluteus pellitus. Until a type

has been located, we find it difficult to interpret this taxon.

Pluteus depauperatus Romagn. (Kthner & Romagnesi 1956: 181) was

described with an 18-50 mm broad, velvety dull pale brown or brownish

grey-ochre pileus, distinctly striate at the margin and sometimes granulate

in the centre. The stipe was described as white when young, but striped

pruinose brown-saffron to brick-saffron in mature specimens (Kihner

& Romagnesi 1953). The holotype of P depauperatus (PC!) contains two

envelopes with material. The envelope with the larger basidiocarp marked

as the type is in a rather poor condition, with deformed or collapsed

cystidia and irregular, probably deformed, basidiospores (7.0-8.0 x 4.0-5.0

um). The envelope with the smaller basidiocarp is better preserved and its

microcharacters agree with Romagnesi’s original description (Ktihner &

Romagnesi 1953) and spores measuring (5.0-) 6-6.5 (—7.0) x 5.0-6.0 um,

avl x avw = 6.33 x 5.43 um. Pilat (1968), who mentioned the similarity of

P. dianae with P. depauperatus, distinguished the latter by its pale brown,

grey-brown or saffron-brown pileus, smaller basidiospores (see above), and

an odor resembling Lepiota cristata (Bolton) P. Kumm. or Scleroderma Pers.

The saffron-tinged mature stipe should also help separate P. depauperatus

from both P dianae and P. punctatus. Moreover, in all observed collections,

P. punctatus lacks a striate pileus margin. Pluteus depauperatus seems to

represent a good species, but verification by holotype DNA sequencing is

needed.

Pluteus hiatulus Romagn. (Kiihner & Romagnesi 1953: 421, as nom. nov.,

validly published in 1956: 182 with the addition of the Latin diagnosis)

was originally described as having a dirty brown, soon fading pileus with

a Coprinus-like membranaceous striate margin, a satin white to pale brown

(more at the base) stipe, and refractive utriform or lageniform pleurocystidia

with pale brown vacuoles Our holotype revision (PC!) confirmed that

the pleurocystidia are refractive, narrowly to broadly (sub)lageniform to

fusiform (rarely subutriform to utriform), and only rarely with very pale

brown guttules. However, it is possible that brown guttulae fade with time

268 ... Sevéikova & al.

in exsiccates. Romagnesi’s measurements (in Kuhner & Romagnesi 1953) of

subglobose basidiospores at 5.7-6.2(-7.5) x 5-6.5 um agree with our type

study. Pluteus dianae and P. punctatus differ from P. hiatulus by having a

thicker-fleshed pileus with a less striate and non-Coprinopsis-like margin

and possessing hyaline non-refractive pleurocystidia.

Pluteus punctipes Orton (1960: 361) was described as having relatively

small to moderately large basidiomata with a sepia or umber to vandyke

brown coloured pileus and sepia to dirty yellowish brown minute fibrillose

scales on both pileus and stipe. Its possession of large (<100 x 30 um)

lageniform pleurocystidia with a short to long neck and with a brown

vacuole is macroscopically expressed in some places as slightly dark brown

lamellae. Orton (1960) also mentioned narrow clamped hyphae below

cylindrical-fusiform pileipellis elements. The species seems to be close

to P. plautus s. str. Pluteus dianae and P. punctatus differ in their hyaline

and shorter pleurocystidia. Further studies and sequencing of the type are

needed to clear up the taxonomic position and delimitation of P punctipes.

Pluteus dryophiloides Orton (1969: 115) was originally described as

having colours similar to Gymnopus dryophilus (Bull.) Murrill with a

whitish or buff coloured stipe; a pileipellis with relatively short (60-152

um) terminal cells; and vesiculose-fusiform, broadly pyriform, or broadly

lageniform pleurocystidia sometimes terminating in solitary short pimples.

Pluteus dianae and P. punctatus differ by their duller pileus colours, longer

pileipellis terminal elements, and different pleurocystidia.

Pluteus granulatus Bresadola (1881: 10) was described as having a

brown-reddish granulate pileus and a whitish, finely furfuraceous stipe

with a yellowish furfuraceous base; the species was described as growing on

coniferous wood, and that substrate may indeed be an important character

for separating P granulatus from other species in the P plautus group. We

consider collections HS11 (BRNM 807610) and HS18_2 (BRNM 807611)

(Fic. 1) to represent P. granulatus in its original sense. Unfortunately, Kuhner

& Romagnesi (1956) cited P granulatus as growing on Fagus sylvatica, which

caused subsequent taxonomic confusion. In GenBank, several collections

assigned to Pluteus granulatus were indicated as growing on Fagus orientalis.

Our ITS sequences (HS9 and HS18_6), molecularly similar to other

collections named P. granulatus from Fagus sylvatica, do not represent

P. granulatus. These collections are here referred to Pluteus sp. VI (Fic. 1)

pending further clarification of the P plautus complex. Pluteus dianae and

P. punctatus differ from P. granulatus by growth on deciduous wood and the

Pluteus dianae & P. punctatus in eastern & northern Europe ... 269

lack of a reddish tinge and distinct granules on the pileus, with the pileus of

Pluteus dianae usually paler. Microscopically, the lectotype (S F14380!) of

P. granulatus possesses lageniform (rarely fusiform) pleurocystidia that lack

the apical excrescences, which are relatively frequent in P dianae. Further,

the pileipellis terminal elements are shorter and more uniform in size than

in P punctatus and P. dianae.

Pluteus puberulus Velenovsky (1921: 607) was described as having a

leathery, ochre, brown-streaked pileus that in places is almost squamulose,

scaly, entirely ochre, or covered by small grains and a stipe that is grooved,

densely velvety pubescent, and dirty yellowish. Velenovsky (1921) also

described yellowish cheilocystidia. The holotype is preserved in a bottle of

Velenovsky’s solution (Fassatiova & al. 1994) in which yellow colours were

said to fade (Velenovsky (1921: 608). As the holotype (PRC, Velenovsky

collection, bottle 78 f!) contains only a part of the pileus in bad condition,

determination of the identity of the preserved fungus is impossible and the

taxonomic position of P puberulus remains unclear.

Pluteus opponendus (Britzelm.) Sacc., originally described with a white

pileus and stipe, might resemble P. dianae except for the significantly different

basidiospores (8-9 x 4 um; Britzelmayr 1881: 136). Pluteus opponendus has

an unclear taxonomical position.

Pluteus tiliaceus Velenovsky (1921: 607), which was described with

white to whitish colours similar to P opponendus, is unique within the

P. plautus complex by its small (~4 um diam.) globose basidiospores,

smooth, glabrous, radially fibrillose pileus, and clavate cystidia (Velenovsky

1921). The original material does not exist, and the taxonomical value and

position of P. tiliaceus remains unclear.

Pluteus praestabilis (Britzelm.) Sacc. was originally characterized

by a blackish brown velvety pruinose pileus, a white stipe with brown

floccules, basidiospores measuring 6 x 4-5 um, and growth on soil near

Fagus sylvaticus (Saccardo 1887: 672). Although it probably belongs to

the P plautus complex, P praestabilis may also represent a species related

to P. podospileus Sacc. & Cub. Pluteus dianae and P. punctatus have larger

basidiospores, with P dianae further distinguished by a stipe without brown

floccules and a paler pileus.

Vellinga & Schreurs (1985) cited Pluteus inflatus Velen. as a synonym

of P plautus. However, our study of the holotype (PRM 154569!) showed

that its pileipellis is composed of sphaeropedunculate and long elements,

and pleurocystidia are absent. Our microscopical examination combined

270 ... Sevéikova & al.

with the macroscopic description by Velenovsky (1921: 609) suggests that

P. inflatus is closer to P. podospileus than to P. plautus.

The North American Pluteus longistriatus (Peck) Peck is characterized

by a cinereous or whitish pileus, with striations that reach the darker,

minutely squamulose-hairy disc, and a glabrous white stipe (Peck 1885).

All these characters separate P. longistriatus from P. dianae and P. punctatus.

Molecular data (Fic. 1) also support recognition of P longistriatus as a

separate taxon. Morphologically, P longistriatus resembles the European

P. hiatulus. Further phylogenetic and morphological research is needed to

clarify the relationship between P longistriatus and P. hiatulus.

As part of a project aiming to clarify the taxonomy of the North American

Pluteus species, A. Justo revised the type collections of the 46 species described

by Murrill (1911, 1917, 1939, 1943, 1946). Six belong to the P plautus

complex: P. atriavellaneus Murrill, P avellaneus Murrill, P. compressipes

Murrill, P fibrillosus Murrill, P fuliginosus Murrill, and P latifolius Murrill.

Pluteus dianae differs microscopically from them mainly by differently

shaped pleurocystidia (only P latifolius possesses pleurocystidia commonly

bearing apical excrescences similar to P dianae). Pluteus punctatus differs

microscopically from the North American taxa mainly by its predominantly

clavate to narrowly utriform cheilocystidia. Pluteus latifolius was originally

described from Washington state, USA, growing on dead Alnus wood

(Murrill 1917). Its pileus is avellaneous-isabelline, tomentose with a radiate-

rugose centre, and <4 cm broad; the stipe is concolorous with the pileus and

densely tomentose (Murrill 1917). Pluteus latifolius differs from P. dianae in

its smaller basidiomata, smaller basidiospores (5.2-—7.2 x 4.5-5.5 um, avg.

= 6.2 x 4.8 um, Q = 1.18-1.39, avQ = 1.29), and lageniform caulocystidia.

Pluteus atriavellaneus was described from Tennessee, USA, growing

solitary on humus (Murrill 1917). In contrast to P dianae, it is a small

species (pileus <2 cm) with a dark fuliginous to avellaneous, hygrophanous

and finely pubescent pileus and white, smooth stipe (Murrill 1917). Pluteus

punctatus differs from P. atriavellaneus by the presence of squamules on

pileus and stipe.

Pluteus avellaneus was described from New York, USA, growing on

unidentified dead wood (Murrill 1917). The pileus is described as small,

hazel coloured, paler at centre, glabrous, hygrophanous and <3.5 cm broad;

the stipe is white and smooth (Murrill 1917). Its smaller pileus and stipe

dimensions and hazel color of the smooth pileus distinguishes P avellaneus

from P. dianae. Pluteus punctatus differs from P. avellaneus by the presence

Pluteus dianae & P. punctatus in eastern & northern Europe... 271

of squamules on pileus and stipe. Neither P punctatus nor P. dianae have a

hygrophanous pileus.

Pluteus compressipes is a tropical species from Jamaica, found growing

on unidentified dead wood (Murrill 1917). Its pileus is rosy-isabelline with

a darker centre, glabrous, and <4 cm broad and its stipe is white, smooth,

and laterally compressed (Murrill 1917). Its occurrence in tropical areas and

a smooth compressed stipe are significant differences from Pluteus dianae

and P. punctatus.

Pluteus fibrillosus, collected in Louisiana, USA, from wet soil was

characterized by a dark, innately fibrillose, <3 cm broad pileus and smooth

white stipe (Murrill 1917). Pluteus dianae and P. punctatus differ by growth

on decaying wood and a (sub)flocculose stipe. The pileus of P punctatus is

not innately fibrillose but floccose-squamulose, and the pileus of P dianae

is paler and larger.

Pluteus fuliginosus, from New York, USA and collected from a decayed

Pinus strobus stump, was described with a uniformly coloured (fuliginous)

pileus clothed with white hairs and <4 cm broad and a white to pale yellow

or pale hazel and slightly squamulose stipe (Murrill 1917). The P fuliginosus

stipe was, as in P. punctatus, originally described as squamulose, although

we observed no caulocystidia on the type collection. The cheilocystidia of

P, fuliginosus are mostly lageniform or fusiform, and commonly provided

with an apical excrescence (A. Justo, pers. obs.). Its habitat on conifer wood

may be a reliable character to separate P fuliginosus from other species in

this complex, but additional collections are necessary to confirm this.

Conclusions

Our analyses confirm Pluteus dianae and P. punctatus as separate,

molecularly supported species. We characterize P dianae by rather large

basidiomata, a whitish to greyish-brown pileus that may turn pale after drying,

and a striate granulose-pruinose whitish stipe. Its striking microscopic

features are its lageniform and fusiform pedicellate pleurocystidia with a

narrowed apex often bearing a small apical excrescence, variable sized

pileipellis elements, and caulocystidia in tufts.

Pluteus punctatus is characterized by medium sized basidiomata, a pale

grey-brown pileus with ochraceous or (grey-) brown floccose squamules,

and a whitish stipe with brown tomentose squamules. Microscopically

striking are the rather short pleurocystidia, the long terminal elements in

the pileipellis and caulocystidia that are isolated or occur in only small tufts.

272 ... Sevéikova & al.

Collections of P dianae from central, eastern, and northern Europe

show that this species is widespread in Europe. Moreover, it has also been

documented from Turkey. The P. punctatus material of from Sweden is the

only collection besides the holotype from Czechia. The distribution of both

species should be further investigated.

Acknowledgments

The authors thank J. Holec (National Museum, Herbarium PRM, Prague, Czech

Republic) and N. Menolli Jr. (Instituto Federal de Educagao, Ciéncia e Tecnologia

de Sao Paulo (IFSP), Brazil) for their pre-submission reviews of the manuscript. The

authors also thank A. Anderberg (S); E. Bloch (NY); A. Bond (K(M)); B. Buyck (PC);

J. Holec (PRM), O. Koukol (PRC), and M. Smith (FLAS) for the possibility to study

herbarium collections, M. Micka and J. Her¢cik for their collections, and M. Sochor

for sequencing some collections. We also thank J.W. Jongepier who helped improve

the language of the manuscript. The studies by Hana Sevéikova were enabled by

support provided to the Moravian Museum by the Ministry of Culture of the Czech

Republic as part of its long-term conceptual development programme for research

institutions (DKRVO, ref. MK000094862). The studies of the Ekaterina F Malysheva

were enabled by support provided to project AAAA-A19-119020890079-6. Ekaterina

F. Malysheva's research was funded by a project of the Komarov Botanical Institute of

the Russian Academy of Sciences, and Michal TomSovsky’s research was supported

by the European Regional Development Fund, Project Phytophthora Research

Centre, Reg. No. CZ.02.1.01/0.0/0.0/15_003/0000453.

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MYCOTAXON

April-June 2020—Volume 135, pp. 275-280

https://doi.org/10.5248/135.275

Aecidium peristrophes, a new record for Pakistan

N.S. AFSHAN”, M. Riaz, A. SAQIB

Department of Botany, University of the Punjab, Quaid-e-Azam Campus-54590, Lahore, Pakistan

“CORRESPONDENCE TO: najamulsehar.botany@pu.edu.pk, pakrust@gmail.com

AsBsTRAcT—During a survey of plant pathogenic fungi of Khanspur, Thandiani,

and adjacent areas, two plant species—Hedera helix (Araliaceae) and Peristrophe sp.

(Acanthaceae)—were found to be infected with rust fungi. After careful morphological

and anatomical observations, these rusts were identified as Aecidium hederae and

A. peristrophes, with A. peristrophes reported as a new record for Pakistan.

Key worps— Abbottabad district, aeciospores, Pucciniales

Introduction

Aecidium Pers. is an asexual genus of rust fungi that possesses non-repeating

spores. Following dikaryotization of spermogonia, aeciospores are produced.

Catenulate spores are the characteristic feature of this genus. These spores are

surrounded by special structures called peridia (Cummins & Hiratsuka 2003).

Although the asexual morph generic name is now considered a synonym of

Puccinia and no longer recognized (Aime & al. 2018), a number of species

previously described in this genus have not been placed in valid genera and thus

must continue to be recognized in Aecidium. Seventeen species of this asexual

morph genus are reported from Pakistan (Ahmad & al. 1997; Ishaq & al. 2018).

During the 2016-18 surveys of pathogenic fungi, we collected plants of Hedera

helix and Peristrophe infected with rust fungi. Based on their morphology these

rusts were identified as Aecidium hederae and A. peristrophes.

Materials & methods

Plants infected with rust fungi were collected from Khanspur and Thandiani

forest, Khyber Pakhtunkhwa province, Pakistan. Healthy plants along with fruits

276 ... Afshan, Riaz, Saqib

PLATE 1: Aecidium peristrophes, A. Infected host plant, Peristrophe sp., B. Infection under

stereomicroscope, C. Aeciospores, D. Peridial cells. Scale bars: A= 5 mm, B= 1 mm, C, D= 10 um.

and inflorescence were also collected for identification. Host plants were identified

by comparing with plants in the herbarium of Department of Botany, University of

the Punjab, Lahore (LAH).

Scratch mounts and free hand sections of infected portions were made in

lactophenol. Partial permanent slides were made by cementing glass cover slips with

nail lacquer and slides were examined under a biological microscope (LABOMED,

Aecidium peristrophes new for Pakistan ... 277

Labo America, Inc. USA). Line drawings of different spore stages were made using

a Camera Lucida (Ernst Leitz, Wetzlar, Germany). Dimensions of spores were taken

using Scope Image 9.0(5X) with 40X objective. Forty spores were measured for each

species.

Taxonomy

Aecidium peristrophes Syd. & P. Syd., Ann. Mycol. 10(3): 272 (1912) PLs 1-2

SPERMOGONIA, UREDINIA, and TELIA not found. AEcIA epiphyllous, loosely

arranged, yellowish brown, irregular. AECIOSPORES ovoid to angular, hyaline to

subhyaline, 1-3 germ pores per cell, 16—23 x 20-24 um; peridial cells hyaline

to subhyaline, angular to hexagonal-pentagonal, 17-30 x 19-28 um, wall

verrucose, 2—5 um thick.

MATERIAL EXAMINED: PAKISTAN, KHYBER PAKHTUNKHWA, Abbottabad District,

Thandiani, at 2750 m asl, on Peristrophe sp. (Acanthaceae), 14 August 2018, N.S. Afshan,

NSA#01 (LAH1408).

PLATE 2: Line drawings of Aecidium peristrophes,

A. Peridial cells, B. Aeciospores. Scale bars: A, B = 5 um. (Drawings by Maria Riaz)

ComMMENTS—Aecidium peristrophes was collected on Peristrophe sp. and

represents a new record for Pakistan. It was previously reported on the same

278 ... Afshan, Riaz, Saqib

host genus from China and India (Sydow & al. 1912, Laundon 1963, Zhuang

2001). Puccinia peristrophes Petr. was also reported on the same host plant

from Malaysia (Petrak 1954), but aeciospores were not cited in the description.

Peristrophe sp. is also reported here as a new host for rust fungi in Pakistan.

Aecidium hederae Wakef., Bull. Misc. Inf., Kew 1931: 202 (1931) PLs 3-4

SPERMOGONIA, UREDINIA and TELIA absent. AEcIA amphigenous, cup-

shaped, pale yellow to yellowish orange spots, mostly grouped, sometimes

scattered, 1-3 x 1-1.5 mm. AEcIosPorEs pale yellow to honey colored with

yellowish orange granules, subglobose or ovoid—obovoid, 11-16 x 15-23 um;

wall hyaline, verrucose, 1-3 um thick; peridial cells angular to subangular

or rhomboidal, hyaline to pale yellow, 11-19 x 15-26 um, wall verrucose to

striolate verrucose, 2—4 um thick; germ pores obscure.

MATERIAL EXAMINED: PAKISTAN, KHYBER PAKHTUNKHWA, Abbottabad District,

Khanspur, at 2250 m asl, on Hedera helix L. (Araliaceae), 3 May 2017, Najam ul

Sehar Afshan NSA#02 (LAH1409); June 2018, N.S. Afshan, A.N. Khalid, NRPU#02

(LAH14096).

3 5 ee

{ee

oe ” '°

Ss,

PLaTE 3: Line drawings of Aecidium hederae,

A. Peridial cells, B. Aeciospores. Scale bars: A, B = 5 um. (Drawings by Maria Riaz)

ComMMENTS—Aecidium hederae has previously been reported from Pakistan

on Hedera helix and H. nepalensis K. Koch from Murree and on H. helix

Aecidium peristrophes new for Pakistan ... 279

PLatTeE 4: Aecidium hederae, A. Infected host plant, Hedera helix; B. Detail of infection; C. Peridial

cells; D. Aeciospores; E. SEM photograph of peridial cells; F SEM photograph of aeciospores.

Scale bars: A = 2 cm, B=5 mm, C, D= 10 um, E, F=5 um.

280 ... Afshan, Riaz, Saqib

from Malkandi, Kaghan (Wakefield 1931, Ahmad 1956, Iqbal & Khalid 1996,

Ahmad & al. 1997). The rust has also been reported on Hedera himalaica Tobler

[= H. nepalensis] and H. helix from India (Farr & Rossman 2018). This is a new

record for Khanspur, Pakistan.

Acknowledgments

We thank Prof. Dr. Abdul Nasir Khalid (Department of Botany, University of

the Punjab, Lahore, Pakistan) for his help in field work. The authors also wish to

thank Dr. Amy Y. Rossman (Oregon State University, Corvallis, USA) and Dr. Omar

Paino Perdomo (Instituto Tecndlogico de Santo Domingo, Dominican Republic) for

presubmission reviews of this manuscript. This work was financially supported by

Higher Education Commission (HEC), Islamabad, Pakistan for Research Project No.

7531/Punjab/NRPU/R&D/HEC/2017 under the “NRPU Scheme’.

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Cummins GB, Hiratsuka Y. 2003. Illustrated genera of rust fungi. 3" ed., The American

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Retrieved 4 April 2019 from https://nt.ars-grin.gov/fungaldatabases/

Iqbal SH, Khalid AN.1996. Material for the fungus flora of Pakistan. II. An updated check list

of rust fungi (Uredinales) of Pakistan. Sultania 1: 39-67.

Ishaq A, Saleem S, Afshan NS, Khalid AN, Niazi AR. 2018. New reports of the plant pathogens

Aecidium viburni and Erysiphe viburni on Viburnum grandiflorum from Pakistan.

Mycotaxon133: 551-557. https://doi.org/10.5248/133.551

Laundon GF.1963. Rust fungi 1: On Acanthaceae. Mycological Papers 89: 1-89.

Petrak F. 1954. Beitrage zur Pilzflora von Britisch Nord-Borneo. Sydowia 8: 12-26.

Sultan MA, Haq I, Khalid AN, Mukhtar H. 2008. Two new anamorphic rust fungi from

northern areas of Pakistan. Mycotaxon 105: 23-27.

Sydow H, Sydow P, Butler EJ. Fungi Indiae orientalis, pars IV. Annales Mycologici 10: 243-280.

Wakefield EM. 1931. Fungi exotici: XXVII. Bulletin of Miscellaneous Information, Kew

1931(4): 201-206. https://doi.org/10.2307/4102578

Zhuang WY. 2001. Higher fungi of tropical China. Mycotaxon, Ltd., Ithaca, NY. 485.

MYCOTAXON

April-June 2020—Volume 135, pp. 281-292

https://doi.org/10.5248/135.281

Westerdykella aquatica sp. nov., producing phytase

Hat-YAN SONG’, ALY FARAG EL SHEIKHA”?*> ©, PING-AN ZHONG’,

JIANG-LIN L1Ao', ZHAO-HAI WANG’, YING-JIN HUANG’,

DIAN-MING Hu??4°?

' Key Laboratory of Crop Physiology, Ecology and Genetic Breeding

(Jiangxi Agricultural University), Ministry of Education of the PR. China,

1101 Zhimin Road, Nanchang 330045, China

’ Bioengineering and Technological Research Centre for Edible and Medicinal Fungi,

Jiangxi Agricultural University, 1101 Zhimin Road, Nanchang 330045, China

> Jiangxi Key Laboratory for Conservation and Utilization of Fungal Resources,

Jiangxi Agricultural University, 1101 Zhimin Road, Nanchang 330045, China

* College of Bioscience and Bioengineering, Jiangxi Agricultural University,

1101 Zhimin Road, Nanchang 330045, China

° Department of Food Science and Technology, Minufiya University,

Shibin El Kom, Minufiya Government, Egypt

° Forestry Department, Jiangxi Environmental Engineering Vocational College,

State road 105, Economic- Technological Development Area, Ganzhou 341002, China

" CORRESPONDENCE TO: “ yjhuang_cn@126.com; * hudianming1@163.com;

° elsheikha_aly@yahoo.com

ABSTRACT— Westerdykella aquatica, isolated from rice field mud and as an endophyte from

Acorus calamus, is described as a new species. The fungus is characterized by its globose

cleistothecia, globose to subglobose, persistent, 8-spored asci, and cylindrical, 3-septate,

light brown ascospores. A phylogenetic tree based on ITS, LSU, and B-tubulin sequences

was constructed to infer the phylogenetic relationship between W. aquatica and other

Westerdykella species in the genus. The new species was found to produce phytase, an

important enzyme to release organic phosphorus in the soil.

Key worps—Ascomycota, Dothideomycetes, freshwater fungi, phylogeny, Sporomiaceae

Introduction

Westerdykella Stolk was established to accommodate W. ornata, isolated

from mangrove mud on the island of Inhaca, Mozambique, East Africa.

282 ... Song &al.

The genus is characterized by its globose, black and astomatous ascomata,

membranous cleistothecium wall consisting of one layer of brown to black

thick-walled cells, subglobose to elliptical multi-spored asci, and globose to

subglobose, brown ascospores (Stolk 1955).

Currently, 13 additional species are accepted in Westerdykella:

W. angulata (A.C. Das) Kruys, W. aurantiaca (J.N. Rai & J.P. Tewari) Kruys,

W. capitulum (Panwar & al.) Gruyter & al., W. centenaria Crous & al.,

W. cylindrica (Malloch & Cain) Arx, W. dispersa (Clum) Cejp & Milko,

W. globosa (J.N. Rai & J.P. Tewari) Tad. Ito & Nakagiri, W. minutispora

(P.N. Mathur) Gruyter & al., W. multispora (Cain) Cejp & Milko, W. nigra

(Routien) Arx, W. ornata Stolk, W. purpurea (Cain) Arx, and W. reniformis

Ebead & Overy (von Arx 1973, 1975, Cejp & Milko 1964, Crous & al. 2017,

Ebead & al. 2012, Gruyter & al. 2013, Ito & Nakagiri 1995, Kruys & Wedin

2009, Stolk 1955).

Molecular phylogenies inferred from miulti-loci showed that

Westerdykella is monophyletic (Kruys & Wedin 2009; Ebead & al. 2012;

Crous & al. 2017). However, morphologically some species do not fit well

within the original generic concept (Stolk 1955). Therefore, a revision is

needed to make the generic concept consistent with the species characters,

and the asexual concept of the genus should be added.

Westerdykella species are an important bioresource, producing various

bioactive substances such as antibiotics (Ebead & al. 2012; Xu & al. 2017b),

cytotoxics (Xu & al. 2017a), enzyme inhibitors (Lee & al. 1997), and

apoptosis inhibitors (Lee & al. 1999). During a long-term investigation of

aquatic fungi (Hu & al. 2007, 2012a,b, 2017; Huang & al. 2018; Song & al.

2018a,b), a new Westerdykella species producing phytase was encountered

in the mud of a rice field and also isolated as an endophyte of an Acorus

plant in Jiangxi Province, China.

Phosphorus, an essential nutrient for the growth and development of

living organisms, plays a vital role in virtually every plant process that

involves energy transfer. Most organic phosphorus is in the form of phytic

acid, an unavailable form of phosphorus. Phytase is an enzyme that releases

the phosphorus from the phytate complex to make it available (Sandhya

& al. 2015). The enzymatic degradation of phytic acid, which does not

produce toxic by-products, is an environmentally friendly process (Ciofalo

& al. 2003). In view of increasing demand, the production of phytase in

a cost-effective manner using microorganisms is an important field of

study (Shivanna & Venkateswaran 2014, Song & al. 2019). To the best of

Westerdykella aquatica sp. nov. (China) ... 283

our knowledge, this is the first paper assessing the phytase activities of the

genus Westerdykella, a new bioresource of this enzyme.

Materials & methods

Sample collection and fungal isolation

Soil and plant samples were collected in various freshwater habitats in Jiangxi

Province, China. The samples were placed in sterile ziplock plastic bags (Supin™) and

brought back to the laboratory.

TABLE 1. Strains used in the phylogenetic analyses.

GENBANK ACCESS. NO.

SPECIES STRAIN No.

ITS 28S B-tubulin

Forliomyces uniseptatus MFLUCC 15-0765 KU721772 KU721762 _

Preussia funiculata Huhndorf 2577 GQ203762 GQ203722 GQ203685

P. typharum CBS 107.69 GQ203766 GQ203726 GQ203689

P. vulgaris Strid 18884 GQ203767 GQ203727 GQ203690

Sparticola juncei MFLU 16-0242 KY659562 KY659565 _

Sporormia fimetaria Lundqvist 2302-c GQ203768 GQ203728 GQ203691

UPS: Dissing Gr.81.194 GQ203769 GQ203729 GQ203692

Sporormiella irregularis Lundqvist 16568-f GQ203780 GQ203739 GQ203700

S. leporina MJR93/04 GQ203782 GQ203741 GQ203702

S. vexans UME23 GQ203793 GQ203751 GQ203712

Westerdykella angulata CBS 610.74 GQ203757 DQ384105 —

IMI 090323 GQ203758 GQ203720 GQ203680

W. aquatica JAUCC 1788 [T] MH411093 MH411090 MH424907

JAUCC 0138 MH411092 MH411091 MH424906

W. aurantiaca FNBR-03 JN118571 — —

IMI 086825 AY943057 _— —

W. centenaria CBS 142400 KY979734 KY979790 KY979908

CBS 272.74 KY979735 — —

W. cylindrica ATCC 24077 AY943056 AY004343 Jx235707

W. dispersa CBS 156.67 DQ468016 — —

CBS 297.56 GQ203797 GQ203753 GQ203716

CBS 508.75 GQ203798 DQ384099 —

W. globosa IFO 32588 AY943046 — —

SWC5 KY065369 _— —

SWC6 KY260681 — —

W. minutispora CBS 509.91 - GU238108 _

W. multispora CBS 383.69 GQ203799 GQ203754 GQ203717

W. nigra ATCC 12756 AY943049 — —

CBS 416.72 GQ203800 GQ203755 GQ203718

W. ornata CBS 379.55 GQ203801 AY853401 GQ203719

W. reniformis DAOM 242243 Jx235700 Jx235704 Jx235706

Stemphylium vesicarium ATCC 11681 AF229479 AF382386 AY749032

Bold font indicates novel sequences generated in this study.

284 ... Song &al.

The soil samples were serially diluted up to 10° with sterile distilled water and

spread on the surface of potato dextrose agar media (PDA) in plates, which were

then incubated at 25 °C for 1-2 days. Stems of Acorus calamus were sliced into small

lengths and cleaned 3-5 times with distilled water (Nonaka & al. 2013). Plant tissue

pieces (c. 0.5 x 0.5 cm) were soaked in 75% alcohol for 30 seconds and immediately

washed 3-5 times in sterile water, dried in a sterile fume hood for c. 5 minutes, and

placed on PDA medium. Mycelium growing around the tissue was transferred to

new PDA plates (Sati & al. 2009). Colonies with different morphological characters

were transferred to new PDA plates. Cultures were dried as specimens and

deposited in the Herbarium of Fungi, Jiangxi Agricultural University, Nanchang,

China (HFJAU). Living cultures were deposited in the Culture Collection of Jiangxi

Agricultural University, Nanchang, China (JAUCC).

Morphological studies

Strains were grown on PDA at 25 °C and their growth rates and colony

morphology were evaluated after 7 and 14 days of incubation. The diameters of

colonies were measured in five directions and the average values were applied to

reflect the dimensions. For micro-morphological measurements and photographs,

fungal structures from 30-day-old cultures were mounted in water on glass slides;

and examined and photographed using a Nikon (Ni) compound microscope with

differential interference contrast (DIC) and a dissecting microscope (Hu & al. 2017).

DNA extraction and PCR amplification

Genomic DNA of strains was extracted by CTAB (hexadecyl trimethyl

ammonium bromide) method (Wu & al. 2001). Fragments of the partial large

subunit (LSU), internal transcribed spacer (ITS) rDNA, and £-tubulin gene (Bt2)

were amplified by the polymerase chain reaction (PCR). For PCR amplification,

primer pairs LROR/LR6 were used for LSU (Rehner & Samuels 1995; Vilgalys &

Hester 1990), ITS4/ITS5 for ITS (White & al. 1990), and T1/Bt2b for B-tubulin

(Glass & Donaldson 1995, O’Donnell & Cigelnik 1997). The amplification was

performed following the method described by Hu & al. (2012b). The PCR products

were purified and sequenced with the same primers used for PCR in a sequencer

(ABI-PRISM3730) at Tsingke Biological Technology Company, Beijing.

Phylogenetic analyses

Six novel sequences from the two strains of the new taxon (JAUCC 1788, JAUCC

0138) and reference sequences obtained from GenBank (TABLE 1), were aligned with

MAFFT v.7 (https://mafft.cbrc.jp/alignment/server/index.html; Katoh & Standley

2013). The concatenated aligned dataset was analyzed separately using maximum

likelihood (ML) and Bayesian inference (BI). The best-fit models of evolution for

the three loci tested were estimated by MrModeltest V.2.2.

The ML analyses were conducted with RAxML v.7.2.6 (Stamatakis & Alachiotis

2010) using a GFRGAMMA substitution model with 1,000 bootstrap replicates.

The robustness of the analyses was evaluated by bootstrap support (MLBS).

Westerdykella aquatica sp. nov. (China) ... 285

Posterior probabilities (PP) (Rannala & Yang 1996; Zhaxybayeva & Gogarten

2002) were determined by Markov chain Monte Carlo sampling (BMCMC) in

MrBayes 3.0b4 (Huelsenbeck & Ronquist 2001). Six simultaneous Markov chains

were run for 1,000,000 generations and trees were sampled every 100 generations

(resulting 10,000 total trees). The first 25% of trees were removed as burn-in phase

and the remaining trees were used to calculate posterior probabilities. Posterior

probabilities values of the BI analyses (BPP) >0.95 were considered significant.

Sequences generated in this study were deposited in GenBank (TaBLE 1) and the

final matrices and trees in TreeBASE (www.treebase.org; http://purl.org/phylo/

treebase/phylows/study/TB2:S22878?x-access-code=7f62703b04d4f5d2876667f65

4bd0398&format=html).

Phytase activity assessment

Mycelia obtained from fungal strains were inoculated to selective media (formula:

calcium phytate 0.1%, dextrose 3.0%, NH,NO, 0.5%, KCl 0.05%, MgSO,°7H,O

0.05%, MnSO ,°4H,,O 0.003%, FeSO ,°7H,O 0.003%, agar 2%, pH 5.5) in a petri dish,

and incubated in dark at 25 °C. Strains with transparent circles, indicating phytase

activity, were selected for further phytase activity evaluation.

The selected fungi were inoculated in 50 mL volume of liquid medium (formula:

dextrose 1.5%, peptone 0.3%, soluble starch 2%, (NH,),SO, 0.2%, KCI 0.05%,

MgSO,,.7H,O 0.05%, MnSO,+4H,O 0.003%, FeSO,+7H,O 0.003%, pH 5.5) in a 250

mL flask with two repeats. The fungi were fermented in a shaker at 28 °C, 160 rpm

for 120 hours. The fermentation broth was sampled three times, each sample was

centrifuged, and 10 mL of the supernatant were used to evaluate the phytase activity.

Phytase activity was evaluated by spectrophotometry according to the protocol of

the Determination of Feed Phytase Activity (GB/T 18634-2009) provided by China

National Standardization Administration (CNSA). One unit of phytase activity

(U) was expressed as the amount of enzyme that liberates 1 umol of inorganic

phosphorus from a 5.0 mM sodium phytate solution per minute at 37 °C and pH

5.5, while enzyme production was expressed as phytase activity U/mL.

Results

Fungal strain isolation

Two fungal strains representing what proved to be a new species were

obtained in this study. One strain JAUCC 1788 was isolated from rice field mud

as a saprophyte, and another strain JAUCC 0138 was isolated from the stems of

Acorus calamus as an endophyte.

Phylogenetic analyses

A multigene phylogeny, based on three loci, was used to infer the

relationships between the new taxon and its allied species (Fic. 1). The resulting

concatenated aligned dataset comprised 31 isolates representing six genera of

286 ... Song &al.

-/99{ Westerdykella aurantiaca FNBR-03

Westerdykella aurantiaca IMI 086825

-/82|-Westerdykella dispersa CBS 508.75

Westerdykella dispersa CBS 156.67

1/100 Westerdykella dispersa CBS 297.56

Is| Westerdykella angulata CBS 610.74

Westerdykella angulata IMI 090323

Westerdykella multispora CBS 383.69

100| Westerdykella globosa SWCS

|! Westerdykella globosa SWC6

Westerdykella globosa \FO 32588

Westerdykella ornata CBS 379.55

Westerdykella reniformis RKGE35

/100|Westerdykella centenaria C.

194 Westerdykella centenaria

1/99

-/93 1/100|Westerdykella aquatica

1/100 Westerdykella aquatica

0.99/100 }$ ————————————Westerdykella cylir d

Westerdykella nigra ATCC 12756

Westerdykella nigra CBS 416.72

0.99/96 Westerdykella minutispora CBS 509.91 =

Preussia funiculata Huhndorf 2577

1/-| |_1/100|-Preussia typharum CBS 107.69 Preussia

Sp orormiaceae Preussia vulgaris Strid 18884

Sporormia fimetaria Lundqvist 2302-c A i

a Be seas HG aS seek ae ‘Sporormia

VW of —— Forliomyces uniseptatus MFLUCC 15-0765 ” Q

Sparticola juncei MFLU 16-0242 ‘Sparticola

0.96/-| 1/100 Sporormiella pecans JN \ yn

SS Sporormiella leporina S:Ri

Sporormiella irregularis Lundqvist

Stemphylium vesicarium ATCC 11681

0.1

Fic. 1 Consensus tree inferred from a maximum likelihood analysis of ITS, LSU, and 6-tubulin

gene sequences. The RAxML bootstrap support values (MLBS) and Bayesian posterior probabilities

(BPP) are given at the nodes (MLBS/BPP). The tree is rooted with Stemphylium vesicarium (ATCC

11681).

Sporormiaceae: Forliomyces, Preussia, Sparticola, Sporormia, Sporormiella, and

Westerdykella; and an isolate of Stemphylium vesicarium (Wallr.) E.G. Simmons

(Pleosporaceae) as outgroup. The dataset consisted of 1952 characters (448

for ITS, 815 for LSU, and 689 for B-tubulin, including alignment gaps). The

trees generated from ML and Bayesian analyses of the individual loci (data not

shown) and the combined dataset showed essentially congruent topologies.

The ML tree based on the combined dataset is presented, with bootstrap

support values (MLBS) and Bayesian posterior probabilities (BPP) indicated

for well supported clades in Fic. 1. Two isolates of the new taxon ( Westerdykella

aquatica) together with other Westerdykella species formed a well-supported

clade (BPP = 1, MLBS = 100%).

Westerdykella aquatica sp. nov. (China) ... 287

Phytase activity

The phytase activities of the two Westerdykella aquatica isolates are presented

in TABLE 2.

TABLE 2. Phytase activity of Westerdykella aquatica strains.

PHYTASE ACTIVITIES (U/mL)

STRAIN NO.

Test 1 Test 2 Test 3 Average Combined average

JAUCC 0138 Treat 1 0.079 0.114 0.096 0.096+0.014 0.085

Treat 2 0.070 0.079 0.070 0.073+0.004

JAUCC 1788 Treat 1 0.079 0.071 0.079 0.076+0.005 0.083

Treat 2 0.079 0.176 0.018 0.091+0.065

Taxonomy

Westerdykella aquatica H.Y. Song & D.M. Hu, sp. nov. FIG. 2

MB 825645

Differs from Westerdykella angulata and W. purpurea by its cylindrical and 3-septate

ascospores; from W. cylindrica by its light brown ascospores; and from other

Westerdykella species by its 8-spored asci.

Type—China, Jiangxi Province, Jingan County, isolated from mud in a rice field, 15

July 2017, Jun-Bo Zhang (Holotype, HFJAU 0676; ex-type living culture, JAUCC 1788;

GenBank MH411090, MH411093, MH424907).

EryMOLOGY—aquatica, referring to the aquatic habitat of the fungus.

CoLonigs on PDA medium reaching 90 mm diam after 3 wk at 25 °C, spreading,

flat, velvety, with sparse aerial mycelium, yellow, reverse dark yellow, slightly

zonate, with several white rings, surface and margins smooth. Hyphae 1.4-4 um

diam, septate, branched, hyaline to light brown. SEXUAL sTATE: Cleistothecia

scattered, superficial, glabrous, globose, black, c. 100-260 um diam. Peridium

thin, membranous, composed of a single layer of globose to ellipsoidal, angular

cells. Asci initials somewhat ovoid, later becoming globose to subglobose,

14-17 x 12-16 (mean = 15.3 x 14.1 um, n = 30), persistent, 8-spored. Ascospores

17-38 x 2.5-4 um (mean = 27 x 3 um, n = 30), cylindrical, with rounded ends,

3-septate, slightly constricted at the septa, mostly separated into four segments

at the very early stage of spore-formation in asci. Ascospore segments 4.5-11

x 2.5-3.5 um (mean = 6.5 x 2.9 um, n = 50), ellipsoidal, subhyaline, smooth-

walled, light brown, mostly with a big guttule at each end, no germ-slits

observed. ASEXUAL STATE: not observed.

ADDITIONAL SPECIMEN EXAMINED—CHINA, JIANGXI PROVINCE, Jiangxi Agricultural

University, endophytic in Acorus calamus L. (Acoraceae) in a pond, 21 March 2014,

Guan-Xiu Guan (HFJAU 0677; living culture, JAUCC 0138; GenBank MH411091,

MH411092, MH424906).

288 ... Song &al.

Discussion

Westerdykella aquatica is characterized by its globose cleistothecia,

globose to subglobose, persistent and 8-spored asci, and cylindrical,

3-septate, light brown ascospores. These characters fit well within the

generic concept of Westerdykella. Furthermore, in the phylogenetic tree

inferred from ITS, LSU, and 6-tubulin genes, W. aquatica and other 19

reference strains of 11 species of Westerdykella form a well-supported clade

(BPP = 1, MLBS = 100%).

Westerdykella aquatica resembles W. angulata, W. cylindrica and W. purpurea

in having 8-spored asci. However, W. aquatica differs from W. angulata and

W. purpurea in the shape of its ascospores. The ascospores of W. aquatica

are cylindrical and 3-septate, while those of W. angulata and W. purpurea

are angular (Das 1962; von Arx 1975; Kruys & Wedin 2009). W. cylindrica

differs from W. aquatica in its reddish brown ascospores (Malloch & Cain

1972). Furthermore, the strains of W. cylindrica and W. aquatica are well

separated in the phylogenetic tree (Fic. 1).

Although various chemical compounds have been reported from species

of Westerdykella (Lee & al. 1997, 1999; Ebead & al. 2012; Xu & al. 2017 ,b),

our study is the first report of phytase production by the genus. Richardson

(1994) reported that 20-80% of P in soils is found in the organic form, of

which phytate (inositol hexaphosphate) is usually a major component. The

benefits of phytase are its double effects on reducing the use of expensive

inorganic phosphorus as fertilizer and its reduction of environmental

pollution from excessive manure phosphorus runoff (Wang & Yang 2007).

Westerdykella aquatica JAUCC 1788 was isolated from the mud of rice field,

and its presence may help liberate P from the soil and reduce the use of

P fertilizer.

Acknowledgments

The authors express their sincere gratitude to Dr. Eric H.C. McKenzie (Landcare

Research, Auckland, New Zealand) and Dr. Huang Zhang (Kunming University of

Science & Technology, China) for their critical review of the manuscript, Dr. Shaun

Pennycook for nomenclatural review, and Dr. Lorelei L. Norvell for editorial review.

This study was funded by the National Natural Science Foundation of China (NSFC)

(No. 31500021 & 31460009), the Key Research and Development Projects of Jiangxi

Provincial Department of Science and Technology, China (No. 20161BBF60078), and

the Graduate Innovation Fund of Jiangxi Province (No. YC2017-B035).

