IV ... MYCOTAXON 134(2)

MYCOTAXON

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

134-2: CONTENTS, NOMENCLATURAL UPDATES, ERRATA, PEERS, EDITORIAL

Nomenclatural novelties Ot ypifications ... .. sah. osx poles He ke ees vii

PVT CES Mme Oh 5 ean Sa rags citer AM yah ACO Pea e eet «athe Oe ix

EPR a5 are ah hei eo Ek otek ark ah Lik ati hele + Riri eet EARL toot te EE x

TROVE ESO REOR © co. Red aecgs ving Bes ernest coat echeseaic A la Th dee Hari ean as xi

LAWL SUOHAESSTONSPTOCCIULE a5 Sy satis, alcklack Sethayd hs PUG Deneck efechnd tate AI xiii

TAXONOMY

Volvate Macrolepiota from Brazil: Macrolepiota dunensis sp. nov.,

M. sabulosa var. velistellaris var. nov., and observations on

M. pulchella DALINE SOARES FREITAS & NELSON MENOLLI JR. 223

Inocybe caroticolor from oak forests of Pakistan

A. NASEER, S. GHANI, A. R. Nrazi, A.N. KHALID 241

Sporidesmium horizontale sp. nov. from China

CoNnG-ConcG AI, JI-WEN XIA, XIU-GUO ZHANG, YUN GENG, L1-Guo Ma 253

Sporidesmiella corniformis sp. nov. from China

CoNnG-CongG AI, JI-WEN XIA, XIU-GUO ZHANG, YUN GENG, LI-Guo Ma 257

New records of Amandinea and Buellia from China

ZUN-TIAN ZHAO, XIAO ZHANG, JIE-MENG Fu, LING Hu 261

Introduction of subfamily names for four clades in

Cladoniaceae and Peltigeraceae (Lecanoromycetes)

H. THORSTEN LUMBSCH & STEVEN D. Leavitt 271

Cacumisporium fusiforme sp. nov. from Jiangxi, China

ZHAO-HuAN Xu, Kal ZHANG, XIU-GUO ZHANG,

RAFAEL F. CASTANEDA-RUIZ, JIAN Ma 275

Acarospora squamulosa, the correct name for A. peliocypha

KERRY KNUDSEN, LINDA IN ARCADIA, JANA KocourKovA 281

Megalaria yunnanensis sp. nov. from Yunnan, China

CHUN-XIAO WANG, XIAO ZHANG, CHUAN-FENG ZHENG, LING Hu 289

The conserved type of Lichen fuscatus [= Acarospora fuscata]