Westerdykella aquatica sp. nov. (China) ... 289

oof,

k Ss eee — a

Fic. 2 Westerdykella aquatica (holotype, HFJAU 0676). a. Colony on PDA; b. Ascoma on PDA;

c. Ascoma; d. Squashed ascoma; e. Peridium; f. Immature ascus; g-l. Asci; m. Ascospores;

n. Ascospore segments. Scale bars: b = 1 mm; c, d = 100 um; e-n = 10 um.

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MYCOTAXON

April-June 2020—Volume 135, pp. 293-298

https://doi.org/10.5248/135.293

Ramiphialis ronuroensis gen. and sp. nov.,

a hyphomycete from the Amazonian rainforest

FLAVIA RODRIGUES BARBOSA’, PATRICIA OLIVEIRA FIUZA?,

RAFAEL FE. CASTANEDA-RUuIZ?

"Instituto de Ciéncias Naturais, Humanas e Sociais, Universidade Federal de Mato Grosso,

Av. Alexandre Ferronato, 1200, 78557-267, Sinop, Brazil

? Programa de Poés-graduacdo em Sistematica e Evolugao,

Universidade Federal do Rio Grande do Norte, Campus Universitario,

Av. Senador Salgado Filho, 3000, Lagoa Nova, Natal-RN, 59078-970, Brazil

° Instituto de Investigaciones Fundamentales en Agricultura Tropical Alejandro de Humboldt

(INIFAT), Académico Titular de la Academia de Ciencias de Cuba,

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

“ CORRESPONDENCE TO: faurb10@yahoo.com. br

ABSTRACT—A new asexual ascomycete genus and species, Ramiphialis ronuroensis,

was discovered on decaying leaves from Amazon rainforest in Brazil and is described

and illustrated here. Morphologically, the fungus is distinguished by macronematous,

monophialidic and multibranched, discrete, and terminal and intercalary conidiogenous

cells that produce filiform to falcate, unicellular, hyaline conidia.

Key worps—biodiversity, taxonomy, tropical fungi

Introduction

The discovery of new species is important to improve understanding of

which and how many species will extinct (Costello & al. 2013). Although

the Amazon occupies a prominent position in relation to global biodiversity,

many species are disappearing due to the human actions. Studies on asexual

fungi from Brazil have received more attention in Atlantic forest and Caatinga

biomes (Barbosa & al. 2013, Fiuza & al. 2015, Santa-Izabel & Gusmao 2018).

According to Sotao & al. (2004), little is known about asexual fungi in

294 ... Barbosa, Fiuza, Castaheda

Amazon. The northern region of the Amazon has more studies (Carmo &

al. 2014, Castro & al. 2012, Hernandez-Gutierrez 2013, Monteiro & al. 2014)

while in the southern region the investigations are still initial (Barbosa & al.

2015, 2017).

A survey of asexual ascomycetes on plant debris was carried out in

the southern Amazon forest and an interesting fungus with branched

conidiophores and phialidic conidiogenous cells is proposed here as a new

genus and species.

Materials & methods

Samples of plant debris were collected in November 2016 at Rio Ronuro Ecological

Station, Nova Ubirata, Mato Grosso state (9°52’24”S 58°13’17”W) and placed in paper

bags. In the laboratory, samples were washed and stored in moist chambers following

Castafieda-Ruiz & al. (2016). After 72 h, the reproductive structures were mounted in

PVL resin (polyvinyl alcohol, lactic acid, and phenol) and examined and photographed

using a microscope equipped with phase contrast. Drawing was produced according

Almeida & Gusmao (2015). The type specimen was deposited in the Herbario Centro-

Norte Mato-Grossense, Sinop, Mato Grosso State, Brazil (CNMT).

Taxonomy

Ramiphialis E.R. Barbosa, Fiuza & R.F. Castafieda, gen. nov.

MB 831423

Differs from Atrosetaphiale by its branched conidiophores, terminal phialides, and

falcate conidia; from Veramycina by the absence of setae; and from Jayarambhatia by its

conidiophores with extensions, discrete phialides, and falcate conidia.

TYPE SPECIES: Ramiphialis ronuroensis ER. Barbosa & al.

Erymotoey: Latin, rami, referring to the branched conidiophores; and phialidis,

referring to the type of conidiogenous cells.

COLONIES scattered, dark brown. Mycelium superficial and immersed,

composed of slightly branched, septate, smooth, brown hyphae.

CONIDIOPHORES macronematous, mononematous, erect, straight or

flexuous, branched, septate, with percurrent extensions, smooth, brown.

CONIDIOGENOUS CELLS monophialidic, terminal and intercalary, discrete,

determinate, pale brown. Conidial secession schizolytic. Conrp1A falcate,

unicellular, hyaline accumulating in a white mucilaginous mass.

Ramiphialis ronuroensis E.R. Barbosa, Fiuza & R.F. Castafieda, sp. nov. Fies 1, 2

MB 831424

Differs from Atrosetaphiale flagelliformis by its branched conidiophores, terminal

phialides, and falcate conidia; from Veramycina elegans by the absence of setae; and from

Ramiphialis ronuroensis gen. & sp. nov. (Brazil) ... 295

Fic. 1. Ramiphialis ronuroensis (holotype, CNMTf 80). A. Conidium; B. Conidia in mucilaginous

mass; C, D. Conidiogenous cells and conidia; E-G. Conidiophores. Scale bars: A = 5 um; B = 10

um; C, D = 20 um; E = 40 um; F, G= 50 um.

296 ... Barbosa, Fiuza, Castaheda

Fic. 2. Ramiphialis ronuroensis (holotype, CNMTf 80).

A. Conidia; B. Conidiophore, conidiogenous cells, and conidia; C. Conidiophore.

Scale bar = 20 um.

Jayarambhatia rhizophorae by its conidiophores with extensions, discrete phialides, and

falcate conidia.

Type: Brazil, Mato Grosso State, Nova Ubirata, Estagao Ecolégica Rio Ronuro, on

decaying leaves, 11.XX.2016, coll. ER. Barbosa (Holotype, CNMTf 80).

EryMo .oey: Latin, ronuroensis, refers to the place where the species was collected.

COLONIES scattered, dark brown. Mycelium superficial and immersed,

composed of slightly branched, septate, smooth, brown hyphae.

CONIDIOPHORES macronematous, mononematous, erect, straight or flexuous,

Ramiphialis ronuroensis gen. & sp. nov. (Brazil) ... 297

multibranched, 5-12 septate, with percurrent extensions, smooth, brown,

130-200 x 6-9 um. The branches are dichotomous starting near the lobed

basal cell. CoNIDIOGENOUS CELLS monophialidic, terminal and intercalary,

discrete, sometimes forming subpenicillate clusters, evident, determinate,

pale brown, cylindrical to slightly ovoid, 12.5-17.5 x 5 um. Conidial secession

schizolytic. Conrp1a filiform, falcate, hyaline, unicellular, smooth, 10-18(-25)

x 0.5-0.7 um, accumulating in a white mucilaginous mass.

CoMMENTS—Atrosetaphiale Matsush., Jayarambhatia J. Pratibha, and

Veramycina Subram. superficially resemble Ramiphialis in possessing

phialidic conidiogenous cells that produce narrow conidia. Atrosetaphiale,

however, is distinguished by its unbranched conidiophores and lateral and

terminal phialides; its type species, A. flagelliformis Matsush., produces

cylindrical-fusiform to flagellate conidia (Matsushima 1995), in contrast to

the filiform conidia characterizing R. ronuroensis. Jayarambhatia differs in

its determinate conidiophores, integrated phialides with elongated narrow

necks, and narrowly obclavate conidia, while the generic type, J. rhizophorae

J. Pratibha, produces much larger conidia (30-50 x 1 um; Pratibha 2013).

Veramycina, the synanamorph of Oedemium Link, is distinguished by the

production of setae, conidiophores bearing terminal short branches, terminal

and lateral phialides, and cylindrical to bacillar conidia; the generic type,

V. elegans Subram., produces much shorter conidia (1.5-3 um; Subramanian

1993). However, Ramiphialis does not produce setae and has dichotomously

branched conidiophores.

Acknowledgments

The authors express their sincere gratitude to Dr. De-Wei Li (The Connecticut

Agricultural Experiment Station, Windsor, CT, USA) and Dr. Taimy Cantillo-

Pérez (Universidade Estadual de Feira de Santana, Novo Horizonte, Brazil) for

their critical review of the manuscript. FR. Barbosa thanks SEMA-MT (Secretaria

de Estado do Meio Ambiente), FUNBIO (Fundo Brasileiro para a Biodiversidade),

PPGCAM/UFMT (Programa de Pés-Graduacgéo em Ciéncias Ambientais), and

PPBIO (Programa de Pesquisa em Biodiversidade) for financial support. Patricia

Fiuza thanks CAPES-PNPD (Coordenacao de Aperfeigoamento de Pessoal de

Nivel Superior - Programa Nacional de Pés-doutorado) for a scholarship (Proc.

88882.306016/2018-01). Dr. Lorelei Norvell’s editorial review and Dr. Shaun

Pennycook’s nomenclature review are greatly appreciated.

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fungi. Mycosphere 6(5): 630-633. https://doi.org/10.5943/mycosphere/6/5/12

298 ... Barbosa, Fiuza, Castaheda

Barbosa FR, Raja HA, Shearer CA, Gusmao LFP. 2013. Some freshwater fungi from the Brazilian

semi-arid region, including two new species of hyphomycetes. Cryptogamie Mycologie 34(3):

243-258. https://doi.org/10.7872/crym.v34.iss2.2013.243

Barbosa FR, Machiner M, Barbosa GCK, Alexandre FS, Ribeiro ML. 2015. Fungos conidiais

decompositores de substratos vegetais. 41-52, in: DJ Rodrigues, JC Noronha, VF Vindica, FR

Barbosa (eds.). Biodiversidade do Parque Estadual Cristalino. Attema Editorial, Sao Paulo.

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hyphomycetes from the Brazilian Amazon rainforest. Mycotaxon 132 (3): 643-647.

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Carmo LT do, Monteiro JS, Gusmao LFP, Sotao HMP, Hernandez-Gutiérrez A, Castafieda-Ruiz

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127: 11-15. https://doi.org/10.5248/127.11

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Castro CCC, Hernandez-Gutiérrez A, Sotao HMP. 2012. Fungos conidiais em Euterpe oleracea

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Fiuza PO, Gusmao LFP, Castafeda-Ruiz RF. 2015. Conidial fungi from the semiarid Caatinga

biome of Brazil: a new species of Selenosporella from submerged leaves. Mycotaxon 130:

601-605. https://doi.org/10.5248/130.601

Hernandez-Gutiérrez A. 2013. New or rare fungi from eastern Amazonia. 1. Circinoconiopsis

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Matsushima T. 1995. Matsushima mycological memoirs 8. Published by the author, Kobe.

Monteiro JS, Gusmao LFP, Castafieda-Ruiz RF. 2014. Two new microfungi from Brazilian

Amazon Forest: Atrogeniculata submersa and Nigrolentilocus amazonicus. Mycotaxon 127:

39-45. https://doi.org/10.5248/127.39

Pratibha J. 2013. Jayarambhatia rhizophorae gen. et sp. nov., an asexually reproducing fungus

from Goa, India. Mycotaxon 125: 139-144. https://doi.org/10.5248/125.139

Santa-Izabel TS, Gusmao LFP. 2018. Richness and diversity of conidial fungi associated with

plant debris in three enclaves of Atlantic Forest in the Caatinga biome of Brazil. Plant Ecology

and Evolution 151 (1): 35-47. https://doi.org/10.5091/plecevo.2018.1332

Sotao HMP, Campos EL, Costa SPS. 2004. Micologia; diversidade de fungos na Amazonia. Série

cadernos de alfabetizacao cientifica. Vol. 1. 27 p.

Subramanian CV. 1993. Veramyces, a new hyphomycete genus from Kumaon Himalayas. Czech

Mycology 47(1): 3-5. https://doi.org/10.33585/cmy.47102

MYCOTAXON

April-June 2020—Volume 135, pp. 299-308

https://doi.org/10.5248/135.299

Phylogenetic placement of Acrospeira

DrE-WEI LI’, RAFAEL F. CASTANEDA-RuIz’, NEIL P. SCHULTES?*

' The Connecticut Agricultural Experiment Station, Valley Laboratory,

153 Cook Hill Road, Windsor, CT 06095, USA

? Instituto de Investigaciones Fundamentales en Agricultura Tropical Alejandro de Humboldt

(INIFAT), Académico Titular de la Academia de Ciencias de Cuba,

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

° The Connecticut Agricultural Experiment Station, Department of Plant Pathology & Ecology,

123 Huntington Street, New Haven, CT 06511, USA

* CORRESPONDENCE TO: neil.schultes@ct.gov

ABSTRACT —The morphology of Acrospeira mirabilis is described and illustrated based on

a collection from Florida, USA. Phylogenies generated from ITS and LSU DNA sequence

analyses place the genus Acrospeira in Ceratostomataceae. Based on molecular data,

Sphaerodes inferior (= Microthecium retisporum var. inferius) is proposed as Microthecium

inferius comb. nov.

KEY WORDS — asexual fungi, hyphomycete, synanamorph

Introduction

Acrospeira Berk. & Broome was erected and typified by A. mirabilis

(Berkeley 1857, Berkeley & Broome 1861). Six out of eight species described

in this genus have been transferred to other genera (Index Fungorum 2019,

MycoBank 2019). With the seventh invalid because it lacks a Latin description

(Art. 39.1, Shenzhen Code; Turland & al. 2018), Acrospeira has been

reduced to its original monotypic status, containing only the type species,

A. mirabilis. Its current phylogeneticand taxonomic placement is incertae sedis,

Pezizomycotina (Index Fungorum 2019, MycoBank 2019, Wijayawardene &

al. 2018).

300 ... Li, Castafieda-Ruiz, Schultes

A culture of Acrospeira mirabilis from a Florida 2018 collection allowed us

to conduct a phylogenetic study using two loci (ITS and LSU) and determine

its taxonomic placement using both morphological and molecular methods.

Materials & methods

Acrospeira mirabilis was obtained during a March 2018 collection trip in Largo,

Florida. The fungus was plated on malt extract medium (MEA) (20 g malt, 20 g

agar, 1 L distilled water), purified and subsequently grown at 25°C for 7 days for

morphological observation and molecular work. The fungus was mounted in 85%

lactic acid for microscopic observation and measurements with a Zeiss Imager.

M2 compound microscope using differential interference contrast (DIC) and

photographed with a Zeiss Axiocam 506 color camera. Fungal structures were

measured using 40-100x objectives and statistically analyzed to determine standard

deviations with 95% confidence interval of means.

The fungus culture has been deposited in the UAMH Centre for Global

Microfungal Biodiversity at University of Toronto (UAMH), Canada.

DNA extraction, amplification, and sequencing

Genomic DNA was extracted from colonies grown in Petri plates according to

the procedure in ZR Fungal/Bacterial DNA MicroPrep Kit. The oligonucleotides

ITS5 and LR7 (Vilgalys and Hester 1990, White et al. 1990) were used to amplify a

DNA fragment containing the internal transcribed spacer 1 and 2 region (ITS) anda

portion of the large subunit rDNA region (LSU) by polymerase chain reaction (PCR)

as described in Li & al. (2017). PCR products were purified using QIAquick PCR

Purification columns; DNA concentration was determined on a ThermoScientific

NanoDrop Lite Spectrophotometer. The purified PCR products were sequenced using

oligonucleotides ITS2, ITS3 and ITS5, LROR, LR3R, LR3B, LR5 & LR7 (Vilgalys &

Hester 1990, White & al. 1990, Li & al. 2017). The DNA was sequenced at the W.M.

Keck Biotechnology Resource Laboratory, Yale School of Medicine (New Haven, CT,

USA). New sequence information was deposited in GenBank (TABLE 1).

The ITS and LSU sequences were used in a BLASTn analysis of GenBank

(http://blast.ncbi.nlm.nih.gov). To identify DNA sequences from allied taxa,

BLASTn analysis was carried out with exclusion of uncultured/unidentified samples,

environmental samples and samples with questionable identifications (Wheeler & al.

2003). Fungal DNA sequences linked to verified fungal voucher cultures that showed

significant sequence similarity with the query sequences were chosen for phylogenetic

analysis. Additional ITS and LSU sequences from allied taxa were obtained totaling 44

sequences from 32 fungi for phylogenetic analyses (TABLE 1).

Phylogenetic analysis

ITS and LSU sequences were aligned independently using MUSCLE (Edgar 2004)

and manually corrected. The aligned sequences were trimmed and concatenated

with FABOX sequence alignment jointer (http://users-birc.au.dk/palle/php/fabox/

alignment_joiner.php).

Acrospeira (Ceratostomataceae) ... 301

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The sequence dataset with 44 nucleotide sequences and 1039 positions was

phylogenetically analysed using the maximum likelihood procedures, Tamura-Nei

model, and uniform rates with MEGA7 (Kumar & al. 2016, Tamura 1992); all sites

were treated equally. All sites were equally weighted, and gaps were treated as missing

data. A bootstrap was calculated with 1000 replicates and posed statistical support at

270%. Xylaria hypoxylon ATCC42768 was designated as an outgroup.

Harzia acremonioides NBRC5937

Harzia acremonioides NBRC8881

Harzia acremonioides NRRL 54327

Harzia acremonioides NRRL 54328

Harzia patula CBS 379.88

Harzia sphaerospora UAMH 11865

8: Harzia tenella CBS108.78

Harzia macrospora CBS 122807

Harzia patula CBS 121524

Harzia cameroonensis CBS 136420 T

a Harzia palmara CBS 995.69

Harzia palmara CBS 158.49

83 Harzia palmara NBRC8861

Harzia verrucosa CBS 101.24

- Harzia acremonioides CBS 101,17

Harzia acremonioides CBS 101.42

Harzia verrucosa CBS 113456

Harzia tenella CBS 121.81

Microthecium brevirostre ATCC 42427

Gonatobotryum parasiticum NBRC9010

Melanospora verrucispora NRBC 31375 T

Melanospora zamiae NBRC7902 Ceratostomataceae

1 Melanospora damnosa NBRC100318

75 Melanospora mycoparasitica SMCD2220 T

93 Dactylidispora ellipsospora NBRC 31376 T

Dactylidispora singaporensis ATCC 38286

Melanospora kurssanoviana NBRC 8098

Acrospeira mirabilis UAMH 12078

Melanospora zamiae NBRC32042

Microthecium zobelii NBRC9442

Microthecium fimbriatum NBRC8615

Microthecium fimicola NBRC8354

Microthecium ciliatum NBRC 9829 T

Microthecium fimicola NBRC9555

Microthecium fusisporum NBRC8806

Sphaerodes inferior NBRC8366 T

Microthecium quadrangulare CBS 112763 T

Melanospora ‘inaequalis' NBRC8426

Microthecium tenuissimum CBS112764 T

Melanospora pascuensis IMI 378527 T

Vittatispora coorgii BICC 7817 T

Melanospora tiffanyae SMCD2222

92 + Pseudomicrothecium subterraneum BJTC fan1001

Xylaria hypoxylon ATCC 42768

Fic. 1. Maximum Likelihood analysis of Acrospeira mirabilis and allied taxa, based on 44 sequences.

Xylaria hypoxylon ATCC42768 is included as outgroup. The bootstrap test was conducted with 1000

replicates; bootstrap values >70% are indicated at the nodes. The scale bar indicates the number of

expected changes per site. Annotations “T’ indicate the ex-type sequences included in the analysis.

304 ... Li, Castafieda-Ruiz, Schultes

Results

The ITS and LSU phylogenetic analyses place Acrospeira mirabilis in the

same clade as genera in Ceratostomataceae (Fic. 1). Acrospeira mirabilis

defines a sister clade to the clade including Microthecium zobelii (the

generic type), M. ciliatum, M. fimbriatum, M. fimicola, M. fusisporum,

M. quadrangulare, M. tenuissimum, Sphaerodes inferior, Melanospora zamiae

NBRC32042, Me. “inaequalis; and Me. zamiae (NBRC32042) (Fic. 1).

Taxonomy

Acrospeira mirabilis Berk. & Broome, Intr. Crypt. Bot.: 305 (1857) Fie.2:

This fungus is dimorphic. COLONIEs attained 35 mm in 7 days on MEA at

25°C, cottony, chocolate brown. Mycelia immersed or aerial. Hyphae septate,

hyaline, smooth, 3-6.8 um diam.

DOMINANT STATE: CONIDIOPHORES semi-macronematous,

mononematous, terminally or laterally arising from the hyphae and branched,

hyaline, 15-35 um long. CONIDIOGENOUS CELLS monoblastic integrated,

terminal, determinate: branch ends enlarged and coiled, hyaline. Conrp1a

solitary, dry, 3-celled, developed by dividing enlarged and coiled portion

of conidiogenous cell with 2 transverse septa—the terminal cell enlarged

greatly into a (sub)sphere, golden brown, tuberculate, (19.4—)21.9-27.1

(-30.5) um diam. (mean + SD = 24.5 + 2.6 um, n = 30), thick-walled; the

median cell enlarged, smooth, hyaline to golden (paler than terminal cell),

tuberculate, hemispherical, or dome-shaped, ornamented; and the basal cell

hyaline to yellow and smooth—the whole conidia (23.7—)25.4—30.4(-33.6) x

(19.4—)21.9-27.1(-30.5) um (27.9 + 2.5 x 24.5 + 2.6 um, n = 30), often with

a visible germ pore on the enlarged (sub)spherical terminal cell. Conidial

secession schizolytic.

SYNANAMORPH: phialidic conidial ontogeny. CONIDIOPHORES

micronematous, mostly reduced to conidiogenous cells, arranged primarily

on aerial hyphae, rarely on assimilative hyphae. CONIDIOGENOUS CELLS

monophialidic, discrete, lageniform or ampulliform, rarely integrated,

subulate, (5.5—)6.4-11.6(-14.7) x (3.4-)3.6-4.4(-4.9) um (9.0 + 2.6 x 4.0

+ 0.4 um, n = 21), hyaline, smooth. Conrpia basipetal, ellipsoid, ovoid,

lacrimiform, subglobose or globose, 1-celled, hyaline, smooth, (2.4—)2.7-3.5

(-4.0) x (2-)2.2-2.8(-3.1) um (3.1 + 0.4 x 2.5 + 0.3 um, n = 30).

SPECIMENS EXAMINED: UNITED STATES, Fioripa: Largo, 27°53’02”N 82°48’31”W,

on unknown leaf litter, 3.1I.2018, coll. De-Wei Li (UAMH 12078).

Acrospeira (Ceratostomataceae) ... 305

Fic. 2. Acrospeira mirabilis (UAMH 12078): a. colony grown on MEA for 7 days at 25°C; b. conidia

at coiled stage and conidiophores; c. conidiophores, conidiogenous cells. and conidia; d. conidia;

e. surface ornament of conidium with a germ pore (arrowed); f. bimorphic states at different

developmental stages; g. synnanamorphic state, conidia and phialides.

306 ... Li, Castafieda-Ruiz, Schultes

Discussion

Conidia of our collection are smaller than those of those described in

previous reports (Berkeley 1857, Berkeley & Broome 1861, Ellis 1971). In

USA, A. mirabilis has previously been reported from Florida, Massachusetts,

and Michigan (Donis Gonzalez & al. 2010, Farr & Rossman 2019).

Acrospeira has a phialidic synanamorph, while Harzia Costantin

develops a proteophiala synanamorph. The proteophiala-like synanamorph

is a key morphological character for merging the formerly accepted genera

Olpitrichum G.F. Atk. and Chlamydomyces Bainier into Harzia (supported by

phylogenetic analyses) and placing an emended Harzia in Ceratostomataceae

(Li & al. 2016, Schultes & al. 2017). Whether the phialidic synanamorph

plays a similar role in Ceratostomataceae, it is necessary to conduct further

morphological and phylogenetic studies including other anamorphic and

related genera. Seifert & al. (2011) suggested that Acrospeira belongs to

Ceratostomataceae, but there is no phylogenetic study or discovery of the

sexual state of the type species to support their statement. Our present

phylogenetic results support the opinion of Seifert & al. (2011) and firmly

demonstrate its taxonomic placement in Ceratostomataceae.

Microthecium was resurrected by Marin-Felix & al. (2018) for the M-S

(Melanospora-Sphaerodes) 2 clade in Schultes & al. (2017). Marin-Felix &

al. (2018), who examined the ex-type CBS 994.72 of Sphaerodes retispora

var. retispora, suggested that it was contaminated by a host fungus and that

S. inferior and S. retispora var. retispora were conspecific. They demoted

S. inferior to a synonym of S. retispora based on their opinion that there

were not enough morphological differences between the two species.

However, although Marin-Felix & al. (2018) used the sequences of ex-type

NBRC 8366 of S. inferior in their phylogenetic analysis, they did not include

the sequences of ex-type S. retispora var. retispora. The sequence from the

S. retispora ex-type fell into a clade with Hypocrea schweinitzii NBRC9063-T

and Trichoderma viridescens CBS 433.34-T (Schultes & al. 2017), unrelated

to Fusarium (the host fungus contaminant). We re-examined ex-type (CBS

994.72) of S. retispora var. retispora: the sexual stage did not develop on

MEA, although the monophialidic anamorph was present. The culture

was pure and we found no indication that the ex-type was contaminated.

In our opinion, the relationship between S. inferior and S. retispora

remains unsettled, and S. inferior should be retained as a separate taxon in

Microthecium.

Acrospeira (Ceratostomataceae) ... 307

Microthecium inferius (Udagawa & Cain) D.W. Li, R.F. Castafieda &

N.P. Schultes, comb. nov.

MB 831435

= Microthecium retisporum var. inferius Udagawa & Cain, Canad.

J. Bot. 47(12): 1928. 1970 [“1969”], as “inferior”.

= Sphaerodes retispora var. inferior (Udagawa & Cain) P.F. Cannon

& D. Hawksw., Bot. J. Linn. Soc. 84: 149. 1982.

= Sphaerodes inferior (Udagawa & Cain) D.W. Li & N.P.

Schultes, Fung. Biol. 121(10): 901. 2017.

Our phylogeny places the unpublished Melanospora ‘inaequalis’ NBRC 8366 =

TRIC 41540 in the Microthecium clade. The conserved strains of Melanospora

‘inaequalis’ need to be studied in the future. Further research of genera in

Ceratostomataceae is also required, especially Melanospora s.l. and Sphaerodes

s.L., given the polyphyletic characters of their representatives.

Acknowledgments

The authors express their sincere gratitude to Dr. Luis FP. Gusmao (Universidade

Estadual de Feira de Santana, Brazil) and Dr. Patricia Fiuza (Universidade Federal do

Rio Grande do Norte, Brazil) for their critical review of the manuscript, and Noelle

Strzalkowski and Sumeyra Yavuz for technical assistance. We would like to express

our gratitude to Dr. Carole Cheah for providing access to some literature. This work

is supported by Hatch Grant [grant no. CONH00813] from the USDA National

Institute of Food and Agriculture. Dr. Lorelei Norvell’s editorial review and Dr. Shaun

Pennycook’s nomenclature review are greatly appreciated.

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cameroonensis Crous & Jol. Roux, sp. nov. Persoonia 31: 212-213.

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Donis Gonzalez IR, Fulbright DW, Ryser ET, Guyer D. 2010. Shell mold and kernel decay of

fresh chestnuts in Michigan. Acta Horticulturae 866: 353-362.

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Edgar RC. 2004. MUSCLE: multiple sequence alignment with high accuracy and high

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Ellis MB. 1971. Dematiaceous hyphomycetes. CABI publishing, Wallingford, UK

Fan L, Hou Cl, Cannon PF, Li Y. 2012. A new species of Melanospora on truffles from China.

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Retrieved April 2, 2019, from https://nt.ars-grin.gov/fungaldatabases/

Index Fungorum. 2019. Index Fungorum. http://www.indexfungorum.org. Accessed April 2, 2019

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Li DW, Schultes NP, Chen JY, Wang YX, Castafeda-Ruiz RF. 2017. Circinotrichum

sinense, a new asexual fungus from Hubei, China. Botany 95(12): 1099-1108.

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MYCOTAXON

April-June 2020—Volume 135, pp. 309-313

https://doi.org/10.5248/135.309

Golovinomyces asperifolii: first record in China and

Bothriospermum chinense as a new host

LuCHAO Bat’”, HUIYAN XIONG”*, BAOGUO SHI?

' State Key Laboratory of Plateau Ecology and Agriculture &

’ College of Agriculture and Animal Husbandry:

Qinghai University, Xining, Qinghai 810016, China

° Forestry and Grassland Bureau of Qinghai Province,

Xining, Qinghai 810000, China

“ CORRESPONDENCE TO: 1723189938@qq.com

ABSTRACT—Golovinomyces asperifolii is described from the Qilian mountains in Qinghai

province as a new record in China. The identification is supported by morphological

characteristics and phylogenetic analysis of rDNA ITS. The fungus was collected on

Bothriospermum chinense, which represents a new host for G. asperifolii.

Key worps—Ascomycota, Boraginaceae, Erysiphaceae, powdery mildew. taxonomy

Introduction

The Qinghai-Tibet Plateau in China, known as “the ridge of the world,” has

very special vegetation. Bothriospermum chinense is a Chinese endemic plant

used as a traditional Chinese medicine for treating urinary tract infections.

In autumn 2018, it was found to be heavily infected by a powdery mildew in

the Qilian mountains, Qinghai. Based on the morphological characteristics

of the asexual morph, the mildew was classified as a Golovinomyces sp. (Braun

& Cook 2012). Molecular analyses were conducted to resolve the species

identity. The internal transcribed spacer (ITS) regions were sequenced

and compared with sequences in depositories. Based on sequence analysis

of the rDNA ITS region, this species has been identified as Golovinomyces

asperifolii.

310... Bai & al.

This species has not previously been reported in China (Zheng & Yu

1987, Wu & Wu 1991, Chen 1993, Wang & al. 2002, Liu 2010, Bai 2015); our

collection constitutes the first Chinese record of this species.

Materials & methods

Living leaves of Bothriospermum chinense bearing the anamorph of a powdery

mildew were collected in autumn 2018 in the Qilian Mountains in China. The

herbarium specimens were kept in Herbarium of Plant Pathology at Qinghai

University, Xining, Qinghai Province, China (QHU).

A sample was mounted in distilled water and examined using an Olympus

CX31RTSF light microscope. A JEOL JSM-6360LV scanning electron microscope

was used to observe the anamorph ultrastructure, particularly conidial surface

features (according to Cook & al. 1997) and hyphal appressoria.

Genomic DNA was extracted from the mycelium and the asexual morph using

Chelex-100 (Walsh & al. 1991, Hirata & Takamatsu 1996). The nuclear rDNA ITS

region (including the 5.8S gene) was amplified via polymerase chain reaction (PCR);

PM5 and PM6 (Takamatsu & Kano 2001) were used to amplify the ITS region.

PCR assays were conducted in a 50 uL final volume (Hirata & Takamatsu 1996)

containing 27 uL of 2x BoisTaq PCR MasterMix, 1 uL of each primer, 1 uL of the

extracted DNA, and 20 uL of ddH,O (Hirata & Takamatsu 1996). Thermal cycling

in a BioRad PTC-200 thermal cycler comprised an initial denaturation step at 95°C

for 5 min, 35 cycles at 94°C for 1 min + 60°C for 1 min + 72°C for 1 min, and a

final elongation step at 72°C for 8 min. A negative control for each set of reactions

replaced the template DNA with ddH,O. The PCR products were separated by

electrophoresis on a 2% agarose gel in TAE buffer and purified using the Zymoclean™

Gel DNA Recovery Kit, according to the manufacturer's instructions. The purified

DNA products were ligated into the pMD18-T vector (Takara) and transformed

into E. coli DH5a cells. The cloned fragments were sequenced by Sangon Biotech

(Shanghai) Co., Ltd.

All DNA sequences were aligned using Clustal X 1.81 (Thompson & al. 1997),

and the alignments were adjusted following Nei & Kumar (2000). All positions

containing gaps or missing data were eliminated from the dataset. Cladistic trees

were constructed using the neighbor-joining method with the Kimura 2-parameter

substitution model in MEGA 4.0 (Tamura & al. 2007). Branch robustness was

assessed by bootstrap analysis with 1000 replicates.

Taxonomy

Golovinomyces asperifolii(Erikss.) U. Braun & H.D. Shin,

Mycobiology 46(3): 198. 2018. PLATE 1

Mycelium on leaves and stems, amphigenous, persistent, irregular

white patches, thin, effuse; hyphae hyaline, smooth, 3-6um wide; hyphal

appressoria nipple-shaped; conidiophores erect, 100-250 um long; foot-cells

Golovinomyces asperifolii on Bothriospermum chinense (China) ... 311

S-4800 10.0kV 9.0mm x600

PLaTE 1. Golovinomyces asperifolii (QHU2018043): A. Bothriospermum chinense infected by a

powdery mildew; B. Chasmothecia and appendages; C. Ascus and ascospores; D. Conidiophore,

conidia, and hyphae; E. Conidia.

straight, cylindrical, 40-130 x 8-14 um, followed by 1-3 shorter cells; conidia

ovoid to doliiform, with irregular longitudinal or reticulate ridges under

the SEM, 22-38x12-20um, length/width ratio 1.4-2.4, Chasmothecia

gregarious or sometime scattered, subglobose to globose, 80-110 um diam;

peridium cells irregularly polygonal, 8-23 um diam; appendages more or less

equatorial and in the lower half of the chasmothecia, numerous, mycelioid,

simple, unbranched, interlaced with each other and with the mycelium,

0.4—2 x times as long as the chasmothecial diam, 3-6 um wide, septate, walls

thin and smooth, hyaline or brown below and paler or colorless towards

the tips; asci 6-12, with oil drops, ellipsoid-obovoid, clavate-saccate, 34-52

x 20-31 um, short-stalked, 2-4-spored; ascospores ellipsoid-ovoid to almost

globose, 10-19 x 10-18 um, colorless.

SPECIMEN EXAMINED: , 38°07’59”N 100°08’38”E, alt. 3280 m, on living leaves of

Bothriospermum chinense Bunge (Boraginaceae), autumn 2018, L.C. Bai (QHU2018043;

GenBank MK425651).

Phylogeny

The ITS rDNA sequence comprising 474 total characters was deposited

in GenBank. The new sequence was aligned with sequences from eleven

312... Bai & al.

56 Golovinomyces arabidis KR048081

Golovinomyces riedlianus AB430819

Sat 95 Golovinomyces macrocarpus AB077685

Golovinomyces artemisiae KM260733

Golovinomyces asperifolii MH189714

100 Golovinomyces asperifolii MK425651

Golovinomyces sordidus KM260734

Golovinomyces cynoglossi MH189715

Golovinomyces orontii AB430818

se Golovinomyces cichoracearum JQ010848

Golovinomyces depressus AB077675

69 Golovinomyces montagnei AB077656

Arthrocladiella mougeotii MF496139

0.01

PLaTE 2. A neighbor-joining tree based on distances derived from sequences of ITS1, ITS2, and

the 5.8S rRNA gene from 12 Golovinomyces sequences and an Arthrocladiella mougeotii outgroup

sequence. The bar indicates a distance of 0.01.

Golovinomyces spp. and an outgroup sequence of Arthrocladiella mougeotii

(Lév.) Vassilkov (PLATE 2). The ITS phylogenetic tree placed the Chinese

specimen and Golovinomyces asperifolii MH189714 (from Germany) in a

strongly supported clade with 100% bootstrap support, indicating that these

two sequences are conspecific.

Discussion

Golovinomyces asperifolii was described worldwide on hosts of the

Boraginaceae in 2018. The chasmothecium we sampled agrees with the

descriptions of G. asperifolii published by Braun & al. (2018), and its

sequence was highly consistent with those reported. According to the host

plant records for this powdery mildew (Braun & al. 2018), Bothriospermum

chinense represents a new host for G. asperifolii.

Acknowledgments

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

(No. 31600513). We thank U. Braun (Institute of Biology, Martin Luther University,

Halle, Germany) for valuable suggestions and critical comments on the manuscript

and Prof. Zhimin Cao (Northwest A&F University, Yangling, China) and Prof. Susumu

Takamatsu (Mycologist Emeritus, Tsu, Mie, Japan) for their expert reviews.

Golovinomyces asperifolii on Bothriospermum chinense (China) ... 313

Literature cited

Braun U, Cook RTA. 2012. Taxonomic manual of the Erysiphales (powdery mildews). CBS

Biodiversity Series 11. 707 p.

Braun U, Bradshaw M, Zhao TT, Cho SE, Shin HD. 2018. Taxonomy of the Golovinomyces

cynoglossi complex (Erysiphales, Ascomycota) disentangled by phylogenetic analyses and

reassessments of morphological traits. Mycobiology 46(3): 192-204.

https://doi.org/10.1080/12298093.2018.1509512

Chen ZX. 1993. Fujian Erysiphaceae. Fuzhou, Fujian Science and Technology Press. (in

Chinese).

Cook RTA, Inman AJ, Billings C. 1997. Identification and classification of powdery mildew

anamorphs using light and scanning electron microscopy and host range data. Mycological

Research 101: 975-1002. https://doi.org/10.1017/S095375629700364X

Hirata T, Takamatsu S. 1996. Nucleotide sequence diversity of rDNA internal transcribed

spacers extracted from conidia and cleistothecia of several powdery mildew fungi.

Mycoscience 37: 283-288. https://doi.org/10.1007/BF02461299

Liu TZ. 2010. Powdery mildew in Inner Mongolia. Hohhot, Inner Mongolia Science and

Technology Press. (in Chinese).

Nei M, Kumar S. 2000. Molecular evolution and phylogenetics. Oxford University Press. New

York.

Takamatsu S, Kano Y. 2001. PCR primers useful for nucleotide sequencing of rDNA of the

powdery mildew fungi. Mycoscience 42: 135-139. http://dx.doi.org/10.1007/BF02463987

Tamura K, Dudley J, Nei M, Kumar S. 2007. MEGA4: Molecular Evolutionary Genetics

Analysis (MEGA) software version 4.0. Molecular Biology and Evolution 24: 1596-1599.

https://doi.org/10.1093/molbev/msm092

Thompson JD, Gibson TJ, Plewniak KF, Jeanmougin KF, Higgins DG. 1997. The

CLUSTAL X_ windows interface: flexible strategies for multiple sequence

alignment aided by quality analysis tools. Nucleic Acids Research 24: 4876-4882.

https://doi.org/10.1093/nar/25.24.4876

Walsh PS, Metzger DA, Higuchi R. 1991. Chelex 100 as a medium for simple extraction

of DNA for PCR-based typing from forensic material. Biotechniques 10: 506-513.

https://doi.org/10.2144/000114018

Wang RS, Dou YX, Zhu KG. 2002. The notes on Erysiphales in the Guiqing Mountain of

Gansu Province. Journal of Gansu Agricultural University 37(4): 416-420. (in Chinese).

https://doi.org/10.3969/j.issn.1003-4315.2002.04.004

Wu MX, Wu MQ. 1991. Guizhou plant powdery mildew. Guizhou, Guizhou Science and

Technology Publishing House. (in Chinese).

Zheng RY, Yu YN. 1987. Flora Fungorum Sinicorum. Volume 1 (Erysiphales). Beijing, Science

Press. (in Chinese).

MYCOTAXON

April-June 2020—Volume 135, pp. 315-320

https://doi.org/10.5248/135.315

Cercosporella bundelkhandae comb. nov.

from India

RAGHVENDRA SINGH’, SANJEET KUMAR VERMA},

SANJAY YADAV’, SHAMBHU KUMAR?

' Centre of Advanced Study in Botany, Institute of Science, Banaras Hindu University,

Varanasi, Uttar Pradesh, India 221005

’ Forest Pathology Department, KSCSTE-Kerala Forest Research Institute,

Peechi, Thrissur, Kerala, India 680653

* CORRESPONDENCE TO: drsinghtaxon@gmail.com, singhr.bot@bhu.ac.in

ABSTRACT—The new combination Cercosporella bundelkhandae (= Pseudocercosporella

bundelkhandae) is proposed, based on critical morphological re-examination of the holotype

specimen and fresh topotypic material and comparison with closely related species of

cercosporoid hyphomycetes. The species was originally collected on living leaves of Tinospora

sinensis from Jhansi, Utttar Pradesh, India.

Key worps—anamorph, Capnodiales, Mycosphaerellaceae, nomenclature, taxonomy

Introduction

The hyphomycetous ramularioid complex (Mycosphaerellaceae,

Capnodiales) comprises genera with colourless conidiophores and conidia.

The morphologically most similar genera of this complex are Cercosporella

Sacc., Pseudocercosporella Deighton, and Ramularia Unger. The taxonomic

problems presented by this complex were discussed by Braun (1990, 1991a,b,

1994, 1995, 1998), Kirschner (2009), Crous & al. (2001a,b), Verkley & al.

(2004), and Videira & al. (2016). In Cercosporella the conidial scars and

hilum are thickened while in Ramularia they are only slightly thickened.

These criteria however are somewhat subjective and, in many descriptions,

become merged or modified as ‘slightly thickened and darkened’ or

316 ... Singh & al.

‘refractive. These difficulties explain the frequent transfers of Cercosporella

species to Ramularia and vice versa. Scanning electron microscopy has

shown that conidiogenous loci are smooth (a flat conidiogenous locus

shaped as a truncated cone) in Cercosporella but resemble Cladosporium-

type loci (consisting of a circular rim and a central dome) in Ramularia

(Kirschner 2009, Bensch & al. 2012). Braun (1995) separated Cercosporella

into two subgenera—C. subg. Cercosporella and C. subg. Pseudovellosiella

U. Braun—based on the presence or absence of secondary superficial

mycelium and development of conidiophores from stromata or secondary

superficial mycelia or both. Cercosporella are primarily hyaline, although in

tropical and subtropical countries, some Cercosporella species occur with

olivaceous or pale olivaceous-brown stromata and coloured conidiophore

bases (Braun 1995).

A large number of ramularioid fungi have been recombined in the genus

Cercosporella from all over the world (Braun 1992, 1993; Davis 1919, 1924;

Deighton 1973; Héhnel 1924; Magnus 1906; Saccardo 1895). Recently,

additional ramularioid species have been described from India (Awasthi &

al. 2016, Singh & al. 2008), suggesting that the diversity of such fungi is still

insufficiently known in this region.

During a survey of the Jhansi region of Uttar Pradesh in 2018, a colourless

foliicolous hyphomycete found on leaves of Tinospora sinensis, was identified

as species of Cercosporella subg. Cercosporella due to presence of prominent

thickened conidial scars and hilum. A literature survey revealed that this

fungus had been described earlier as Pseudocercosporella bundelkhandae

(Shrivastava & al. 2009). As the fungus is characterized by thickened scars

and colourless conidiophores and conidia and the absence of secondary

superficial mycelium and solitary conidiophores, it is taxonomically

worthwhile to transfer the species into Cercosporella subg. Cercosporella.

Materials & methods

The holotype specimen [conserved in the Herbarium, Division of Mycology &

Plant Pathology, Indian Agricultural Research Institute, New Delhi, India (HCIO)]

and a fresh specimen collected from living T’ sinensis leaves from the type locality

[conserved in the Mycological Herbarium, Banaras Hindu University, Varanasi,

India (MH-BHU)] were examined in lacto-glycerol-cotton blue using an Olympus

Magnus CH20i-TR light microscope. At least 50 measurements of each fungal

structure were taken for taxonomic diagnoses. Histological sections were prepared

in order to observe the relationship between the fungus and the host tissue. The

fungus was compared with closely related cercosporoid taxa by the help of current

literature.

Cercosporella bundelkhandae comb. nov. (India) ... 317

Taxonomy

Cercosporella bundelkhandae (S. Shrivast., N. Verma & A.N. Rai) Raghv. Singh,

S.K. Verma, S. Yadav & Sh. Kumar, comb. nov. FIG. 1

MB 831083

= Pseudocercosporella bundelkhandae S. Shrivast., N. Verma &

A.N. Rai, J. Mycol. Pl. Pathol. 39(2): 305 (2009)

(Description based on type material): INFECTION spoTs amphigenous,

circular to irregular, 3-10 mm in diam., initially hyaline, then becoming very

light- to mid-brown. COLONIES amphigenous, effuse. Mycelium internal.

STROMATA well developed, immersed, pseudo-parenchymatous, hyaline to

olivaceous or light olivaceous-brown, 35-50 um in diam. CONIDIOPHORES

macronematous, 5-10 fasciculate, sometimes synnematous, unbranched, erect

to procumbent, straight to flexuous, geniculate, smooth, 2—5-septate, thick-

walled, hyaline, (18—)50-80(-85) x 2.5-5(-7) um. CONIDIOGENOUS CELLS

integrated, terminal, polyblastic, sympodial, mostly conspicuously geniculate,

cylindrical, usually swollen at apex; loci thickened, darkened, outwardly

bulging, 2-3.5 um wide. Conip1a simple, solitary, acropleurogenous,

obclavate-cylindrical, ellipsoid, straight to slightly curved, apex subacute,

base subconic-truncate, dry, smooth, thin walled, 1-4-septate, hyaline,

(10-)20-55(-65) x (2-)3-6(-8.5) um, hilum thickened, 2-3.5 um wide.

SPECIMEN EXAMINED: INDIA, UTTAR PRADESH, Jhansi, on leaves of Tinospora cordifolia

(Willd.) Miers [= T. sinensis (Lour.) Merr. (Menispermaceae)], 10 December 2010, leg. S.

Jain (HCIO 44091, holotype); Jhansi, 25.4484°N 78.5685°E, on living leaves of Tinospora

sinensis, December 2018, leg. Raghvendra Singh (MH-BHU 9).

Discussion

Cercosporoid fungi previously reported on Tinospora hosts include:

Cercospora madhauliensis Kamal & al.; Cercosporella dioscoreophylli

(Henn.) Deighton; Passalora menispermi (Ellis & Holw.) U. Braun & Crous;

Pseudocercospora cocculi (Syd.) Deighton; Ps. tinosporae A.N. Rai & Kamal; Ps.

tinosporicola U. Braun & Bagyan.; and Ps. tinosporigena U. Braun (Deighton

1973, 1976; Braun & Crous 2005; Crous & Braun 2003; Bagyanarayana &

Braun 1992; Rai & Kamal 1989; Kamal & al. 1987).

Molecular sequence data have already confirmed that the presence/

absence of thickened and darkened conidiogenous loci, slightly thickened

and darkened or refractive scars (refractivity indicating the absence of scar),

presence/absence of conidiophore and conidial pigmentation, and planate/

pileate thickened dark conidiogenous loci are taxonomically important

318 ... Singh & al.

Fic. 1. Cercosporella bundelkhandae (holotype, HCIO 44091):

a. Symptoms; b. Stromata with fascicle of conidiophores and conidiogenous cells; c. Conidia.

Cercosporella bundelkhandae comb. nov. (India) ... 319

features (Crous 1999; Untereiner 2000; Crous & al. 2000, 2001a,b; Pretorius

& al. 2003; Stewart & al. 1999; Taylor & al. 2003).

Cercosporella bundelkhandae differs from Cercospora madhauliensis,

P. menispermi, Ps. cocculi, Ps. tinosporae, Ps. tinosporicola, and Ps. tinosporigena

because they all have either coloured conidiophores or coloured conidia or

both.

Due to its hyaline nature, Cercosporella bundelkhandae shows a close

resemblance to C. dioscoreophylli, distinguished by its longer (180 x 3-5.5 um)

branched conidiophores and longer, pluriseptate conidia (25-80 x 3-7.5 um,

1-6-septate).

Acknowledgments

Sincere thanks are due to Curator of the herbarium HCIO for make it possible to

examine the holotype collection of P bundelkhandae. We express our deep gratitude

to Dr. R.E Castafieda Ruiz (INIFAT, Tropical Alejandro de Humboldt, Havana,

Cuba) and Dr. Flavia Rodrigues Barbosa (Universidade Federal de Mato Grosso,

Sinop, Brazil) for critical review. We express our deep appreciation to Prof. Dr. Kamal

(Emeritus Scientist (DST), D.D.U. Gorakhpur University, Gorakhpur) for his valuable

suggestions and kind help. We are also thankful to the Head of the Department of

Botany of Banaras Hindu University, Varanasi, U.P., for providing necessary facilities.

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MYCOTAXON

April-June 2020—Volume 135, pp. 321-332

https://doi.org/10.5248/135.321

Coprinellus ovatus sp. nov.

from Pakistan

MUHAMMAD KAMRAN & SANA JABEEN’

Department of Botany, Division of Science and Technology, University of Education,

Township, Lahore, Pakistan

“CORRESPONDENCE TO: sanajabeenue@gmail.com; sanajabeen@ue.edu.pk

ABSTRACT—A Coprinellus specimen collected in Islamabad, Pakistan, showed characters

that were distinct from those of all other Coprinellus spp.: an ovoid shaped pileus; a caespitose

stipe base; utriform cheilocystidia bearing crystals; a pileipellis of loosely arranged clavate to

mucronate hyphae; and basidiospores (9.5-11.6 x 6.0-7.8 x 5.6-6.2 um) with shapes ranging

from mitriform or triangular to ellipsoid in face view, amygdaliform to ellipsoid in side view.

Combination of all these characters and molecular analysis of ITS sequences support its

identification as a new species.

Key worps—coprinoid mushrooms, Micacei, Psathyrellaceae, taxonomy

Introduction

Coprinoid mushrooms are an important group of macrofungi that can be

readily distinguished from normal agarics by having gills that liquify at maturity.

As a result, coprinoid mushrooms are often called inky caps. Coprinoid

mushrooms, which are characterized by thin fragile pilei that become plicate

on opening, deliquescent gills, and dark brown to blackish basidiospores with

germ-pores (Schafer 2014), spread across Agaricales in evolutionary lineages

that are either fully or partially coprinoid (Toth & al. 2013). Fully coprinoid

genera include: Coprinus Pers., Coprinellus P. Karst., Coprinopsis P. Karst., and

Parasola Redhead & al. Some species of Leucocoprinus Pat., Conocybe Fayod,

Bolbitius Fr., and Galerella Earle show coprinoid combinations of characters

(Matheny & al. 2006, Nagy 2011, Nagy & al. 2011, Toth & al. 2013).

322 ... Kamran & Jabeen

Coprinellus, with approximately 84 described species, represents an

independent lineage in Psathyrellaceae (Redhead & al. 2001; Walther & al.

2005; Padamsee & al. 2008; Vasutova & al. 2008; Nagy & al. 2011, 2012, 2013;

Orstadius & al. 2015; Hussain & al. 2018a). These mushrooms are commonly

found on bare soil, wood chips, grassy debris, leaf-litter, and herbivore dung

(Schafer 2010). Coprinellus species are divided into three sections based on

veil anatomy and the presence or absence of cap pileocystidia. Coprinellus

sect. Domestici (Singer) D.J. Schaf. is characterized by a pileus veil comprising

floccose scales consisting of chains of fusiform or subglobose cells, often with

encrusted walls. In C. sect. Micacei (Fr.) D.J. Schaf., veil remnants are present

in the form of scattered (often disappearing) granulose flocks of globose

cells arising from a matrix of narrow branched hyphae. In C. sect. Setulosi

(J.E. Lange) DJ. Schaf., the veil may be present or absent, but the pileus and

stipe are covered with thin-walled pileocystidia and caulocystidia, respectively

(Schafer 2010). However, Nagy & al. (2012) showed that molecular phylogeny

did not entirely support these sections in that clades corresponding to C. sect.

Micacei and C. sect. Domestici each included some setulose species.

Twenty-two coprinoid species have been reported from Pakistan (Ahmad

& al. 1997; Hussain & al. 2016, 2017, 2018a,b), of which eight belong to

Coprinellus: C. campanulatus S. Hussain & H. Ahmad, C. disseminatus (Pers.)

J.E. Lange, C. disseminatisimilis S. Hussain, C. marculentus (Britzelm.) Redhead

& al., C. micaceus (Bull.) Vilgalys & al., C. pakistanicus Usman & Khalid,

C. radians (Desm.) Vilgalys & al. C. tenuis S. Hussain (Ahmad & al. 1997;

Hussain & al. 2018b).

During surveys of basidiomycetous fungi of Pakistan in 2018, one

interesting collection presenting a Coprinellus morphology was encountered.

Closer inspection revealed that this collection presented a unique combination

of features compared with other Coprinellus species. Here we propose a new

species based on morphological characters and molecular phylogenetic analysis

of rDNA internal transcribed spacer (ITS).

Materials & methods

The sample was collected from the capital territory Islamabad, Pakistan, along the

Murree Express Road. Islamabad (33.43°N 73.04°E) lies at the edge of the Pothohar

Plateau at the base of the Margalla Hills, occupies an area of 906 km’ with elevation of

507 ma.s.l., and has a humid subtropical climate (K6ppen climate classification). The

territory recognizes winter (November—February), spring (March-April), summer

(May-June), rainy monsoon (July-August), and autumn (September—October),

with June the hottest month with the average temperature routinely exceeding 38°C

Coprinellus ovatus sp. nov. (Pakistan) ... 323

TABLE 1. Coprinellus and Psathyrella species and specimens used for the molecular

phylogenetic analysis.

SPECIES VOUCHER COUNTRY ITS #

C. aff. eurysporus E

ist ais i Pein NL-3418 Hungary JN159540

C. aff. xanthothrix B

(as C. xanthothrix in GenBank) ee Sys eae

C. angulatus NL-1934 Hungary HQ846994

Arnolds99-22 Netherlands JN159536

NL-0906 Hungary JN159535

C. bisporiger WU7403 Hungary HQ846974

Daams71-98 Hungary JN159520

C. bisporus NL-0158 Sweden GU227705

NL-2512 Sweden FN396107

WU6011 Sweden JN159517

C. brevisetulosus NL-1445 Hungary GU227710

C. callinus NL-1931 Hungary FN396105

Ulje1204 Netherlands JN159518

C. campanulatus Coprinellus _sp._SHP144 Pakistan MH753666

Coprinellus _sp._SH144T Pakistan MH753667

C. cinereopallidus NL-0177 Hungary HQ847001

Ulje1163 Netherlands JN159524

C. congregates NL-0588 Sweden FM878013

NL-2138 Hungary JN159554

NL-1221 Hungary JN159550

NL-1433 Hungary JN159552

C. curtus NL-1490 Hungary JN159569

NL-2339 Sweden FM878016

NL-1023 Hungary JN159568

C. deminutus NL-0761 Hungary JN159572

C. disseminatisimilis Coprinellus _sp._SH15 Pakistan MH753668

Coprinellus _sp._Cr_3wT Pakistan MH753670

Coprinellus _sp._Cr_3b Pakistan MH753669

C. disseminates NL-2337 Sweden FM878017

NL-0786 Hungary JN159560

NL-3401 Hungary JN159561

C. domesticus Cervenka286 Hungary HQ847043

TOK21808 Netherlands JN159580

C. eurysporus NL-1761 Hungary HQ846992

C. flocculosus NL-0838 Hungary JN159573

NL-1567 Hungary JN159575

NL-1661 Norway FN396138

Caffocgiaens: ae FVDB1743 Hungary JN159574

(as Coprinus “maysoidisporus”)

C. fuscocystidiatus NL-2720 Norway HQ846977

NL-3622 Sweden JN159514

C. hiascens NL-1349 Hungary JN159525

NL-2536 Norway FM878018

NL-1350 Hungary GU227720

NL-2598 Hungary JN159526

C. impatiens NL-1164 Hungary FM163177

NL-0568 Hungary JN159511

TABLE 1, continued on p. 324

324 ... Kamran & Jabeen

TABLE 1, continued

SPECIES VOUCHER COUNTRY ITS #

C. micaceus NL-3888 Hungary GU227721

NL-3656 Hungary JN159567

NL-1939 Hungary FN396104

NL-4253 Hungary JN159566

NL-2739 Hungary JN159564

NL-2744 Hungary JN159565

[as C. sp. 10 LGN-2011 in GenBank &

C. “xylophilus” in Nagy & al. 2011] EN BI 26 HnNgary US tee eee

*C. ovatus LAH35939 Pakistan MK764023

C. pakistanicus LAH35322 Pakistan MH366735

LAH35323 Pakistan MH366736

LAH35324 Pakistan MH366737

C. pallidus NL-1556 Hungary JN159521

NL-0625 Hungary HQ846989

NL-4218 Hungary JN159522

C. pellucidus NL-1076 Hungary GU227713

NL-2928 Slovakia GU227714

NL-2344 Hungary FM878023

C. plagioporus NL-1365 Hungary HQ846981

NL-1086 Hungary JN159512

C. radians NL-1373 Sweden JN159579

NL-3896 Sweden HQ847045

C. radicellus NL-3168 Sweden GU227719

NL-0594 Sweden GU227716

NL-0957 Norway GU227718

NL-2121 Sweden GU227717

Coprinellus sabulicola NL-1560 Hungary JN159557

NL-2906 Hungary JN159558

NL-1027 Hungary JN159559

NL-1763 Hungary HQ847007

C. sassii NL-1495 Hungary FN396101

NL-1237 Hungary JN159509

C. sclerocystidiosus NL-0797 Hungary JN159539

NL-1022 Hungary JN159538

NL-1761 Hungary HQ846992

NL-1444 Hungary JN159537

C. silvaticus NL-3035 Hungary HQ846986

C. subimpatiens ToK 27/8-08 Hungary HQ846993

NL-0162 Hungary JN159542

Uljé 1191 Netherlands HQ846995

C. tenuis Coprinellus _sp._SHP10 Pakistan MH753663

Coprinellus _sp_SH10T Pakistan MH753664

C. truncorum NL-1101 Hungary JN159562

NL-1294 Sweden FM878007

C. uljei NL-2492 Sweden JN159513

NL-0157 Sweden HQ846982

NL-3985 Slovakia JN159515

C. verrucispermus NL-2146 Hungary JN159577

C. xanthothrix TOK12808 Netherlands JN159578

NL-3417 Hungary HQ847044

Psathyrella candolleana NFP-MU1 India KT443864

* Species information in bold face represents data generated during this study.

Coprinellus radicellus GU227719

91| Coprinellus radicellus GU227716

45 Coprinellus radicellus GU227718

Coprinellus radicellus GU227717

Coprinellus pallidus JN159521

Coprinellus pallidus HQ846989

Coprinellus pallidus JN159522

Coprinellus callinus FN396105

Coprinellus callinus JN159518

96 | Coprinellus cinereopallidus HQ847001

— Coprinellus cinereopallidus JN159524

r— Coprinellus hiascens JN159525

96 Coprinellus hiascens FM878018

Coprinellus hiascens GU227720

“ Coprinellus hiascens JN159526

73, Coprinellus bisporiger HQ846974

ag mai Coprinellus bisporiger JN159520

al) Coprinellus sassii FN396101

Coprinellus sassii JN159509

nial Coprinellus fuscocystidiatus HQ846977

Coprinellus fuscocystidiatus JN159514

93) Coprinellus impatiens FM163177

Coprinellus impatiens JN159511

Coprinellus plagioporus HQ846981

Coprinellus plagioporus JN159512

Coprinellus uljei JN159513

Coprinellus uljei HQ846982

Coprinellus uljei JN159515

;~ Coprinellus bisporus GU227705

99 D Coprinellus bisporus FN396107

Coprinelius bisporus JN159517

Coprinellus congregatus FM878013

99 | Coprinellus congregatus JN159554

Coprinellus congregatus JN159550

Coprinellus congregatus JN159552

Coprinellus sabulicola JN159557

4 65

99 | Coprinellus sabulicola JN159558

Coprinellus sabulicola JN159559

Coprinellus sabulicola HOSA7007.

97 6) Coprinellus angulatus HQ846994

Coprinellus angulatus JN159536

80 | Coprinellus angulatus JN159535

100 ; Coprinelius pakistanicus MH366735

—_—_—"I

0.05

|__46|

Ce Coprinellus pakistanicus MH366736

Coprir

prinellus pakistanicus MH366737

Coprinellus deminutus JN159572

g9| Coprinellus curtus JN159569

-—*t Coprinellus curtus FM878016

Coprinellus curtus JN159568

Coprinellus tenuis MH753663

— Coprinellus tenuis MH753664

Coprinellus subimpatiens HQ84699

Coprinellus subimpatiens JN15954

Coprinellus ovatus sp. nov. (Pakistan) ... 325

a Coprinellus pellucidus GU227713

Coprinellus pellucidus GU227714

Coprinellus pellucidus FM878023

Coprinellus brevisetulosus GU227710

Coprinellus aff. eurysporus JN159540

Coprinellus eurysporus HQ846992

Coprinellus aff. eurysporus HQ846995

Coprinellus sclerocystidiosus JN159539

Coprinellus sclerocystidosus JN159538

Coprinellus sclerocystidioss HQ846992

Coprinellus sclerocytidiosus JN159537

100, Coprinellus campanulatus MH753666

Coprinellus micaceus JN159567

Coprinellus micaceus FN396104

Coprinellus micaceus JN159566

Coprinellus micaceus JN159564

63' Coprinellus micaceus JN159565

= 8 © Coprinellus ovatus MK764023

Coprinellus micaceus GU227721

Coprinellus truncorum JN159562

— Coprinellus truncorum FM878007

\ Coprinellus campanulatus MH753667

78|| Coprinellus micaceus as C. "xylophilus” JN159563

Coprinellus disseminatus FM878017

; Coprinellus disseminatus JN159560

98 Coprinellus disseminatus JN159561

400/~ Coprinellus disseminatisimilis MH753668

Coprinellus silvaticus HQ846986

97 '- Coprinellus flocculosus JN159573

Coprinellus flocculosus JN159575

- Coprinellus flocculosus FN396138

96) Coprinellus disseminatisimilis MH753670

Coprinellus disseminatisimilis MH753669

ree Coprinellus verrucispermus JN159577

r Coprinellus flocculosus as C. "maysoidisporus” JN159574

99) Coprinellus domesticus HQ847043

74 Coprinellus domesticus JN159580

——— Coprinellus aff. xanthothrix FM87008

— Coprinellus radians JN159579

94 Coprinellus radians HQ847045

77 y Coprinellus xanthothrix JN159578

Coprinellus xanthothrix HQ847044

Psathyrella candolleana KT443864

C. sect. Setulosi

C. sect. Micacei

C. sect. Domistici

J Out group

Figure. 1. Molecular phylogenetic analysis of Coprinellus spp. based in ITS sequences. The tree with

the highest log likelihood (—5095.2205) is shown. The percentage of trees in which the associated

taxa clustered together is shown next to the branches. Initial trees for the heuristic search were

obtained by applying the Neighbor-Joining method to a matrix of pairwise distances estimated

using the Maximum Composite Likelihood (MCL) approach. A discrete Gamma distribution was

used to model evolutionary rate differences among sites (5 categories; +G, parameter = 0.2939).

The tree is drawn to scale, with branch lengths measured in the number of substitutions per site.

The analysis involved 96 nucleotide sequences. The sequence generated from LAH35939 is marked

with®.

326 ... Kamran & Jabeen

and July the wettest (https://en.wikipedia.org/wiki/Climate_of_Islamabad; Peel & al.

2007, Belda & al. 2014).

Basidiomata were tagged, photographed, and dried. Morphological features

(size, shape, and color) of basidiomata were noted in the field. Color was coded

following Munsell (1975). Tissues were mounted in 2% aqueous KOH (w/v) solution

for anatomical study and examined using a China XSZ-107-BN light microscope.

At least 20 measurements were recorded for each anatomical feature. Basidiospore

measurements were taken as: length range x width range x breadth range, with

outliers shown in parentheses; Q values designate length/width ratios, with Qav =

the average Q value. The specimens are deposited at the Herbarium, Department of

Botany, University of the Punjab, Lahore, Pakistan (LAH).

DNA was extracted using Extract-N-Amp™ Plant Tissue PCR Kit following

manufacturer's instructions. PCR amplification and sequencing were commercially

performed by TsingKe, P.R. China.

For phylogenetic analysis, the ITS sequences generated from the Pakistani

collections were compared with other sequences in GenBank using Basic Local

Alignment Search Tool (BLAST) at NCBI (https://www.ncbi.nlm.nih.gov/). Closely

matching sequences and those cited in Hussain & al. (2018b) were downloaded

from the GenBank for phylogenetic analysis. Psathyrella candolleana (Fr.) Maire

(HQ847040) was chosen as an outgroup (Hussain & al. 2018b). Sequences were

aligned using online MUSCLE tool (http://www.ebi.ac.uk/). Best model selection

approach was employed on the final data set and Maximum Likelihood (ML)

analysis was performed by using Tamura-Nei model (Tamura & Nei 1993) in MEGA

6 (Tamura & al. 2013). The phylogenetic tree was constructed at one thousand rapid

bootstrap replicates.

Molecular phylogenetic results

The sequences from specimens used for the molecular phylogenetic analysis are

given TABLE 1. The final ITS dataset comprised 96 nucleotide sequences, with 697

positions after trimming from both ends. The resultant phylogenetic tree (FIGURE 1)

places the Coprinellus sequences in several distinct clades that more or less correspond

to previous studies (Nagy & al. 2012, Hussain & al. 2018b). The sequence generated

from LAH35939 clustered in C. sect. Micacei.

Taxonomy

Coprinellus ovatus Kamran & Jabeen, sp. nov. FIGURES 2, 3

MB 830529

Differs from Coprinellus campanulatus by its ovate fibrillose pileus, brownish black

hymenium, caespitose stipe base, larger mitriform, amygdaliform, and ellipsoid

basidiospores, crystal-bearing utriform cheilocystidia, and loosely arranged clavate to

mucronate pileipellis hyphae.

Coprinellus ovatus sp. nov. (Pakistan) ... 327

Figure. 2. Coprinellus ovatus (holotype, LAH35939). Basidiomata.

Scale bars: A = 1 cm; B, C = 0.2 cm. Photographs by Muhammad Kamran.

Type: Pakistan, Islamabad, on soil under broadleaf trees, 1 May 2018, Muhammad

Kamran K4 (LAH35939).

Erymo.oey: The specific epithet (L.) refers to oval shape of the pileus.

328 ... Kamran & Jabeen

PiLEus 9-13 x 14-17 mm, ovoid to parabolic, with plane center, orange yellow

color (10YR8/8), surface fibrillose, deeply plicate from center to margin.

LAMELLAE 1.5-2.5 mm wide, free, entire, crowded, brownish black (10YR2/3).

LAMELLULAE absent. STIPE 1.4—2.7 x 2-4 mm, central, smooth radicating, base

3-5 mm wide, yellowish brown (10YR7/8), caespitose, dry, fibrillose. ANNULUS

absent. ODoR not observed.

BASIDIOSPORES (8.6—)9.6-11.4(-11.6) x (5.7-)6.0-7.5(-7.8) x (5-)5.6-6.

(-7.2) um, average = 10.5 x 6.7 x 6.0 um, Qav = 1.5, mitriform or triangular

to ellipsoid in face view, amygdaliform to ellipsoid in side view, dark brown to

blackish brown in KOH, apiculus prominent, with central germ-pore (2.2-3.0

um). BAsrpIa (17.3—)21.8-36.9(-41.7) x (7-)8.25-10.6(-11) um, cylindrical,

clavate to subclavate, hyaline. CHEILocysTIp1a 20-29.3 x 9.2-12.1 um,

utriform, and apex usually bears crystals. PLEUROCYSTIDIA absent. PILEIPELLIS

an epithelium of loosely arranged hyphae with clavate to mucronate terminals

(51-)55.5-68(-94) x (14-)20-28(-67) um. UNIVERSAL VEIL ELEMENTS

19.4-44.2 um wide, globose to subglobose with filament 9.4-26.2 um.

STIPITIPELLIS (3.2—)11.9-22.9(-29.5) um, filamentous with frequent septa,

with rare clamp connections.

ECOLOGY & DISTRIBUTION—Among broadleaf trees including Morus sp.

Discussion

Coprinellus ovatus is phylogenetically most closely related to C.

campanulatus and C. truncorum. Coprinellus campanulatus is separated

morphologically by a campanulate pileus, pale orange-yellow to dark

yellowish-brown lamellae, the presence of lamellulae, smaller basidiospores,

larger cheilocystidia, and a pileipellis that is an epithelium of loosely arranged

globose to subglobose pileocystidia (Hussain & al. 2018b). Coprinellus

truncorum differs in the presence of lamellulae and smaller ellipsoid

basidiospores that are not distinctly lentiform (Keirle & al. 2004, Uljé 2005).

Coprinellus micaceus differs in its closely crowded adnexed to free lamellae

and smaller basidiospores that are lacriform to submitriform or mitriform in

face view, and conical towards the base (Keirle & al. 2004, Uljé 2005).

Coprinellus ovatus also differs from three recently described species

from Pakistan. Coprinellus disseminatisimilis is diagnosed by its pruinose

parabolic to umbo-campanulate pileus, sinuate to uncinate lamellae, the

presence of lamellulae, ellipsoid to cylindrical/obovoid basidiospores, much

larger narrowly clavate to utriform cheilocystidia, and a loosely arranged

euhymeniderm pileipellis with more or less utriform pileocystidia (Hussain

Coprinellus ovatus sp. nov. (Pakistan) ... 329

a9 IQ

OY ‘

E —_ PF —

Figure. 3. Coprinellus ovatus (holotype, LAH35939). A. Basidiospores; B. Cheilocystidia;

C. Universal veil elements; D. Basidia; E. Pileipellis hyphae with clavate to mucronate terminals;

FE. Stipitipellis. Scale bars: A-D = 6 um; E, F = 15 um. Drawing by Sana Jabeen.

330 ... Kamran & Jabeen

& al. 2018b). Coprinellus pakistanicus differs by its convex to plane pileus, the

presence of lamellulae, slightly smaller basidiospores, larger cheilocystidia;

and the presence of true lageniform/cylindrical pileocystidia (Hussain &

al. 2018b). Coprinellus tenuis, which produces similar basidiospores, is

separated by its convex to plane pileus, sinuate to uncinate lamellae, presence

of lamellulae, equal cylindrical stipe, and the presence of lageniform to

cylindrical pileocystidia (Hussain & al. 2018b).

TABLE 2. Comparison of Coprinellus ovatus and related species.

Coprinellus sp. PILEUs (mm); LAMELLAR BASIDIOSPORES PILEIPELLIS type CHEILO-

Shape; ATTACHMENT (um) (terminal cells) CYSTIDIA

Surface (lamellulae) F = face view (um)

S = side view

ovatus 9-13 x Free, 9.6-11.4 x 6-7.5 Loose epithelium —§ 20-29.3 x

14-17; (absent) F: mitriform (clavate / 9.2-12.1

Ovoid to / ellipsoid; mucronate)

parabolic; S: amygdaliform /

Fibrillose ellipsoid

campanulatus <40 x 15 Adnexed, 8-10.5 x 5.5-6.5 Loose epithelium 36-47 x

Yg: ovoid / (present, F: mitriform (globose to 35-45

parabolic, later 1-4 series) / ellipsoid; subglobose

campanulate; S: cylindrical, elements)

Pruinose to amygdaliform /

granulose ellipsoid

disseminati- <20 x 20; Sinuate to 8-9 x 5-5.5 Loose 70-165 x

similis Parabolic then uncinate, F: ellipsoid to euhymeniderm; 11-15

campanulate; (present, cylindrical; (utriform

Pruinose to 0-2 series) S: amygdaliform / pileocystidia)

pulverulent ellipsoid

micaceus *<24 mm Adnexed or 6.5-10 x 4.5-7 * Epithelium * 30-40 x

diam; Obtuse free F: lacriform (globose cells) 20-25

cylindrical, (absent) to mitriform;

then convex / conical towards

campanulate; the base

white mica-

like; granular,

detersile

pakistanicus <35mm;Convex _ Free (present, 8.5-11.5 x Irregular 42-75 x

to plane, disc 0-2 series) 6.5-8; epithelium 14-25

depressed; F: obovoid to (lageniform

Smooth, phaseoliform; to cylindrical;

pulverulent, S: ellipsoid to tapering neck,

granulose obovoid obtuse apex)

tenuis <20 mm; Sinuate to 10.5-14.5 x Loose epithelium 22-30 x

Pulvinate, uncinate 8-9.5; (lageniform 19-28

convex, plane; (present, F: ellipsoid to to obtuse long

Glabrous, 0-2 series) ovoid cylindrical)

furred S: sl. pyriform to

ellipsoid

truncorum *<14 mm; Oval * Adnexed or 8.5-9 x 5.5-6; * Loose * 25-40 x

then convex; free (present, Ellipsoid in all epithelium 25-30

finely plicate, 1-4 series) views, not truly ([sub] globose to

granular or with lentiform broadly clavate)

mica-like flecks

* Reference for C. micaceus and C. truncorum: Keirle & al. 2004

Coprinellus ovatus sp. nov. (Pakistan) ... 331

Full dimensions and comparative characters of the Coprinellus species

cited above are given in TABLE 2. Although C. ovatus is in C. sect. Micacei,

it is sister to C. micaceus and forms a subclade with C. truncorum and C.

campanulatus. Coprinellus micaceus sequence (GU227721) from Hungarian

collection (NL-3888) also clustered in the same clade with C. ovatus, this

specimen showed 99.2% sequence similarity and 0.6% divergence from

C. ovatus. No morphological data is available from this specimen. Since

C. ovatus showed remarkable morphological differences from C. micaceus

(Keirle & al. 2004, Uljé 2005), we conclude that LAH35939 represents a new

species in C. sect. Micacei.

Acknowledgments

Authors are thankful to Dr. Shah Hussain (Center for Plant Sciences and

Biodiversity, University of Swat, Pakistan) for valuable comments and suggestions to

improve the manuscript. Sincere thanks to Dr. Chang-Lin Zhao (Forestry College,

Southwest Forestry University, Kunming, P. R. China) and Mr. Muhammad Usman

(University of the Punjab, Lahore, Pakistan) for acting as presubmission reviewers.

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Hussain S, Afshan NS, Ahmad H. 2016. First record of Parasola lilatincta from Pakistan. Mycotaxon

131(2): 317-323. https://doi.org/10.5248/131.317

Hussain S, Afshan NS, Ahmad H, Khalid AN, Niazi AR. 2017. Parasola malakandensis

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https://doi.org/10.1016/j.myc.2016.09.002

Hussain S, Ahmad H, Ullah S, Afshan NS, Pfister DH, Sher H, Ali H, Khalid AN. 2018a. The

genus Parasola in Pakistan with the description of two new species. MycoKeys 30: 41-60.

https://doi.org/10.3897/mycokeys.30.21430

Hussain S, Usman M, Afshan NS, Ahmad H, Khan J, Khalid AN. 2018b. The genus Coprinellus

(Basidiomycota; Agaricales) in Pakistan with the description of four new species. MycoKeys 39:

41-61. https://doi.org/10.3897/mycokeys.39.26743

Keirle MR, Hemmes DE, Desjardin DE. 2004. Agaricales of the Hawaiian Islands. 8. Agaricaceae:

Coprinus and Podaxis; Psathyrellaceae: Coprinopsis, Coprinellus and Parasola. Fungal Diversity

15(3): 33-124.

Matheny PB, Curtis JM, Hofstetter V, Aime MC, Moncalvo JM, Ge ZW, Yang ZL, Slot JC, Ammirati

JE, Baroni TJ, Bougher NL. 2006. Major clades of Agaricales: a multilocus phylogenetic overview.

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Nagy GL. 2011. An investigation of the phylogeny and evolutionary processes of deliquescent

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Nagy GL, Hazi J, Vagvélgyi C, Papp T. 2012. Phylogeny and species delimitation in the

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Nagy GL, Vagvolgyi C, Papp T. 2013. Morphological characterization of clades of the Psathyrellaceae

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https://doi.org/10.1017/S0953756205002868

MYCOTAXON

April-June 2020—Volume 135, pp. 333-337

https://doi.org/10.5248/135.333

An update of G.K. Merrill’s 1909 “Lichen notes no. 14”

STEVEN B. SELVA! & R. TROY MCMULLIN?*

' University of Maine at Fort Kent, 23 University Drive, Fort Kent, ME 04743 USA

? Canadian Museum of Nature, PO Box 3443, Station D, Ottawa, ON K1P 6P4 Canada

*CORRESPONDENCE TO: tmcmullin@nature.ca

ABSTRACT—G.K. Merrill proposed three new Calicium taxa that have largely been absent

from North American lichen literature since their publication in THE BRYOLOGIST in 1909.

Calicium obscurum [= Chaenotheca obscura], which colonizes the basidiocarps of Trichaptum

abietinum, predates the use of Chaenotheca balsamconensis. Calicium minutissimum was

reintroduced as a member of the North American calicioid biota in 1999. Calicium curtisii

var. splendidulum [as “splendidula”| is within the range of morphological variation of

Phaeocalicium curtisii and does not warrant varietal status.

Key worps — Caliciales, Maine, pin fungi, stubble lichens

In his Lichen Notes No. 14, G.K. Merrill (1909) described three new Calicium

taxa: two new species, C. obscurum and C. minutissimum, and one new variety,

C. curtisii var. splendidulum [as “splendidula”]. All three were collected within

30 miles of Penobscot Bay in Knox County, Maine. Calicium obscurum was

included in Merrill’s (1909-13) Lichenes Exsiccati as #92, and C. curtisii var.

splendidulum as #24.

Calicium obscurum G. Merr., Bryologist 12: 107. 1909 Fic. 1A,B

= Chaenotheca obscura (G. Merr.) Nadv., Stud. Bot. Cech. 5: 123. 1942

= Chaenotheca balsamconensis J.L. Allen & McMullin, Bryologist 118: 55. 2015

Calicium obscurum was collected “on dead Polyporei,’ which we have

identified as Trichaptum abietinum (Pers. ex J.E Gmel.) Ryvarden, growing

on young pines in Rockland, Maine (Fic. 1A). Two species of algae were

reported to be growing with it: Cystococcus, better known today as Trebouxia,

334 ... Selva & McMullin

the photobiont of C. obscurum, and “a cylindrical articulated form that I

am unable to name” (Merrill 1909). This unknown alga is Stichococcus,

which commonly grows on the surface of T: abietinum. Merrill refers to a

second Calicium, also growing “on a Polyporus on Hemlock Spruce,” that

was collected by Willey (1892) “and identified by Tuckerman as Calicium

trichiale” [= Chaenotheca trichialis (Ach.) Th. Fr.]|. He wondered if the

Willey specimen might prove to represent C. obscurum, which differs from

C. trichiale by its lack of a thallus, two forms of algae, and the absence of

pruina.

One species that has been erroneously reported from Trichaptum in

North America is Chaenotheca brunneola (Ach.) Mull. Arg. Tibell (1980,

1999) reported finding it occasionally “on old polypores” in Europe, and

Fries (1865) noted a Trichaptum-dwelling variant which he named as

Ch. brunneola f cilians Th. Fr. This variety, which may be the earliest valid

name for Ch. obscura, has no priority at species rank; moreover, according

to Tibell (1981) and Suija & al. (2016, and personal communication), the

type material of the variety has not been located. Hutchison (1987, p. 789)

erroneously cited Calicium obscurum as a synonym of Ch. brunneola, and

Keissler (1938, p. 597) erroneously cited Calicium obscurum as a synonym of

Phaeocalicium polyporaeum. Keissler stated that Muraschinskij had sent him

a specimen collected on Polystictus pergamenus (Fr.) Cooke [= Trichaptum

biforme (Fr.) Ryvarden] from Siberia, “which is identical to Calicium

polyporaeum Nyl? [= Phaeocalicium polyporaeum (Nyl.) Tibell] “that is also

found in North America under the synonym Calicium obscurum Merr.’ In

North America, P. polyporaeum is most commonly found on T. biforme,

while Ch. obscura has, to date, been collected only on T! abietinum.

According to Selva (2014), four species of calicioid lichens and fungi have

been collected on T. abietinum in northeastern North America: Chaenotheca

ferruginea (Turner) Mig., Ch. trichialis, Chaenothecopsis pusiola (Ach.)

Vain., and Phaeocalicium polyporaeum. Allen & McMullin (2015) argued

that Ch. ferruginea is a corticolous and lignicolous species that does not

colonize Trichaptum abietinum. They proposed the name Chaenotheca

balsamconensis for the species that Selva (2014) recorded as Ch. ferruginea

growing on T: abietinum. The two species are remarkably similar, but

Ch. balsamconensis differs in having smooth rather than ornamented to

fissured spores, in often having a KOH+ magenta compound in the stalk,

and in having a predominantly endosubstratic thallus. After reviewing an

isotype of C. obscurum (Merrill exsiccati #92 [CANL]), we found it to be

Calicium taxonomy updated ... 335

Fic. 1. Chaenotheca obscura: A. Apothecia (Calicium obscurum isotype, Merrill exsicc. #92,

CANL); B. Apothecia (McMullin 16823, CANL). Phaeocalicium curtisii. C. Apothecia (Calicium

curtisii var. splendidulum isotype, Merrill exsicc. #24, CANL); D. Ascospores (Merrill exsicc.

#24, CANL); E. Apothecia (McMullin 14102, CANL); E Ascospores (McMullin 20101, CANL).

Scale bars: A, B = 0.5 mm; C, E = 0.2 mm; D, F = 20 um

conspecific with Ch. balsamconensis (Fic. 1A-B), the chemotype lacking the

KOH+ magenta stalk. Since C. obscurum is a validly published name for this

taxon, it should be resurrected. After its publication by Merrill in 1909, the

name has never appeared on any of the North American lichen checklists

336 ... Selva & McMullin

and seems to have been lost from North American lichen literature.

Coincidentally, the same fate befell the two other Calicium taxa published

in Merrill (1909).

Calicium minutissimum G. Merr., Bryologist 12: 107. 1909

= Phaeocalicium minutissimum (G. Merr.) Selva, Bryologist 102: 390.

Calicium minutissimum was not recognized as a member of the North

American calicioid biota until it was reintroduced by Selva & Tibell (1999).

They transferred it to Phaeocalicium and, because the type specimen was no

longer recognizable, designated a neotype. The original type was first collected

by Merrill (1909) on the smooth bark of Quercus rubra L. at the summit of

Mt. Battie in Camden Hills State Park, Knox County, Maine. The type locality

remained the only known location for 90 years until Selva & Tibell (1999)

expanded the range to include additional locations in New Brunswick, Maine,

New Hampshire, and Vermont. It was later reported from Ontario, Quebec,

and Massachusetts by Selva (2010).

Calicium curtisii var. splendidulum G. Merr., Bryologist 12: 107. 1909

[as “splendidula” | FIG. 1c-F

= Calicium curtisii Tuck., Amer. J. Sci. Arts, Ser. 2, 28: 201. 1859 [var. curtisii]

= Phaeocalicium curtisii (Tuck.) Tibell, Symb. Bot. Upsal. 21(2): 58. 1975

Calicium curtisii var. splendidulum was collected on the main stems and larger

branches of Rhus typhina L. in Rockland, Maine, and on the same substrate in

Ottawa, Ontario (Merrill 1909) (Fic. 1C). Tibell (1975) transferred the species

to Phaeocalicium. The description does not clearly state the characteristics that

distinguish C. curtisii var. splendidulum from the autonomous variety, and the

average height of the apothecia and the ascospore measurements fall within

those known for the species (Fic. 1C-F). Merrill reported that the ascospores

are “simple or indistinctly bilocular;” however, all ascospores observed in an

isotype (Merrill exsiccati #24 [CANL]) were clearly 2-celled (Fic. 1D). Merrill

also reported that the “capitula are turbinate to subglobose,” a character

common in young apothecia (Fic. 1C, E). We propose that C. curtisii var.

splendidulum does not significantly differ from Phaeocalicium curtisii and does

not warrant varietal status. Fink (1935), who also came to a similar conclusion,

did not provide any discussion or reason for synonymizing the variety.

Acknowledgments

We gratefully acknowledge Otto Gockman (Midwest Natural Resources, USA)

and Rikke Naesborg (Santa Barbara Botanic Garden, USA) for helpful reviews of our

manuscript.

Calicium taxonomy updated ... 337

Literature cited

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Trichaptum abietinum from North America that is benefiting from widespread conifer

fatalities. Bryologist 118: 54-58. https://doi.org/10.1639/0007-2745-118.1.054

Fink B. 1935. The Lichen Flora of the United States. University of Michigan Press, Ann Arbor.

Fries TM. 1865. Nya Skandinaviska Laf-arter. Botaniska Notiser 1865(2): 38-40.

Hutchison LJ. 1987. Studies on Phaeocalicium polyporaeum in North America. Mycologia 79:

786-789. https://www.jstor.org/stable/3807832

Keissler K. 1938. Pyrenulaceae bis Mycoporaceae, Coniocarpineae. Rabenhorst's Kryptogamen-

Flora von Deutschland, Osterreich und der Schweiz 9: 1-846.

Merrill GK. 1909. Lichen notes no. 14. Three new forms of Calicium. Bryologist 12: 107-108.

Selva SB, Tibell L. 1999. Lichenized and non-lichenized calicioid fungi from North America.

Bryologist 102: 377-397. https://www.jstor.org/stable/3244225

Selva SB. 2010. New and interesting calicioid lichens and fungi from eastern North America.

Bryologist 113: 272-276. https://doi.org/10.1639/0007-2745-113.2.272

Selva SB. 2014. The calicioid lichens and fungi of the Acadian Forest ecoregion of northeastern

North America, II. The rest of the story. Bryologist 117: 336-367.

https://doi.org/10.1639/0007-2745-117.4.336

Suija A, Suu A, Lohmus P. 2016. Substrate specificity corresponds to distinct phylogenetic

lineages: the case of Chaenotheca brunneola. Herzogia 29: 355-363.

https://doi.org/10.13158/heia.29.2.2016.355

Tibell, L. 1975. The Caliciales of Boreal North America. Symbolae Botanicae Upsalienses 21(2).

128 p.

Tibell L. 1980. The lichen genus Chaenotheca in the northern hemisphere. Symbolae Botanicae

Upsalienses 23(1). 65 p.

Tibell L. 1981. Comments on Caliciales Exsiccatae II. Lichenologist 13: 51-64.

https://doi.org/10.1017/S002428298 1000066

Tibell L. 1999. Calicioid lichens and fungi. Nordic Lichen Flora 1: 20-94.

Willey H. 1892. Enumeration of the lichens found in New Bedford, Massachusetts and its

vicinity, from 1862-1892. E. Anthony and Sons, Inc., Printers, New Bedford.

MYCOTAXON

April-June 2020—Volume 135, pp. 339-344

https://doi.org/10.5248/135.339

Elaphomyces citrinus and E. cyanosporus,

new for Turkey

YASIN UzuN! & ABDULLAH Kaya”

' Department of Biology, Science Faculty, Karamanoglu Mehmetbey University,

70100, Karaman, Turkey

? Department of Biology, Science Faculty, Gazi University, 06500, Ankara, Turkey

“ CORRESPONDENCE TO: kayaabd@hotmail.com

ABSTRACT— he first records of two black Elaphomyces, E. citrinus and E. cyanosporus, are

reported from Turkey. Short descriptions, photographs, and comments are provided.

Key worps — biodiversity, Elaphomycetaceae, hypogeous ascomycetes, sequestrate, truffle-

like fungi

Introduction

Elaphomyces T. Nees, “the deer fungus,’ is a sequestrate genus in

Elaphomycetaceae. Its species are characterized by globose to subglobose or

irregular hypogeous ascomata that have a fleshy to leathery and smooth to

variously warty brown to dark blue or black peridium; a single chambered

gleba comprising a powdery slate blue, purple-brown, dark brown, grey-black,

or black spore mass; globose to subglobose 1-8-spored asci; and subglobose

to globose spiny to verrucose spores (Trappe 1979, Castellano & al. 2012,

Castellano & Stephens 2017).

Index Fungorum (2018) accepts 73 Elaphomyces taxa, of which four—

E. granulatus Fr., E. leucocarpus Vittad., E. muricatus Fr., E. septatus Vittad.—

have been reported from Turkey (Turkoglu & al. 2015, Uzun & Kaya, 2019a,b).

We report Turkish collections of two black Elaphomyces species, E. citrinus

and E. cyanosporus. Current checklists (Sesli & Denchev 2014, Solak & al.

2015) and recent research papers (Isik & Turkekul 2017, Kasik & al. 2017,

340 ... Uzun & Kaya

Kaya 2015, Kaya & al. 2016, Kaya & Uzun 2018, Sesli 2018, Selem & al. 2019;

Turkekul 2017, Uzun & Kaya 2019c) have not yet reported E. citrinus and

E. cyanosporus from Turkey. This study aims to contribute additional knowledge

to the mycobiota of Turkey.