KERRY KNUDSEN, JIRi MALICEK, JANA KOCOURKOVA 295

Beltraniopsis cyclobalanopsidis sp. nov. from Guizhou, China

ZHONG-JIU XIAO, XIAO-XIA LI, CHANG-HoNG CHu,

TinG Liu, ZHENG-ZHENG Lu, PEI- YONG SonG 301

APRIL-JUNE 2019... V

Perichaena acetabulifera sp. nov. from Juarez City, Mexico

Marcos LizARRAGA, GABRIEL MORENO, IRAIS FLORES-ROMERO 307

Gliocladiopsis wuhanensis sp. nov. from China

NI-PinG ZHAI, ZI-QUAN SUN, YA-LONG ZHANG, RUI ZANG,

CuaAo Xu, YUE-HuaA GENG, MENG ZHANG 313

Four new records of Haematomma from Southern China

RONG TANG, XIAO ZHANG, CHUN-XIAO WANG, Lu-LU ZHANG 321

Cordana meilingensis and C. lushanensis spp. nov. from Jiangxi, China

CoNnG-ConcG AI, JIAN Ma, KAI ZHANG,

RAFAEL F. CASTANEDA-Ruiz, X1U-GuUO ZHANG 329

Coprophilous fungi from Brazil: new records for the Neotropics

ROGER ER. MELO, DANIEL B.P. Do Monte, NICOLE H.B. GONDIM,

LEONOR C. Mata, ANDREW N. MILLER 335

Two Heterostelium species newly recorded from China

Pu Liu, SHUNHANG ZHANG, XIAOYAN ZHOU, JIANJUN ZHAO, Yu Li 353

Pseudocercospora meliosmicola sp. nov. and three new Pseudocercospora

records from China Qian ZHAO, Bao-Ju Li, YAN-X1A SHI,

XUE-WEN XIE, A-L1 CHAI, YING-LAN Guo 359

Heteroplacidium compactum reported as a genus new to China

XIANG-MIN CHENG, Da-LE Liu, XIN-LI WEI, JIANG-CHUN WEI 369

Geopora ahmadii sp. nov. from Pakistan

M. Sasa D. HAELEWATERS, T. ITURRIAGA,

T. ASHRAB, A.N. KHALID, D.H. PFISTER 377

Seychellomyces sinensis sp. nov. from China

MIN Qiao, Hua ZHENG, ZHE ZHANG, ZE-FEN Yu 391

Ramaria flavescentoides sp. nov. with clamped basidia from Pakistan

MUHAMMAD Hanlr, ABDUL NASIR KHALID, RONALD L. EXETER 399

MycosBioTa (FUNGA) NEW TO THE MYCOTAXON WEBSITE

Aphyllophoroid fungi in Teide National Park

(Tenerife, Canary Islands) [summary]

ESPERANZA BELTRAN-TEJERA, JESUS LAURA RODRIGUEZ-ARMA,

MIGUEL JONATHAN Diaz ARMAS, LUIS QUIJADA 407

BOOK REVIEWS AND NOTICES ELSE VELLINGA 409

vi ... MYCOTAXON 134(2)

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

MYCOTAXON for JANUARY-MARCH 2019 (I-xII + 1-222)

was issued on March 27, 2019

APRIL-JUNE 2019... VII

NOMENCLATURAL NOVELTIES AND TYPIFICATIONS

PROPOSED IN MYCOTAXON 134(2)

Beltraniopsis cyclobalanopsidis Z.J. Xiao & Xiao X. Li

[FN 570598], p. 306

Cacumisporium fusiforme Z.H. Xu, Jian Ma, X.G. Zhang & R.F. Castaneda

[MB 829269], p. 276

Cordana lushanensis C.C. Ai, Jian Ma, X.G. Zhang & R.E Castaneda

[LIF 555814], p. 332

Cordana meilingensis C.C. Ai, Jian Ma, X.G. Zhang & R.F Castafieda

[IF 555813], p. 330

Gliocladiopsis wuhanensis Meng Zhang, N.P. Zhai & Y.H. Geng

[MB 825569], p. 316

Geopora ahmadii Saba, T. Ashraf, Khalid & Pfister

[MB 822666], p. 382

Lichen squamulosus Schrad. 1797 (lectotypified), p. 282

Lobarioideae Lumbsch & S.D. Leav.,

[MB 830563], pe272

Macrolepiota dunensis D.S. Freitas & Menolli

[MB 823088], p. 228

Macrolepiota sabulosa var. velistellaris D.S. Freitas & Menolli

[MB 823089], p.234

Megalaria yunnanensis C.X. Wang & L. Hu

[MB 829259], p. 290

Nephromatoideae Lumbsch & S.D. Leav.,

[MB 830564], p. 272

Perichaena acetabulifera Lizarraga, G. Moreno & Flores-Rom.

[MB 829449], p. 308

Pseudocercospora meliosmicola Y.L. Guo, Qian Zhao & Y.X. Shi

[MB 823728], p. 364

Ramaria flavescentoides Hanif & Khalid

[MB 823965], p. 402

Seychellomyces sinensis Z. F. Yu & Hua Zheng

[MB 829084], p. 395

Sporidesmiella corniformis L.G. Ma, C.C. Ai & X.G. Zhang,

[MB 830544], p. 258

Vul ... MYCOTAXON 134(2)

Sporidesmium horizontale L.G. Ma, C.C. Ai & X.G. Zhang

[[MB 830543], p. 254

Squamarinoideae Lumbsch & S.D. Leav.,

[MB 830561], p. 272

Stereocauloideae Lumbsch & S.D. Leav.,

[MB 830562], p. 272

APRIL-JUNE 2019...

REVIEWERS — VOLUME ONE HUNDRED THIRTY-FOUR (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.

Laise de Holanda Cavalcanti Andrade

Ditte Bandini

Annarosa Bernicchia

Uwe Braun

Irwin M. Brodo

Rafael FE. Castahteda-Ruiz

Maria Martha Dios

John A. Elix

Ana Esperanza Franco Molano

Masoomeh Ghobad-Nejhad

Shouyu Guo

Rosanne Healy

Brendan Hodkinson

Sana Jabeen

Ze-Feng Jia

Alfredo Justo

Gintaras Kantvilas

John C. Landolt

Steve Leavitt

James C. Lendemer

De-Wei Li

Jian Ma

Li-Guo Ma

Josiane Santana Monteiro

Lorelei L. Norvell

Shaun R. Pennycook

Christian Printzen

Jun-Zhi Qiu

Leif Ryvarden

Matthew E. Smith

Steven L. Stephenson

Andrei Tsurykau

Nadja Santos Vitoria

Ping Zhang

Ying Zhang

x ... MYCOTAXON 134(2)

ERRATA

Mycotaxon now includes within this section not only errata from past volumes but

also corrections of mistakes in the current issue that were present in approved author

input files but not detected by authors until after PDF conversion. PDFs will be corrected

at deadline for no charge only when they result from editorial error or when authors

agree to pay $40 per correction. This section accommodates all other corrections.