Materials & methods

The Elaphomyces fruit bodies were collected from Rize and Trabzon provinces

during 2015-2018. The ascomata were photographed and the ecological data were

recorded in the field; macroscopic and microscopic features were measured in the

laboratory. The specimens were examined microscopically using a Nikon Eclipse

Ci-S trinocular light microscope and Hitachi SU5000 scanning electron microscope

and identified by consulting the literature (Pegler & al. 1993; Montecchi & Sarasini

2000; Arroyo & al. 2005; Binion & al. 2008; Saitta & al. 2009; Castellano & al. 2018;

Hobart 2012; Paz & al. 2012, 2017). The specimens are preserved in the fungarium

of Department of Biology, Science Faculty, Karamanoglu Mehmetbey University,

Karaman, Turkey (KMU).

Taxonomy

Elaphomyces citrinus Vittad. Monogr. Tuberac.: 65 (1831) FIG. 1

AscomMa 10-40 mm in diameter, subglobose, usually covered with adherent

lemon-yellow mycelium, not easily separating and binding soil particles

to form a crust. CorTEx thin, composed of heavily carbonized dark brown

hyphae. PERrp1um 1-2 mm thick, with a thinner black outer cortex and a dark

grey layer below, with a tissue of filamentous hyphae. GLEBA a grey-dark brown

powder of thin-walled hyphae. Ascr globose, 8-spored. AScosPpores 10.5-12.5

um in diameter, globose, dark brown when ripe, lightly aculeate-roughish,

ornamented with fine anastomosed 0.5-1.5 um high rods or ridges that form

an irregular labyrinthine pattern.

SPECIMENS EXAMINED—TURKEY, TRABZON: Tonya, Hosarli village, 40°56’N

39°10’E, 1100 m, mixed forest, in soil under Castanea sativa Mill. and Fagus orientalis

Lipsky, 17.09.2015, Y.Uzun 4537 (KMU); K6priibasi, Yagmurlu village, 40°49’N

40°07’E, 420 m, mixed forest, under Corylus avellana L. and E orientalis, 11.09.2017,

Y.Uzun 5821 (KMU).

CoMMENTS—In general, the macro- and micromorphological characteristics

of our Turkish materials agree with Elaphomyces citrinus as described in the

literature (Pegler & al. 1993, Saitta & al. 2009, Paz & al. 2012). The bright

yellow mycelial crust is reported to be diagnostic of this species (Pegler &

al. 1993, Montecchi & Sarasini 2000, Arroyo & al. 2005, Paz & al. 2017).

Elaphomyces cibulae Castellano & al., also possesses a carbonized outer

peridium and similar spore characters, but its white to ivory mycelial crust

Elaphomyces citrinus & E. cyanosporus in Turkey ... 341

KMU-BILTEM 15.0kV M-x12,000 SE(L)

Fic 1. Elaphomyces citrinus (KMU, Uzun 4537).

a. Ascomata; b. Ascospores; c. SEM image of an ascospore. Scale bar: b = 10 um.

and thicker peridial layer distinguish E. cibulae from E. citrinus (Castellano

& al. 2018).

Elaphomyces cyanosporus Tul. & C. Tul, Fungi Hypog.: 113 (1851) FIG. 2

AscoMA 15-30 mm in diameter, globose to ovoid, dark brown to black,

covered with blackish, thin, irregularly warty cortex. PERrp1um thick and

double layered, the outer layer light brown with whitish and reddish tones,

the darker inner layer almost equal in thickness with the outer layer. GLEBA

felty and grey when young, firm, turquoise to blackish blue at maturity. Asc1

globose 60-80 um in diameter with thin hyaline walls. AscospoREs 22-26 um

including ornamentation, globose, turquoise to black, ornamentation almost

2.5 um high, forming a fine reticulum with coarse ridges.

SPECIMENS EXAMINED—TURKEY, Rize: Ardesen, Kirazlik village, 41°07’N 41°05’E,

in soil under Fagus orientalis and Rhododendron ponticum L., 600 m, 08.08.2016,

Y.Uzun 5207 (KMU); TRABZON: Siirmene, Ormanseven village, 40°50’N 40°09’E,

in soil under F. orientalis and Carpinus betulus L., 695 m, 07.07.2018, Y.Uzun

6660 (KMU); Yomra, [kisu village, 40°55’N 39°47’E, in soil under EF. orientalis and

R. ponticum, 280 m, 08.07.2018, Y.Uzun 6667 (KMU); Besikdiizii, Oguz village,

41°01’N 39°09’E, in soil under Castanea sativa and R. ponticum, 740 m, 26.08.2018,

Y.Uzun 6687 (KMU).

CoMMENTS—Elaphomyces cyanosporus is a distinctive species with its blue-

green tinged inner peridium and spores (Montecchi & Sarasini 2000, Binion &

al. 2008, Hobart 2012). It is one of the reticulate-spored members of the genus

342 ... Uzun & Kaya

KMU-BILTEM 15.0kV M-x6,000 SE(L)

Fic 2. Elaphomyces cyanosporus (KMU, Uzun 6667).

a. Ascomata; b, c. Asci and ascospores; d. SEM image of an ascospore.

Scale bars: b, c = 20 um.

(Zhang 1991, Castellano & al. 2016), and the reticulate spore ornamentation

of our samples fits well with Castellano & al. (2018). Elaphomyces persoonii

Vittad., which also has a black warty peridium and similar reticulate spore

ornamentation, differs from E. cyanosporus by its taller alveoli walls and

larger spores (Castellano & al. 2018).

Acknowledgments

The authors would like to thank Karamanoglu Mehmetbey University Research

Fund (02-M-15 and 16-M-16) for its financial support, Dogancan Kuduban

and Yiicel Uzun for their kind help in field, and Dr. Vladimir Antonin (Moravian

Museum, Brno, Czechia), Prof. Dr. Ertugrul Sesli (Trabzon University, Turkey), and

Dr. Shaun Pennycook (Manaaki Whenua Landcare Research, Auckland) for their

helpful comments and careful review.

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MYCOTAXON

April-June 2020—Volume 135, pp. 345-354

https://doi.org/10.5248/135.345

New records of graphidoid and thelotremoid lichens

from India

P. Gupta’, P. RANDIVE?, S. NAYAKA' , R. DAIMARI,

S. JOSEPH’, M.K. JANARTHANAM?

' Lichenology laboratory, CSIR-National Botanical Research Institute,

Rana Pratap Marg, Lucknow, Uttar Pradesh - 226 001

? Department of Botany, Goa University, Taleigao Plateau, Goa - 403 206

° Department of Botany, Bodoland University, Kokrajhar, Assam - 783370

* CORRESPONDENCE TO: nayaka.sanjeeva@gmail.com

AxBsTRACT—Chapsa cinchonarum, C. farinosa, Diorygma sticticum, Fissurina albocinerea,

Graphis bungartzii, G. discarpa, G. nigririmis, Ocellularia alba, Phaeographis pseudostromatica,

Sarcographa verrucosa, and Thelotrema crassisporum are described and illustrated as new

records for India.

Key worps —lichenized fungi, Graphidaceae, Ostropales, taxonomy

Introduction

Graphidaceae (sensu Rivas Plata & al. 2012) is the largest family of tropical

crustose lichens with about 2700 accepted species world-wide (Licking &

al. 2014, 2017a,b); it includes the previously separated, well-known families

Graphidaceae and Thelotremataceae. Kraichak & al. (2018) resurrected

Diploschistaceae and Thelotremataceae which were previously synonymized

under Graphidaceae and accepted the family Fissurinaceae proposed by

Hodkinson (2012).

Here we use Graphidaceae sensu Rivas Plata & al. (2012). The family is

characterized by: crustose thallus with a trentepohlioid photobiont;lirellate to

rounded ascomata; carbonized or non-carbonized proper exciple; inspersed

or non-inspersed hymenium; presence or absence of periphysoids; hyaline

to brown, transversely septate to muriform ascospores often with amyloid

346 ...Gupta & al.

septa, lens-shaped to rectangular lumina, and the presence or absence of

lichen compounds. A total of 570 graphidoid and 131 thelotremoid species

were known from India (Singh & Sinha 2010, Sinha & al. 2018). During the

study of Graphidaceae specimens available in the herbarium of the National

Botanical Research Institute, Lucknow, India (LWG), eleven more species

were identified as new to India. They are described below.

Material & methods

Morphological observations were made using the Leica SSAPO stereomicroscope

with image analyzer software. Thin, hand-cut sections of the thalli and ascomata were

mounted in water, lactophenol cotton blue, 10% KOH, or Lugol's iodine solution

and observed under the Leica DM2500 compound microscope. All anatomical

measurements were taken in water mounts. Secondary metabolites were identified

by thin layer chromatography (TLC) following Orange & al. (2010). The examined

specimens are preserved in the herbarium LWG.

New records

Chapsa cinchonarum (Fée) Frisch, Biblioth. Lichenol. 92: 95 (2006) FIG. 1A

Thallus crustose, corticolous, white to pale greenish-grey, continuous,

matte to mealy or faulty, ecorticate; apothecia angular to elongated and often

slightly branched, 0.5-0.8 mm long; margin jagged to lobed, slightly incurved

to recurved, with a white to pale fawn, compact to felty inner surface; disc

pale brown, covered by thick white felty to crystalline pruina; exciple fused,

uncarbonized; hymenium hyaline, clear, 70-110 um high, I-; ascospores 8/

ascus, hyaline, fusiform, 8-14-locular, 18-35 x 4-6 um, I-; lichen substances

absent.

SPECIMEN EXAMINED: INDIA. Goa, Cotigao Wildlife Sanctuary, 14°57’11”N

74°09’21”E, on bark, 12 September 2017, Pallavi P. Randive GU L575 (LWG-35879).

ComMENtTs—Morphologically Chapsa cinchonarum closely resembles C.

alborosella (Nyl.) Frisch, which differs in having more rounded apothecia with

a rim-like margin and 5-9-locular, smaller (17-22 x 4.5-5 um) ascospores. It

somewhat resembles Astrochapsa platycarpella (Vain.) Parnmen & al., which

possesses 4—7-locular, smaller (12-16 x 3-5 um) ascospores.

Previously known from tropical Africa and the Neotropics (Frisch & al.

2006, Rivas Plata & al. 2010).

Chapsa farinosa Liicking & Sipman, Phytotaxa 55: 37 (2012) FIG. 1B

Thallus crustose, corticolous, white-grey, distinctly farinose, ecorticate;

apothecia erumpent, angular-rounded, 0.2-0.5 mm diam.; disc exposed,

Graphidoid and thelotremoid lichens new for India ... 347

flesh-coloured to pale brown, white-pruinose; margin lobulate to recurved,

pale brown, felty white-pruinose; exciple fused, uncarbonized; hymenium

hyaline, clear, I-; ascospores 8/ascus, hyaline, fusiform-oblong, with slightly

thickened septa and rectangular lumina, 8-10-locular, 20-32 x 4-6 um, I-;

lichen substances absent.

SPECIMENS EXAMINED: INDIA. Goa, Ta Rivana, Quinamol, 15°09’20”N 74°06’21’E, 2

December 2016, Pallavi P. Randive GU 400, 404 (LWG-35882, 35883).

CoMMENTS—Chapsa farinosa is characterized by its whitish, farinose thallus

lacking any traces of a cortex. Chapsa cinchonarum can be easily confused

with C. farinosa but has a compact thallus with loose cortex and often elongate

lirellae. Anatomically, it resembles C. albida (Nyl.) Liicking & Sipman but

differs in the distinctly verruculose thallus with irregular cortex (Rivas Plata

& al. 2010).

Previously known from Costa Rica (Sipman & al. 2012).

Diorygma sticticum Sutjar., Kalb & Liicking, Phytotaxa 18: 49 (2011) FIG. 1C

Thallus crustose, corticolous, light green, smooth to uneven, continuous,

ecorticate; apothecia lirellate in dense clusters, stellately branched, erumpent,

with thick, complete, white thalline margin, 1-2.5 x 0.15-0.3 mm; disc concealed

to partly exposed, pale brown but thickly white-pruinose; exciple conspicuous,

uncarbonised, entire, basally orange-brown, upper part hyaline; epihymenium

granulose, yellow-brown; hymenium hyaline, clear, I-; ascospores 8/ascus,

hyaline, fusiform, submuriform, 10-15 x 6-8 um, I+ blue, rarely found; stictic

acid present.

SPECIMEN EXAMINED: INDIA. Goa, Cotigao Wildlife Sanctuary, 15°01’07”N

74°12’33”E, on bark, 15 December 2017, Pallavi P. Randive GU 193 (LWG-35880).

ComMMENTS—Diorygma sticticum is characterized by its small submuriform

ascospores and the presence of stictic acid. Diorygma nothofagi (A.W. Archer)

A.W. Archer and D. poitaei (Fée) Kalb & al. are morphologically similar but

D. nothofagi differs by larger (18-23 um long) ascospores and the presence of

norstictic acid while D. poitaei is distinguished by the larger (40-65 um long)

ascospores and presence of hypostictic and hypoconstictic acids.

Previously known from Thailand (Kalb & al. 2004, Feuerstein & al. 2014).

Fissurina albocinerea (Vain.) Staiger, Biblioth. Lichenol. 85: 126 (2002) FIG. 1D

Thallus crustose, corticolous, greyish-green, smooth, glossy, corticate;

apothecia lirellate, immersed to slightly raised, simple to very rarely branched,

0.4-2 mm long, straight to curved, terminally acute; disc sunken, slit-like,

348 ...Gupta & al.

epruinose; exciple entire, uncarbonized, orange-brown; hymenium hyaline,

clear, I-; ascospores 8/ascus, hyaline, 4-locular, 16-18 x 6-9 um, I-; psoromic

acid present.

SPECIMEN EXAMINED: INDIA. Goa, Cotigao Wildlife Sanctuary, 14°58’23”N

74°09’36’E, on bark, Pallavi P. Randive, GU 99, 108 (LWG-35884).

ComMENTS—Fissurina albocinerea belongs to the group of species having

Dumastii-type ascomata and is clearly distinguished from the other species of

this group in having small ascospores and psoromic acid in the thallus.

Previously known from the Philippines (Staiger 2002).

Graphis bungartzii A.B. Pefia, Licking, Herrera-Camp. & R. Miranda, Lichenologist

46: 71 (2014) FIG. 1E

Thallus crustose, corticolous, pale greenish-brown, smooth, continuously

corticate; apothecia lirellate, flexuous, branched, erumpent with lateral

thalline margin, 1-2.5 mm long, 0.1-0.3 mm wide; labia concealed, striate,

black; exciple thick, laterally carbonized, 15-45 um wide; epithecium brown,

granulose; hymenium hyaline, clear, I-; ascospores 8/ascus, hyaline, ellipsoid

to fusiform, 4-10-locular, 25-32 x 5-8 um, I+ blue; norstictic and connorstictic

acids present.

SPECIMENS EXAMINED: INDIA. Assam, Sonitpur district, Panigaon, Miranda, 75 m, 25

February 2013, on Syzygium cumini bark, R. Daimari 14-020543, 14-022465, 14-022467

(LWG); Pithakowa, 70 m, 18 March 2012, on Bombax sp., R. Daimari 14-022460 (LWG);

New Siliguri, 118 m, 21 March 2013, R. Daimari 14-022463 (LWG); Borbhogia, 64 m,

on Lannea grandis bark, 26 March 2013, R. Daimari 14-022464 (LWG); Ratanjuli, on

Artocarpus heterophyllus bark, 21 March 2013, R. Daimari 14-020556 (LWG).

ComMENTS—Graphis bungartzii differs from G. elegans (Sm.) Ach. by the

presence of erumpent lirellae with lateral thalline margin and the smaller

ascospores. Graphis filiformis Adaw. & Makhija is also similar to G. bungartzii

but differs in having longer, irregularly to radiately branched lirellae with entire

labia and in lacking connorstictic acid.

Previously known from Mexico (Barcenas-Pena & al. 2014).

Graphis discarpa A.W. Archer, Mycotaxon 89: 325 (2004) FIG. 1F

Thallus crustose, corticolous, pale olive-green, dull, corticate; apothecia

lirellate, numerous, scattered, immersed, simple, straight, curved or sinuous,

rarely branched, concealed, 1-3 x 0.15-0.3 mm, visible as a thin black line with

white margins, separated from the thallus; exciple entire, completely carbonized;

hymenium hyaline, clear, I-; ascospores 8/ascus, hyaline, narrowly ellipsoid to

fusiform, 8-11-locular, 32-40 x 8-10 um, I+ blue; stictic acid present.

Graphidoid and thelotremoid lichens new for India ... 349

vt fuy a?

SE ae . ids she

“4 Cay ee ov

* ; Abe

Fic. 1. A. Chapsa cinchonarum (LWG-35879); B. Chapsa farinosa (LWG-35882); C. Diorygma

sticticum (LWG-35880); D. Fissurina albocinerea (LWG-35884); E. Graphis bungartzii (LWG-

14-020543); E Graphis discarpa (LWG-14-024670). Scale bars = 1 mm.

350 ...Gupta & al.

SPECIMEN EXAMINED: INDIA. WEsT BENGAL, West Medinipur district, Mohunpur,

IISER-Kolkata campus, January 2014, T. Hembram, S. Nayaka & S. Asthana 14-024670

(LWG).

ComMENtTs—Graphis discarpa is closely related to G. glaucescens Fée with

respect to external morphology but distinguished by its entire to striate exciple,

ecorticate, glaucous grey thallus without lichen substances. Graphis immersella

Mill. Arg., which is also morphologically similar, can be distinguished from

G. discarpa in having stictic acid and a laterally carbonized exciple.

Previously known from Papua New Guinea (Archer 2004).

Graphis nigririmis (Nyl.) Mull. Arg., Bull. Herb. Boissier 3: 320 (1895) FIG. 2A

Thallus crustose, corticolous, pale olive green, thin, smooth and shiny;

apothecia lirellate, inconspicuous, immersed, visible only as a thin black or

dark brown line, sometimes becoming conspicuous as black immersed lirellae,

simple to branched, 1-3 mm long, 0.05-0.1 mm wide; disc concealed or slightly

open; exciple entire, pale orange-brown, weakly apically carbonised; hymenium,

hyaline, clear, I-; ascospores 8/ascus, ellipsoid to fusiform, 4—7-locular, 18-25

x 8-16 um, I+ blue; lichen substances absent.

SPECIMEN EXAMINED: INDIA. Tamit Napbu, Namakkal district, Kolli Hills, 800 m, on

bark, 20 March 2016, S. Nayaka & party 16-029294 (LWG).

COMMENTS—Graphis nigririmis resembles G. glaucescens, which is

distinguished by the larger ascospores and the corticated, farinose thallus.

Previously known from Australia (Archer 2009).

Ocellularia alba (Fée) Mill. Arg., Mém. Soc. Phys. Genéve 29(8): 6 (1887) Fic. 2B

Thallus crustose, corticolous, white-grey to pale olive, smooth to uneven,

with prosoplectenchymatous cortex; apothecia erumpent, rounded, 0.2-0.3

mm diam., ecolumellate; disc exposed, pale brown, white-pruinose; margin

white pruinose; pores usually small, occasionally broad, to c. 0.2 mm wide,

mostly +rounded to somewhat irregular; exciple usually thin, hyaline

internally, pale yellowish to yellowish brown marginally, fused; hymenium

hyaline, clear; ascospores 8/ascus, hyaline, fusiform, with slightly thickened

septa and rectangular lumina, 4-6-locular, 18-24 x 4-8 um, I-; lichen

substances absent.

SPECIMEN EXAMINED: INDIA. Goa, Cotigao Wildlife Sanctuary, 14°57’11”N

74°09'21”E, on bark, 15 January 2018, Pallavi P. Randive GU 676 (LWG-35881).

ComMENtTS—Ocellularia alba is morphologically resembles O. papillata

(Leight.) Zahlbr. and differs in the dark brown exciple and columella.

Previously known from Australia, Brazil, the Philippines (Hale 1978).

Graphidoid and thelotremoid lichens new for India ... 351

Phaeographis pseudostromatica Seavey & J. Seavey, Bull. Florida Mus. Nat. Hist.

53: 228 (2017) FIG. 2C

Thallus crustose, corticolous, brownish-grey, shiny, continuous; apothecia

lirellate, thin, brownish-black, immersed to erumpent, partly branched,

0.2-1 mm long, arranged in broad confluent pseudostromatic areas; disc partly

open, brownish-black, epruinose; exciple brown, apically slightly thickened,

20-25 um wide; epihymenium brown; hymenium hyaline, inspersed;

ascospores 8/ascus, brown, fusiform, submuriform, 4-6 x 1—2-locular, 22—30

x 7-10 um; norstictic acid present.

SPECIMEN EXAMINED: INDIA. AssaM, Sonitpur district, Arimora, on Korsun bark, 54

m, 13 March 2012, R. Daimari 14-022470 (LWG).

COMMENTS—Phaeographis pseudostromatica is morphologically similar

to P quadrifera (Nyl.) Staiger but differs by its lirellae arrangement in

pseudostromal tissue. It is also similar to P nylanderi (Vain.) Zahlbr. which

shares the inspersed hymenia and the presence of norstictic acid but differs in

the transversely septate ascospores and the absence of pseudostromata.

Previously known from Florida (Seavey & al. 2017).

Sarcographa verrucosa (Vain.) Zahlbr., Cat. Lich. Univ. 2: 467 (1924) Fic. 2D

Thallus crustose, corticolous, olive-brown to yellow-brown, verruculose,

corticated; apothecia lirellate, grouped together in white stromata at the thallus

level or sunken; thalline margin at edge of the stroma or with extended lirellae;

stromata irregularly ovoid, flattened; black, with crowded, open, not confluent

or branched, sub-circular, 0.2-0.4 mm long lirellae; disc black, white-pruinose;

exciple thin, carbonized or dark brown, 10-15 um; epihymenium grey to

brownish, granular; hymenium hyaline, inspersed; ascospores 8/ascus, brown,

4-6-locular, 19-26 x 6-8 um; stictic acid present.

SPECIMEN EXAMINED: INDIA. Assam, Sonitpur district, Choibari, on Chukrasia

tabularis bark, 81 m, 25 February 2013, R. Daimari 14-022477 (LWG).

COMMENTS—Sarcographa verrucosa differs from all other Sarcographa

species by its olive brown, verruculose thallus, highly inspersed hymenium,

submuriform ascospores, and stictic acid chemistry. In having white

stroma, white pruinose disc and muriform, brown ascospores, Sarcographa

subtorquescens (Nyl.) Zahlbr. resembles S. verrucosa but differs in having

a clear hymenium and larger (2-27 x 9-11 um) ascospores. Sarcographa

labyrinthica (Ach.) Mull. Arg. and S. glyphiza (Nyl.) Kr.P. Singh & G.P. Sinha

are also morphologically very similar, but S. labyrinthica differs in having

transversely septate, 4-6-locular ascospores and a K+ reddish thallus, while

352 ...Gupta & al.

ve eS .

Eye >.

MEE E

‘

Fic. 2. A. Graphis nigririmis (LWG-16-029294); B. Ocellularia alba (LWG-35881); C. Phaeographis

pseudostromatica (LWG-14-022470); D. Sarcographa verrucosa (LWG-14-022477); E. Thelotrema

crassisporum (LWG-35878). Scale bars = 1 mm.

Graphidoid and thelotremoid lichens new for India ... 353

S. glyphiza has larger (25-31 x 9-13 um) ascospores and a K+ red thallus with

stictic and constictic acid.

Previously known from Australia, Indonesia, and the Philippines (Archer

2009).

Thelotrema crassisporum Mangold, Flora of Australia 57: 657 (2009) FIG. 2E

Thallus crustose, corticolous, pale yellowish grey to pale greenish grey,

smooth, continuous, ecorticate; apothecia c. 0.5-0.6 mm diam., +rounded,

solitary to slightly marginally fused, emergent, mostly hemispherical; disc

pale greyish, slightly pruinose, invisible from above; pores c. 0.1 mm diam.,

irregular; exciple completely free, thin, hyaline internally, yellowish brown

marginally, apically dark brown; hymenium hyaline, clear; ascospores 8/ascus,

hyaline, submuriform, subglobose to ellipsoidal, with +trounded ends, 4-7 x

1-2-locular, 14-20 x 8-10 um, I-; constictic and stictic acids present.

SPECIMEN EXAMINED: INDIA. Goa, Cotigao Wildlife Sanctuary, 14°57’11”N

74°09'21”E, on bark, 06 February 2016, Pallavi P. Randive, GU 581 (LWG-35878).

ComMENtTs— Thelotrema crassisporum is similar to T! myriocarpum Fée, which

is distinguished by a shiny, smooth, corticated, non-fissured thallus and larger

(<40 um long) ascospores.

Previously known from Australia (Mangold 2009).

Acknowledgments

We are thankful to the Director, CSIR-NBRI, Lucknow, for providing the

facilities to carry out the research and to Dr D.K. Upreti (National Botanical

Research Institute, Lucknow) for encouragement. We thank Drs. T.A.M. Jagadeesh

Ram (Botanical Survey of India, ANRC, Port Blair) and Harrie Sipman (Botanical

Museum, Berlin) for reviewing the manuscript. Authors PR and MKJ thank Vice

Chancellor, Goa University, Goa for facilities and the Forest Department of Goa for

permission to survey Cotigao Wildlife Sanctuary. PR is also thankful to DST for

financial assistance under the Women Scientist Scheme (SR/WOS-A/LS-354/2016).

Author RD is thankful to Dr. Raza R. Hoque, Tezpur University for his guidance.

One of the authors (SJ) is thankful to SERB for financial assistance under the N-PDF

scheme (PDF/2016/002054).

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Ascomycota) from the Solomon Islands. Mycotaxon 89: 321-329.

Archer AW. 2009. Graphidaceae. Flora of Australia 57: 84-194.

Barcenas-Pefa A, Licking R, Miranda Gonzalez R, Herrera Campos MA 2014. Three new

species of Graphis (Ascomycota: Ostropales: Graphidaceae) from Mexico, with updates to

taxonomic entries for 41 species described between 2009 and 2013. Lichenologist 46: 69-82.

https://doi.org/10.1017/s0024282913000637

354 ...Gupta & al.

Feuerstein SC, Cunha Dias IPR, Aptroot A, Eliasaro S, Caceres MES. 2014. Three new Diorygma

(Graphidaceae) species from Brazil, with a revised world key. Lichenologist 46: 753-761.

https://doi.org/10.1017/s002428291400036x

Frisch A, Kalb K. Grube M. 2006. A monograph of Thelotremataceae with a complex structure

of the columella. Bibliotheca Lichenologica 92: 371-539.

Hale ME. 1978. A revision of the lichen family Thelotremataceae in Panama. Smithsonian

Contribution to Botany 38: 1-60.

Hodkinson, BP. 2012. An evolving phylogenetically based taxonomy of lichens and allied fungi.

Opuscula Philolichenum 11: 4-10.

Kalb K, Staiger B, Elix JA. 2004. A monograph of the lichen genus Diorygma - a first attempt.

Symbolae Botanicae Upsalienses 34(1): 133-181.

Kraichak E, Huang J-P, Matthew N, Leavitt SD, Lumbsch HT. 2018. A revised classification of

orders and families in two major subclasses of Lecanoromycetes (Ascomycota) based on a

temporal approach. Botanical Journal of the Linnean Society 188(3): 233-249.

Liicking R, Johnston MK, Aptroot A, Kraichak E, Lendemer JC & al. 2014. One hundred and

seventy-five new species of Graphidaceae: closing the gap or a drop in the bucket? Phytotaxa

189: 7-38. https://doi.org/10.11646/phytotaxa.189.1.3

Liicking R, Hodkinson BP, Leavit SD. 2017a. The 2016 classification of lichenized fungi in the

Ascomata and Basidiomycota — approaching one thousand genera. Bryologist 119: 361-416.

https://doi.org/10.1639/0007-2745-119.4.361

Liicking R, Hodkinson BP, Leavit SD. 2017b. Corrections and amendments to the 2016

classification of lichenized fungi in the Ascomycota & Basidiomycota. Bryologist 120: 58-69.

https://doi.org/10.1639/0007-2745-120.1.058

Mangold A, Elix JA, Lumbsch HT. 2009. Thelotremataceae. Flora of Australia 57: 195-420.

Orange AP, James W, White FJ. 2010. Microchemical methods for the identification of lichens.

2nd Ed. British Lichen Society, London.

Rivas Plata E, Licking R, Sipman HJM, Mangold A, Kalb K, Lumbsch HT. 2010. A worldwide

key to the thelotremoid Graphidaceae, excluding the Ocellularia-Myriotrema-Stegobolous

clade. Lichenologist 42: 139-185. https://doi.org/10.1017/s0024282909990491

Rivas Plata E, Lticking R, Lumbsch HT. 2012. A new classification for the family

Graphidaceae (Ascomycota: Lecanoromycetes: Ostropales). Fungal Diversity 52: 107-121.

https://doi.org/10.1007/s13225-011-0135-8

Seavey F, Seavey J, Gagnon J, Guccion J, Kaminsky B, Pearson J, Podaril A, Randall B. 2017. The

lichens of Dagny Johnson Key Largo Hammock Botanical State Park, Key Largo, Florida,

USA. Bulletin of the Florida Museum of Natural History 53(5): 201-268.

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Sipman HJM, Licking R, Aptroot A, Chaves JL, Kalb K, Umafia Tenorio L. 2012. A first

assessment of the Ticolichen biodiversity inventory in Costa Rica and adjacent

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MYCOTAXON

April-June 2020—Volume 135, pp. 355-363

https://doi.org/10.5248/135.355

Carbonea assimilis and Rinodina aspersa, new to Poland

KATARZYNA SZCZEPANSKA

Department of Botany and Plant Ecology, Wroctaw University of Environmental and Life Sciences,

pl. Grunwaldzki 24a, PL-50-363 Wroclaw, Poland

* CORRESPONDENCE TO: katarzyna.szczepanska@upwr.edu.pl

ABSTRACT—Two saxicolous species of crustose lichens new to Poland, Carbonea assimilis

and Rinodina aspersa, were recorded from the SW part of the country where there is an

abundance of different rock formations, including natural outcrops of volcanic rocks. The

characteristics of these species, as well as their ecology and geographical distribution, are

provided and briefly discussed.

Key worps—Ascomycota, Lecanoraceae, Physciaceae, taxonomy

Introduction

The lichen biota of Poland (now estimated at c. 1600 species; Faltynowicz

2003) continues to be enlarged by newly reported taxa, most of which are

corticolous, crustose, and especially sorediate lichens, the determination of

which has been made possible through the use of thin layer chromatography or

phylogenetic analysis (Kukwa & Kubiak 2007; Czarnota & al. 2009; Kukwa &

Jabtonska 2009; Kukwa & al. 2012; Guzow-Krzeminska & al. 2016, 2018). Less

frequently new records are foliose (Ossowska & al. 2014, Ossowska & Kukwa

2016) or saxicolous species (Wilk & Flakus 2006; Flakus 2007, 2014; Sliwa &

Flakus 2011; Wilk 2011).

In this paper, new records of saxicolous species collected from natural

outcrops of volcanic rock formations in SW Poland are presented. Volcanic

rocks, including basalt rocks, are often habitats for rare lichens and

consequently rare lichenicolous fungi (Kossowska 2000; Szczepanska 2012a,

2012b, 2015; Szczepanska & al. 2013) due to specific conditions, usually warm

356 ... Szczepariska

and dry, as well as a substrate rich in minerals, such as calcium and magnesium

compounds.

As the two presented species possess inconspicuous, crustose thalli that

could be confused with similar taxa, their diagnostic features are provided as

well as habitat requirements and distribution in Central Europe.

Materials & methods

Specimens were collected by the author in 2012 and 2013 during lichenological

investigations of natural outcrops of volcanic rocks in the Polish Sudety Mountains.

Descriptions of the species are based on the personal observations, measurements,

and TLC analyses. The morphology of the species was studied with dissecting and

light microscopes following routine techniques; for light microscopy, free-hand

sections were made with a razor blade and mounted in water. Tissue measurements

were made in water, and ascospore measurements in 10% KOH (K). The TLC analyses

were performed in solvent systems A, B, and C, using the standardized method of

Culberson (1972) and following Orange & al. (2001). Localities of the species are

mapped according to the ATPOL grid square system (Zajac 1978) modified by

Cieslinski & Faltynowicz (1993). Herbarium material is housed in the private herbaria

of the author (Hb. Szczepanska), with duplicates donated to the Herbarium, Wroclaw

University, Wroclaw, Poland (WRSL) and to the Herbarium, Gdansk University,

Gdansk, Poland (UGDA).

The species

Carbonea assimilis (Hampe ex K6rb.) Hafellner & Hertel,

Flecht. Baden-Wirttembergs, Verbreitungsatlas: 511, 1987. Fig. 1

THALLUS lichenized, crustose, small and thin, areolate, forming irregular

patches between other lichens thalli. PROTHALLUS invisible. AREOLES

flat to slightly convex, 0.2-1.0 mm diam., angular, matte to glossy, olive-

brown. APOTHECIA sessile, small, 0.1-0.3 mm diam., dispersed, rounded,

proper margin persistent, glossy and rather thick, disc black, flat and

glossy. HyMENIuM colourless, 40-50 um tall, epithecium greenish-blue,

HYPOTHECIUM colourless, exciple thick, dark, brownish-black, paraphyses

simple to slightly branched. Ascr 8-spored, Lecanora-type with non-amyloid

apical cushion, ocular chamber visible. Ascosporss: hyaline, simple,

ellipsoid, 8-10 x 5-6 um.

CHEMISTRY—No chemical substances were detected in the analyzed

material.

SPECIMEN EXAMINED—POLAND, SupETY Mountains, Pogérze Zachodniosudeckie

Foothills, Ostrzyca Proboszczowicka hill, alt. 450 m, ATPOL grid square Eb-40, on

basanite rocks, 26 Sept. 2012, K. Szczepanska 939 (Hb. Szczepanska; duplicate in WRSL).

Carbonea & Rinodina spp. new to Poland... 357

Fic. 1. Carbonea assimilis (WRSL — Szczepanska 939): A. thallus; B. apothecia; C. cross section

through apothecium; D. ascospores in the ascus. Scale bars: A = 1 mm, B = 0.5 mm, C = 50 um,

D=20um.

EcoLtocy—tThe species is known as an obligate parasite of crustose lichens.

It initially parasitizes various lichen taxa growing on siliceous rocks, such as

Aspicilia spp., Lecanora frustulosa, Lecidea lapicida, Pertusaria pseudocorallina,

Rhizocarpon spp., and Tephromela atra, before developing independent thalli

(Andreev 2003, Hafellner 2006, Pirogov & al. 2014). Typical habitats for

Carbonea assimilis are neutral to weakly basic, mineral-rich rocks (e.g. basalt,

gneiss, and slate), in sunny and warm places, in mountain regions (Wirth

1995, Chambers & al. 2009).

The Polish locality of Carbonea assimilis was on a volcanic hill in SW

Poland. The hill is one of the relics of ancient volcanic activity in this region,

and during the last glaciation it was nunatak (free of ice) (Staffa 2002). In

this locality C. assimilis was found in a non-parasitic state, growing directly

on the rock in an exposed and insolated place. Associated lichen species

included Candelariella vitellina, Lecanora soralifera, Lecidea fuscoatra, and

Xanthoparmelia loxodes.

358 ... Szczepariska

DIsTRIBUTION—Carbonea assimilis is considered a widely distributed but

rare species, occurring in Europe, Asia, and North America (Andreev 2003).

In Europe it has been found in Austria (Hafellner 1997), Czech Republic

(Vézda & Liska 1999), Denmark (Sochting & Alstrup 2002), Germany (Wirth

1995), Britain (Smith & al. 2009), France (Roux 2012), Norway (Santesson

& al. 2004), Portugal (Boom 1999), Romania (Costache & al. 2007), Spain

(Llimona & Hladun 2001), Sweden (Santesson & al. 2004), and Ukraine

(Pirogov & al. 2014).

CoMMENTS—'The genus Carbonea (Hertel) Hertel, which was separated

from Lecidea s. lat. by Hertel (1983), is represented by about 20 species

of lichenized and lichenicolous fungi worldwide (Pirogov & al. 2014), of

which five were previously known from Poland (Faltynowicz 2003). Most of

these species are exceedingly rare in the country, having scattered localities

almost exclusively in the mountainous areas.

The Polish specimen of Carbonea assimilis presents this species’ typical

brown areolate thallus and very small black apothecia (Fic. 1A, B) with

characteristic greenish-blue epithecium, dark exciple and colourless

hypothecium (Fic. 1C). However, some small differences may be observed

in the size of spores (Fic. 1D), which are shorter than cited in the literature

data (see Chambers & al. 2009, Pirogov & al. 2014).

Carbonea assimilis can be confused with members of Lecidella Korb.

Both Carbonea and Lecidella species usually have black apothecia with

a lecideine margin, a blue-green epithecium and Lecanora-type asci.

Carbonea differs mainly in its narrower spores and black true exciple,

which in Lecidella species is usually pale within. Among Carbonea species

developing independent, distinct thalli, C. assimilis may be confused

primarily with C. vorticosa (Florke) Hertel. However, C. vorticosa has a grey

to whitish thallus, dark brown-black hypothecium, and narrower (4-5 um

diam.) ascospores and mainly grows in high montane regions (Pirogov &

al. 2014).

Rinodina aspersa (Borrer) J.R. Laundon, Lichenologist 18: 175. 1986. Fic. 2

THALLUS small, more or less circular, <13 mm diam., composed of

aggregated or (more rarely) dispersed, areoles, delimited by a clearly visible,

wide black prothallus, <0.2 mm. AREOLES discrete, rounded to angular,

slightly convex, pale to dark grey, matte. SoraLiA discrete, punctiform to

circular, <0.3 mm diam., elevated, arising on the surface of the areoles, mainly

in the centre of the thallus, concolorous with areoles or slightly grey-green

Carbonea & Rinodina spp. new to Poland ... 359

Fic. 2. Rinodina aspersa (WRSL - Szczepanska 941): A. prothallus. B. soralia; C. thallus.

Scale bars: A= 0.5 mm, B= 1mm, C =5.0 mm.

in shaded places and dark-grey in more exposed habitats. SorEDIA farinose.

APOTHECIA not observed in the studied material.

[Additional data from Mayrhofer & Moberg 2002: Apothecia very rare,

<1 mm diam., lecanorine, disc brown to black, plane. Hymenium <100

um tall, epihymenium brown, with fine granules. Hypothecium colourless.

Ascospores 15-20 x 13-20 um, Pachysporaria-type.]

CHEMISTRY—Rinodina aspersa contains atranorin in the cortex (K+

yellow) and additional substances in the medulla, such as gyrophoric (C+

red; usually accompanied by lecanoric and orsellinic acids), umbilicaric,

ovoic, and 5-O-methylhiascic acids (Mayrhofer & Moberg 2002). Substances

detected in the Polish specimens are consistent with the literature data.

SPECIMEN EXAMINED—POLAND, SUDETY MOUNTAINS, Gory Kamienne Mts, Krucza

Skala Mt., alt c. 680 m, ATPOL grid square Eb-91, on volcanic rocks - porphyries, 17

Sept. 2012, K. Szczepanska 941 (Hb. K. Szczepanska; duplicates in UGDA-L and WRSL).

EcoLtoGcy—Typical habitats of R. aspersa in lichenological literature mention

humid, foggy, and shaded places (Mayrhofer & Moberg 2002, Motiejtnaité

360 ... Szczepariska

& Grochowski 2014), including coastal sites (Giralt & Barbero 1995, Wirth

1995, Giavarini & al. 2009, Sheard 2010). Lime-free, hard silicate rocks (such

as basalt, flint, gneiss, granite, shale, and rhyolite) are mentioned as substrata

for this species. Very rarely R. aspersa has been noted in well-lit or sunny

habitats (Vondrak & al. 2006) or on anthropogenic substrata, such as granitic

megalithic monuments (Giralt & al. 1997).

The Polish locality of Rinodina aspersa lies in SW Poland on the rocky

outcrops of a small mountain. Numerous R. aspersa thalli were observed

on rocks and on stones forming a scree. Associated lichens were Acarospora

fuscata, Amandinea punctata, Aspicilia cinerea, and Lecanora polytropa.

Habitat conditions here are rather specific, with high insolation and low

moisture of the substrates. This newly reported record of Rinodina aspersa

may indicate a wider ecological spectrum, including not only shaded and

humid habitats, but also warm, sunny and dry ones.

DISTRIBUTION—Rinodina aspersa is considered a rare species, distributed

from temperate Europe to south Portugal and the Mediterranean islands. It

is found in the Czech Republic (Vézda & Liska 1999) Denmark (Sochting

& Alstrup 2002), Finland (Vitikainen 1997), France (Mayrhofer & Moberg

2002, Roux 2012), Germany (Wirth 1995), Italy (Mayrhofer & Moberg

2002), Lithuania (Motiejunaité & Grochowski 2014), Montenegro (Knezevic

& Mayrhofer 2009), Netherlands (Aptroot & al. 1999), Portugal (Giralt &

Barbero 1995), Spain (Llimona & Hladun 2001), Sweden (Santesson & al.

2004), and U.K. (Smith & al. 2009). It has also been reported from North

America (Glew 1999) and Taiwan (Aptroot & Sparrius 2003).

COMMENTS—Rinodina (Ach.) Gray is represented by c. 200 species (Giavarini

& al. 2009), of which c. 30 are known in Poland (Faltynowicz 2003). Rinodina

aspersa adds a fifth sorediate Rinodina species to the four previously recorded

from Poland (Czarnota & Kukwa 2007, Kukwa & Kubiak 2007). However,

unlike the other four species, which are epiphytic, R. aspersa occurs only

on rocky substrates. In comparison with the other Rinodina species noted

in Poland, it can be distinguished by its thallus chemistry, especially by the

presence of gyrophoric acid, which is absent in other sorediate Rinodina

species. The chemistry of the thallus and the occurrence of soralia are the

most significant characters allowing one to differentiate R. aspersa from

other species of Rinodina. All of the morphological features, as well as the

chemistry of the Polish specimen are typical for this taxon and consistent

with the descriptions given in the literature (Giralt & al. 1997, Giavarini & al.

2009, Sheard 2010).

Carbonea & Rinodina spp. new to Poland... 361

Due to the presence of gyrophoric acid in the thallus and soralia (C+ red),

R. aspersa may be confused with the saxicolous species of Trapelia M. Choisy,

especially T: obtegens that additionally possess a similar thallus composed of

scattered, convex areoles. The features that allow one to distinguish the two

taxa are the clearly visible, black prothallus (Fic. 2A), discrete soralia (Fic.

2B), and more or less circular thallus of Rinodina aspersa (Fic. 2C).

Acknowledgments

I am very grateful to Prof. Mark R.D. Seaward (University of Bradford, UK) and

Dr hab. Adam Flakus (W. Szafer Institute of Botany, Krakow) for reviewing the

manuscript and valuable comments on a previous version of the paper, to Prof. dr

hab. Martin Kukwa (University of Gdansk) for confirming the identity of Rinodina

aspersa by thin layer chromatography, and to Karina Wilk (W. Szafer Institute of

Botany, Krakéw) for helpful assistance with photos.

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MYCOTAXON

April-June 2020—Volume 135, pp. 365-370

https://doi.org/10.5248/135.365

Badhamia versicolor and Trichia subfusca,

new records for Belarus

EvGENy Moroz! & ANDREI TSURYKAU 7”

' Laboratory of Mycology, V.E. Kuprevich Institute of Experimental Botany

of the National Academy of Sciences of Belarus,

Akademicheskaja str. 27, Minsk 220072 Belarus

? Department of Biology, Francisk Skorina Gomel State University,

Sovetskaja str. 104, Gomel 246019 Belarus

° Department of Ecology, Botany and Nature Protection, Institute of Natural Sciences,

Samara National Research University,

Moskovskoye shosse 34, Samara 443086 Russia

* CORRESPONDENCE TO: tsurykau@gmail.com

ABSTRACT—Two myxomycetes, Badhamia versicolor and Trichia subfusca, are reported for

the first time for Belarus. Descriptions and illustrations of both species are provided.