VOLUME 134 (1)

p- 120, line 2-3 DELETE: Mains 1954

VOLUME 134 (2) [CURRENT ISSUE]

p. 225, line 17 FOR: “F The nrITS region was amplified ...”

READ: “The nrITS region was amplified ...”

p. 314, line 8 FOR: wheat-straw.; READ: wheat-straw;

p- 316, line 26 for: COLONIES read: COLONIES

(the first letter in ‘Colonies’ should be in black font)

APRIL-JUNE 2019... XI

FROM THE EDITOR-IN-CHIEF

NEW TO THE EDITORIAL ADvIsORY BOARD—We are pleased to announce the

appointment of two new editorial board members. Former Mycotaxon Book

Review Editor and University of California researcher, Dr. ELSE C. VELLINGA (who

still consents to deliver reviews on a gratifyingly regular basis), is well known to

agaricologists through her numerous contributions to the taxonomy of ‘lepiotaceous

fungi and as contributing editor to the FLORA AGARICINA NEERLANDICA series.

Well-schooled in phylogenetic analysis as well as biodiversity and field-work, Else

is an active member of the North American Mycoflora Project. Mycologist Dr.

Topp OsmMuUNDSsON, Associate Professor of Biology at the University of Wisconsin

Lacrosse, also participates in the North American Mycoflora Project. He offers

expertise in systematics, biogeography, bioinformatics, DNA barcoding, and

molecular phylogenetics and consistently receives high ratings from his students.

The new appointees join Dr. P. BRANDON MATHENY (University of Tennessee,

Knoxville USA) and Dr. KAREN HANSEN (Curator, Swedish Museum of Natural

History (S), Stockholm, Sweden). The newly reconstituted board, which advises

MycoTaxon on editorial matters, will elect an two additional members this year.

ELIMINATION OF ‘MyCOBANK, “‘INDEXFUNGORUM, “FUNGAL NAME’ PRECEDING

IDENTIFIERS—Members of the Nomenclature for Fungi Committee have

discussed the custom of associating numerical fungal identifiers with the name of

the assigning database. Because each number is unique, it is not really necessary

to designate the issuing nomenclatural registry. Even though your editors and

many researchers find such database designations helpful, we agree that it is not

necessary to precede an identifier with both name and initials. MycoTAxon has

decided to compromise, and henceforth we will cite only the abbreviations FN, IF,

and MB, replacing the redundancies FUNGAL NAME FN, INDEXFUNGORUM IF, and

MycoBank MB. This allows us to focus properly on the number—the formal (and

unique) identifier.

NOT NECESSARILY “FIRST COME, FIRST SERVE’—Your volunteer two-person

editorial team has long tried to review manuscripts in the order in which they

are received. Although we view ‘fast-tracking’ manuscripts as inherently unfair,

the Ep1ror-IN-CHIEF has recently begun urging the NOMENCLATURE EDITOR—

our first line of defense against incompetent submissions—to ‘cherry-pick’ well-

prepared work from later submissions to fill out an issue in a timely manner. The

situation became even more urgent last year: after we eliminated required page

charges, our submissions trebled. For that reason, Shaun, MycoTaxon’s primary

defensive lineman, announces our revised review policy:

“We attempt to review manuscripts in the order in which they were accessioned.

However, in order to meet publication deadlines, badly written and poorly

prepared manuscripts that do not follow MycotTaxon’s formatting instructions

xl ... MYCOTAXON 134(2)

(and thus require time-consuming editorial reviews) may be set aside in favour of

well-presented manuscripts that can be reviewed quickly and easily,’

THE ERRATUM AND HOW IT BREEDS — We have all experienced the embarrassment

of not seeing an error until it pops out on the page of a published manuscript. We

all (including this Ep1ror-1N-CHIEF) make mistakes, and it seems only reasonable

to expect them to be corrected when reported prior to publication. From 2004-2009

your Editor spent considerable time at the close of each issue reprocessing PDFs

when authors ‘suddenly’ discovered an error that had been in their own text files

throughout the entire review process. In 2011, after publication had sometimes been

delayed by a week or more while she opened six or more files, corrected mistakes and

entries, re-processed PDF proofs, and corresponded with the authors and the press,

she announced that we would EITHER publish corrections of errors the authors

should have seen in the errata of later volumes for no charge OR correct editorial

conversion error prior to publication, also at no charge. Having encountered a

number of rather major author errors, this year we initiated a third alternative:

authors may pay $40 to have any major author error corrected at press time.