Key worps—biodiversity, Mycetozoa, myxobiota, Protozoa, slime moulds

Introduction

Published accounts of the myxomycetes of Belarus date from the end of

the 19th century (Blonski 1888, 1889, 1890; Twardowska 1885). During the

ensuing years, the studies of slime moulds were accidental. Important research

during the 20th century by Moroz & Novozhilov (1988, 1994) and Moroz

(1996) reported 139 species including literature citations and new records.

Since then and until very recently (Moroz & Novozhilov 2018; Tsurykau 2017),

myxomycetes have been neglected in Belarus.

Recent revision of herbarium material revealed two globally rare myxomycete

species new to Belarus, Badhamia versicolor and Trichia subfusca. The present

contribution details their phenotypic characters, distribution, and ecological

preferences.

366 ... Moroz & Tsurykau

Material & methods

The material was examined using Nikon SMZ745 and Olympus SZ61 dissecting

microscopes and an Olympus BX51 compound microscope. Tissues were examined

and measured in water. Voucher specimens are deposited in the herbarium of V.L.

Komarov Botanical Institute of RAS, Saint Petersburg, Russia (LE) and the Scientific

Herbarium of Belarusian Polesie of Francisk Skorina Gomel State University, Gomel,

Belarus (GSU). Duplicate specimens are stored in the Herbarium of the Institute of

Experimental Botany of the National Academy of Sciences of Belarus, Minsk, Belarus

(MSK). Nomenclature follows Lado (2019).

Taxonomy

Badhamia versicolor Lister, J. Bot. 39: 81. 1901. Fic. 1

Mainly clustered, grey to dingy white, rugulose, sessile sporocarps with

a narrowed base, 0.3-0.5 mm diam. Capillitium white. Spores ovoid, dull

purple, minutely warted, 10-14 x 9-11 um, arranged in spherical or elliptic

clusters (frequently hollow) with 10-40 spores in each cluster. Plasmodium

colourless.

SPECIMEN EXAMINED - BELARUS. MINSK REGION, Miadzel district, close to

Konstantinovo village, 54°56’35”N 26°26’48’E, black alder forest, on wood of Alnus

glutinosa (L.) Gaertn. (Betulaceae): 20.VII.1995, E. Moroz (LE 320679, MSK-F 42108).

ECOLOGY & DISTRIBUTION. Badhamia versicolor is a facultatively corticolous

species usually found in forest communities in temperate zone, although it

can also thrive in arid areas (Abdel-Azeem & Salem Fatma 2013, Estrada-

Torres & al. 2009, Wellman 2015). The bark of living trees, mosses, and

lichens are the typical substrates (Pliszko & Bochynek 2017).

The species is known from all continents except Antarctica (Abdel-

Azeem & Salem Fatma 2013, Macbride 1922, Moreno & al. 2013, Pliszko &

Bochynek 2017, Ranade & al. 2012; Wellman 2015). In Europe B. versicolor

has been reported from France, Germany, Great Britain, Italy, Poland, Russia

(Leningrad region and the Republic of Karelia), Spain, Switzerland, Turkey,

and Ukraine (Krzemieniewska 1960; Lado 1994; Martin & Alexopoulos

1969; Novozhilov 1993, 2005).

Despite its cosmopolitan range, B. versicolor is uncommon and often

considered a rare species (Ing, 1999, Martin & Alexopoulos 1969). In Poland,

B. versicolor is known as one of the rarest slime mould species, reported from

only two localities (Pliszko & Bochynek 2017). Currently it is included in

Polish national red list of myxomycetes (Drozdowicz & al. 2006).

In Belarus, the species was found within Narochansky National Park

at the border of Blue Lakes landscape wildlife sanctuary, one of the most

Badhamia versicolor & Trichia subfusca in Belarus ... 367

Fic. 1. Badhamia versicolor (LE 320679): spore clusters. Scale bar = 20 um.

undisturbed areas in the country. The forest area is dominated by Alnus

glutinosa (black alder) with a high degree of waterlogging.

Discussion. The most similar species are B. capsulifera (Bull.) Berk. and

B. dubia Nann.-Bremek., which differ from Badhamia versicolor by smaller

spore clusters and larger sporocarps (Pliszko & Bochynek 2017, Poulain

& al. 2011). Our Belarusian voucher has a white capillitium and therefore

resembles B. papaveracea Berk. & Ravenel, which differs by having stalked

sporocarps (Poulain & al. 2011).

Trichia subfusca Rex, Proc. Acad. Nat. Sci. Philadelphia 42: 192. 1890. Fic. 2

SPOROCARPS are scattered stalked sporangia, total height 0.8-1.5 mm.

Sporotheca subglobose, brown, 0.4-0.8 mm diam. Stipe dark brown,

reaching about half the sporocarp height. Peridium double, with an inner

membranous and an outer cartilaginous layer. Dehiscence occurring at the

top of the sporotheca. Capillitium formed by yellow elaters of 4.5-5.5 um

diam. with 3-4 smooth spirals and with free, short, acute, rarely curved ends.

Spores yellow in mass, pale yellow by transmitted light, 11-15 um diam.,

regularly warted. Plasmodium not seen.

SPECIMEN EXAMINED — BELARUS. GOMEL REGION, Buda-Koshelevo district, close

to Rudnja-Olhovka village, 52°32’N 30°22’E, Scots pine forest, on thallus of lichen

Cladonia uncialis, 7.V11.2005, A. Tsurykau (GSU 00266, MSK-F 42461).

ECOLOGY AND DISTRIBUTION. Like other Trichia species, T: subfusca inhabits

decaying wood (Schirmer & al. 2015).

Trichia subfusca is a very rare species (e.g., Schirmer & al. 2015),

occasionally reported from Europe (Austria, Italy, Poland, Russia [Moscow

368 ... Moroz & Tsurykau

Fic. 2. Trichia subfusca (GSU 00266): capillitium and spores. Scale bar = 10 um.

and Murmansk regions, Perm Territory, Republic of Karelia, and Komi

Republic], Spain, Sweden), Asia (India, Japan, Kazakhstan, Russia, Sri-

Lanka), and North America (Canada, USA) (Fefelov 2005; Gmoshinskiy

2014; Lado 1994; Novozhilov 1993, 2005; Schirmer & al. 2015; Vasyagina &

al. 1977).

In Belarus the species was found in well-lit dry Pinus sylvestris L. (Scots

pine) forest at the border of a sand quarry. The sporocarps were found

on the terricolous lichen Cladonia uncialis (L.) EH. Wigg. infected with

lichenicolous fungus Taeniolella beschiana Diederich.

Discussion. The Trichia botrytis group includes five morphologically related

species: T: botrytis (J.E. Gmel.) Pers., T: erecta Rex, T. flavicoma (Lister) Ing,

T: munda (Lister) Meyl., and T: subfusca (Schirmer & al. 2015). The species

most similar to T: subfusca is T. botrytis, which differs in larger (2-3 mm.),

areolate sporocarps with paler lines of dehiscence, elaters that gradually

taper to long slender points, and smaller (9-11 mm.) spores. Also similar to

T: subfusca is T: macrospora B. Zhang & Yu Li, distinguished by thicker (5-6

um diam.) elaters and larger (17-19 um diam.) densely verrucose spores

(Zhang & Li 2016).

Badhamia versicolor & Trichia subfusca in Belarus ... 369

Acknowledgments

We thank our reviewers, Kerry Knudsen (Czech University of Life Sciences

Prague, Prague, Czechia) and Grazina Adamonyté (Research Council of Lithuania,

Vilnius, Lithuania), Mycotaxon Nomenclature Editor Shaun B. Pennycook for helpful

comments and improvements, and Yuri K. Novozhilov (Komarov Botanical Institute

of RAS, Saint Petersburg, Russia) for confirming the identification of Trichia subfusca.

The work of Evgeny Moroz was financially supported by the grant 14.W03.31.0015

from the A.O. Kovalevsky Institute of Marine Biological Research of RAS.

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MYCOTAXON

April-June 2020—Volume 135, pp. 371-382

https://doi.org/10.5248/135.371

Vanderbylia cinnamomea sp. nov.

from southwestern China

JuN-ZHU CHEN?}, XIONG YANG", CHANG-LIN ZHAO???

' Key Laboratory of State Forestry and Grassland Administration on

Highly Efficient Utilization of Forestry Biomass Resources in Southwest China,

Southwest Forestry University, Kunming 650224, PR. China,

’ Key Laboratory for Forest Resources Conservation and Utilization in

the Southwest Mountains of China, Ministry of Education,

Southwest Forestry University, Kunming 650224, PR. China

° College of Biodiversity Conservation, Southwest Forestry University,

Kunming 650224, PR. China

“ CORRESPONDENCE TO: fungichanglinz@163.com

AxsstTRAcTt—Vanderbylia cinnamomea is proposed as a new wood-inhabiting polypore

species based on morphological characters supported by phylogenetic analysis. The species is

characterized by an annual, pileate basidiocarp, a brown to black pileal surface, an olivaceous

buff to smoke grey pore surface, a dimitic hyphal system with clamped generative hyphae, and

subglobose to amygdaliform, hyaline, thick-walled, smooth, dextrinoid, and cyanophilous

basidiospores. ITS+nLSU sequence analyses place V. cinnamomea in Vanderbylia within a

subclade with a high support (100% BS, 100% BP, 1.00 BPP) sister to a subclade comprising

V. fraxinea and V. robiniophila.

Key worps—Perenniporia, Polyporaceae, Polyporales, taxonomy, Yunnan Province

Introduction

Vanderbylia D.A. Reid (Polyporaceae, Polyporales) is typified by V. vicina

(Lloyd) D.A. Reid (Reid 1973). Ryvarden & Johansen (1980) reduced the

genus to a synonym under Perenniporia Murrill for a long time, but recent

phylogenetic research of a combined sequence dataset of the internal

transcribed spacer (ITS) and the large subunit nuclear ribosomal RNA

gene (nLSU) supports Vanderbylia as an independent genus (Robledo &

372 ... Chen, Yang, Zhao

al. 2009, Zhao & al. 2013, Cui & al. 2019). Morphologically Vanderbylia is

characterized by pileate basidiocarps, a dimitic hyphal system with clamped

generative hyphae and variably dextrinoid skeletal hyphae, and subglobose

to amygdaliform, non-truncate and variably dextrinoid basidiospores (Reid

1973, Zhao & Cui 2013). Currently, nine species are accepted in Vanderbylia

worldwide: V. borneensis Corner, V. delavayi (Pat.) B.K. Cui & Y.C. Dai,

V. devians (Bres.) D.A. Reid, V. fraxinea (Bull.) D.A. Reid, V. nigroapplanata

(Van der Byl) D.A. Reid, V. robiniophila (Murrill) B.K. Cui & Y.C. Dai,

V. subincarnata Corner, V. ungulata D.A. Reid, and V. vicina (Bresadola 1920,

Reid 1975, Corner 1987, Gilbertson & Ryvarden 1987, Nunez & Ryvarden

2001, Ryvarden & Melo 2014, Cui & al. 2019).

Robledo & al. (2009), who revealed the relationship between Perenniporia

and Perenniporiella Decock & Ryvarden, grouped two Vanderbylia species

together—the generic type V. vicina and V. fraxinea. In their phylogenetic

overview of Perenniporia s.l., Zhao & al. (2013) and revealed that Vanderbylia

fraxinea, V. robiniophila, and V. vicina form a single lineage. Subsequently,

Cui & al. (2019) investigated the species diversity, taxonomy, and phylogeny

of Polyporaceae in China and proposed two new combinations, Vanderbylia

delavayi and V. robiniophila.

During investigations on wood-inhabiting fungi in Yunnan Province,

an additional taxon was found that could not be assigned to any described

species. In examining the taxonomy and phylogeny of this new species,

we employed a two-gene molecular phylogenetic approach using internal

transcribed spacer (ITS) and long subunit (nLSU) plus an expanded

sampling of Vanderbylia isolates that support the existence of a new species,

Vanderbylia cinnamomea.

Materials & methods

The specimens studied are deposited at the herbarium of Southwest Forestry

University, Kunming, PR. China (SWFC). Macromorphological descriptions are

based on field notes. Color terms follow Petersen (1996). Micromorphological data

were obtained from the dried specimens and observed under light microscopy

following Dai (2012). The following abbreviations are used: KOH = 5% potassium

hydroxide, CB = Cotton Blue, CB- = acyanophilous, IKI = Melzer’s reagent, IKI- =

both 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 the L/W ratios between the specimens studied, n (a/b) = number of spores (a)

measured from given number (b) of specimens.

We extracted genomic DNA from dried specimens using Magen Biotech HiPure

Fungal DNA Mini Kit II according to the manufacturer's instructions with some

Vanderbylia cinnamomea sp. nov. (China) ... 373

TABLE 1. Species, specimens, and sequences used in this study.

(New sequences in bold).

GENBANK NO.

SPECIES NAME SAMPLE NO. —ToO__—.._ REFERENCES

ITS nLSU

Donkioporia expansa MUCL 35116 FJ411104 FJ393872 Robledo & al. (2009)

Pyrofomes demidoffii MUCL 41034 FJ411105 FJ393873 Robledo & al. (2009)

Vanderbylia cinnamomea = CLZhao 8952 T MT372778 MT372788 Present study

CLZhao 8959 MT372779 MT372789 Present study

CLZhao 8961 MT372780 MT372790 Present study

CLZhao 8962 MT372781 MT372791 Present study

CLZhao 8963 MT372782 MT372792 Present study

CLZhao 9001 MT372783 MT372793 Present study

V. fraxinea DP 83 AM269789 AM269853 Robledo & al. (2009)

Cui 7154 HQ654095 HQ654110 Zhao & al. (2013)

Cui 8871 JE706329 JE706345 Zhao & al. (2013)

V. robiniophila Cui 5644 HQ876609 —«JF706342 Zhao & al. (2013)

Cui 7144 HQ876608 JF706341 Zhao & al. (2013)

Cui 9174 HQ876610 JF706343 Zhao & al. (2013)

V. vicina MUCL 44779 FJ411095 AF518666 Robledo & al. (2009)

modifications. A small piece (about 30 mg) of dried fungal material was ground

to powder with liquid nitrogen, 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 the

addition of 0.4 mL phenol-chloroform (24:1) to each tube, 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. That mixture

was 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 30 s. After washing twice with 0.5 mL washing buffer, the AC was

transferred to a clean centrifuge tube, and 0.1 mL elution buffer was added to the

middle of adsorbed film to elute the genomic DNA. The ITS region was amplified

with primer pairs ITS5 and ITS4 (White & al. 1990). Nuclear LSU region was

amplified with primer pairs LROR and LR7 (https://sites.duke.edu/vilgalyslab/rdna_

primers_for_fungi). The PCR procedure for ITS was optimized as follows: initial

denaturation at 95 °C for 3 min, followed by 35 cycles at 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 condition

for nLSU was as follow: initial denaturation at 94 °C for 1 min, followed by 35 cycles

at 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. The six new sequences from specimens

of Vanderbylia cinnamomea were aligned with additional Vanderbylia sequences

downloaded from GenBank (TABLE 1).

Sequences were aligned in MAFFT 7 (http://mafft.cbrc.jp/alignment/server/)

using the “G-INS-I” strategy, and manually adjusted in BioEdit (Hall 1999).

374 ... Chen, Yang, Zhao

Alignment datasets were deposited in TreeBase (submission ID 24081). GenBank

sequences of Donkioporia expansa (Desm.) Kotl. & Pouzar and Pyrofomes demidoffii

(Lév.) Kotl. & Pouzar obtained from GenBank were used as an outgroup to root trees

following Zhao & al. (2013) in the ITS+nLSU analysis.

Maximum parsimony analysis was applied to the ITS+nLSU dataset sequences.

The tree construction procedure was performed in PAUP* version 4.0b10 (Swofford

2002). All characters were equally weighted, and gaps were treated as missing data.

Trees were inferred using the heuristic search option with TBR branch swapping

and 1000 random sequence additions. Max-trees were set to 5000, branches of zero

length were collapsed and all parsimonious trees were saved. Clade robustness was

assessed using a bootstrap (BP) analysis with 1000 replicates (Felsenstein 1985).

Descriptive tree statistics tree length (TL), consistency index (CI), retention index

(RI), rescaled consistency index (RC), and homoplasy index (HI) were calculated for

each Maximum Parsimonious Tree generated. Maximum Likelihood (ML) analysis

with RAxML-HPC2 was conducted for ITS datasets on Abe through the Cipres

Science Gateway (www.phylo.org; Miller & al. 2009). Branch support (BS) for ML

analysis was determined by 1000 bootstrap replicate.

MrModeltest 2.3 (Nylander 2004) was used to determine the best-fit evolution

model for each data set for Bayesian inference (BI). Bayesian inference was

calculated with MrBayes_3.1.2 with a general time reversible (GTR+I+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 2 million generations and trees were sampled every 100 generations. The

first one-fourth generations were discarded as burn-in. A majority rule consensus

tree of all remaining trees was calculated. Branches that received bootstrap support

for greater than or equal to 80% (maximum likelihood, BL), 50% (maximum

parsimony, BP), and 0.95 (Bayesian posterior probabilities, BPP) were considered as

significantly supported.

Phylogenetic results

The ITS dataset included sequences from 15 fungal specimens or isolates

representing 6 species. The dataset had an aligned length of 1844 characters,

of which 1690 characters were constant, 65 were variable and parsimony-

uninformative, and 89 were parsimony-informative. Maximum parsimony

analysis yielded 6 equally parsimonious trees (TL = 178, CI = 0.921, HI

= 0.787, RI = 0.944, RC = 0.870). Best model for the ITS+nLSU dataset

estimated and applied in the Bayesian analysis was GTR+I+G. Bayesian and

ML analyses produced a similar topology as MP analysis, with an average

standard deviation of split frequencies = 0.001512 (BI).

The phylogeny (Fic. 1) inferred from ITS+nLSU sequences obtained for

related taxa of Vanderbylia support the new species as independent in one

sub-clade and sister to the /fraxinea-robinophila subclade.

Vanderbylia cinnamomea sp. nov. (China) ... 375

Vanderbylia fraxinea DP 83

Vanderbylia fraxinea Cui 7154

Vanderbylia fraxinea Cui 8871

Vanderbyliarobiniophila Cui 7144

Vanderbyliarobiniophila Cui 5644

Vanderbyliarobiniophila Cui 9174

Vanderbylia cinnamomea CLZhao 8963

Vanderbylia cinnamomea CLZhao 8961

Vanderbylia cinnamomea CLZhao 8962

100/100/1.00 | Vanderbylia cinnamomea CLZhao 8959

Vanderbylia cinnamomea CLZhao 9001

Vanderbylia cinnamomea CLZhao 8952 T

Vanderbyliavicina MUCL 44779

Pyrofomes demidoffii MUCL 41034

Donkioporia expansa MUCL 35116

100/100/1.00

Fic. 1. Maximum Parsimony strict consensus tree illustrating the phylogeny of Vanderbylia

cinnamomea and related species in Vanderbylia based on ITS+nLSU sequence analyses. Branches

are labeled with maximum likelihood bootstrap >70%, parsimony bootstrap proportions >50%

and Bayesian posterior probabilities >0.95.

Taxonomy

Vanderbylia cinnamomea C.L. Zhao, sp. nov. Figs 2-4

MB 831788

Differs from Vanderbylia vicina by its annual basidiocarps and its smaller basidiospores.

Type: China. Yunnan Province: Kunming, Tanhuasi Park, on living tree of

Cinnamomum camphora (L.) J. Presl (Lauraceae), 13 October 2018, CL Zhao 8952

(Holotype, SWFC 008952; GenBank MT372778, MT372788).

Erymovoey: The specific epithet cinnamomea (Lat.) refers to the host of Cinnamomum

camphora.

BASIDIOMATA annual, pileate, imbricate, without odor or taste and corky

when fresh, becoming hard corky upon drying. Pilei dimidiate, projecting

<10 cm, 18 cm wide, 4.5 cm thick at centre. Pileal surface pale brown to

brown when fresh and brown to black from the base upon drying, smooth,

gently concentrically sulcate, glabrous. Pore surface pure buff to olivaceous

buff when fresh, olivaceous buff to smoke grey upon drying; pores round,

5-7 per mm; dissepiments thick, entire. Sterile margin narrow, buff, <0.5

376 ... Chen, Yang, Zhao

mm wide. Context cream, cottony, <4 cm thick. Tubes concolorous with pore

surface, <5 mm long.

HYPHAL STRUCTURE dimitic; generative hyphae with clamp connections,

strongly dextrinoid, CB+; tissues unchanging in KOH.

CONTEXT generative hyphae infrequent, hyaline, thin-walled, unbranched,

2-3 um in diam; skeletal hyphae dominant, hyaline, thick-walled with a wide

to narrow lumen, branched, interwoven, 4.5—-5.5 um in diam.

TUBES generative hyphae infrequent, hyaline, thin-walled, unbranched,

1.5-2.5 um in diam; skeletal hyphae dominant, hyaline, thick-walled with

a wide to narrow lumen, branched, interwoven, 4-5 um in diam. Presence

of arboriform hyphae, thick-walled to almost solid, with few branches and

short, lateral or terminal processes, tortuous, 25-40 x 2-6 um. Cystidia and

cystidioles not seen; basidia clavate, with 4 sterigmata and a basal clamp

connection, 20-24 x 6-9.5 um; basidioles dominant, clavate-shaped to pear-

shaped.

BasIDIOSPORES subglobose to amygdaliform, hyaline, thick-walled,

smooth, strongly dextrinoid, cyanophilous, 5-6(-6.5) x 4-5(-5.5) um,

L = 5.67 um, W = 4.73 um, Q = 1.17-1.21 (n = 180/6).

TYPE OF ROT: white.

ADDITIONAL SPECIMENS EXAMINED: CHINA. YUNNAN PROVINCE. Kunming: Tanhuasi

Park, on living tree of Cinnamomum camphora, 15 October 2018, CLZhao 8959

(SWFC 008959; GenBank MT372779, MT372789); CLZhao 8961 (SWFC 008961;

GenBank MT372780, MT372790); CLZhao 8962 (SWFC 008962, GenBank MT372781,

MT372791); CLZhao 8963 (SWFEC 008963; GenBank MT372782, MT372792); CLZhao

9001 (SWFC 009001; GenBank MT372783, MT372793).

Discussion

We describe here a new species Vanderbylia cinnamomea based on

phylogenetic analyses and morphological characters.

Phylogenetically, Vanderbylia cinnamomea grouped with species

V. fraxinea, V. robiniophila, and V. vicina in the rDNA-based phylogeny

(Fic. 1). Morphologically V. fraxinea differs in its perennial basidiocarps

and larger basidiospores (6-8 x 5-6.5 um, Ryvarden & Melo 2014), while

Vanderbylia robiniophila is distinguished by its growth on Robinia L., its

white to pale brown pore surface, and its larger basidiospores (5-8 x 5-7 um,

Gilbertson & Ryvarden 1987; Nufez & Ryvarden 2001); Vanderbylia vicina

differs in its perennial basidiocarps and larger basidiospores (8-8.9 x 6.9-7.5

um, Reid 1973).

Four species are morphologically similar to Vanderbylia cinnamomea:

V. borneensis, V. devians, V. subincarnata, and V. ungulata. Vanderbylia

Vanderbylia cinnamomea sp. nov. (China) ... 377

Fic. 2. Vanderbylia cinnamomea (holotype, SWFC 008952).

Basidiocarps. Scale bars = 5 cm.

378 ... Chen, Yang, Zhao

ine

| O

oltte

ile

Se 4, 8, 2

ISG

= — SBE

Gs SINS

SK eee

tae ey.

will

TESS

Fic. 3. Vanderbylia cinnamomea (holotype, SWFC 008952).

A. Basidiospores; B. Basidia and basidioles; C. Hyphae from trama; D. Hyphae from context.

Scale bars = 10 um.

Vanderbylia cinnamomea sp. nov. (China) ... 379

Fic. 4. Vanderbylia cinnamomea (holotype, SWFC 008952).

Arboriform hyphae. Scale bar = 10 um.

380 ... Chen, Yang, Zhao

borneensis differs from V. cinnamomea in its smaller pores (8-10 per mm) and

the presence of skeletal hyphal endings in the dissepiments (Corner 1987);

V. devians is distinguished by its substipitate basidiocarps with alutaceous

to isabelline pore surface and larger pores (3-4 per mm, Bresadola 1920);

V. subincarnata is diagnosed by its pinkish pore surface and smaller pores

(7-10 per mm, Corner 1987); and V. ungulata has ungulate basidiocarps with

an ochraceous pore surface and hyaline to yellow basidiospores (Ryvarden &

Johansen 1980).

When Decock & Ryvarden (1999) examined the holotype of Polyporus

delavayi Pat., they proposed that it be transferred to Perenniporia based on

its strongly dextrinoid, cyanophilous skeletal hyphae and dextrinoid, thick-

walled basidiospores; they also examined the type specimen of Perenniporia

formosana 'T.T. Chang and synonymized it under P. delavayi (Pat.) Decock

& Ryvarden. Cui & al. (2019) subsequently transferred Polyporus delavayi

to Vanderbylia based on molecular data and morphological characters on

finding P delavayi nested within the Vanderbylia lineage and manifesting

morphological features consistent with the concept of Vanderbylia.

Morphologically, V. delavayi differs from V. cinnamomea by its dirty greyish

orange to greyish brown pore surface, larger pores (4-5 per mm), and larger

basidiospores (5.7-7 x 4.7—5.9 um, Decock & Ryvarden 1999).

Wood-rotting basidiomycetes have been extensively studied (Gilbertson

& Ryvarden 1987, Nunez & Ryvarden 2001, Bernicchia & Gorjon 2010, Dai

2012, Ryvarden & Melo 2014), but Chinese wood-rotting fungi diversity is

still not well known, especially in the subtropics and tropics. The new species

Vanderbylia cinnamomea is from the Chinese subtropics, where many new

taxa in Polyporales have been described (Cui & al. 2007, 2011; Cui & Dai

2008; Du & Cui 2009; Li & Cui 2010; He & Li 2011; Jia & Cui 2011; Yu & al.

2013; Yang & He 2014; Chen & al. 2015; Zhao & Wu 2017; Shen & al. 2018;

Zhao & Ma 2019). We anticipate that more new polypore taxa will be found

in China after further investigations and molecular analyses.

Acknowledgments

Special thanks are due to Drs. Shah Hussain (University of Swat, Pakistan) and

Yuan-Yuan Chen (Henan Agricultural University, PR. China) who reviewed the

manuscript. The research was supported by the National Natural Science Foundation

of China (Project No. 31700023) and the Science Foundation of Yunnan Department

of Education (2018JS326) and the Key Laboratory of State Forestry Administration

for Highly Efficient Utilization of Forestry Biomass Resources in Southwest China

(Southwest Forestry University) (Project No. 2019-KF10).

Vanderbylia cinnamomea sp. nov. (China) ... 381

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MYCOTAXON

April-June 2020—Volume 135, pp. 383-404

https://doi.org/10.5248/135.383

G.H. Cunningham's use of te reo Maori in fungal epithets

SHAUN R. PENNYCOOK

Manaaki Whenua - Landcare Research

Private Bag 92 170, Auckland, New Zealand

CORRESPONDENCE TO: PennycookS@LandcareResearch.co.nz

ABSTRACT—New Zealand mycologist G.H. Cunningham (1892-1962) frequently used the

indigenous language of New Zealand, te reo Maori, when creating epithets for new species

and replacement names. All of these epithets are listed, and nomenclatural treatments

(including etymologies) of the taxa are presented. In addition, tables cite all te reo Maori and

te reo-derived generic names and epithets published by mycological authors. Guidelines are

discussed for the appropriate orthography for names and epithets derived from te reo Maori

and other non-latinate vernacular languages.

Key worps—ta re Moriori, undeclinable epithets, nouns in apposition, diacritical marks,

host species

Introduction

Gordon Herriot Cunningham (1892-1962), who was New Zealand's first

resident professional mycologist (McKenzie 2004; https://teara.govt.nz/en/

biographies/4c47/cunningham-gordon-herriot), is regarded as the “Father

of New Zealand Plant Pathology and Mycology.’ After about 17 years of

employment in the Department of Agriculture, during which he gained B.Sc.,

M.Sc., and Ph.D. degrees, Cunningham served the Department of Scientific and

Industrial Research [DSIR] as Director of the Plant Diseases Division [PDD]

from its foundation in 1936 until his death; his numerous dried specimens

formed the foundation of the current New Zealand Fungarium (PDD). His

initial focus was mainly on plant pathology, but taxonomic mycology soon

became a passion, and he published numerous papers and monographic books

on rust fungi, gasteromycetes, polypores, and thelephoraceous fungi.

384 ... Pennycook

A feature of Cunningham’s new taxa was his frequent use of fungal epithets

drawn from te reo Maori [the indigenous language of Aotearoa/New Zealand]

(Pennycook & McKenzie 2002). A survey across all biological disciplines (Veale

& al. 2019) identified Cunningham as the fourth most prolific proponent of

this practice (after an arachnologist and two malacologists) but did not discuss

his contribution. A search of Cunningham’s taxonomic publications has

identified 56 species (proposed as either sp. nov. or nom. nov.) with 49 epithets

either drawn directly, or derived, from te reo Maori. These are presented here,

with nomenclatural, taxonomic, and etymological details. In addition, tables

are presented of te reo Maori and te reo-derived genus names and species/

subspecies/variety epithets published by all mycological authors. Guidelines

are discussed for the appropriate orthography for names and epithets derived

from te reo Maori and other non-latinate vernacular languages.

Materials & methods

All of Cunningham's taxonomic publications were searched; 22 of them proposed

names with te reo Maori or te reo-derived epithets (Cunningham 1923, 1924a,b,

1928, 1930, 1931, 1938, 1944, 1945a,b, 1947, 1948a,b,c, 1949, 1953a,b, 1954, 1955,

1956, 1963, 1965).

For a comprehensive overview of te reo Maori or te reo-derived fungal

names (genera, species, subspecies, and varieties), a careful search was made of

comprehensive modern listings of New Zealand fungi (Buchanan & Ryvarden 2000;

Gadgil & al. 2005; Galloway 2007; Gordon 2012; Horak 2008, 2018; Pennycook

1989, 2004; Pennycook & Galloway 2004; Petersen 1988; Stephenson 2003; Vanky

& McKenzie 2002), and these were cross-checked and augmented by searches of

the major mycological nomenclature websites (Index Fungorum 2020, MycoBank

2020, Nga Harore 2020). Epithets derived from other Polynesian languages

(e.g., Hawaiian, Tahitian) and epithets with a misleading similarity to te reo Maori

(e.g., wainioi referring to the Finnish lichenologist Vainio/Wainio; waikerieanum

from the Australian locality Waikerie) were deleted from the search results.

Cunningham's epithets

The 49 te reo Maori or te reo-derived fungal epithets proposed by

Cunningham are listed alphabetically. Presented beneath each epithet are the

new species or replacement names in which the epithet was first published,

together with all subsequent homotypic synonyms in which the epithet was

recombined. The currently accepted name for each species is indicated with

bold font. The etymology of each epithet is explained, together with the type

host and the type locality when these are relevant to the etymology; in addition,

the host species of the type specimen is listed for all rust species, even if not

involved in the etymology. Maori names of host plants were sourced from

Williams (1971) and Beever (1991). Localities are referred to the “Crosby

Cunningham & te reo Maori (New Zealand) ... 385

districts” defined by Crosby & al. (1998). Host nomenclature and synonymy

follows Nga Tipu (2020).

akiraho

Puccinia akiraho G. Cunn., Trans. & Proc. New Zealand Inst. 61: 411. 1930.

= Puccinia novae-zelandiae G. Cunn., Trans. & Proc. New Zealand

Inst. 54: 686. 1923. [non P. novozelandica Bubak 1901].

TyPE HOST: Olearia forsteri (Compositae) [= Olearia paniculata (J.R. Forst. & G.

Forst.) Druce].

ErymMo oey: akiraho, Maori name for Olearia paniculata.

aorangi

Puccinia aorangi G. Cunn., Trans. & Proc. New Zealand Inst. 61: 411. 1930.

TYPE LOCALITY & HOST: 1600-2000 m altitude on Taranaki/Mt Egmont, North

Island; Celmisia major Cheeseman (Compositae) [= Celmisia major var. brevis Allan,

the variety endemic to Taranaki/Mt Egmont].

EryMo.ocy: aorangi/aoraki [“cloud-piercer”], Maori name (and common name)

for Mt Cook, South Island; perhaps used in a general sense, referring to the type

locality on a North Island cloud-piercer.

= Puccinia egmontensis G. Cunn., Trans. & Proc. New Zealand Inst. 54: 681. 1923.

aotearoa

Lachnella aotearoa G. Cunn., New Zealand Dept. Sci. Industr. Res.,

Bull. 145: 331. 1963.

= Flagelloscypha aotearoa (G. Cunn.) Agerer, Mitt.

Bot. Staatssamml. Miinchen 19: 252. 1983

ETYMOLOGY: aotearoa, Maori name (and common name) for New Zealand.

Stereum aotearoa G. Cunn., Trans. Roy. Soc. New Zealand 84(2): 212. 1956.

ETYMOLOGY: aotearoa, Maori name (and common name) for New Zealand.

aroha

Poria aroha G. Cunn., New Zealand Dept. Sci. Industr. Res., Pl. Dis. Div.,

Bull. 72: 39. 1947.

= Flaviporus aroha (G. Cunn.) G. Cunn., New Zealand

Dept. Sci. Industr. Res., Bull. 164: 150. 1965.

EryMo ocy: aroha, Maori, referring to the type locality, Mt Te Aroha, Bay of Plenty.

= Australoporus tasmanicus (Berk.) PK. Buchanan & Ryvarden, Mycotaxon 31: 5. 1988.

awhitu

Fomes awhitu G. Cunn., New Zealand Dept. Sci. Industr. Res., Pl. Dis. Div.,

Bull. 79: 16. 1948.

EryMo.ocy: awhitu, Maori, referring to the type locality, Awhitu Peninsula,

Auckland.

= Abundisporus roseoalbus (Jungh.) Ryvarden, Belg. J. Bot. 131: 154. 1999.

386 ... Pennycook

haumata

Uredo haumata G. Cunn., Trans. & Proc. New Zealand Inst. 59: 499. 1928.

Correct name will combine this epithet in Uromyces (E.H.C. McKenzie, in prep.).

TyPE Host: Danthonia cunninghamii (Gramineae) [= Chionochloa conspicua subsp.

cunninghamii (Hook.f.) Zotov].

EryMo.ocy: haumata, Maori name for broad-leaved snowgrass, Chionochloa spp.

= Uromyces macnabbii Cummins, Rust Fungi Cereals, Grasses and Bamboos: 483. 1971.

hautu

“Hysterangium hautu” G. Cunn., Trans. & Proc. Roy. Soc. New Zealand 67: 409.

1938, nom. inval. [no Latin description].

= Protubera hautuensis Castellano & Beever, New Zealand J. Bot. 32: 322. 1994.

EryMo oey: hautu, Maori name for the type locality, Waimarino, Taupo.

heketara

Puccinia heketara G. Cunn., Trans. & Proc. New Zealand Inst. 55: 393. 1924.

TyPE HOST: Olearia cunninghamii (Compositae) [= Olearia rani (A. Cunn.) Druce].

EtyMo.oGcy: heketara, Maori name for Olearia rani.

= Puccinia atkinsonii G. Cunn., Trans. & Proc. New Zealand Inst. 54: 675. 1923.

hoheriae

Puccinia hoheriae G. Cunn., Trans. & Proc. New Zealand Inst. 54: 661. 1923,

nom. illeg. [non Wakefield 1917].

TyPE HOST: Hoheria populnea A. Cunn. (Malvaceae).

ETYMOLOGY: genitive case of generic name (and common name) Hoheria, derived

from hohere, Maori name for Hoheria spp.

= Puccinia hoheriae Wakef., Bull. Misc. Inform. Kew. 1917: 312. 1917.

= Puccinia plagianthi McAlpine, Proc. Roy. Soc. Victoria 7: 218. 1895.

horopito

Uredo horopito G. Cunn., Trans. & Proc. Roy. Soc. New Zealand 75: 326. 1945.

TYPE HOST: Pseudowintera axillaris (J.R. Forst. & G. Forst.) Dandy (Winteraceae).

EtryMo.ocy: horopito, Maori name (and common name) for Pseudowintera spp.

huia

Solenia huia G. Cunn., Trans. Roy. Soc. New Zealand 81(2): 179. 1953.

= Porotheleum huia (G. Cunn.) W.B. Cooke, Mycologia 49: 691. 1957.

= Stigmatolemma huia (G. Cunn.) W.B. Cooke, Beih. Sydowia 4: 128. 1961.

= Stromatoscypha huia (G. Cunn.) G. Cunn., New Zealand

Dept. Sci. Industr. Res., Bull. 145: 305. 1963.

= Resupinatus huia (G. Cunn.) Thorn & al., Mycologia 97: 1148. 2006.

EryMo ocy: huia, Maori, referring to the type locality, Huia, Auckland.

Cunningham & te reo Maori (New Zealand) ...

hunua

Poria hunua G. Cunn., New Zealand Dept. Sci. Industr. Res., Pl. Dis. Div.,

Bull. 72: 39. 1947.

= Antrodiella hunua (G. Cunn.) Ryvarden, Prelim. Polyp. Fl. E. Africa: 257. 1980.

= Flaviporus hunua [as “hunuus”] (G. Cunn.) Ginns, Mycotaxon 21: 326. 1984.

EryMo ocy: hinua, Maori, referring to the type locality, Hunua Range, Auckland.

hupiro

Aecidium hupiro G. Cunn., Trans. & Proc. New Zealand Inst. 55: 36. 1924.

TYPE HOST: Coprosma foetidissima J.R. Forst. & G. Forst. (Rubiaceae).

EtryMotoey: hipiro, Maori name for Coprosma foetidissima.

kaiku

Caeoma kaiku G. Cunn., Trans. & Proc. New Zealand Inst. 61: 416. 1930.

TyPE Host: Parsonsia capsularis (G. Forst.) DC. x heterophylla A. Cunn.

(Apocynaceae).

Erymo .ocy: kaiki, Maori name for Parsonsia spp.

kamahi

Fuscoporia kamahi G. Cunn., New Zealand Dept. Sci. Industr. Res.,

Bull. 164: 263. 1965.

387

= Phellinus kamahi (G. Cunn.) P.K. Buchanan & Ryvarden, Mycotaxon 31: 15. 1988.

TYPE HOST: Weinmannia racemosa L.f. (Cunoniaceae).

EtryMo.oGcy: kamahi, Maori name (and common name) for Weinmannia racemosa.

karetu

Uredo karetu G. Cunn., Trans. & Proc. New Zealand Inst. 55: 41. 1924.

Type HOST: Hierochloe redolens (Vahl) Roem. & Schult. (Gramineae).

ETyMOLoGy: karetu, Maori name (and common name) for Hierochloe redolens.

kauri

Corticium kauri G. Cunn., Trans. Roy. Soc. New Zealand 82: 303. 1954.

TYPE SUBSTRATE: much decayed wood of Agathis australis (D. Don) Lindl. ex

Loudon (Araucariaceae).

Erymo ocy: kauri, Maori name (and common name) for Agathis australis.

keae

Puccinia keae G. Cunn., Trans. & Proc. New Zealand Inst. 59: 493. 1928.

TyPE HOST: Olearia nummulariifolia (Hook.f.) Hook.f. (Compositae).

ETYMOLOGy: kea, Maori name (and common name) for Nestor notabilis [mountain

parrot]; referring to the type locality, Kea Point, Mackenzie.

388 ... Pennycook

koherika

Puccinia koherika G. Cunn., Trans. & Proc. New Zealand Inst. 61: 408. 1930.

TyPE Host: Angelica rosifolia Hook. (Umbelliferae) [= Scandia rosifolia (Hook.) J.W.

Dawson].

Erymo ocy: koheriki, Maori name for Scandia rosifolia.

konini

Uredo konini G. Cunn., Trans. & Proc. New Zealand Inst. 59: 502. 1928, nom. illeg.

[a superfluous nom. nov., proposed to avoid creating a comb. nov. that would be

an illegitimate later homonym of Uredo fuchsiae Arthur & Holway1918; however,

Cunningham was apparently unaware that the combination had already been

published legitimately in 1903].

= Coleosporium fuchsiae Cooke, Grevillea 14: 129. 1886

= Uredo fuchsiae (Cooke) Henn., Beibl. Hedwigia 42: (73). 1903.

TyPE HosT: Fuchsia excorticata (Vahl) Roem. & Schult. (Onagraceae).

EtyMo.oGcy: konini, Maori name for fruit of Fuchsia excorticata.

= Mikronegria fuchsiae PE. Crane & R.S. Petersen, New Zealand J. Bot. 45: 709. 2007.

kopoti

Puccinia kopoti G. Cunn., Trans. & Proc. New Zealand Inst. 54: 668. 1923.

TYPE HOST: Anisotome aromatica Hook.f. (Umbelliferae).

ErymMo ocy: kopoti, Maori name for Anisotome aromatica.

kowhai

Aecidium kowhai G. Cunn., Trans. & Proc. New Zealand Inst. 55: 35. 1924.

TyPE Host: Edwardsia tetraptera (Leguminosae) [= Sophora tetraptera J.F. Mill.].

EtryMo ocy: kowhai, Maori name (and common name) for Sophora spp.

= Uromyces edwardsiae G. Cunn., Trans. & Proc. New Zealand Inst. 55: 392. 1924.

maire

Laricifomes maire G. Cunn., New Zealand Dept. Sci. Industr. Res.,

Bull. 164: 262. 1965.

= Fomitopsis maire (G. Cunn.) P.K. Buchanan & Ryvarden,

Mycotaxon 31: 15. 1988.

TyPE HOST: Gymnelaea cunninghamii (Oleaceae) [= Nestegis cunninghamii (Hook.f.)

L.A.S. Johnson].

ETyMOLoGy: maire, Maori name (and common name) for Nestegis spp.

mania

Puccinia mania G. Cunn., Trans. & Proc. New Zealand Inst. 61: 404. 1930.

TyPE HOST: Carex wakatipu Petrie (Cyperaceae).

EryMo_LoGy: mania, Maori name for some Carex spp.

Cunningham & te reo Maori (New Zealand) ... 389

manuka

Poria manuka G. Cunn., New Zealand Dept. Sci. Industr. Res., Pl. Dis. Div.,

Bull. 72: 38. 1947.

= Oligoporus manuka (G. Cunn.) P.K. Buchanan & Ryvarden,

Mycotaxon 31: 17. 1988.

= Postia manuka (G. Cunn.) PK. Buchanan & Ryvarden,

New Zealand J. Bot. 38(2): 303. 2000.

TyPE HosT: Leptospermum scoparium J.R. Forst. & G. Forst. (Myrtaceae).

EryMoLocy: manuka, Maori name (and common name) for Leptospermum

scoparium.

maurea

Puccinia maurea G. Cunn., Trans. & Proc. New Zealand Inst. 61: 406. 1930.

TYPE HOST: Carex inversa R. Br. (Cyperaceae).

ETyMOLOoGy: maurea, Maori name for some Carex spp.

namua

Puccinia namua G. Cunn., Trans. & Proc. New Zealand Inst. 55: 3. 1924.

TyPE HosT: Anisotome filifolia (Hook.f.) Cockayne & Laing (Umbelliferae).

ETYMOLOGy: namua, derivation unknown. (Similar te reo Maori words are namu

for sandfly [blackfly] or mosquito; and namunamu for indigenous Geranium spp.

and the exotic weed Geranium molle (Geraniaceae). The Maori name for Anisotome

aromatica is kopoti.)

nikau

Epithele nikau G. Cunn., Trans. Roy. Soc. New Zealand 83: 629. 1956.

= Skeletohydnum nikau (G. Cunn.) Jilich, Persoonia 10: 331. 1979.

Type HosT: Rhopalostylis sapida H. Wendl. & Drude (Palmae).

EryMo oey: nikau, Maori name (and common name) for Rhopalostylis sapida.

Lachnella nikau G. Cunn., New Zealand Dept. Sci. Industr. Res.,

Bull. 145: 331. 1963.

Type HosT: Rhopalostylis sapida H. Wendl. & Drude (Palmae).

ErymMo oey: nikau, Maori name (and common name) for Rhopalostylis sapida.