We now offer a fourth alternative: to issue corrections of minor author errors in

the same issue in which the paper is published. This option both saves considerable

editorial time at deadline and provides the correct text promptly at no charge.

MYCOTAXON 134(2) contains 22 papers by 88 authors (representing 10 countries)

as revised by 32 expert reviewers and the editors.

The 2019 April-June MycoTaxon proposes 4 new subfamilies—Lobarioideae,

Nephromatoideae, Squamarinoideae, Stereocauloideae—in the Lecanoromycetes plus

15 other taxa new to science representing Beltraniopsis, Cacumisporium, Cordana,

Gliocladiopsis, Megalaria, Pseudocercospora, Seychellomyces, Sporidesmiella, and

Sporidesmium from CHINA; Geopora and Ramaria from PAKISTAN; Macrolepiota

from BRAZIL; and Perichaena from Mexico. We also publish the lectotypification

of Lichen squamulosus, basionym of Acarospora squamulosa.

New range extensions reported include Amoebozoa: Heterostelium for

CHINA; Ascomycetes: coprophils for BRAzIL and Pseudocercospora for CHINA;

Basidiomycetes: Inocybe for Paxistan; and the lichens Amandinea, Buellia,

Haematomma, and Heteroplacidium for CHINA.

Also included are keys to Beltraniopsis and Cacumisporium species, a list of new

GenBank sequences from the newly conserved holotype Lichen fuscatus (basionym

of Acarospora fuscata), and a summary of the new funga covering aphyllophoroid

fungi of the Canary Islands, uploaded to the MycoTaxon mycobiota website in May.

Warm regards,

Lorelei L. Norvell (Editor-in-Chief)

Shaun R. Pennycook (Nomenclature Editor)

28 June 2019

APRIL-JUNE 2019 ... XIII

2019 MyYCOTAXON SUBMISSION PROCEDURE

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

and subscription articles are offered.

MY COTAXON

April-June 2019—Volume 134, pp. 223-239

https://doi.org/10.5248/134.223

Volvate Macrolepiota from Brazil: M. dunensis sp. nov.,

M. sabulosa var. velistellaris var. nov.,

and observations on M. pulchella

DALINE SOARES FREITAS*” & NELSON MENOLLI JR.”

' Depto de Ciéncias e Matematica (DCM) / Biologia,

Instituto Federal de Educacdo, Ciéncia e Tecnologia de Sao Paulo (IFSP),

Campus Sao Paulo, Rua Pedro Vicente 625, 01109-010 Sao Paulo, SB, Brazil

? Nucleo de Pesquisa em Micologia, Instituto de Botanica,

Av. Miguel Stefano,3687, Agua Funda, CEP: 04301-012, Sao Paulo, SP, Brazil

“ CORRESPONDENCE TO: menollijr@yahoo.com.br

ABSTRACT—Taxonomic and phylogenetic studies of volvate Macrolepiota specimens

collected from Atlantic Forest in Northeast and Southeast Brazil are presented based

on morphological and molecular (nrITS) data. Macrolepiota dunensis and M. sabulosa

var. velistellaris are proposed as new taxa, and additional observations are presented

about M. pulchella, a species previously known from Brazil. Macrolepiota dunensis and

M. pulchella clustered in the clade corresponding to M. sect. Volvatae. Macrolepiota

sabulosa var. velistellaris was placed together with non-volvate M. sabulosa var. sabulosa,

close to another volvate taxon (M. rhodosperma var. velicopia), and together with species

classified in M. sect. Macrolepiota. Our phylogenetic results provide additional evidence

that the volva, which is not restricted to members of M. sect. Volvatae, evolved more than

once in the genus.