Peniophora nikau G. Cunn., New Zealand Dept. Sci. Industr. Res.,

Bull. 145: 127. 1963.

= Peniophora sororia G. Cunn., Trans Roy. Soc. New Zealand 83(2):

280. 1955, nom. illeg. [non Bourdot & Galzin 1913].

= Subulicystidium nikau (G. Cunn.) Julich, Ber. Deutsch.

Bot. Ges. 81(9): 419. 1969 [“1968”].

Type HosT: Rhopalostylis sapida H. Wendl. & Drude (Palmae).

EryMo oey: nikau, Maori name (and common name) for Rhopalostylis sapida.

390 ... Pennycook

otagensis/—ense

Corticium otagense G. Cunn., New Zealand Dept. Sci. Industr. Res.,

Bull. 145: 331. 1963.

EryMo_oey: referring to the European name Otago (historically the southernmost

province of New Zealand), derived from 6takou, a Kai Tahu [southern Maori]

locality name [see also the otakou epithet entry, below]; the guttural Kai Tahu

pronunciation of ‘k’ was misheard as a ‘g’ by early settlers.

Pellicularia otagensis G. Cunn., Trans. Roy. Soc. New Zealand 81: 324. 1953.

= Sistotrema otagense (G. Cunn.) Stalpers & PK.

Buchanan, New Zealand J. Bot. 29: 331. 1991.

EryMo_oey: referring to the European name Otago (historically the southernmost

province of New Zealand), derived from 6takou, a Kai Tahu [southern Maori]

locality name [see also the otakou epithet entry, below]; the guttural Kai Tahu

pronunciation of ‘k was misheard as a ‘g’ by early settlers.

otakou

Poria otakou G. Cunn., New Zealand Dept. Sci. Industr. Res., Pl. Dis. Div.,

Bull. 72: 38. 1947.

= Ceriporia otakou (G. Cunn.) P.K. Buchanan & Ryvarden, Mycotaxon 31: 22. 1988.

= Gloeoporus otakou (G. Cunn.) Zmitr. & Spirin, Mycena 6: 35. 2006.

EryMoLoecy: otakou, Maori name of the Kai Tahu kaika [village] near Taiaroa

Head, Otago Peninsula, where in 1848 Scottish settlers made their first contact

with tangata whenua [the people of the land]; referring to the type locality, Kinloch,

Otago Lakes.

Uromyces otakou G. Cunn., Trans. & Proc. New Zealand Inst. 54: 627. 1923.

TyPE HOST: Poa caespitosa Spreng. (Gramineae) [= Poa cita Edgar].

EtryMoLoGcy: otakou, Maori name of the Kai Tahu kaika [village] near Taiaroa

Head, Otago Peninsula, where in 1848 Scottish settlers made their first contact with

tangata whenua [the people of the land]; referring to the type locality, Routeburn

Valley, Otago Lakes.

otira

Aecidium otira G. Cunn., Trans. & Proc. New Zealand Inst. 59: 498. 1928.

Correct name will combine this epithet in Puccinia (E.H.C. McKenzie, in prep.).

TyPE HOST: Olearia arborescens (G. Forst.) Cockayne & Laing (Compositae).

ETyMOLoGy: Otira, Maori name for the Otira river flowing westward from the

vicinity of the type locality, Arthurs Pass, North Canterbury.

pirongia

Poria pirongia G. Cunn., New Zealand Dept. Sci. Industr. Res., Pl. Dis. Div.,

Bull. 72: 39. 1947.

= Haploporus pirongia (G. Cunn.) Meng Zhou & al., MycoKeys 54: 92. 2019.

EtyMo_ocy: p irongia, Maori, referring to the type locality, Mt Pirongia, Waikato.

Cunningham & te reo Maori (New Zealand) ... 391

pounamu

Puccinia pounamu G. Cunn., Trans. & Proc. New Zealand Inst. 54: 688. 1923.

TyPE HOST: Senecio southlandicus (Compositae) [= Brachyglottis lagopus (Raoul) B.

Nord. ].

ETYMOLOGy: pounamou, Maori word for greenstone [nephrite jade]; referring to

the type locality Greenstone Valley, Otago Lakes.

rata

Poria rata G. Cunn., New Zealand Dept. Sci. Industr. Res., Pl. Dis. Div.,

Bull. 72: 40. 1947.

= Antrodiella rata (G. Cunn.) P.K. Buchanan & Ryvarden, Mycotaxon 31: 25. 1988.

TyPE HOST: Metrosideros robusta A. Cunn. (Myrtaceae).

EryMo.oey: rata, Maori name (and common name) for some Metrosideros spp.

rautahi

Puccinia rautahi G. Cunn., Trans. & Proc. New Zealand Inst. 61: 405. 1930.

TYPE HOST: Carex gaudichaudiana Kunth (Cyperaceae).

EryMo_ocy: rautahi, Maori name for some Carex spp.

tararua

Puccinia tararua G. Cunn., Trans. & Proc. New Zealand Inst. 54: 671. 1923.

TYPE HOST: Gentiana patula (Gentianaceae) |= Gentianella patula (Kirk) Holub].

ETyMoLoGcy: tararua, Maori; referring to the type locality, Tararua Ranges,

Wellington.

tawa

Trametes tawa G. Cunn., New Zealand Dept. Sci. Industr. Res., Pl. Dis. Div.,

Bull. 80: 9. 1948.

= Metuloidea tawa (G. Cunn.) G. Cunn., New Zealand

Dept. Sci. Industr. Res., Bull. 164: 250. 1965.

FREQUENT SUBSTRATE: bark of dead upright branches of Beilschmiedia tawa (A.

Cunn.) Benth. & Hook.f. ex Kirk (Lauraceae).

ETYMOLOGy: tawa, Maori name (and common name) for Beilschmiedia tawa.

= Metuloidea rhinocephala (Berk.) Miettinen, Ann. Bot. Fenn. 53(3-4): 165. 2016.

tawhai

Fuscoporia tawhai G. Cunn., New Zealand Dept. Sci. Industr. Res., Pl. Dis. Div.,

Bull. 73: 8. 1948.

= Phellinus tawhai (G. Cunn.) G. Cunn., New Zealand

Dept. Sci. Industr. Res., Bull. 164: 229. 1965.

Type Host: Nothofagus cliffortioides (Nothofagaceae) [= Fuscospora cliffortioides

(Hook.f.) Heenan & Smissen].

EtyMoLocy: tawhai/tawai, Maori name for Nothofagus [= Fuscospora and

Lophozonia] spp.

392 ... Pennycook

tiritea

Puccinia tiritea G. Cunn., Trans. & Proc. New Zealand Inst. 54: 654. 1923.

Type HosT: Muehlenbeckia australis (G. Forst.) Meisn. (Polygonaceae).

ETyMOoLoGy: turitea/tiritea, Maori; referring to the type locality, Turitea,

Wellington.

= Puccinia otagensis (Linds.) McKenzie & Padamsee, Pl. Pathol. 66: 1252. 2017.

toa

Puccinia toa G. Cunn., Trans. & Proc. New Zealand Inst. 61: 408. 1930.

= Puccinia haloragis [as “halorrhagidis”] G. Cunn., Trans. & Proc. New

Zealand Inst. 54: 664. 1923, nom. illeg. [pon Sydow & Sydow 1913].

Type Host: Haloragis [as “Halorrhagis”| depressa (Haloragaceae) [= Gonocarpus

aggregatus (Buchanan) Orchard].

ETYMOLOGy: toa/toatoa, Maori name for some Haloragis spp.

toatoa

Tyromyces toatoa G. Cunn., New Zealand Dept. Sci. Industr. Res., Bull. 164: 262.

1965.

Type Host: Phyllocladus alpinus Hook.f. (Podocarpaceae).

ETyMOLoGy: toatoa, Maori name for Phyllocladus spp.

toetoe

Uredo toetoe G. Cunn., Trans. & Proc. New Zealand Inst. 55: 41. 1924.

TyPE HOST: Arundo conspicua (Gramineae) [= Chionochloa conspicua (G. Forst.)

Zotov].

ETyMOLoGy: toetoe, Maori name (and common name) for Chionochloa conspicua.

tongariro

Cyphella tongariro G. Cunn., Trans. Roy. Soc. New Zealand 81: 185. 1953.

= Lachnella tongariro (G. Cunn.) W.B. Cooke, Beih. Sydowia 4: 80. 1961.

= Flagelloscypha tongariro (G. Cunn.) Agerer, Sydowia 32: 9. 1980 [“1979”].

EtryMoLocy: tongariro, Maori; referring to the type locality, Mt Tongariro, Taupo.

totara

Cyphella totara G. Cunn., Trans. Roy. Soc. New Zealand 81: 182. 1953.

= Calyptella totara (G. Cunn.) W.B. Cooke, Beih. Sydowia 4: 43. 1961.

= Lachnella totara (G. Cunn.) G. Cunn., New Zealand Dept.

Sci. Industr. Res., Bull. 145: 318. 1963.

TyPE HOST: Podocarpus totara D. Don (Podocarpaceae).

ETyMOLoGy: totara, Maori name (and common name) for Podocarpus totara and

P. laetus.

Cunningham & te reo Maori (New Zealand) ... 393

Peniophora totara G. Cunn., Trans. Roy. Soc. New Zealand 83: 271. 1955.

= Phlebia totara (G. Cunn.) Stalpers & P.K. Buchanan,

New Zealand J. Bot.29: 338. 1991.

Type HOST: Podocarpus hallii (Podocarpaceae) |= Podocarpus laetus Hooibr. ex Endl.].

ETyMoLoey: totara, Maori name (and common name) for Podocarpus laetus and

P. totara.

Poria totara G. Cunn., New Zealand Dept. Sci. Industr. Res., Bull. 164: 261. 1965

= Ceriporia totara (G. Cunn.) P.K. Buchanan & Ryvarden, Mycotaxon 31: 33. 1988

= Gloeoporus totara [as “totarus”| (G. Cunn.) Zmitr. & Spirin, Mycena 6: 36. 2006.

TyPE HOST: Podocarpus totara D. Don (Podocarpaceae).

ETyMOLoGy: totara, Maori name (and common name) for Podocarpus totara and

P. laetus.

tupare

Uredo tupare G. Cunn., Trans. & Proc. New Zealand Inst. 55: 44. 1924.

Type Host: Olearia colensoi Hook.f. (Oleaceae).

EryMoLoGy: tupare, Maori name for Olearia colensoi.

waiouru

Cintractia waiouru G. Cunn., Trans. & Proc. Roy. Soc. New Zealand 75: 335. 1945.

TyPE HOST: Carpha alpina R. Br. (Cyperaceae).

ETyMoLoGy: waiouru, Maori; referring to the area south of the type locality,

Waihohonu Stream, National Park, Taupo.

= Anthracoidea carphae (Speg.) Vanky, Bot. Not. 132: 230. 1979.

weraroensis

Poria weraroensis G. Cunn., New Zealand Dept. Sci. Industr. Res., Pl. Dis. Div.,

Bull. 72: 40. 1947, nom. dub.

[a name of uncertain application; Buchanan & Ryvarden 2000: 301].

ETyMOLoGy: weraroa, Maori; referring to the type locality, Weraroa, Wellington.

whakatipu

Puccinia whakatipu G. Cunn., Trans. & Proc. New Zealand Inst. 55: 4. 1924.

TYPE Host: Anisotome filifolia (Hook.f.) Cockayne & Laing (Umbelliferae).

EryMo.ocy: whakatipu/wakatipu, Maori; referring to the type locality, Lake

Wakatipu, Otago Lakes.

wharanui

Uredo wharanui G. Cunn., Trans. & Proc. New Zealand Inst. 55: 46. 1924.

Type Host: Olearia insignis (Compositae) [= Pachystegia insignis (Hook.f.)

Cheeseman].

EtryMo_ocy: wharanui, Maori; referring to the type locality, Wharanui, Kaikoura.

394 ... Pennycook

TABLE 1. Fungal generic names based on te reo Maori.

Classification is to family and order; “[L]’= lichenised taxon.

NAME TAXON CLASSIFICATION

Aotearoamyces Aotearoamyces P.R. Johnst. & al. 2018 Tympanidaceae, Helotiales

Harorepupu Harorepupu P.R. Johnst. & al. 2015 Inc. sed., Onygenales

Podotara Podotara Malcolm & Vézda 1996 Pilocarpaceae, Lecanorales [L]

Pureke Pureke PR. Johnst. 1991 Rhytismataceae, Rhytismatales

Weraroa Weraroa Singer 1958 Hymenogastraceae, Agaricales

TABLE 2. Fungal species and variety epithets based on te reo Maori and ta re Moriori;

only new and replacement names are listed.

Classification is to subphylum; [L] = lichenised taxon; [Lc] = lichenicolous taxon.

EPITHET

akatorensis

TAXON

Buellia akatorensis Elix & A. Knight 2017

CLASSIFICATION

Pezizomycotina [L]

akiraho Puccinia akiraho G. Cunn. 1930 Pucciniomycotina

anauensis Cortinarius anauensis Soop 2001 Agaricomycotina

aorangi Puccinia aorangi G. Cunn. 1930 Agaricomycotina

Stephanospora aorangi Beever & al. 2015 Pucciniomycotina

aotearoa Armillaria aotearoa L.A. Hood & Ramsfield 2016 Agaricomycotina

Chalara aotearoa Nag Raj & S. Hughes 1974 Pezizomycotina

Colletotrichum aotearoa B.S. Weir & P.R. Johnst. 2012 Pezizomycotina

Harorepupu aotearoa PR. Johnst. & al. 2015 Pezizomycotina

Lachnella aotearoa G. Cunn. 1963 Agaricomycotina

Neopestalotiopsis aotearoa Maharachch. & al. 2014 Pezizomycotina

Phyllisciella aotearoa Henssen & J.K. Bartlett 1984 Pezizomycotina [L]

Stereum aotearoa G. Cunn. 1956 Agaricomycotina

aotearoae Chaetosphaeria aotearoae S. Hughes 1966 Pezizomycotina

Hypochnicium aotearoae B.C. Paulus & al. 2007 Agaricomycotina

Neotyphodium aotearoae C.D. Moon & al. 2002 Pezizomycotina

Penicillium aotearoae Visagia & Seifert 2016 Pezizomycotina

aotearoana Pannaria aotearoana Elvebakk & Elix 2016 Pezizomycotina [L]

araniiti Cortinarius araniiti Soop 2014 Agaricomycotina

aroha Poria aroha G. Cunn. 1947 Agaricomycotina

awakinoana Chlorociboria awakinoana P.R. Johnst. 2005 Pezizomycotina

awhitu Fomes awhitu G. Cunn. 1948 Agaricomycotina

corokiae Septoria corokiae Henn.1898 Pezizomycotina

hakaroa Gymnopus hakaroa J.A. Cooper & P. Leonard 2013 Agaricomycotina

harakeke Marthamyces harakeke P.R. Johnst. 2019 Pezizomycotina

haumata Uredo haumata G. Cunn. 1928 Pucciniomycotina

hautu Hysterangium hautu G. Cunn. 1938 Agaricomycotina

hautuensis Protubera hautuensis Castellano & Beever 1994 Agaricomycotina

hauturu Cordyceps hauturu Dingley 1953 Pezizomycotina

Nectria hauturu Dingley 1951 Pezizomycotina

Phyllachora hauturu P.R. Johnst. & P.F. Cannon 2004 Pezizomycotina

hauturuanum Lophodermium hauturuanum PR. Johnst. 1989 Pezizomycotina

EPITHET

heketara

hoheriae

horoeka

horopito

huia

huiaensis

hunua

hupiro

iringa

iti

itiiti

kaikawakae

kaiku

kaimanawa

kaitokensis

kamahi

kamaka

kanuka

kanukaneus

kapiti

kapitiae

kapitiana

karaka

karea

karetu

kauri

keae

kiko

kiwiorum

koherika

kohu

konini

kopoti

kopuwaianus

Cunningham & te reo Maori (New Zealand) ... 395

TAXON

Puccinia heketara G. Cunn. 1924

Eichleriella hoheriae McNabb 1969

Trenopsis hoheriae Hansf. 1955

Myxosporium hoheriae J.D. Atk. 1940

Nectria hoheriae Dingley 1989

Puccinia hoheriae G. Cunn. 1923

Puccinia hoheriae Wakef. 1917

Pirottaea horoeka P.R. Johnst. 1998

Uredo horopito G. Cunn. 1945

Solenia huia G. Cunn. 1953

Bullera huiaensis Hamam. & Nakase 1996

Hypocrea hunua Dingley 1952

Poria hunua G. Cunn. 1947

Aecidium hupiro G. Cunn. 1924

Cortinarius iringa Soop 2003

Rhodocybe iti E. Horak 1979

Microsporidium itiiti L.A. Malone 1985

Lophodermium kaikawakae P.R. Johnst. 1989

Caeomo kaiku G. Cunn. 1930

Cortinarius kaimanawa Soop 2003

Strigula kaitokensis Sérus. & Polly 1996

Fuscoporia kamahi G. Cunn. 1965

Clitopilus kamaka J.A. Cooper 2014

Hydnangium kanuka J.A. Cooper 2014

Stephanospora kanuka T. Lebel & Castellano 2015

Marasmius kanukaneus G. Stev. 1964

Anthostomella kapiti Whitton & al. 2000

Stachybotrys kapiti Whitton & al. 2001

Anthostomella kapitiae Whitton & al. 2000

Niesslia kapitiana Whitton & al. 2012

Cercospora karaka G.F. Laundon 1973

Amanita karea G.S. Ridl. 1991

Uredo karetu G. Cunn. 1924

Corticium kauri G. Cunn. 1954

Puccinia keae G. Cunn. 1928

Hymenoscyphus kiko P.R. Johnst. 2013

Austrosmittium kiwiorum M.C. Williams & Lichtw. 1990

Puccinia koherika G. Cunn. 1930

Lecanora kohu Printzen & al. 2017

Uredo konini G. Cunn. 1928

Puccinia kopoti G. Cunn. 1923

Tetramelas kopuwaianus Elix & H. Mayrhofer 2018

CLASSIFICATION

Pucciniomycotina

Agaricomycotina

Pezizomycotina

Pucciniomycotina

Pezizomycotina

Pucciniomycotina

Agaricomycotina

Pezizomycotina

Agaricomycotina

Pucciniomycotina

Agaricomycotina

Microsporidia

Pezizomycotina

Pucciniomycotina

Agaricomycotina

Pezizomycotina [L]

Agaricomycotina

Pezizomycotina

Agaricomycotina

Pucciniomycotina

Agaricomycotina

Pucciniomycotina

Pezizomycotina

Kickxellomycotina

Pucciniomycotina

Pezizomycotina [L]

Pucciniomycotina

Pezizomycotina [L]

396 ... Pennycook

EPITHET

kowhai

kukutae

mahinapua

mahinapuense

mahuianum

maire

makarorae

mamaku

manawaorae

manawatua

mangatepopense

mania

manuka

manukanea

marama

maruiaensis

maungahukae

maungatautari

maungatuensis

maurea

mumura

murihikuana

muritai

muritaiensis

namua

nehuta

ngohengohe

nikau

ohakune

ohauensis

okatina

omahutaense

oratiensis

TAXON

Aecidium kowhai G. Cunn. 1924

Clonostachys kowhai Schroers 2001

Nectria kowhai Dingley 1956

Phaeosphaeria kukutae G.S. Ridl. 1988

Pirottaea mahinapua P.R. Johnst. 1998

Sarcostroma mahinapuense Gadgil & M.A. Dick 1999

Lophodermium mahuianum P.R. Johnst. 1989

Laricifomes maire G. Cunn. 1965

Psilocybe makarorae P.R. Johnst. & P.K. Buchanan 1995

Mycena mamaku Segedin 1991

Ascochyta manawaorae Verkley & al. 2010

Anthostomella manawatua Whitton & al. 2000

Lophodermium mangatepopense P.R. Johnst. 1989

Puccinia mania G. Cunn. 1930

Hypocrea manuka Dingley 1952

Nectria manuka Dingley 1951

Phyllachora manuka P.R. Johnst. & PF. Cannon 2004

Poria manuka G. Cunn. 1947

Astrosporina manukanea E. Horak 1978

Debaryomyces marama di Menna 1954

Lactarius maruiaensis McNabb 1971

Lecidea lapicida var. maungahukae Hertel 2001

Ellisembia maungatautari McKenzie 2010

Solicorynespora maungatautari McKenzie 2010

Buellia maungatuensis Elix & H. Mayrhofer 2017

Puccinia maurea G. Cunn. 1930

Amanita mumura G.S. Ridl. 1991

Umbilicaria murihikuana D.J. Galloway & L.G. Sancho 2005

Lepista muritai G. Stev. 1964

Hygrophorus muritaiensis G. Stev. 1963

Puccinia namua G. Cunn. 1924

Amanita nehuta G.S. Ridl. 1991

Rhodotorula ngohengohe Padamsee & al. 2017

Epithele nikau G. Cunn. 1956

Lachnella nikau G. Cunn. 1963

Peniophora nikau G. Cunn. 1963

Sebacina nikau McNabb 1969

Hymenoscyphus ohakune P.R. Johnst. 2013

Thaxterogaster ohauensis Soop 1998

Anthostomella okatina Whitton & al. 2000

Sporidesmium omahutaense Matsush. 1985

Mycena oratiensis Segedin 1991

CLASSIFICATION

Pucciniomycotina

Pezizomycotina

Agaricomycotina

Pezizomycotina

Pucciniomycotina

Pezizomycotina

Agaricomycotina

Saccharomycotina

Agaricomycotina

Pezizomycotina [L]

Pezizomycotina

Pezizomycotina [L]

Pucciniomycotina

Agaricomycotina

Pezizomycotina [L]

Agaricomycotina

Pucciniomycotina

Agaricomycotina

Pucciniomycotina

Agaricomycotina

Pezizomycotina

Agaricomycotina

Pezizomycotina

Agaricomycotina

EPITHET

orokonuiana

otagensis/-ense

otagoana

otagoensis/-ense

otakou

otira

pahiensis

papakaiensis

papakurae

papanui

paraoniti

paraonui

pareparina

patearoana

pekeoides

pirongia

poropingao

pounamu

poutoensis

puawhananga

pumatona

rakiurae

rangitatensis

rapua

rata

Cunningham & te reo Maori (New Zealand) ... 397

TAXON

Megalaria orokonuiana Fryday & A. Knight 2012

Acarospora otagensis H. Magn. 1943

Aecidium otagense Linds. 1867

Buellia otagensis Zahlbr. 1941

Corticium otagense G. Cunn. 1963

Lecidea otagensis Nyl. 1867

Marasmius otagensis G. Stev. 1964

Melanospora otagensis Linds. 1867

Nectria otagensis Linds. 1867

Parmelia otagensis Zahlbr. 1941

Pellicularia otagensis G. Cunn. 1953

Porina otagensis P.M. McCarthy 1999

Sphaeria otagensis Linds. 1867

Verrucaria otagensis Zahlbr. 1941

Lecidea otagoana Zahlbr. 1941

Pertusaria otagoana D.J. Galloway 1983

Coniocybe otagoensis Js. Murray 1960

Leucopaxillus otagoensis G. Stev. 1964

Ramalina otagoensis W. Martin & J. Child 1972

Poria otakou G. Cunn. 1947

Uromyces otakou G. Cunn. 1923

Aecidium otira G. Cunn. 1928

Lecidea pahiensis Zahlbr. 1941

Russula papakaiensis McNabb 1973

Circinotrichum papakurae S. Hughes & Piroz. 1971

Buellia papanui Elix & H. Mayrhofer 2017

Caloplaca papanui D.J. Galloway 2004

Cortinarius paraoniti Soop 2013

Cortinarius paraonui Soop 2005

Amanita pareparina G.S. Ridl. 1991

Buellia patearoana Elix & A. Knight 2017

Amanita pekeoides G.S. Ridl. 1991

Poria pirongia G. Cunn. 1947

Stephanospora poropingao T. Lebel & Castellano

Hispidula pounamu P.R. Johnst. 2003

Puccinia pounamu G. Cunn. 1923

Stephanospora pounamu T. Lebel & Castellano 2015

Chlorociboria poutoensis P.R. Johnst. 2005

Uredo puawhananga G.T.S. Baylis 1954

Amanita pumatona G.S. Ridl. 1991

Parmeliella rakiurae P.M. Jorg. & D.J. Galloway 2004

Amandinea rangitatensis Elix & H. Mayrhofer 2017

Microsporidium rapua J.B. Jones 1981

Poria rata G. Cunn. 1947

CLASSIFICATION

Pezizomycotina [L]

Pucciniomycotina

Pezizomycotina [L]

Agaricomycotina

Pezizomycotina [L]

Agaricomycotina

Pezizomycotina [L]

Pezizomycotina

Pezizomycotina [L]

Agaricomycotina

Pezizomycotina [L]

Pezizomycotina

Pezizomycotina [L]

Pezizomycotina

Agaricomycotina

Pezizomycotina [L]

Agaricomycotina

Pucciniomycotina

Pezizomycotina [L]

Agaricomycotina

Pezizomycotina

Pezizomycotina [L]

Agaricomycotina

Pezizomycotina [L]

Agaricomycotina

Pezizomycotina

Pucciniomycotina

Agaricomycotina

Pezizomycotina

Pucciniomycotina

Agaricomycotina

Pezizomycotina [L]

Microsporidia

Agaricomycotina

398 ... Pennycook

EPITHET

rautahi

rekohu

rimuphilus

rimutaka

rongomai-

pounamu

rotoruaensis

ruapehu

taiepa

takapauensis

takoropuku

tapawera

tararua

tararuana

tararuensis

taupoensis

tawa

tawai

tawhai

tekapo

tiritea

toa

toatoa

toetoe

tokerau

tongariro

toro

totara

totarae

totaranuiensis

tuapekensis

TAXON

Puccinia rautahi G. Cunn. 1930

Phyllachora hauturu subsp. rekohu P.R. Johnst. &

P.E Cannon 2004

Marasmius rimuphilus Desjardin & E. Horak 1997

Collybia rimutaka G. Stev. 1964

Candida rongomai-pounamu Padamsee & al. 2017

Chytriomyces rotoruaensis Karling 1970

Nectria ruapehu Dingley 1951

Amanita taiepa G.S. Ridl. 1991

Nosema takapauensis I.M. Hall & al. 1976

Elsinoe takoropuku G.S. Ridl. & Ramsfield 2006

Macowanites tapawera T. Lebel 2002

Puccinia tararua G. Cunn. 1923

Lecanora tararuana Zahlbr. 1941

Lecidea tararuensis Zahlbr. 1941

Sporobolomyces taupoensis Hamam. & Nakase 1995

Hypocrea tawa Dingley 1952

Nectria tawa Dingley 1951

Sebacina tawa McNabb 1969

Trametes tawa G. Cunn. 1948

Tremella tawa McNabb 1990

Trichoderma tawa P. Chaverri & Samuels 2003

Lactarius tawai McNabb 1971

Russula tawai McNabb 1973

Fuscoporia tawhai G. Cunn. 1948

Puccinia tekapo McNabb 1962

Colletotrichum ti B.S. Weir & P.R. Johnst. 2012

Puccinia tiritea G. Cunn. 1923

Puccinia toa G. Cunn. 1930

Tyromyces toatoa G. Cunn. 1965

Uredo toetoe G. Cunn. 1924

Hispidula tokerau P.R. Johnst. 2003

Cyphella tongariro G. Cunn. 1953

Hypocrea toro Dingley 1952

Cyphella totara G. Cunn. 1953

Peniophora totara G. Cunn. 1955

Poria totara G. Cunn. 1965

Auricula totarae Lloyd 1920

Hysterium pulicare subsp. totarae Sacc. 1908

Lecanactis totarae Zahlbr. 1941

Scutellinia totaranuiensis J. Moravec 1996

Buellia tuapekensis Elix & A. Knight 2017

CLASSIFICATION

Pucciniomycotina

Pezizomycotina

Agaricomycotina

Saccharomycotina

Chytridiomycotina

Pezizomycotina

Agaricomycotina

Microsporidia

Pezizomycotina

Agaricomycotina

Pucciniomycotina

Pezizomycotina [L]

Pucciniomycotina

Pezizomycotina

Agaricomycotina

Pezizomycotina

Agaricomycotina

Pucciniomycotina

Pezizomycotina

Pucciniomycotina

Agaricomycotina

Pucciniomycotina

Pezizomycotina

Agaricomycotina

Pezizomycotina

Agaricomycotina

Pezizomycotina

Pezizomycotina [L]

Pezizomycotina

Pezizomycotina [L]

EPITHET

tuku

tupare

umerensis

ura

urewera

waikaia

waikanaensis

waikaremoana

waiouru

waiporianus

waiporiensis

waipoua

waitakere

waitemataensis

wakatipu

weraroa

weraroensis

whakapapaensis

Cunningham & te reo Maori (New Zealand) ... 399

TAXON

Ramomarthamyces tuku PR. Johnst.

Uredo tupare G. Cunn. 1924

Lactarius umerensis McNabb 1971

Russula umerensis McNabb 1973

Mycena ura Segedin 1991

Zanclospora urewera J.A. Cooper 2005

Hymenoscyphus waikaia P.R. Johnst. 2013

Hygrophorus waikanaensis G. Stev. 1963

Phaeosphaeria waikanaensis G.S. Ridl. 1988

Entoloma waikaremoana E. Horak 2008

Cintractia waiouru G. Cunn. 1945

Cortinarius waiporianus Soop 2013

Leucopaxillus waiporiensis G. Stev. 1964

Parmelia waiporiensis Hillmann 1938

Uromyces waipoua McNabb 1966

Stachybotrys waitakere Whitton & al. 2001

Niesslia waitemataensis W. Gams & al. 2019

Anthracoidea wakatipu Vanky 2000

Psilocybe weraroa Borov. & al.

Poria weraroensis G. Cunn. 1947

Sphaerophorus whakapapaensis Wedin 1991

CLASSIFICATION

Pezizomycotina

Pucciniomycotina

Agaricomycotina

Pezizomycotina

Agaricomycotina

Pezizomycotina

Agaricomycotina

Ustilaginomycotina

Agaricomycotina

Pezizomycotina [L]

Pucciniomycotina

Pezizomycotina

Ustilaginomycotina

Agaricomycotina

Pezizomycotina [L]

whakatipae Lecidea whakatipae C. Knight 1876 Pezizomycotina [Lc]

whakatipu Puccinia whakatipu G. Cunn. 1924 Pucciniomycotina

wharanui Uredo wharanui G. Cunn. 1924 Pucciniomycotina

wheroparaonea Limacella wheroparaonea G.S. Ridl. 1993 Agaricomycotina

Discussion

Pre-Cunningham use of te reo Maori

The earliest te reo-derived fungal epithet was otagensis/otagense, used for

five specimens (two lichens, two unlichenised ascomycetes, and a spectacular

rust affecting puawhananga [Clematis paniculata] blossoms) collected by

Scottish lichenologist William Lauder Lindsay during an extended visit to

Otago Province, and later described by Lindsay (1867) and Nylander (1867).

A lichenicolous fungus was described by New Zealand-resident lichenologist

Charles Knight (1876) as Lecidea whakatipae; the epithet refers to Wakatipu,

one of the Otago Lakes, probably the vicinity of the unlisted type locality. The

only other 19" century te reo-derived epithet was a pathogen of a korokio

tree [Corokia buddleioides] growing in Germany and described by German

mycologist Paul Hennings (1899) as Septoria “corockeae” (correctly corokiae;

400 ... Pennycook

Turland & al. 2018: Art. F9.1). In the early 20" century, three more te reo-

derived epithets were published: Hysterium pulicare subsp. totarae on a totara

tree [Podocarpus totara] growing in Italy (Saccardo 1908); Puccinia hoheriae

on a specimen of hohere [Hoheria populnea] sent to Kew, England for fungal

identification (Wakefield 1917); and Auricula totarae on dead totara wood

sent to USA for fungal identification (Lloyd 1920).

Use of te reo Maori by Cunningham and his successors

Cunningham began proposing te reo Maori epithets in his first paper

on rust fungi in New Zealand (Cunningham 1923) and continued to do

so until his two posthumously published monographs (Cunningham

1963, 1965). His example has inspired numerous subsequent New Zealand

mycologists (and others working on New Zealand fungi) to use te reo Maori

and ta re Moriori [the indigenous language of Rékohu/Chatham Islands,

New Zealand] as the basis of new fungal names. A total of 231 such fungal

names: five genera (TABLE 1); and 226 species/subspecies/varieties (TABLE

2), have been located, and are listed either as published or with minor

orthographic corrections required to comply with the International Code

of Nomenclature for algae, fungi, and plants (ICN; Turland & al. 2020).

Although Veale & al. (2019: Appendix S1) published a “List of taxa recorded

with te reo / ta re epithets’, this included only one fungal genus and 140 fungal

species/subspecies (i.e., 90 names, including 14 of Cunningham's 56 species,

were omitted); their list contained several misspellings and miscitations, as

well as three entries that have no connection with te reo Maori (ackamae

refers to the host genus Ackama, which is an anagram; horakii honours

Austrian mycologist Egon Horak, who has been collecting and describing

New Zealand agarics for more than 50 years; and Waitea refers to the Waite

Research Institute, Adelaide, Australia).

Nomenclatural use of te reo Maori and other non-latinate languages

The majority of the published te reo Maori epithets have been treated as

undeclinable adjectives or nouns in apposition, preserving the authentic te

reo Maori orthography (apart from the problem of “long” vowels indicated

by macron diacritical marks; see next section, below). The presentation

of vernacular epithets as nouns in apposition has a tradition dating

from the birth of botanical nomenclature: Linnaeus (1753) published

numerous examples, e.g., Calamus rotang L., rattan; Piper betle L., betel

nut; Theobroma cacao L., cacao [cocoa]. Other te reo Maori epithets

have been modified in what appear to be misguided attempts to convert

Cunningham & te reo Maori (New Zealand) ... 401

them into “Latin” adjectives or genitive nouns. Like the majority of non-

European vernacular languages, te reo Maori does not conform to the

latinate word-model of a linguistic stem + a declinable inflection that

indicates grammatical case, gender, number, etc. Attempts to modify

vernacular words according to latinate grammatical conventions result in

the creation of chimaeras that convey no Latin meaning and mangle the

vernacular word. Every syllable in te reo Maori words consists of a vowel

(with or without a preceding consonant); removal of the final vowel from

a word does not create a “linguistic stem’, but rather it deletes either the

whole of the final syllable or leaves a final consonant deprived of its vowel,

distorting the pronunciation and meaning of the word and diminishing its

mana [status; prestige; spirituality].

Nomenclatural use of te reo Maori macron-vowels

In modern orthography, te reo Maori vowels are differentiated as short

[“a/e/i/o/u”] or long [either “aa/ee/ii/oo/uu’, or “a/é/i/6/a"]. ICN Art.

60.7 (Turland & al. 2020) requires the suppression of all diacritical letters;

therefore the macron-vowel orthography is not permitted, but the double-

vowel orthography would be acceptable as “necessary transcriptions” of the

macron-vowels. Veale & al. (2019) reached the same conclusion, in spite of

having misinterpreted and mistranscribed Art. 60.7. It is diacritical signs

(including macrons) that are forbidden, because the diacritical letters are

additions to the basic Latin alphabet; the diaeresis, two dots placed over

the second of two adjacent vowels, is permitted as an optional phonetic

device to indicate the separate pronunciation of the two vowels. (Confusion

often arises because the typographies of the diacritical umlaut and the non-

diacritical diaeresis are identical.)

Place name epithets in te reo Maori and other non-latinate languages

There is no reason why epithets derived from vernacular place names

cannot also be presented as nouns in apposition; as discussed above, they

should not be presented as “Latin” genitive nouns. However, place names in

all languages are routinely formed into adjectival epithets by adding one of

the geographical Latin suffixes recommended by ICN Rec. 60D.1 (Turland

& al. 2020). For vernacular place names, this is an acceptable alternative

to the preferred noun in apposition epithets, provided the place names are

not abridged to create spurious “linguistic stems”; for te reo Maori place

names, this means that the final vowel should always be retained before the

geographical sufhx is added, e.g., awakinoana from Awakino; huiaensis from

Huia; and weraroaensis from Weraroa.

402 ... Pennycook

Acknowledgments

Thanks to my Manaaki Whenua - Landcare Research mycological colleagues

Peter Buchanan, Jerry Cooper, Peter Johnston, Eric McKenzie, Mahajabeen

Padamsee, and Bevan Weir (all authors of te reo Maori epithets) for their help in

searching for and supplying overlooked names; and to Geoff Ridley (an ardent

advocate for, and author of, te reo Maori epithets) and Holden Hohaia (both of

Manaaki Whenua — Landcare Research, Wellington) for acting as referees.

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MYCOTAXON

April-June 2020—Volume 135, pp. 405-414

https://doi.org/10.5248/135.405

Re-collection of secotioid Entoloma calongei in Europe

IVONA KAUTMANOVA’ , VACLAV KAUTMAN?,

VIKTOR KUCERA}, DARINA ARENDT?

*Slovak National Museum-Natural History Museum,

Vajanskeho nab. 2, PO.Box 13, 81006 Bratislava, Slovakia

*Mierovd 16, 82105, Bratislava, Slovakia

Plant Science and Biodiversity Centre, Slovak Academy of Sciences,

Dubravska cesta 9, 84523 Bratislava, Slovakia

* CORRESPONDENCE TO: ivona.kautmanova@snm.sk

AxBsTRACT—In 2017, the secotioid Entoloma calongei was recorded in Slovakia, the first

report since its Spanish holotype collection. Morphological and ecological characteristics

are presented and compared with the type collection. LSU, mSSU, and ITS sequences

were obtained and a combined maximum likelihood tree of SSU and LSU sequences was

constructed.

Key worps—Entolomataceae, taxonomy, Rhodogaster, sequestrate fungi, genetic distance

Introduction

In Entoloma sporocarp morphology is highly variable: ranging in size from

tiny to large and in form as pleurotoid, omphalinoid, collybioid, mycenoid,

tricholomatoid, and sequestrate. In the past, three smaller non-agaricoid

genera have been distinguished based on habit: Rhodocybella TJ. Baroni &

R.H. Petersen (cyphelloid), Rhodogaster E. Horak (secotioid), and Richoniella

Costantin & L.M. Dufour (gasteroid).

The first record of a secotioid entolomatoid species was published by

Horak (1963) from Chile, introducing a new monotypic genus for the species

Rhodogaster chilensis E. Horak. In 1995, a similar collection found in Spain

was initially misidentified as R. chilensis (Calonge & Pasaban 1995) but later

recognized as a new species, Rhodogaster calongei (Horak & Moreno 1998).

406 ... Kautmanova & al.

Molecular phylogenetic studies of Co-David & al. (2009) placed the sequestrate

genera Richoniella and Rhodogaster within Entoloma; and they transferred the

two Rhodogaster species to Entoloma.

Entoloma calongei was recorded during field research in Lubovnianska

Upland in NE Slovakia in 2017, and its identification has been confirmed by

DNA sequence analyses. The Slovak collection represents only the second

published record of this species.

Material & methods

SPECIMENS: Morphological and anatomical studies were conducted on fresh and

dried basidiomata. Colours were determined according to Kornerup & Wanscher

(1974). Microscopic features were studied in water or 3% KOH. All light micrographs

of microscopic structures were made with a Nikon Eclipse Ni light microscope using

bright field or differential interference contrast without staining, and images were

captured with a Nikon DS-Ri2 digital camera. Images were processed and descriptive

statistical analysis of basidiospore dimensions was produced using NIS-Elements BR

4.40 software. The voucher specimen was deposited in the herbarium of the Slovak

National Museum-Natural History Museum, Bratislava, Slovakia (BRA).

DNA EXTRACTION, AMPLIFICATION AND SEQUENCING: DNA from the Slovakian

E. calongei sporocarp was obtained from fresh material preserved in acetyl

trimethylammonium bromide (CTAB) buffer and ground by using a plastic

pestle and isolated by JENA Bioscience Blood-Animal-Plant DNA Preparation

Kit following manufacture protocol. Target regions of the partial mitochondrial

small subunit (mtSSU) rDNA and the nuclear large ribosomal subunit (LSU) were

amplified using primers MS1/MS2 (White & al. 1990) and NL1/NL4 (Vilgalys &

Hester 1990). The reaction was performed in 25ul of reaction mixture containing

10 pmol of both primers, Crystal Hot Start Master, and 20 ng of template DNA.

The cycling programme for mtSSU consisted of an initial denaturation at 94°C for

3 min, followed by 40 cycles of 94°C 30 s, 55°C for 30 s, 72°C for 1 min, and a final

extension at 72°C for 5 min and for LSU of an initial denaturation at 95°C for 3 min,

followed by 32 cycles of 95°C 30 s, 55°C for 30 s, 72°C for 1 min + increasing time

2 sec per cycle, and a final extension at 72°C for 10 min. The PCR products were

purified using Qiagen columns (QIJAquick PCR Purification Kit, Qiagen) according

to the manufacturer’s recommendations and sequenced commercially by Eurofins

Genomics GmbH (Cologne, Germany).

ALIGNMENT AND PHYLOGENETIC ANALYSIS: DNA sequences of Entolomataceae anda

selected Clitocybe outgroup were downloaded from NCBi on 18 January 2019. These

included LSU and mtSSU sequences associated with Baroni & Matheny (2011). In

total 15 LSU, and 15 mtSSU sequences were retrieved, including E. calongei type

(TABLE 1). To these, we added newly generated LSU and mtSSU sequences from

Entoloma calongei (BRACR30482). The tree was reconstructed by the maximum-

Entoloma calongei new to Slovakia ... 407

TABLE 1. Entoloma and Clitocybe sequences used for phylogenetic analysis

(newly obtained sequences in bold).

SPECIES COUNTRY LSU SSU ITS

C. dealbata Unknown AF223175 DQ825431 —

E. albidoquadratum [holotype] India GQ289151 GQ289291 —

E. calongei [holotype] Spain GQ289158 GQ289298 —

E. calongei BRACR 30482 Slovakia MK531556 MK530244 MK907407

E. cocles Finland GQ289159 GQ289299 —

E. costatum Netherlands GQ289161 GQ289301 —

E. excentricum Germany GQ289163 GQ289303 —

E. gasteromycetoides New Zealand GQ289164 GQ289304 —

E. phaeomarginatum Tasmania GQ289179 GQ289319 —

E. porphyrescens Tasmania GQ289182 GQ289322 _

E. procerum Tasmania GQ289183 GQ289323 —

E. serricellum Belgium GQ289190 GQ289330 —_

E. serrulatum Tasmania GQ289192 GQ289332 —

E. tectonicola [holotype] India GQ289196 GQ289336 =

E. transmutans Tasmania GQ289200 GQ289340 —

likelihood method in MEGA-X with 1000 bootstrap replicates. The General Time

Reversible model (Nei & Kumar 2000) with gamma-distributed evolutionary rates

(G) and invariable sites (I) was chosen by MEGA as the best-fitting model. A matrix

of inter-species distances was constructed using Kimura-2 parameter distances

(Kimura 1980) in MEGA-X. We also produced an ITS barcode for Entoloma calongei

(BRACR30482), now stored as SKDNA023-19 in BOLD v4 database (Barcode of Life

Datasystems http://v4.boldsystems.org/).

Taxonomy

Entoloma calongei (E. Horak & G. Moreno) Noordel. & Co-David,

Persoonia 23: 166 (2009) Fics 1-3

= Rhodogaster calongei E. Horak & G. Moreno, Sydowia 50: 188 (1998)

PILEUS <25 mm diam., globose to subglobose (pyriform), margin involute

and mostly attached to stipe, not expanding, greyish brown (8B1) with dark

spots, silky and fibrillose, dry. Stipe short, with columella, <20 mm long,

hollow, attenuated at base, silver grey (8B1), smooth, silky, fibrillose, dry.

GLEBA irregularly labyrinthiform of reduced lamellae, non-gelatinized,

lamellae greyish-white in section and pale pink (8B3) on surface. SMELL

AND TASTE not distinctive. SPORE PRINT pink. BAsIpIospoREs cuboid or

rhomboid (“pseudocuboid” sensu Karstedt & al. 2019), rarely pentagonal,

(5.6-)6.3(-7.1) x (5.4-)6.8(-9.0) um (Q = 0.80-1.01), often distorted,

smooth, thin walled with distinct apiculus. Basidia 30 x 10 um cylindrical to

subclavate, 4 spored, clampless. Tramal hyphae short, cylindrical, clampless,

408 ... Kautmanova & al.

Fic. 1. Entoloma calongei (BRACR30482): a. In situ, 20.X.2017. (Photo: F. Fuljer); b. Sporocarps

in cross section (Photo: A. Polhorsky). Scale bar = 10 mm.

10-30 x 2-10 um. PILEIPELLIS a cutis of cylindrical, narrow (80-100 x 5-6

um), parallel clampless hyphae, subcutis elements wider and shorter (40-80

x 15-20 um).

SPECIMEN EXAMINED: SLOVAKIA, PrESov REGION, Lubovnianska Upland, Obrucné

village, 49.3080°N 20.9877°E, alt. 650 m, in grass in pasture, close to mixed forest, 20. X.

2017, leg. V. Kucera, det. V. Kautman (BRACR30482).

Entoloma calongei new to Slovakia ... 409

Fic. 2. Entoloma calongei (BRACR30482): a. Detail of gleba in cross section. (Photo: A. Polhorsky);

b. Spores (Photo: I. Kautmanova). Scale bar: a= 1 mm; b = 10 um.

410 ... Kautmanova & al.

HABITAT—Seminatural meadow mowed and/or grazed, southern slope; soil

flysch sandy-silty, slightly basic but not calcareous. The specimens grew in

grass, herbs (Plantago, Trifolium, Taraxacum), and mosses, ca 50 m from the

edge of mixed forest (Picea abies, Carpinus betulus, Fagus sylvatica, Corylus

avellana, Cornus sanguinea, Betula, Acer). The species was found in a rich

grassland-fungi community together with various earth-tongues, waxcaps,

and clavarioid fungi.

Discussion

Vidal & al. (2016) reported a total of 11 sequestrate Entoloma species,

including four from Europe/Spain. Nine species were gasteroid and only

two secotioid: E. calongei and E. chilense (E. Horak) Noordel. & Co-David.

These two species differ from each other in a number of basic characters,

including basidiospore morphology and pileipellis structure.

Basidiospores of E. chilense were described as “ellipsoidnodular”

(9.5-11.5 x 7.5 um; Horak 1963), and those of E. calongei as cuboid (side

lengths 6-10 um; Horak & Moreno 1998). Basidiospores of our collection

were cuboid or rhomboid, often broader than long and side length is

6-8 um.

Although Horak (1963) described clamp connections as absent on

basidia and tramal and cortical hyphae in the type description of E. chilense,

Horak & Moreno (1998) later reported regularly observing clamp

connections in E. chilense, contrasting with the clampless E. calongei. No

clamp connections were observed in our specimens.

The pileipellis in the Slovak collections is a cutis composed of cylindrical

parallel hyphae, the same as cited for E. calongei in the protologue and

differing from the trichoderm of E. chilense (Horak 1963).

Ecology of the type localities of the two taxa was distinctly different.

Entoloma chilense occurs among deep litter and humus in the coastal

anectotrophic rain forest of Central Chile; the vegetation around the

type locality was dominated by trees in Lauraceae and Myrtaceae, with

Chusquea sp. (Bambuseae) in the understory (Horak 1963). By comparison,

E. calongei was found in northern Spain in a colline-temperate mixed

forest of conifers (Larix, Pinus) and broadleaf trees (Fagus, Acer, Quercus,

Fraxinus, Corylus) at about 700 m a.s.l. (Calonge & Pasaban 1995). Our

Slovakian collection site differed in that the basidiomata grew among

grass and mosses in a pasture; however, the site was only 50 m from a

forest composed of conifers and broadleaf trees similar to those in Spain.

Entoloma calongei new to Slovakia ... 411

100 - Entoloma calongei BRACR 30482

Entoloma calongei

Entoloma costatum

Entoloma albidoquadratum

Entoloma procerum

Entoloma cocles

Entoloma porphyrescens

Entoloma tectonicola

Entoloma excentricum

Entoloma phaeomarginatum

Entoloma serrulatum

Entoloma transmutans

Entoloma sericellum

Entoloma gasteromycetoides

Clitocybe dealbata

-—4

0.02

Fic. 3. Maximum likelihood phylogenetic tree constructed from the combined dataset

(LSU+mtSSU). Red clade represents well-delimited species Entoloma calongei. Clitocybe

dealbata was used as outgroup. Numbers above branches represent bootstrap support (only

values >50% are shown).

It should be added that many Entoloma species occur both in semi-open

forests and in grasslands without trees (Noordeloos 1992, 2004).

The sequence of E. calongei from Slovakia is almost identical with

the type and the identity of Slovak collection has also been confirmed

by using Kimura-2 parameter distances model. Our phylogeny (Fic. 3),

like that of Co-David & al. 2009, places E. calongei close to E. costatum

(Fr.) P. Kumm. with c. 3% dissimilarity (TABLE 2). Entoloma costatum

is an extremely rare, rather fleshy, clitocybeoid species with rhomboid-

412 ... Kautmanova & al.

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cuboid spores (Noordeloos 2012). However, according to Karstedt & al.

(2019), E. calongei is closer to Entoloma asterosporum (Coker & Crouch)

TJ. Baroni & Matheny [= Richoniella asterospora]. These three species

form one of many independent clades of Entoloma species possessing

cuboid spores (Karstedt & al. 2019). However, the three differ from the

others in having a special “pseudocuboid” spore-shape (Karstedt & al.

2019). Co-David & al. (2009) and Karstedt & al. (2019) place these species

basal to the Inocephalus—Cyanula superclade, with weak support.

Acknowledgments

The authors express appreciation to Olga Morozova (Komarov Botanical Institute,

St. Petersburg) and Tor Erik Brandrud (Norwegian Institute for Nature Research,

Trondheim) for their critical revision, comments, and pre-submission manuscript

review. We would like to thank Adam Polhorsky, Filip Fuljer and Jan Cervenka

for cooperation in field and for providing pictures and Lance Arendt for English

corrections of the manuscript. The research of I. Kautmanova and D. Arendt was

funded by the Operational Program of Research and Development and co-financed

with the European Fund for Regional Development (EFRD): ITMS 26230120004:

Building of research and development infrastructure for investigation of genetic

biodiversity of organisms and joining IBOL initiative. This study was also supported

by grant VEGA 2/0061/19 to V. Kuéera.

Literature cited

Baroni TJ, Matheny PB. 2011. A re-evaluation of gasteroid and cyphelloid species of

Entolomataceae from Eastern North America. Harvard Papers in Botany 16: 293-310.

https://doi.org/10.3100/0.25.016.0205

Calonge FD, Pasaban PM. 1995. Rhodogaster chilensis E. Horak (Gasteromycetes), encontrado

en Espafla y nuevo para Europa. Boletin de la Sociedad Micoldgica de Madrid 20: 277-281.

Co-David D, Langeveld D, Noordeloos ME. 2009. Molecular phylogeny and spore evolution of

Entolomataceae. Persoonia 23: 147-176. https://doi.org/10.3767/003158509X480944

Horak E. 1963. Fungi austroamericani. III. Rhodogaster gen. nov. - a new link from Chile

towards the Rhodophyllaceae. Sydowia 17(1-6): 190-192.

Horak E, Moreno G. 1998. Rhodogaster calongei sp. nov. (Basidiomycota), first European record

of the secotiaceous entolomatoid genus from northern Spain. Sydowia 50(2): 187-191.

Karstedt F, Capelari M, Baroni TJ, Largent DL, Bergemann SE. 2019. Phylogenetic and

morphological analyses of species of the Entolomataceae (Agaricales, Basidiomycota) with

cuboid basidiospores. Phytotaxa 391 (1): 1-27. https://doi.org/10.11646/phytotaxa.391.1.1

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

through comparative studies of nucleotide sequences. Journal of Molecular Evolution 16(2):

111-120. https://doi.org/10.1007/BF01731581

Kornerup A, Wanscher JH. 1978. Methuen handbook of colour. 3rd ed., Eyre Ltd., London.

Kumar S, Stecher G, Li M, Knyaz C, Tamura K. 2018. MEGA X: Molecular Evolutionary Genetics

Analysis across computing platforms. Molecular Biology and Evolution 35(6): 1547-1549.

https://doi.org/10.1093/molbev/msy096

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Nei M, Kumar S. 2000. Molecular evolution and phylogenetics. Oxford University Press,

New York.

Noordeloos ME. 1992. Entoloma s.1. Fungi Europaei, vol. 5. Giovanna Biella, Saronno, Italy.

Noordeloos ME. 2004. Entoloma s.]. Fungi Europaei, vol. 5a. Edizione Candusso, Italy.

Noordeloos ME. 2012. Entoloma. 517-576, in: H Knudsen, JH Vesterholt (eds). Funga

Nordica: agaricoid, boletoid, clavarioid, cyphelloid and gastroid genera, 2nd ed.

Copenhagen.

Vidal JM, Bellanger JM, Moreau PA. 2016. Tres nuevas especies gasteroides del género

Entoloma halladas en Espana. Boletin Micol6gico de FAMCAL 11: 53-78.

Vilgalys R, Hester M. 1990. Rapid genetic identification and mapping of enzymatically

amplified ribosomal DNA from several Cryptococcus species. Journal of Bacteriology

172(8): 4238-4246. https://doi.org/10.1128/jb.172.8.4238-4246.1990

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

MYCOTAXON

April-June 2020—Volume 135, pp. 415-423

https://doi.org/10.5248/135.415

Xerula setulosa,

a new Neotropical agaric record for Argentina

Maria M. ALBERTI’, NICOLAS NIVEIRO?, EDGARDO O. ALBERTO?

"Instituto Tecnologico Chascomus INTECH (UNSAM-CONICET),

Intendente Marino Km 8.2, CP 7130, Chascomus, Buenos Aires, Argentina

? Instituto de Botanica del Nordeste (UNNE-CONICET),

Sargento Cabral 2131, CC 209, CP 3400, Corrientes Capital, Argentina

" CORRESPONDENCE TO: ealberto@intech.gov.ar

ABSTRACT—A collection of Xerula setulosa is recorded for the first time from Argentina. This

species was previously reported from Central America and Brazil. A complete description of

the specimens, photographs in the natural habitat, and illustrations of diagnostic microscopic

characters are presented. Comments regarding the distribution and related species are

provided.

Key worps—Agaricales, fungi, mushroom, Physalacriaceae, taxonomy

Introduction

Maire (1933) proposed Xerula to accommodate Agaricus longipes Bull.

[= Xerula pudens (Pers.) Singer]. The genus has undergone many successive

taxonomic reclassifications including genus, subgenus, and section, but always

associated in some way with the genera Oudemansiella Speg. and Mucidula

Pat. (Moser 1955; Singer 1962a,b, 1964, 1986; Cleémen¢gon 1979; Dorfelt 1979;

Pegler & Young 1987; Halling & Mueller 1999; Petersen 2008; Yang & al. 2009).

Morphological and molecular phylogenetic analyses now support Xerula as

an independent genus within Physalacriaceae (Binder & al. 2006, Petersen &

Hughes 2010).

Xerula is characterized by its collybioid basidiomata with dark colors

(gray, brown to black); white distant and broad lamellae; rooting stipe, with

416... Alberti, Niveiro, Alberté

velutinous, strigose or hispid surface and presence of caulosetae; white spore-

print; and hyaline, broadly ellipsoid to globose spores (Wang & al. 2008).

Xerula is related with Paraxerula R.H. Petersen, Protoxerula R.H. Petersen,

and Hymenopellis R.H. Petersen, differing from these in the thick-walled, stiff,

brittle, blond to rufous brown pileocystidia (Petersen & Hughes 2010).

There are no previous Argentinean records of species currently accepted in

Xerula. Xerula radicata (Relhan) Déorfelt, which was recorded from Argentina

(Niveiro & Alberté 2012; Raithelhuber 1974, 1987, 1991, 1995; Singer 1964;

Singer & Digilio 1952), is currently accepted as Hymenopellis radicata (Relhan)

R.H. Petersen (Petersen & Hughes 2010). The aim of this work is to describe

the first Argentinean Xerula collection.

Materials & methods

The material, which was collected in the province of Salta, Argentina, was

photographed and described macroscopically in situ. The specimens were identified

through morphological examination following Largent (1986), Vellinga (1988), and

Lodge & al. (2004). Color terminology is according to Kornerup & Wanscher (1978).

Specimens were sectioned by freehand and mounted in 5% KOH (v/w) with 1%

phloxine aqueous solution for microscopical examination. Melzer’s reagent (Wright

& Alberté 2002) was used to verify amyloid reaction. Microscopic structures were

measured directly with 1000x immersion objective or through photographs taken

with a Leica EC3 built-in camera using Image Pro Plus” 4.0 software. The minimum-

maximum intervals were provided for the different microscopic structures. For

basidiospores, n = number of spores measured, x = mean values, Q = length/diameter

coefficient, and Qx = mean value of Q. Authors of scientific names follow Index

Fungorum (2019), and herbarium acronyms follow Thiers (2019). The collection

was dried, kept in the freezer for a week, and deposited as reference in the CTES

herbarium.

Taxonomy

Xerula setulosa (Murrill) R.H. Petersen & T.J. Baroni,

Mycotaxon 101: 114. 2007. FIGs 1, 2

= Gymnopus setulosus Murrill, N. Amer. Fl. 9: 373. 1916.

= Lentinus pilosus Rick, Lilloa 2: 310. 1938, nom. illegit. [non (Fr.) Fr. 1838].

= Xerula pilosa Singer, Lilloa 26: 86. 1954.

= Oudemansiella pilosa (Singer) Singer, Sydowia 15: 59. 1962.

BasipIoMATA collybioid, solitary, radicating, growing on humus. PILEus

<35 mm diam., convex to depressed plane, with a truncate to mammilate

umbo, surface dry, roughened, sometimes delicately arched, on the disc

light brown (5D5), pale yellow (4A3) and greyish yellow (4C4), or yellowish

Xerula setulosa new to Argentina ... 417

Fic. 1. Xerula setulosa (CTES - Niveiro & al. 2198) A. General aspect of basidiomata; B. Detail

of pileus; C. Detail of lamellae; D. Detail of stipe pubescence. Scale bars = 20 mm. Photographs

by N. Niveiro.

brown (5D6), towards the margin olive brown (5E5) to dark brown (6F5),

strongly pubescent with brown hairs, margin somewhat incurved, undulated,

not striate. CONTEXT fleshy but very thin, <1 mm, white, unchanging when

cut. Smell and taste not checked. LAMELLAE distant to subdistant, white

(1A1), adnexed to adnate, ventricose, margin entire, lamellulae two-tiered.

STIPE central, 150 x 4-8 mm, compressed, attenuated base and apex, hollow,

grayish orange (6B6) to brownish orange (7C8), strongly pubescent, hairs <1

mm, greyish-yellow (4B4) to golden (4C6); leathery pseudorhiza attenuating

quickly, stipe concolorous with the stipe, pubescent. ANNULUs and other velar

structures absent. SPORE-PRINT white (1A1).

BASIDIOSPORES 10-13.8 x 10-12.5 um; x = 12.2 x 11 um; Q = 0.97-1.3;

Qx =1.1; n = 25; globose to subglobose, thin-walled, smooth, chestnut,

lacking evident germ pore, with a prominent apiculus. Basip1a 30-60

418 ... Alberti, Niveiro, Alberté

Fic. 2. Xerula setulosa (CTES — Niveiro & al. 2198): A. Basidia; B. Spores; C. Cheilocystidia;

D. Pleurocystidia; E. Pileipellis elements; F. Pileipellis pileosetae.

Scale bars: A~D = 10 um, F = 40 um.

x 10-17 um, claviform to ventricose, 4-spored, thin-walled, hyaline.

PLEUROCYSTIDIA60-130 x 12-35 um, broadly lageniform, with a truncate

apex, thin to thick-walled mainly at the base. CHEILocysTIpIA 50-80 x

10-24 um, fusiform to utriform, thin-walled to thickened at the base, hyaline.

HYMENOPHORAL TRAMA bilateral with a thin half-layer and divergent

gelatinous hyphae. PILEIPELLIS hymeniform, composed of two types of

elements: 1) Pileocystidia fusiform, 45-60 x 15-18 um, thick-walled,

pigmented with olive content; 2) Pileosetae <700 um long, 10-17 um diam.,

dark brown with sharp ends and thickened (<4 um) walls. Stipe surface

composed of two elements, similar to the pileocystidia and pileosetae. CLAMP

CONNECTIONS absent.

SPECIMEN EXAMINED—ARGENTINA. SALTA PROVINCE. Santa Victoria Dept., Baritu

National Park, road to Bariti town, 22°26’41”S 64°44’37”W, 1421 m as.L, 22.11.2011,

leg. Niveiro & al. 2198 (CTES).

Xerula setulosa new to Argentina ... 419

Discussion

Xerula setulosa is characterized by its strongly pubescent basidioma,

yellowish brown to dark brown pileus surface, orange brown stipe, pure

white lamellae, globose smooth spores, and numerous cystidia (Wang &

al. 2008). It was originally described from Brazil by Rick (1938) under

the illegitimate name Lentinus pilosus [= Xerula pilosa (Singer 1954);

= Oudemansiella pilosa (Singer 1962b)]; however, Petersen & Baroni (2007)

synonymized these names under Xerula setulosa.

The only Xerula species previously reported from Argentina (as

X. radicata) is currently named as Hymenopellis radicata, which differs

morphologically from X. setulosa by the absence of pubescence (Putzke &

Pereira 1988, Raithelhuber 1995).

Six species are currently accepted in Xerula: X. hispida Halling &

G.M. Muell., X. melanotricha Dorfelt, X. pudens, X. setulosa, X. sinopudens

R.H. Petersen & Nagas., and X. strigosa Zhu L. Yang & al. They can be

distinguished by basidiome color, basidiospore morphology, setae, and

other microscopic characters (Wang & al. 2008, Petersen & Hughes 2010).

Distribution is also significant in that species are generally limited to certain

geographical areas; so far, species reported from Europe and Asia are not

found in the Americas.

Xerula setulosa seems closely related to X. hispida because of its strongly

pubescent basidioma and the presence of a pseudorhiza (Petersen & Baroni

2007, Rodriguez-Alcantar & Herrera-Fonseca 2016). However, X. hispida is

distinguished by microscopic characters, such as bisporic basidia and the

presence of clamp connections. Petersen & Baroni (2007) hypothesized that

X. hispida might represent an asexual state of X. setulosa, but unpublished

molecular studies by Wang & al. (2008) placed the two species in different

clades. On the other hand, Halling & Mueller (1999) reported absence of

clamp connections in X. hispida. Both species have a similar distribution in

Mexico, Central America, and South America.

Xerula melanotricha is clearly separated by its distribution (Europe,

western Asia and northern Africa) and the presence of its very long, almost

black setae <3 mm that are usually visible to the naked eye. However, in

some younger X. melanotricha specimens possess hairs that barely exceed

1 mm; these can be difficult to measure, especially hairs on the cap of dried

herbarium material which are often broken (Ronikier 2003).

Xerula pudens (the current name of the generic type Agaricus longipes

Bull.) is widely distributed throughout Europe and in southwestern Russia;

420 ... Alberti, Niveiro, Alberté

0 500 1000km

Fic. 3. Distribution of Xerula setulosa in America. Map adapted by N. Niveiro.

it is distinguished from X. setulosa by its longer basidioma with a small

pileus in relation to the stipe.

Xerula sinopudens, distributed in tropical to temperate Asia, is characterized

by its carrot-shaped pseudorhiza and interspersed long (0.55 mm) and

short pileosetae (Petersen & Hughes 2010).

Xerula strigosa of southwest China and Southeast Asia, which is

macroscopically remarkably similar to X. setulosa, can be distinguished with

difficulty microscopically by its longer (11-5 x 9-11.5 um) basidiospores,

occasional clamp connections on the septa of basidia and lamellar hyphae,

and thin-walled pleurocystidial apices (Wang & al. 2008).

Xerula setulosa new to Argentina ... 421

As regards pileus size, our material differs from previous descriptions.

The pileus is larger (35 mm) than those reported by Putzke & Pereira (1988;

<10 mm) and Petersen & Baroni (2007; <20 mm), although it is close to

or matches material from Parana, Brazil (23-40 mm; Meijer 2008) and

Jalisco, Mexico (20-30 mm; Rodriguez-Alcantar & Herrera-Fonseca 2016).

Our Argentinean basidiospore size is similar to those in Meijer (2008) and

Petersen and Baroni (2007), larger than in Singer (1954, 1964; 10 x 6.9 um)

and smaller than in Dorfelt (1985; 10-16 x 9.5-14 um) and Wang (2008;

11-16 x 9.5-12 um). Taking into account pileus colour and size, the spores,

the pleurocystidia, and the caulocystidia size, the Argentinean specimen

described here matches the descriptions by Meijer (2008) and Rodriguez-

Alcantar & Herrera-Fonseca (2016).

Xerula setulosa has a Neotropical distribution (Fic. 3) and has been

reported from Mexico, Belize, Jamaica, Puerto Rico (Petersen & Baroni

2007), Brazil (Putzke & Pereira 1988, Meijer 2002, 2006, 2008), and Costa

Rica (Wang & al 2008). The present record represents the first report from

Argentina.

Acknowledgments

The authors thank V.G. Cortez (Universidade Federal do Parana, Brazil) and

G. Moreno Horcajada (Universidad de Alcala, Madrid) for the critical revision of the

manuscript. This research was supported by the National Scientific and Technical

Research Council (CONICET) from Argentina (PIP 714).

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96: 235-240. https://doi-org/10.1002/fedr.49 109603 13

Halling RE, Mueller GM. 1999. A new species and a new record for the genus Xerula

(Agaricales) from Costa Rica. Mycotaxon 71: 105-110.

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Meijer AAR de. 2002. Mycological work in the Brazilian state of Parana. Nova Hedwigia 72:

105-159.

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Boletim do Museu Botanico Municipal, Curitiba 68: 1-55.

Meijer AAR de. 2008. Notable macrofungi from Brazil's Parana Pine forest. Embrapa Florestas,

Colombo. 431 p.

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Niveiro N, Alberté E. 2012. Checklist of the Argentine Agaricales 4. Tricholomataceae and

Polyporaceae. http://www.mycotaxon.com/resources/checklists/Niveiro-v121-checklist.pdf

[Abstract: Mycotaxon 121: 499.]

Pegler DN, Young TWK. 1987. Classification of Oudemansiella (Basidiomycota:

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Petersen RH. 2008. The genus Xerula (Agaricales; Fungi) in Australia and New Zealand. Nova

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MYCOTAXON

April-June 2020—Volume 135, pp. 425-429

https://doi.org/10.5248/135.425

Haematomma rubidum sp. nov. from China

RONG TANG *” 4, AN-CHENG YIN*»3, ZUN-TIAN ZHAO 7?

' School of Life Science, Yunnan University, Kunming, 650091, China

? Key Laboratory of Plant Stress Research, College of Life Sciences, Shandong Normal University,

Jinan, 250014, China

° Key Laboratory for Plant Diversity and Biogeography of East Asia, Kunming Institute of Botany,

CAS, Kunming, 650201, Yunnan, China

‘ The First High School of Liangshan, Jining, 272600, China

“CORRESPONDENCE TO: ztzhao@sohu.com

ABSTRACT—Haematomma rubidum from southern China is described as new species

characterized by convex apothecia with thin whitish margins that become hidden in age;

submuriform ascospores with 15-21 transverse and 0-3 longitudinal septa; and a thallus

containing atranorin and russulone but lacking dibenzofurans or xanthones. Photographs of

the new species accompany a detailed taxonomic description.

Keyworps—East Asia, Haematommataceae, lichenized fungi, taxonomy

Introduction

Haematomma A. Massal. is a cosmopolitan genus characterized by blood-

red lecanorine apothecia and transversely septate, submuriform to muriform

ascospores. Three species that produce submuriform spores are H. parda

Aptroot, H. staigeriae Nelsen & al., and H. wattii (Stirt.) Zahlbr. (Staiger &

Kalb 1995, Nelsen & al. 2006, Aptroot 2007), while only one, H. gallowayi

Brodo, is characterized by muriform spores (Brodo 2007).

During our studies, we discovered a new Haematomma species in Yunnan

Province (21°08’—29°15’N, 97°31’-106°12’E) in southwestern China. The

natural conditions and dry and wet climate in the province provide excellent

*RONG TANG & AN-CHENG YIN contributed equally to this research.

426 ... Tang, Yin, Zhao

habitat for many rare species and Yunnan is the most species-rich region for

lichens in China (Wang 2012).

Here we describe a new submuriform-spored species of Haematomma

and present a brief diagnosis, together with discussion on the differences

between (sub)muriform-spored species of Haematomma.

Materials & methods

The voucher specimens were examined morphologically using an Olympus SZ51

stereomicroscope and Olympus CX21 polarizing microscope and photographed

with Olympus SZX16 and BX61with DP72 cameras. Both thallus and medulla were

tested with K (a 10% aqueous solution of potassium hydroxide) and C (a saturated

solution of aqueous sodium hypochlorite). Lichen substances were identified using

standardized thin layer chromatography techniques (TLC) with solvent systems A

and C (Orange & al. 2010). The collection is deposited in the lichen section of the

Herbarium, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming,

Yunnan, China (KUN).

Taxonomic description

Haematomma rubidum R. Tang & Z.T. Zhao, sp. nov. Fic. 1

MB 835897

Differs from Haematomma staigeriae by its convex apothecia, its lack of soralia, and

the absence of lichexanthone.

Type: China. Yunnan Province, Lijiang City, Mt. Maanshan, Jiushijiulongtan Lake,

26°39’N, 99°46’E, alt. 3400 m, on Rhododendron, 16 Aug. 2000, LS Wang 00-20099

(Holotype, KUN).

Erymo .oey: rubidum refers to both the red Rhododendron blooms of its substrate and

the red disc of the lichen.

THALLUS crustose, corticolous, smooth, slightly cracked, without isidia

or soralia, whitish, thin, 0.05 mm thick; prothallus not seen. APOTHECIA

0.5-1.0 mm diam., constricted at the base, dispersed, disc cinnabar-red

to brownish-red, primarily plane, soon becoming convex, epruinose,

margin thin, initially flush with disk but soon becoming hidden with age;

AMPHITHECIUM 75-95 um thick; EprayMENIUM orange-red, K+ red,

5-7.5(-10) um thick; Hymenium hyaline, 75-90 um tall; HyPOTHECIUM

hyaline or slightly brown; Parapuyszs branched and anastomosing. AscI

clavate, containing 8 spores; AscosPorEs persistently colorless, fusiform,

submuriform with 15-21 transverse and 0-3 longitudinal septa per segment

in optical view, 75-100 x 10-12.5 um, the last cell of the spore, 20-25

um long. The upper half of some spores muriform, while the lower half

transversely septate. PYCNIDIA: not observed.

Haematomma rubidum sp. nov. (China) ... 427

Fic.1: Haematomma rubidum (holotype, KUN 00-20099). A. Thallus; B. Thallus with apothecia;

C. Section through apothecium; D-F. Ascospores. Scale bars: A = 2 mm; B = 1 mm; C = 50 um;

D, E= 10 um; F = 5 um.

CHEMISTRY—Cortex and medulla K+ yellow, C-, KC-, IKI-. Atranorin

and russulone detected by TLC.

HasBiTat—Growing with Lecidella spp. on the trunk of Rhododendron on

a sunny slope, above 3000 m elevation.

COMMENTS—Haematomma rubidum is characterized by its convex apothecia,

submuriform ascospores, and K+ red epihymenium. When young the

apothecial margin is usually even with disk but eventually becomes hidden

or covered in the lower part due to the gradual bulging of disk. Haematomma

identification is still based on classical taxonomy. Nonetheless, its unique

morphology and chemistry significantly separate H. rubidum from other

Haematomma species. These differences support our proposing a single

collection as a new species, but future DNA sequence analyses are needed to

confirm its taxonomic disposition.

Haematomma staigeriae, which also possesses submuriform spores and

a K+ red epihymenium, can be distinguished by its narrower (6-10 um)

428 ... Tang, Yin, Zhao

TaBLE1. Distinguishing characteristics of (sub)muriform-spored Haematomma species

CHARACTER H. rubidum H. gallowayi H. parda H. staigeriae H. wattii

SORALIA Absent Present Partly present Present Absent

APOTHECIUM

Disc Cinnabar- to Cinnabar red, = Brown, Cinnabar-red, | Cinnabar- red,

brownish-red, | smooth flat smooth smooth

convex

Margin Thin Thick Thick Thick Thick

EPIHYMENIUM K+ red K+ purple K- K+ red K+ purple

ASCOSPORE

Type Submuriform Muriform Submuriform Submuriform Submuriform

Long. septa 0-3 1-3 0-2 0-1 0-1

Transverse septa. 15-21 Phare 14-17 11-23 15-25

Length (um) 75-100 x (46-)51-78 x 55-75 x 50-90 x 60-95 x

Width (um) 10-12.5 10-15.5 5-7.5 6-10 6.5-10

CHEMISTRY

Atranorin + + +

Haematommone — _— _—

Lichexanthone — — — + —

Placodiolicacid — + — —_— +

Russulone + — — + —

Zeorin —_— + — — —

Unk. substance = — — + (2) + ‘a

REFERENCE This study Brodo 2007 Aptroot 2007 Nelsen & al. Staiger & Kalb

2006 1995

ascospores, flat discs, and a thallus with soralia and lichexanthone. In China,

another species of Haematomma with submuriform spores, H. wattii, has

been reported. However, H. wattii contains haematommone rather than

russulone. The main differences among (sub)muriform-spored species of

Haematomma are shown in TABLE 1.

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, China) for presubmission review. We are

grateful to Dr. Li-song Wang and Dr. Xin-yu Wang (Kunming Institute of Botany,

China) for sending specimens. 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).

Haematomma rubidum sp. nov. (China) ... 429

Literature cited

Aptroot A, Saipunkaew W, Sipman HJM, Sparrius LB, Wolseley PA. 2007. New lichens from

Thailand, mainly microlichens from Chiang Mai. Fungal Diversity 24: 75-134.

Brodo IM. 2007. Notes on the genus Haematomma from Sabah, Malaysia. Bibliotheca

Lichenologica 95: 147-153.

Nelsen MP, Liicking R, Chaves JL, Sipman HJM, Umafia L, Navarro E. 2006. A first assessment of

the Ticolichen biodiversity inventory in Costa Rica: the genus Haematomma (Lecanorales:

Lecanoraceae). Lichenologist 38(3): 251-262. https://doi.org/10.1017/S0024282906005573

Orange A, James PW, White FJ. 2010. Microchemical methods for the identification of lichens.

2nd edition. British Lichen Society, London.

Staiger B, Kalb K. 1995. Haematomma-Studien: I. Die Flechtengattung Haematomma.

Bibliotheca Lichenologica 59: 3-198.

Wang LS. 2012. Lichens of Yunnan Province, China. Shanghai Science and Technology

Publishing House, Shanghai.

MYCOTAXON

April-June 2020—Volume 135, pp. 431-441

https://doi.org/10.5248/135.431

Armillaria xiaocaobaensis sp. nov. from China

Jia-Hua PENG’ & CHANG-LIN ZHAO”

'Gastrodia Tuber Research Institute of Zhaotong,

Zhaotong 657000, Yunnan Province, P.R. China

’ Key Laboratory for Forest Resources Conservation and Utilization in the

Southwest Mountains of China, Ministry of Education, Southwest Forestry University,

Kunming 650224, Yunnan Province, P.R. China

* CORRESPONDENCE TO: fungichanglinz@163.com

ABSTRACT—A new wood-inhabiting fungal species from Yunnan Province, China, is

proposed based on morphological and molecular characters. Armillaria xiaocaobaensis is

characterized by a central stipe, striate to furrowed orange yellow to pinkish brown pileus

surface, membranous annulus, and ellipsoid, a monomitic hyphal system with generative

hyphae bearing simple septa, and slightly thick-walled basidiospores averaging 7.7 x 4.9 um.

TEF1 sequence analyses support A. xiaocaobaensis as a distinct taxon in Armillaria.

Key worps—Agaricales, phylogenetic, Physalacriaceae, pathogenic fungi, taxonomy

Introduction

Species in Armillaria (Fr.) Staude (Physalacriaceae, Basidiomycota) are

well known as important plant pathogens that can cause serious root diseases

on diverse trees and woody plants, resulting in huge economic losses (Wargo

& Shaw 1985, Baumgartner 2004, Lalande & al. 2018).

Armillaria includes approximately 40 species (Volk & Burdsall 1995, Kirk

& al. 2008, Brazeel & al. 2012, Hood & Ramsfield 2016, Elias-Roman & al.

2018) worldwide, and ten Armillaria species are currently recognized in

China (Qin & al. 2000, 2007; Sun & al. 2003; Zhao & al. 2008; Coetzee & al.

2015; Guo & al. 2016).

Recent phylogenetic studies of Armillaria based on sequences of the

internal transcribed spacer (ITS) region, the large subunit of nuclear

432 ... Peng & Zhao

ribosomal RNA gene (nLSU), translation elongation factor 1-a (TEF1), and

B-tubulin have greatly increased the number of identified fungal species and

revealed the relationships among species in this genus (e.g., Maphosa & al.

2006, Hasegawa & al. 2010, Ross-Davis & al. 2012, Tsykun & al. 2013, Coetzee

& al. 2015, Park & al. 2018). Lima & al. (2008) described one new South

American species, Armillaria paulensis Capelari, based on morphology and

ITS sequences that showed its closeness to A. luteobubalina Watling & Kile,

while Brazee & al. (2012) introduced A. altimontana Brazee & al. in North

America based on morphological and molecular phylogenetic evidence. In

New Zealand, Hood & Ramsfield (2016) revealed A. aotearoa I.A. Hood &

Ramsfield using morphology, interfertility cultures, and DNA sequence data.

Guo & al. (2016) analyzed Chinese Armillaria samples using the sequences

of ITS, TEF1 and B-tubulin gene, which revealed 15 phylogenetic lineages

of Armillaria from China and showed that TEF1 was the most parsimony

informative. Koch & al. (2017) resolved phylogeny and biogeography of

Armillaria, Desarmillaria (Herink) R.A. Koch & Aime, and Guyanagaster

T.W. Henkel & al. Elias-Roman & al. (2018) described A. mexicana Elias-

Roman & al. based on morphology, DNA-sequence data (ITS, nLSU and

TEF1), and phylogenetic analyses.

Our research on macrofungi in southern China revealed specimens

that could not be assigned to any described species within Armillaria. The

authors sampled previous research using the translation elongation factor-1

alpha gene (TEF1) sequences to explore the taxonomy and phylogeny of this

undescribed species, proposed below as Armillaria xiaocaobaensis.

Materials & methods

Macroscopical descriptions were based on field notes. Color terms follow

Petersen (1996). Dried material was examined microscopically using a compound

microscope following Dai (2012). The following abbreviations were used:

KOH = 5% potassium hydroxide, CB = cotton blue, CB- = acyanophilous,

IKI = Melzer’s reagent, IKI- = both inamyloid and nondextrinoid, L = mean spore

length (arithmetic average of all spores), W = mean spore width (arithmetic average

of all spores), Q = variation in the L/W ratios between the specimens studied, n

(a/b) = number of spores (a) measured from given number (b) of specimens. The

specimens studied are deposited at the herbarium of Southwest Forestry University,

Kunming, China (SWFC).

Genomic DNA was extracted from dried specimens using the EZNA HP Fungal

DNA Kit according to the manufacturer’s instructions with some modifications. A

small piece (about 30 mg) of dried fungal specimen was ground to powder with

liquid nitrogen. The powder was transferred to a 1.5 mL centrifuge tube, suspended

Armillaria xiaocaobaensis sp. nov. (China) ... 433

in 0.4 mL of lysis buffer, and incubated in a 65 °C water bath for 60 min. After 0.4

mL phenol-chloroform (24:1) was added to the tube, 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. TEF1 was amplified with

primer pairs EF1-983F and EF1-2218R (Rehner 2001). The PCR procedure for

TEF1 was as follows: initial denaturation at 94 °C for 1 min, followed by 35 cycles

at 94 °C for 30 s, 59 °C for 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 in GenBank (TABLE 1).

GeneCodes Sequencher 4.6 was used to edit the DNA sequence. Sequences were

aligned in MAFFT 7 (https://mafft.cbrc.jp/alignment/server/index.html) using the

“G-INS-I” strategy and manually adjusted in BioEdit (Hall 1999). The sequence

alignment was deposited in TreeBase (submission ID 24610). Oudemansiella

cubensis (Berk. & M.A. Curtis) R.H. Petersen and Strobilurus esculentus (Wulfen)

Singer were used as outgroup to root trees following Koch & al. (2017) in the TEF1

analyses.

Phylogenetic analyses of the TEF1 sequences were performed 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 were set to 5000, branches of zero length were collapsed, and

all parsimonious trees were saved. Clade robustness was assessed using bootstrap

(BT) analysis with 1000 replicates (Felsenstein 1985). Tree length (TL), consistency

index (CI), retention index (RI), rescaled consistency index (RC), and homoplasy

index (HI) were calculated for each Maximum Parsimonious Tree (MPT) generated.

Sequences were analyzed using Maximum Likelihood (ML) with 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 (Nylander 2004) was used to determine the best-fit evolution

model for each data set for Bayesian inference (BI). Bayesian inference was

calculated with MrBayes_3.1.2 using a general time reversible (GTR) 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 5 million generations (TEF1) and trees were sampled every 100

generations. The first 25% of the generations were discarded as burn-in. A majority

434 ... Peng & Zhao

TABLE 1. Sequences used in the phylogenetic analyses; new sequences in bold.

SPECIES

Armillaria altimontana

A. aotearoa

A. borealis

A. calvescens

A. cepistipes

A. fumosa

A. fuscipes

A. gallica

A. gemina

A. heimii

. hinnulea

. limonea

A. luteobubalina

A. mellea

A. mexicana

A. nabsnona

A. novae-zelandiae

A. ostoyae

A. pallidula

A. puiggarii

SAMPLE NO.

D 82

POR 100

NZFRIM 5283

HKAS 56108

HKAS 76263

ST 17

ST 18

ST 3

F 011081

S 20

CMW 4955

CMW 3164

CMW 4953

M70

ST 22

ST 8

ST11A

K59

PH 8724

CMW 4980

CMW 4680

CMW 4991

Plim 8466

CMW 4977

CMW 8876

CMW 4613

G 0405414

MEX 74

MEX 85

MEX 87

MEX 88

C21

HKAS 85523

CMW 4722

CMW 5448

EC4

EC 5

3626

CMW 4971

MCA 3111

TH 9751

GENBANK NO.

TEF1

JN944611

JN944606

KU295542

KT822293

KT822294

JF895836

JF895837

JF895835

KT822416

JF313116

DQ435646

DQ435621

DQ435622

HQ432899

HQ432897

JF313136

JF313133

FJ618644

FJ618651

DQ435648

DQ435655

DQ435656

FJ618654

DQ435657

DQ435658

DQ435637

KT822345

KC111011

KR061313

KRO061314

KR061315

JF313119

KT822411

DQ435652

DQ435653

JE746925

JF746926

FJ618665

DQ435647

KU289104

KU289113

REFERENCE

Brazee & al. 2012

Hood & Ramsfield 2016

Guo & al. 2016

Brazee & al. 2011

Guo & al. 2016

Brazee & al. 2011

Maphosa & al. 2006

Direct Submission

Elias-Roman & al. 2018

Brazee & al. 2011

Elias-Roman & al. 2018

Maphosa& al. 2006

Maphosa & al. 2006

Elias-Roman & al. 2018

Maphosa & al. 2006

Guo & al. 2016

Elias-Roman & al. 2018

Guo & al. 2016

Maphosa & al. 2006

Brazee & al. 2011

Elias-Roman & al. 2018

Maphosa & al. 2006

Koch & al. 2017

Armillaria xiaocaobaensis sp. nov. (China) ... 435

SPECIES SAMPLE NO. GENBANK NO. REFERENCE

TEF1

A. sinapina A 9601539 KT822422 Guo & al. 2016

M50 JF313114 Brazee & al. 2011

ST 12 JF313132 Brazee & al. 2011

ST 13 JF313131 Brazee & al. 2011

A. solidipes P 1404 JF313140 Brazee & al. 2011

ST 2 JF313139 Brazee & al. 2011

A. xiaocaobaensis SWEC 12637 MN298777 This study

SWEC 12638[T] MN298778 This study

SWEC 12639 MN298779 This study

Desarmillaria ectypa 7001113 KT822438 Guo & al. 2016

CMW 15693 FJ875698 Koch & al. 2017

MY 84941 FJ618643 Koch & al. 2017

D. tabescens 90158 KT822439 Koch & al. 2017

19912213 KT822441 Guo & al. 2016

PT 8412 FJ618658 Koch & al. 2017

Guyanagaster necrorhizus G 314 KU289110 Koch & al. 2017

G 352 KU289109 Koch & al. 2017

MCA 3950 KU289107 Koch & al. 2017

RAK 31 KU289108 Koch & al. 2017

Oudemansiella cubensis MCA 5434 KU289105 Koch & al. 2017

Strobilurus esculentus Yang 5027 KF530581 Koch & al. 2017

rule consensus tree of all remaining trees was calculated. Branches that received

bootstrap support for maximum likelihood (BS), maximum parsimony (BT), and

Bayesian posterior probabilities (BPP) greater than or equal to 75% (BS, BT) and

0.95 (BPP) were considered significantly supported.

Molecular phylogeny

The TEF1 dataset included sequences from 62 fungal specimens

representing 29 species (TABLE 1). The alignment comprised 544 characters

of which 312 characters were constant, 29 variable and parsimony-

uninformative, and 203 parsimony-informative. Maximum parsimony

analysis yielded four equally parsimonious trees (TL = 728, CI = 0.496,

HI = 0.504, RI = 0.781, RC = 0.387). Best model for the TEF1 dataset

estimated and applied in the Bayesian analysis: GIR+I+G. Bayesian

analysis and ML analysis resulted in a similar topology as MP analysis, with

an average standard deviation of split frequencies = 0.003738 (BI).

The phylogeny inferred from TEF1 sequences placed Armillaria

xiaocaobaensis in a monophyletic lineage with strong support (BS = 100%;

BT = 100%; BPP = 1).

436 ... Peng & Zhao

90/85/0.96

S591 -

99/95/0.96

100/100/1.00

-/67/-

100/100/1.00

100/100/1.00 , Armillaria fuscipes CMW 4953

Armillaria fuscipes CMW 3164

Armillaria heimii PH 8724

100/100/1.00 '4rmillaria heimii K 59

100/100/1.00, Armillaria pallidula CMW 4971

Armillaria pallidula 3626

Armillariafumosa CMW 4955

Armillaria hinnulea CMW 4980

Armillaria luteobubalina CMW 8876

Armillarialimonea Plim 8466

Armillaria linonea CMW 4680

Armillaria linonea CMW 4991

100/100/1.00;—Armillaria puiggarii TH 9751

100/100/1.00 Armillaria puiggarii MCA 3111

Armillaria novae-zelandiae CMW 4722

-/67/-

“69 Armillaria aotearoa NZFRIM 5283

100/100/1.00 Armillaria mellea CMW 4613

—{ _—— Armillaria mellea G 0405414

Armillaria mellea MEX 74

NOOHGOH OO Armillariamexicana MEX 85

|__oo001.00 __\4rmiftaria mexicana MEX 87

Armillaria mexicana MEX 88

100/100/1.00, Armillaria gemina ST11A

92/89/0.98 Armillaria gemina ST 8

peal Armillaria solidipes ST 2

; Armillaria solidipes P 1404

Armillaria borealis HKAS 76263

Armillaria borealis HKAS 56108

Armillaria ostoyae EC 5

100/100/1.00' Armillaria ostoyae EC 4

Armillaria sinapina A 9601539

Armillaria sinapina ST 12

Armillaria sinapina ST 13

Armillaria sinapina M 50

100/100/1.00, Armillaria cepistipes F 011081

Armillaria cepistipes S 20

Armillarianabsnona HKAS 85523

Armillarianabsnona C 21

80/75/-

100/100/1.00

Armillaria altimontana D 82

100/100/1.00! Armillaria altimontana POR 100

Armillaria luteobubalina CMW 4977

100/100/1.00L Armillaria novae-zelandiae CMW 5448

-/S8/-

—

100/99/1.00 ' Guyanagaster necrorhizus MCA 3950

200/1001.90 Armillaria xiaocaobaensis SWFC 12639

\_s00r1007.00 fs rmillaria xiaocaobaensis SWFC 12638

‘Armillaria xiaocaobaensis SWFC 12637

Desarmillaria tabescens PT 8412

_——0011o01.00__f—“'Desarmillaria tabescens tabescens 90158

Desarmillaria tabescens tabescens 19912213

Desarmillaria ectypa MY 84941

|__1oono0r.00 _f!Desarmiflaria ectypa 7001113

Desarmillaria ectypa CMW 15693

100/100/1.00 ;Guyanagaster necrorhizus G 314

Guyanagaster necrorhizus G 352

Guyanagaster necrorhizus RAK 31

Oudemansiella cubensis MCA 5434

FE)

Strobilurus esculentus Yang 5027

Fic. 1. Maximum parsimony strict consensus tree illustrating the phylogeny of Armillaria

xiaocaobaensis and related species based on TEF1 sequences. Branches are labeled with

maximum likelihood bootstrap >70%, parsimony bootstrap proportions >50%, and Bayesian

posterior probabilities >0.95.

Taxonomy

Armillaria xiaocaobaensis Jia H. Peng & C.L. Zhao, sp. nov.

MB 832231

Fics 2, 3

Differs from Armillaria altimontana by its centrally stipitate basidiomes with striate to

furrowed, orange yellow to pinkish brown pileus surface, membranous annulus, and

smaller, ellipsoid, thin-walled basidiospores.

Type: China. Yunnan Province: Zhaotong, Yiliang County, Xiaocaoba Town, on

angiosperm trunk, 19 April 2019, Jia-Hua Peng (Holotype, SWFC 0012638; GenBank

MN298777).

EryMOLoGy: xiaocaobaensis (Lat.) refers to the locality (Xiaocaoba Town) of the type

specimen.

Armillaria xiaocaobaensis sp. nov. (China) ... 437

Fic. 2. Armillaria xiaocaobaensis (holotype, SWFC 0012638).

Scale bars: A = 4 cm; B, C=1cm;D=5 mm; E=2 mm.

BASIDIOMATA pileate, centrally stipitate, caespitose. PILEUs 30-40 mm diam.,

umbonate, convex to plano-convex, orbicular in apical view, margin deflexed to

lobed, pileus surface striate to furrowed, orange yellow to pinkish brown in the

center, and pale yellow to pale brown at margin. LAMELLAE 5 mm deep, close,

438 ... Peng & Zhao

decurrent, adnate, edges entire, buff to light yellow. LAMELLULAE attenuate,

edges entire, multi-tiered. ANNULUs 4-5 mm wide, superior, persistent,

membranous, appressed to stipe, buff to light yellow. StrpE 35-60 x 4-6 mm,

central, cylindrical, solid, light yellow to pale pinkish brown to brown to black

toward the base in fresh specimens, base bulbous. RHIZOMORPHS absent. TasTE

AND ODOR not tested. SPORE PRINT white.

Basipiosporss ellipsoid, hyaline, slightly thick-walled, smooth, IKI-, CB-,

(6.5—)7-8.5(-9) x 4-5.5(-6) um, L = 7.74 um, W = 4.93 um, Q = 1.53-1.64

(n = 90/3). Basrp1a barrel-shaped to clavate, with 2-4 sterigmata and a

simple basal septum, 20-28 x 5.5-8.5 um; basidioles dominant, mostly

clavate but slightly smaller than basidia. HYMENOPHORAL TRAMA generative

hyphae bearing simple septa, hyaline, thin-walled, unbranched, 5-8 um in

diam. CysTip1A none observed. PILEIPELLIS hyphae hyaline, thin-walled,

unbranched, 8-13 um diam. CONTEXT 100-150 um wide, hyphae hyaline,

thin-walled, unbranched, 6-10 um diam. St1prTiPELLis hyphae hyaline, thin-

walled, unbranched, 3.5-5.5 um diam. CLAMP CONNECTIONS absent.

ADDITIONAL SPECIMENS EXAMINED: CHINA. YUNNAN PROVINCE. Zhaotong: Yiliang

County, Xiaocaoba Town, on angiosperm trunk, 19 June 2019, Jia-Hua Peng (SWFC

0012637; SWFC 0012639).

Discussion

Phylogenetic analyses and morphological characters support Armillaria

xiaocaobaensis as a new species.

In the TEF1 analyses (Fic. 1), A. xiaocaobaensis formed a monophyletic

lineage with strong support (BS = 100%; BT = 100%; BPP = 1) and grouped

with Armillaria altimontana, A. calvescens Bérubé & Dessur., A. cepistipes

Velen., A. gallica Marxm. & Romagn., and A. nabsnona T.J. Volk & Burds.

However, morphologically A. altimontana differs from A. xiaocaobaensis by

its tricholomatoid basidiomata with a reddish brown pileus bearing cream

scales and short fibrils (Brazee & al. 2012) and from A. calvescens by the

presence of pileus scales and fibrils, and larger basidiospores (8.5-10 x 5-7

um; Bérubé & Dessureault 1989). Armillaria cepistipes differs in its yellow

ochraceous to ochraceous brown pileus, longer stipe (100 x 9-11 mm), and

larger basidia (29-45 x 8.5-11 um; Antonin & al. 2009); and A. gallica by

the presence of dark brown to pinkish brown pileus, long fibrillose pileus

scales, and ellipsoid to amygdaloid, larger basidiospores (7.5-11 x5-6.5 um;

Antonin & al. 2009). Armillaria nabsnona has a hygrophanous pileus with a

white context and larger basidiospores (8-10 x 5.5-6.5 um; Volk & al. 1996).

Armillaria xiaocaobaensis sp. nov. (China) ... 439

Same

\ a a

®. KE =

—

(

Fic. 3. Microscopic structures of Armillaria xiaocaobaensis (drawn from the holotype,

SWEC 0012638). A. Basidiospores; B. Section of hymenium; C. Pileipellis; D. Stipitipellis.

Scale bars: A~D = 10 um.

Armillaria xiaocaobaensis is close to Desarmillaria, a member in the sister

genus based on the molecular data (Fic. 1). Desarmillaria is distinguished

morphologically from Armillaria by the absence of an annulus (Koch & al.

2017).

440 ... Peng & Zhao

Acknowledgments

Special thanks are due to Dr. Sana Jabeen (Department of Botany, Division of

Science and Technology, University of Education, Lahore, Pakistan) and Dr. Vladimir

Antonin (Department of Botany, Moravian Museum, Brno, Czech Republic) who

reviewed the manuscript. The research is supported by the Yunnan Agricultural

Foundation Project (2017FG001-042).

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MYCOTAXON

April-June 2020—Volume 135, pp. 443-452

https://doi.org/10.5248/135.443

Phylogeny and morphology of Ellismarsporium parvum

and the new combination E. varium

KAI ZHANG”?, WEIHUA GUO"*, DAYNET SOSA},

FREDDY MAGDAMA;}, LIZETTE SERRANO’, ELAINE MALOSSO4,

DE-WEI LI>, RAFAEL F. CASTANEDA-RUIZ°

' Institute of Ecology and Biodiversity, School of Life Sciences, Shandong University,

Qingdao, 266237, China

? Shandong Agriculture and Engineering University, Jinan, Shandong 250100, China

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

Campus Gustavo Galindo Km. 30.5 Via Perimetral,

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

‘Centro de Biociéncias, Departamento de Micologia, Universidade Federal de Pernambuco,

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

° The Connecticut Agricultural Experiment Station, Valley Laboratory,

153 Cook Hill Road, Windsor, CT 06095, USA

° Instituto de Investigaciones Fundamentales en Agricultura Tropical Alejandro de Humboldt

(INIFAT), Grupo Agricola, OSDE, Académico Titular de la Academia de Ciencias de Cuba,

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

“ CORRESPONDENCE TO: whguo_sdu@163.com

ABSTRACT—LSU DNA sequence analyses place Ellismarsporium parvum in Pleosporales. The

morphology of the species in pure culture is described and illustrated. Helminthosporium

varium is recombined as E. varium, based on its morphology.

Key worps—Ascomycota, asexual fungi, hyphomycetes, phylogeny, taxonomy

Introduction

Ellismarsporium R.F. CastaMeda & X.G. Zhang, typified with

E. hypselodelphyos (M.B. Ellis) R.F. Castaheda & X.G. Zhang is characterized

by macronematous and mononematous unbranched conidiophores that

444 ... Zhang & al.

bear integrated and intercalary polytretic conidiogenous cells (Castafieda-

Ruiz & al. 2017). The phragmosporic distoseptate conidia are produced in

acropetal unbranched or branched chains. Production of conidial chains

is the main diagnostic character distinguishing Ellismarsporium from

Helminthosporium Link typified by H. velutinum Link. Phylogenetic LSU

sequence analysis of strains of E. parvum and Helminthosporium species has

provided additional information on the differences between these genera.

Materials & methods

Pure cultures of Ellismarsporium parvum from INIFAT WDCM 853 were

transferred to sterilized Petri dishes containing V8 agar (125 ml V8 juice, 18 g agar,

1000 ml distilled water, pH 6.3). Plates were incubated at 25°C in a 12 h dark/light

cycle using a box equipped with near UV (black light) VICA FLB-20W T10 lamps.

Colony colors were coded according to Rayner (1970). Mounts were prepared in

PVL (polyvinyl alcohol, lactic acid) and structures were measured at x1000 under

a Nikon Eclipse Ni-U microscope with bright field optics and photographed using

DIC optics with a Nikon DS-Fi2 camera at Universidade Federal de Pernambuco,

Recife, Brazil.

TABLE 1. LSU sequences of Ellismarsporium parvum and allied species used in the

phylogenetic analysis. New sequences are indicated in bold.

SPECIES CULTURE GENBANK REFERENCE

Anteaglonium abbreviatum GKM219N GQ221881 Mugambi & Huhndorf 2009

A. parvulum SMH5210 GQ221907 Mugambi & Huhndorf 2009

Clohesyomyces aquaticus MFLUCC11-0092 JX276950 Zhang & al. 2012

Corynespora cassiicola C13-1 KF590123 Palomares-Rius & al. 2014

SKO1 LC177365 Chairin & al. 2017

C. pseudocassiicola CPC 31708 MH327830 Crous & al. 2018

C. smithii L133 188 KY984299 Voglmayr & Jaklitsch 2017

C. torulosa CPC 15989 KF777207 Crous & al. 2013

Ellismarsporium parvum INIFAT-2457 MN032445 This study

INIFAT-2483 MN032446 This study

CBS 190.95 MH874149 Vu & al. 2019

Flavomyces fulophazii CBS 135761 KP184040 Knapp & al. 2015

Fuscostagonospora cytisi MFLUCC 16-0622 KY770978 Crous & al. 2018

EF. sasae KT 1467 AB807548 Tanaka & al. 2015

Helminthosporium asterinum CBS 203.35 MH867156 Vu & al. 2019

H. caespitosum L151 KY984306 Voglmayr & Jaklitsch 2017

H. dalbergiae H 4628 AB807521 Tanaka & al. 2015

H. genistae L144 KY984312 Voglmayr & Jaklitsch 2017

H. leucadendri CBS 135133 KF251654 Quaedvlieg & al. 2013

H. massarinum KT 838 AB807523 Tanaka & al. 2015

H. magnisporum

H. microsorum

H. oligosporum

Hi. quercinum

Hongkongmyces pedis

H. thailandica

Keissleriella cladophila

K. culmifida

K. gloeospora

Kirschsteiniothelia aethiops

Lentithecium carbonneanum

L. clioninum

L. lineare

L. pseudoclioninum

L. unicellulare

L. voraginesporum

Lindgomyces ingoldianus

L. rotundatus

Magnicamarosporium

iriomotense

Monocillium mucidum

Noosia banksiae

Periconia cookei

P. cyperacearum

P. digitata

P. homothallica

P. igniaria

P. macrospinosa

P. pseudodigitata

P. thailandica

Pleurophoma acaciae

Preussia fleischhakii

P terricola

P. typharum

Pseudodictyosporium

thailandica

Spadicoides bina

S. fuscolutea

S. hyalostoma

Sporidesmium tengii

Sporormia fimetaria

Sporormiella megalospora

H 4627

L96

L93

L107

HKU 35

MFLUCC 16-0406

UTHSC:DI116-326

KT 2308

KT 2642

KT 829

CBS 111.56

CBS 118.59

CBS 111.61

CBS 144076

KT 1149A

IFRD 2008

KT 1113

MD6004

H-22560

ATCC 200398

MAFF 239473

KT 2822

CBS 980.70

CPC:17282

MFLUCC 17-1399

CPC 32138

CBS 510.77

KT 916

CBS 845.96

CBS 135663

KT 1395

MFLUCC 17-0065

CPC 29188

CBS 565.63

CBS 527.84

CBS 107.69

MFLUCC 16-0029

CBS 137794

CBS 141263

CBS 139771

HKUCC 10837

UPS:

Lundqvist 2302-c

UPS:Kruys 305

Ellismarsporium parvum in Ecuador ... 445

AB807522

KY984329

KY984333

KY984336

NG 056287

KY771321

LN907469

AB807591

AB807592

AB807589

MH869065

MH877780

MH869545

MH062991

AB807540

FJ795435

AB807545

KX505376

KX499520

AB521736

KF314114

AB807509

MG826851

JF951167

MG333493

MH327851

AB807561

AB807565

AB807567

KP184038

AB807564

KY753888

KY173524

GQ203721

GQ203724

GQ203726

KX259522

KY931824

KY931826

KY931830

DQ408559

GQ203728

GQ203743

Tanaka & al. 2015

Voglmayr & Jaklitsch 2017

Tsang & al. 2014

Crous & al. 2018

Valenzuela-Lopez & al. 2017

Tanaka & al. 2015

Vu & al. 2019

Crous & al. 2018

Tanaka & al. 2015

Zhang & al. 2009

Tanaka & al. 2015

Crous & al. 2018

Hirayama & al. 2010

Tsang & al. 2014

Tanaka & al. 2015

Crous & al. 2018

Crous & al. 2011

Hyde & al. 2018

Crous & al. 2018

Tanaka & al. 2015

Knapp & al. 2015

Tanaka & al. 2015

Liu & al. 2017

Crous & al. 2013

Kruys & Wedin 2009

Wang & al. 2016

Réblova & al. 2018

Shenoy & al. 2006

Kruys & Wedin 2009

446 ... Zhang & al.

DNA extraction, amplification, alignment, and phylogenetic analysis

The INIFAT-2483 and INIFAT-2457 isolates were cultured on V8 for 10 days at

25°C under near UV ina 12 h dark/light cycle. DNA extraction followed a modified

protocol from Cenis (1992). The primer pair LROR/LR5 was used to amplify the

D1/D3 domain of the LSU nrDNA (White & al. 1990). PCR products were purified

and sequenced at Macrogen Inc., South Korea. Consensus sequences assembled and

edited using Geneious (ver. 10.1.2) were compared with those of the National Center

for Biotechnology Information (NCBI) using BLAST.

Alignments for each data set were made in MEGA 6.0 (Tamura & al. 2013)

using the ClustalW algorithm (Thompson & al. 1994) and refined with MUSCLE

(Edgar 2004). The alignment included our strain sequences and others representing

different genera in Pleosporales available from NCBI (TABLE 1). A LSU sequence

phylogeny was generated using Maximum Likelihood (ML) with the best nucleotide

substitution model found in MEGA 6.0 (Tamura & al. 2013). The best model used

was Tamura-Nei with Gamma distribution. Bootstrap analysis of 1000 replicates

assessed the reliability of the reconstructed phylogenies. ML bootstrap values =70%

were considered significant.

Results

The BLAST query revealed that LSU sequences of E. parvum INIFAT-2483 and

INIFAT-2457 showed a 99.3% similarity with E. parvum [= Helminthosporium

parvum, ex-type] CBS 190.95 and 92.1% with Helminthosporium asterinum

Cooke CBS 203.35. However, they showed sequence identity of <86% with LSU

sequences of other Helminthosporium species.

We carried out a phylogenetic analysis of the LSU locus to assess the

relationships of Ellismarsporium with members of the Pleosporales. The final

analysis encompassed 62 sequences and comprised 552 bp. The ML tree nested

all E. parvum isolates (INIFAT-2483, INIFAT-2457, CBS 190.95) in a well-

supported subclade (bootstrap value = 99%), with H. asterinum CBS 203.35

and Kirschsteiniothelia aethiops (Sacc.) D. Hawksw. as the closest sister taxa

(Fic. 1).

Both morphological and phylogenetic analyses of E. parvum support

Ellismarsporium within Pleosporales.

Taxonomy

Ellismarsporium parvum R.F. Castafieda & W.B. Kendr.,

Mycotaxon 132(4): 763 (2017) FIGS 2, 3

= Helminthosporium parvum R.E. Castaheda & W.B. Kendr., Univ.

Waterloo Biol. Ser. 35: 57 (1991), nom. illeg., non Grove (1886)

CoLonizs on V8 agar at 25°C attaining 24 mm diam after 10 days,

felted, sparsely minutely lanate, greenish olivaceous. CONIDIOPHORES

Ellismarsporium parvum in Ecuador ... 447

/ncertae sedis

Fic. 1. Maximum Likelihood (ML) tree inferred from LSU sequences of Ellismarsporium parvum

and related genera, based on 62 sequences. The bootstrap test was conducted with 1000 replicates;

bootstrap values =>70% are indicated at the nodes. The scale bar indicates the number of expected

changes per site.

448 ... Zhang & al.

Fic. 2. Ellismarsporium parvum (holotype, INIFAT C90/101). Colonies on V8 agar.

macronematous, mononematous, erect, straight or flexuous, unbranched,

cylindrical, sometimes slightly clavate or inflated toward the apex,

4-10-septate, brown, smooth, <220 um long. CONIDIOGENOUS CELLS

polytretic, intercalary and terminal, integrated, cylindrical (terminal cells

somewhat clavate to inflated or slightly curved), brown, 10-40 x 3-8 um.

Conip1A acropleurogenous, cylindrical-ellipsoidal, cylindrical to oblong

(rarely obovoid), 1-2-distoseptate, smooth, brown to smoke-olivaceous-

brown, 14-23 x 6-10 um, in short branched acropetal chains.

SPECIMENS EXAMINED: CUBA, PINAR DEL RIO PROVINCE, San Juan y Martinez, San

Simon de las Cuchillas, 22°16’N 83°50’W, on bark of Quercus oleoides var. sagrana (Nutt.)

C.H. Mull. [= Q. cubana A. Rich.], 24 March 1990, R.F. Castaneda Ruiz (holotype, INIFAT

C90/101; INIFAT-2457, GenBank MN032445 [URM 94323 & CCMCIBE-H500];

INIFAT-2483, GenBank MN032446 [URM 94322 & CCMCIBE-H501]).

Helminthosporium varium, published by Alves-Barbosa & al. (2017), is

distinguished by distoseptate, long obclavate, subcylindrical, oblong, or

navicular, 1-4-septate, irregularly verrucose or verruculose, gray brown to

brown conidia produced in branched and unbranched acropetal chains.

As the species clearly falls within the Ellismarsporium generic concept, we

propose a new combination:

Ellismarsporium varium (Alves-Barb., Malosso & R.F. Castafieda) K. Zhang,

Alves-Barb., Malosso & R.F. Castafieda, comb. nov.

MB 832010

= Helminthosporium varium Alves-Barb., Malosso & R.F. Castaneda,

Nova Hedwigia 105: 67 (2017)

Ellismarsporium parvum in Ecuador ... 449

m\¥

Fic. 3. Ellismarsporium parvum (from culture ex holotype INIFAT C90/101). A-C. Conidia;

D-G. Conidiogenous cells and conidia; H. Hyphae, conidiophores, conidiogenous cells, and

conidia. Scale bars: A-G = 10 um; H = 50 um.

Discussion

The phylogenetic placement of Ellismarsporium was previously unknown,

despite several studies on the phylogenetic relationships of Pleosporales

450 ... Zhang & al.

(Hashimoto & al. 2017, Li & al. 2017). Our phylogenetic analysis places

Ellismarsporium in Pleosporales. However, its placement at the family level

remains unresolved. Our results show Ellismarsporium as sister to the clades

of K. aethiops (Kirschsteiniotheliaceae) and H. asterinum CBS 203.35 (Fic. 1).

As Ellismarsporium was proposed in 2017, it is understandable that the genus

was not included in recent phylogenetic studies on Kirschsteiniotheliaceae (Su

& al. 2016, Mehrabi & al. 2017, Bao & al. 2018). To determine the placement of

Ellismarsporium at family level, a study using multiple loci is necessary in the

future. Additionally, our results place H. asterinum CBS 203.35 distant from the

Helminthosporium clade (Fic. 1), indicating that species also requires further

study.

Acknowledgments

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

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

Monteiro (Museu Paraense Emilio Goeldi, Belém, Brazil) and Dr. Jian Ma (Jiangxi

Agricultural University, Nanchang, China) for their critical review of the manuscript.

Dr. Lorelei Norvell’s editorial review and Dr. Shaun Pennycook’s nomenclature review

are greatly appreciated.

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MYCOTAXON

April-June 2020—Volume 135, pp. 453-463

https://doi.org/10.5248/135.453

Acarospora scottii and Sarcogyne paradoxa spp. nov.

from North America

KERRY KNUDSEN & JANA KOCOURKOVA

Department of Ecology, Faculty of Environmental Sciences, Czech University of Life Sciences,

Prague, Kamycka 129, Praha 6 - Suchdol, CZ-165 00, Czech Republic

* CORRESPONDENCE: knudsen@fzp.czu.cz

ABSTRACT—Acarospora scottii, a facultative lichenicolous lichen on crustose lichens, is

described and typified from Minnesota. Sarcogyne paradoxa, which is described and typified

from California, grows as an endolithic lichen or as a lichenicolous fungus endokapylic in

crustose lichens.

KEY WORDS—Acarospora americana, Acarosporaceae, New Mexico, Polysporina, taxonomy

Introduction

Lichenicolous lichens begin their life cycle as non-lichenized parasites in

the microbiomes of lichens, expropriate the algae of the host, and eventually

morph out of the host thallus, destroying the host (Diederich & al. 2018;

Knudsen & Kocourkova 2018a; Knudsen & al. 2012, 2014). They persist

as lichenized fungi in a new epilithic thallus that differs from the host

form. Two hundred fifty-eight obligatory and facultative species have been

reported worldwide (Diederich & al. 2018, Knudsen & Kocourkova 2018a).

Reports in older literature are not based on this distinction and may not be

parasites but crustose lichens that overgrow and destroy other lichens in

the competition for space (although winning the competition may bestow a

saprobic benefit).

Obligatory lichenicolous lichens always begin their life cycle as juvenile

non-lichenized parasites. Obligatory lichenicolous lichens can be host

specific. For instance, Heteroplacidium transmutans K. Knudsen & al. from

454 ... Knudsen & Kocourkova

the Sonoran Desert has only been found growing out of Acarospora socialis

H. Magn. and has only been found as an independent lichen in association

with populations of its host (Knudsen & al. 2014). Or obligate lichenicolous

lichens can be opportunistic, occurring on different species or genera. For

instance, Diploschistes muscorum (Scop.) R. Sant. parasitizes many different

Cladonia species and (rarely) is parasitic on Lepraria xerophila Tonsberg

and Leprocaulon americana Lendemer & B.P. Hodk. in coastal habitats in

southern California (Kocourkova & al. 2012).

Facultative lichenicolous lichens can begin as a non-lichenized parasite

in the microbiome of a lichen, although they usually begin their life cycle as

non-parasitic lichenized fungi. They are also usually opportunistic, although

records may be based on only one host. A classic example of a facultative

lichenicolous lichen is Acarospora succedens H. Magn. It was described from

a single specimen as a parasite on Dimelaena suboreina (B. de Lesd.) Hale &

W.L. Culb. in New Mexico (Magnusson 1930). The taxon was described again

from New Mexico as the non-parasitic lichen A. interspersa H. Magn., which

is frequent in the Chihuahuan and Sonoran Deserts and is rarely found as a

parasite on crustose lichens (Knudsen 2007, 2011; Magnusson 1933). In this

paper we describe a facultative lichenicolous lichen, Acarospora scottii, from

Minnesota and Ontario, Canada.

In contrast to lichenicolous lichens, we describe Sarcogyne paradoxa

which occurs as either a non-lichenized parasite dwelling in the microbiome

of crustose lichens or as an endolithic lichen. It grows in the Mojave Desert

and other xerothermic habitats in Asia and North America. The implications

of S. paradoxa for a revision of species of the polyphyletic genus Polysporina

is discussed.

Material & methods

A specimen collected by Perry Scott, a citizen lichenologist, and specimens

from ASU & UCR herbaria were studied using standard microscopy and spot

tests. The amyloid reaction of the hymenial gel and subhymenium was tested

with fresh undiluted IKI (Merck’s Lugol; see protocol in Knudsen & Kocourkova

2018b). Measurements were made in water. Macrophotographs were taken with

the Olympus DP74 digital camera mounted on Olympus SZX 16 stereomicroscope

equipped with PRO-SZM1 - Focus Drive Motorization for stacking pictures and

stacked using Olympus DeepFocus 3.4 module of Promicra QuickPhoto Camera

3.2 software. The figure plates were processed with the same software fitted with

Promicra Figure Maker modul and eventually refined with Adobe Photoshop CS4

Extended ver. 11.0.

Acarospora & Sarcogyne spp. nov. (USA & Canada) ... 455

Taxonomy

Acarospora scottii K. Knudsen & Kocourk., sp. nov. Fics 1, 2

MB 831962

Similar to Acarospora americana but differing in having dark blue hymenial gel in

Lugol’s (euamyloid), in having a continuous polysaccharide epicortex giving the

areoles a dark shiny appearance, and in having a cortex of vertical hyphae.

Type: USA, Minnesota, Cook. Co., Superior National Forest, Caribou Rock Trail,

on vertical east-facing side of Caribou Rock, 48.06380°N 90.45655°W, 560 m, on

unknown crustose lichen and Aspicilia species and independent on diabase, 6 August

2017, P.A. Scott 5416 (Holotype, PRM!)

Erymo .oey: Named in honour of the collector, citizen lichenologist Perry Scott, who

collected the new species.

HyYPoTHALLus endolithic, continuous with medulla, no algae observed,

broadly and firmly attached or becoming loosely attached and squamulose,

replicating by division. THaLLus of areoles <1.3 mm wide, round to angular,

flat to bullate, 300-500 um thick, broadly attached, replicating by division.

Upper surface dark brown, shiny, epruinose, black edged. Lower surface

usually dark brown. Epicortex 5 um thick, continuous. Cortex 30-50 um

thick, of distinct vertical hyphae, 2-3 um wide, breaking up into irregular

cells, terminating in expanded apices, 4-5 um wide in dark brown gel cap,

upper layer of cortex dark brown, c. 10 um high, lower layer hyaline. Algal

layer continuous without hyphal bundles, 50-100 um thick, algal cells

10-15 um wide. Medulla 150-300 um thick, hyphae thin-walled, 2-3 um

diam.

APOTHECIA usually 1 per areole, rarely 2 to 6, immersed <1 mm wide,

the disc epruinose, rugulose, usually round. Parathecium indistinct or

narrow to 15 um wide, intergrading with cortex. Epihymenium 10-20 um

thick, coherent, brown to yellowish brown, sealed with polysaccharide

hyaline layer c. 5 um, a continuation of the epicortex. Hymenium (including

epihymenium) 100-120(-150) um tall, highest in bullate areole, hymenial

gel IKI+ blue (euamyloid). Paraphyses 1.5-2 um diam. at mid-level, apices

expanded to 3 um in pigment cap with blackish pigment mark, often with

abundant oil drops. Asci 70-90 x (8-)10-12(-15) um, narrow, c. 100

ascospores per ascus. Ascospores 3-4(-5) x 1.5-2 um. Subhymenium 30-50

um thick, IKI+ blue (euamyloid). Hypothecium 10 um wide. Pycnrp14 not

observed. Spot tests negative, K-, KC-, C-, P-.

SELECTED ADDITIONAL SPECIMENS EXAMINED: CANADA. Ontario. Thunder Bay

District, Sleeping Giant Provincial Park, by shore of Lake Superior, 48.31463°N

88.88739°W, 190 m, on diabase bedrock and on Aspicilia species, 7 September 2019,

456 ... Knudsen & Kocourkova

PA. Scott 7030 & J. Hollinger (hb. Scott). USA. MINNEsoTa, Cook Co., Boundary

Waters Canoe Area Wilderness, near west end of Mountain Lake, 48.09710°N,

90.28500°W, 595 m, on Aspicilia species on northwest facing diabase rocks near top of

cliff, 15 August, 2019, P.A. Scott 6837 (hb. Scott), Superior National Forest, southwest-

facing cliff overlooking Moss Lake, 48.0654°N 90.4611°W, on diabase, 5 August 2017,

P.A. Scott 5405A (hb. Scott), Caribou Rock, Caribou Rock Trail, on exposed vertical

east-facing rock near top of cliff, 48.06380°N, 90.45640°W, 560 m, on Aspicilia species

and on diabase, 11 August 2019, P.A. Scott 6767 (NY), 6768 (hb. Scott).

ECOLOGY & DISTRIBUTION. Epilithic on diabase and/or parasitic on crustose

lichens in North America. Diabase sills form the caps of the cliffs in this area

of Minnesota. Expected to be more widespread in United States and Canada

on other intermediate siliceous rocks poor in SiO,, possibly also on more

acid rock. Specimens in herbaria are probably identified as A. americana

H. Magn. or A. veronensis A. Massal.

Discussion. Acarospora scottii is similar to A. americana but differs in

having areoles that are not dull brown but shiny due to a continuous

polysaccharide epicortex and in having a consistent black margin around

the edge of areole and becoming squamulose with dark brown to black

lower surface. It also differs in having euamyloid hymenial gel in Lugol's vs.

hemiamyloid hymenial gel in Lugol's and in having a cortex of vertical or

irregular hyphae vs. distinctly paraplectenchymatous hyphae (Knudsen & al.

2011). Acarospora americana has a parathecium that usually widens around

the apothecial disc, even forming an elevated parathecial crown the same

color as thallus. Acarospora scottii has an indistinct or narrow parathecium,

often intergrading with the cortex, but in some specimens may have a raised

thalline collar surrounding and higher than the apothecial disc.

Acarospora veronensis differs from A. scottii in having a narrow cortex

(c. 15 um vs. 30-50 um thick) and in having hemiamyloid hymenial gel in

Lugol's.

Acarospora scottii is a typical facultative lichenicolous lichen. Its non-

lichenized hypothallus spreads through the upper layer of the rock. In

patches of bare rock, the hypothallus parasitizes algae forming an epilithic

lichen thallus. Spreading under crustose lichens, the hypothallus penetrates

them from below, expropriating the algal photobiont from the host,

eventually destroying the mycobiont. It morphs out of the host forming an

epilithic lichen thallus. In A. scottii, the areoles that emerged from crustose

lichens are much more robust, thicker or bullate than the areoles growing on

bare rock (Fics 1, 2). This suggests that not only are they receiving nutrition

from the photobiont of the host but also from the saprobic absorption of the

Acarospora & Sarcogyne spp. nov. (USA & Canada) ... 457

Fic. 1. Acarospora scottii (holotype, PRM - Scott 5416). A. Areoles emerging from unknown

crustose lichen; B. Thick or bullate areoles emerging from Aspicilia thallus. Scale bars = 1 mm.

mycobiont of the host. Such a difference is not distinctive in other facultative

lichenicolous lichens we have studied, for instance A. schleicheri (Ach.)

A. Massal. Like most facultative lichenicolous lichens it is opportunistic and

458 ... Knudsen & Kocourkova

Fic. 2 Acarospora scottii (holotype, PRM — Scott 5416).

Areolate thallus with flat areoles growing on bare rock. Scale bar = 1 mm.

not host specific (in holotype it occurs on Aspicilia species and unknown

crustose lichen). The paratype was non-lichenicolous.

Areoles in the holotype were often sterile or with only a few mature

asci. The holotype was collected during summer in Minnesota, suggesting

ascospores are released seasonally, possibly early spring, at least in Minnesota.

Sarcogyne paradoxa Kocourk. & K. Knudsen, sp. nov. Fig. 3

MB 831963

Differs from Acarospora subfuscescens by its reddish brown to brown apothecia that

remain brown when hydrated.

Type: USA, California, San Bernardino Co., Joshua Tree National Park, below Belle

Mountain, in unnamed wash, 34.0189°N 116.0025°W, 1382 m, common on granite

boulder on wash, apothecia emerging from rock, 22 August 2005, K. Knudsen 3620

(Holotype, UCR; isotype, ASU).

EtyMo.oecy: Named because it grows as either an endolithic lichen or a lichenicolous

fungus.

THALLUS in non-parasitic specimens endolithic, the algal layer occurring in

the substrate below the apothecia beneath a mycelial base continuous with

hypothecium. In parasitic specimens endokapylic, no algal layer occurring in

substrate below host, apothecia emerging from the host.

Acarospora & Sarcogyne spp. nov. (USA & Canada) ... 459

Fic. 3 Sarcogyne paradoxa (holotype, UCR - Knudsen 3620). A. Habit of the thallus in rock

crevices (with Lichenothelia species); B. Emerging and developed apothecia; C. Apothecia with

conspicuous elevated margins; D. Fully developed apothecium with rugose surface. Scale bars:

A = 1mm; B, C = 500 um; D = 200 um.

460 ... Knudsen & Kocourkova

APOTHECIA superficial on host or occurring on substrate, brown when

dry or wet, 0.5-1.0 mm diam, round or irregular, dispersed or contiguous,

forming clusters of apothecia to 3 mm wide; disc brown to reddish brown,

convex, with furrows, ridges, and umbos, rough and uneven, margin distinct

but becoming fissured. Parathecium to 100 um wide, hyphae mostly 2-3 um

diam, septate, cells mostly 2-3 um wide, ending in expanded apices forming

outer wall of the margin. Outer layer of margin reddish-brown to dull brown,

20-30 um thick; inner layer hyaline. Epihymenium 10-20 um thick, in

reddish brown gel, accretions on surface of disc to 200 um high formed from

the epihymenial gel. Hymenium 80-120 um tall, hyaline, hymenial gel IKI+

deep blue (euamyloid), paraphyses coherent (1-)2-3 um diam, sometimes

constricted at septa, infrequently branching in lower half, unexpanded or

expanded apices sometimes in darker pigment caps. Asci 60-90 x 12-20 um,

ascospores mostly 100 per ascus, ascospores (3.0—)4.0—4.5(-7) x 2.0-3.0 um,

mostly broadly ellipsoid. Subhymenium hyaline, 30-50 um thick, IKI+ blue.

Hypothecium up to 50 um thick, continuous with vegetative vertical hyphae

forming a mycelial base above algal layer in substrate. PYcNIDIA not seen.

Spot tests C-, KC-, K-. P-.

SELECTED ADDITIONAL SPECIMENS EXAMINED: CALIFORNIA, San Bernardino Co.,

Granite Mountains: Sweeney Granite Mountains UC Reserve, Sibyl Allison trail,

34.7892°N 115.6717°W, 1692 m, on granite, 14 December 2007, K Knudsen 8050 &

R. Muertter (UCR). Santa Barbara Co., Channel Island National Park, Santa Rosa

Island, south slope of South Peak, 33.9064°N 120.1283°W, 243 m, on unknown host

and independent on sandstone, 15 August 2007, K. Knudsen 8762.2 (UCR). San Luis

Obispo Co., San Simeon, San Simeon State Park, Molinari Property, exposed slab of

volcanic rock next to ravine along old Highway One dirt road, 35.6039°N 121.1275°W,

41 m, on crustose lichen, 8 January 2007, K. Knudsen 8116 (UCR)

ECOLOGY & DISTRIBUTION. Occurring in siliceous rock, usually granite,

as an endolithic lichen or endokapylic in the thallus of crustose lichens

as a non-lichenized parasite in North America (California) and Asia

(Afghanistan, Xinjiang in China) (Knudsen & Kocourkova 2008, 2009;

L. Nurtai, pers comm.) The specimen from Afghanistan at GZU is a parasite

on an unknown lichen on granite. The specimens from Xinjiang are endolithic

lichens on granite. Based on the distribution of Acarospora gyrocarpa

(H. Magn.) K. Knudsen & M. Westb. [= Polysporina gyrocarpa (H. Magn.)

N.S. Golubk.] in China and southwestern United States, S. paradoxa is expected

like A. gyrocarpa to be widespread but probably rare in the Chihuahuan,

Mojave, and Sonoran Deserts in southwestern North America. (Knudsen &

Kocourkova 2009). The species is easily over-looked in field inventories.

Acarospora & Sarcogyne spp. nov. (USA & Canada) ... 461

Discussion. Sarcogyne paradoxa differs from the lichenicolous fungus

Acarospora subfuscescens (Nyl.) H. Magn. in having brown apothecia with

epihymenial accretions that are brown when hydrated instead of dark

brownish black to black apothecia with epihymenial accretions that are

black when hydrated (Knudsen & Kocourkova 2008). Sarcogyne paradoxa

was treated previously as Polysporina arenacea (H. Magn.) K. Knudsen &

Kocourk. [a misapplication] and was only subsequently recognized as a

lichenicolous fungus (Knudsen & Kocourkova 2008). Acarospora arenacea

H. Magn. is a rare lichen known only from Utah and North Dakota, USA

(Magnusson 1952). In phylogenetic analyses S. paradoxa falls into the

Sarcogyne lineage (Westberg & al. 2015, Knudsen & al. 2019).

Sarcogyne paradoxa has a different lifestyle from lichenicolous lichens

like the obligatory Diploschistes muscorum or the facultative Acarospora

scottii. It can be a lichen; the hyphae of the hypothallus parasitizes algae

occurring in rock in the upper 1 mm of the substrate, forming an endolithic

lichenized thallus that eventually produces epilithic apothecia. Or it can

be a lichenicolous fungus, the endolithic hypothallus apparently entering

the host from beneath the thallus. It does not expropriate the algae of

the host like a lichenicolous lichen, eventually forming an independent

epilithic thallus. Like many lichenicolous fungi, it expropriates a portion of

the carbon produced by the photobiont of the lichen host but maintains a

symbiotic relationship, not destroying the host. In the few specimens seen, it

does appear to suppress the host forming apothecia, which would conserve

for its nutrition carbon produced by the photobiont of the host that would

otherwise nourish the sexual reproduction of the host.

During our work on the genus Polysporina Vézda, we used broad

morphological and anatomical species concept of Polysporina subfuscescens

(Nyl.) Knudsen & Kocourk. for identifying taxa growing on lichens and

a broad concept of Polysporina simplex (Borrer ex Hooker) Vézda for

identifying lichenized taxa growing on rock (Knudsen & Kocourkova 2008;

Westberg & al. 2015). Based on phylogenetic analyses of a large sampling

of specimens, Polysporina was found to be polyphyletic with most taxa

occurring in several different lineages of Acarospora (Westberg & al. 2015). In

that tree, the lineages Polysporina A and Polysporina B contained specimens

identified as P subfuscescens (lichenicolous fungus) and P._ simplex

(endolithic lichen). These two lineages apparently represent single species,

both of which either occur as a lichenicolous fungus or an endolithic lichen

like Sarcogyne paradoxa. Because Polysporina is polyphyletic and negated as

462 ... Knudsen & Kocourkova

a monophyletic genus, we treat the taxa as Acarospora or Sarcogyne based on

their position in the current phylogenetic trees of Acarosporales (Westberg

& al. 2015; Knudsen & al. 2019). Acarospora subfuscescens and Acarospora

simplex (Borrer ex Hooker) Jatta are heterogeneous in these phylogenetic

analyses and obviously in need of a revision (Gueidan & al. 2014, Westberg

& al. 2015).

Acknowledgments

We thank our reviewers, John McCarthy, S.J. (Canada) and Alejandro Delgado

(Mexico). For her assistance we thank J.N. Adams (UCR) and the curators of ASU

& LD. The work of Kerry Knudsen and Jana Kocourkova was financially supported

by the grant of Ministry of Education, Youth and Sports of the Czech Republic, the

program of international cooperation between the Czech Republic and U.S.A. for

research, development and innovations INTER-EXCELLENCE, INTER-ACTION,

no. LTAUSA18188.

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MYCOTAXON

April-June 2020—Volume 135, p. 465

https://doi.org/10.5248/135.465

Regional annotated mycobiota new to the Mycotaxon website

ABSTRACT—We are pleased to announce that we uploaded our 138th free access

funga to the MycoTaxon mycobiota site in May. Everyone is invited to download the

14-page “A checklist of marine fungi from Australia” by Fryar, Hyde, and Catcheside

via: http://www.mycotaxon.com/mycobiota/index.html

AUSTRALASIA

Australia

SALLY C. FryAR, KEvIN D. Hype, Davip E.A. CATCHESIDE. A checklist of

marine fungi from Australia. 14 p.

AssTRACT—A checklist of non-lichenized marine fungi in Australia is

presented. From 1954 to 2020 a total of 121 species in 93 genera were observed.

Most species were ascomycetes, with only three basidiomycetes. Of these

ascomycetes, most were seen in their sexual state (93 species) compared with

their asexual state (21 species).

Kry worps—biodiversity, fungal diversity

MYCOTAXON

April-June 2020—Volume 135, p. 467

https://doi.org/10.5248/135.467

Regional annotated mycobiota new to the Mycotaxon website

ABSTRACT—MYCOTAXON announces the posting of its 139th free access funga to

its mycobiota site in June. The 42-page checklist of Brazilian corticioid fungi by

Chikowski, Lira, Larsson, and Gibertoni may be downloaded from our website via

http://www.mycotaxon.com/mycobiota/index.html

SOUTH AMERICA

Brazil

RENATA S. CHIKOWSKI, CARLA R.S. DE LIRA, KARL HENRIK LARSSON,

TATIANA B. GIBERTONI. A checklist of corticioid fungi (Agaricomycetes,

Basidiomycota) from Brazil. 42 p.

ABSTRACT— This study updates the available data about species of corticioid

fungi from Brazil. We based our compilation on literature, herbarium revisions,

and added information from recent field trips in the Atlantic Rain Forest,

Amazonia, Caatinga, and Cerrado phytogeographic domains. Currently,

there are 367 accepted species of corticioid fungi recorded in Brazil. Those

representing new records include four for the Neotropics, 15 for the country,

18 for Northeast Brazil, and 41 for Brazilian phytogeographic domains.

KEY worps—diversity, Hymenochaetales, Polyporales, resupinate fungi,

Russulales

MYCOTAXON

April-June 2020—Volume 135, p. 469

https://doi.org/10.5248/135.469

Regional annotated mycobiota new to the Mycotaxon website

ABSTRACT—MYCOTAXON announces the posting of its 140th free access funga to its

mycobiota site in June. This excellent 18-page global checklist of aquatic arbuscular

mycorrhizal fungi by Queiroz, Jobim, Vista, Leroy, Gomes, and Goto may be

downloaded from our website via http://www.mycotaxon.com/mycobiota/index.html

GLOBAL

MarIANA Bessa DE QUEIROZ, KHADIJA JOBIM, XOCHITL MARGARITO VISTA,

JULIANA APARECIDA SOUZA LEROY, STEPHANIA RUTH BASILIO SILVA

GomEs, BRUNO TomIo GoTo. Occurrence of Glomeromycota species in

aquatic habitats: a global overview. 18 p.

AxssTRACT—Arbuscular mycorrhizal fungi (AMF) are recognized in terrestrial

and aquatic ecosystems. ‘The latter, however, have received little attention from

the scientific community and, consequently, are poorly known in terms of

occurrence and distribution of this group of fungi. This paper provides a global

list on AMF species inhabiting aquatic ecosystems reported so far by scientific

community (lotic and lentic freshwater, mangroves, and wetlands). A total of

82 species belonging to 5 orders, 11 families, and 22 genera were reported in 8

countries. Lentic ecosystems have greater species richness. Most studies of the

occurrence of AMF in aquatic ecosystems were conducted in the United States

and India, which constitute 45% and 78% reports coming from temperate and

tropical regions, respectively.

Key worps—checkilist, flooded areas, mycorrhiza, taxonomy