Key worps—Agaricaceae, Lepiotella, Neotropics, South America, Volvolepiota

Introduction

Macrolepiota Singer (Agaricaceae) was proposed by Singer (1948) to

accommodate specimens characterized by giant basidiospores that are

metachromatic in Cresyl blue, a white to cream spore print, and hyphae

with clamp connections. Singer (1959) proposed Volvolepiota Singer for the

224 ... Freitas & Menolli

illegitimate genus Lepiotella Rick (Rick 1938), separated from Macrolepiota

by the presence of a distinct volva. At present morphological and molecular

data (Vellinga 2003; Vellinga & al. 2003) treat Macrolepiota and Volvolepiota

as synonyms, with Macrolepiota restricted to specimens that in addition to

Singer’s diagnostic characters (1948, 1986) possess a trichodermal pileus

covering composed of long and often thick-walled elements, a stipe covered

with hymeni-trichodermal patches often arrayed in colored bands, and

basidiospores with a rounded apex and a germ pore covered by a hyaline cap.

The current concept of Macrolepiota also includes sequestrate species (Lebel &

Syme 2012). Supported by nrITS sequence analyses, Ge & al. (2010) proposed

Macrolepiota sect. Volvatae Z.W. Ge & al. to accommodate volvate species with

small amygdaliform-ellipsoid basidiospores and no clamp connections.

Morphological and molecular studies support the existence of only five

volvate Macrolepiota, distributed mainly in tropical regions: M. brunnescens

Vellinga from Argentina (Vellinga & Yang 2003; Singer 1959 as Volvolepiota

albida); M. eucharis Vellinga & Halling from Australia (Vellinga 2003);

M. pulchella from Brazil (Vellinga & Yang 2003, de Meijer 2006; Rick 1938,

1961, as Lepiotella brunnea; Singer 1954, 1959, Heinemann & de Meijer 1996,

Bononi & al. 1981, as V. brunnea); M. rhodosperma var. velicopia Vizzini &

Contu from Italy (Vizzini & al. 2011); and M. velosa Vellinga & Zhu L. Yang

from China (Vellinga & Yang 2003).

Ge & al. (2010), who included the volvate species Macrolepiota velosa

and M. eucharis in M. sect. Volvatae, suggested that M. brunnescens and

M. pulchella might also belong to this section. The description of a volvate

taxon (M. rhodosperma var. velicopia) not phylogenetically related to

M. sect. Volvatae representatives (Vizzini & al. 2011) provoked a new discussion

about recognizing a natural classification of M. sect. Volvatae based on velar

structures.

The only volvate Macrolepiota recorded from Brazil is M. pulchella, known

from the southern and southeastern states of Rio Grande do Sul (Rick 1938,

1961, as L. brunnea; Singer 1954, 1959, as V. brunnea), Sao Paulo (Bononi &

al. 1981, as V. brunnea), and Parana (de Meijer 2006; Heinemann & de Meijer

1996, as V. brunnea).

Here we report morphological and molecular data for three volvate

Macrolepiota taxa based on specimens collected in the Atlantic Forest of Rio

Grande do Norte (Northeast Brazil) and Sao Paulo (Southeast Brazil). Two—

M. dunensis and M. sabulosa var. velistellaris—are described as new, and we

discuss the taxonomy and phylogenetic position of the third, M. pulchella.

Macrolepiota dunensis sp. nov., M. sabulosa var. velistellaris var. nov. (Brazil) ... 225

Materials & methods

Morphological studies

The macroscopic and ecological descriptions are based upon collector's notes

from fresh materials. Color terms follow Kiippers (1979). For microscopic analyses,

sections of lamellae, pileus covering, and stipe were moistened in 70% ethanol and

rehydrated in 5% KOH. Reagents used were Congo red to stain hyaline structures,

Cresyl blue to test metachromasy, and Melzer’s reagent to test basidiospore amyloidy.

Microscopic illustrations were made with the aid of a drawing tube attached

to an Olympus BX50 compound microscope. For basidiospore measurements,

cc 9D

[a/b/c] indicates “a” basidiospores measured from “b” basidioma(ta) taken from

“c’ collection(s); Q = length/width quotients from all measured basidiospores,

Qm = average of all computed Q values; Lm (Wm) = average of lengths (widths)

of all measured basidiospores. At least 20 basidiospores from each basidioma were

measured in profile view. Basidiospore shape terminology follows Bas (1969), and

descriptive terms for other morphological features follow Vellinga (1988) and

Vellinga & Noordeloos (2001). The terminology of vegetation types is according to

Veloso & al. (1991), and the herbarium acronyms follow Thiers (2018).

Molecular methods and phylogenetic analyses

DNA extraction, PCR, and sequencing methods followed Justo & al. (2011a,b).

F The nrITS region was amplified and sequenced using the primer pair ITS-1 and

ITS4 (White & al. 1990; Gardes & Bruns 1993). The newly generated sequences were

phylogenetically analysed with other DNA sequences of the main representatives of

Macrolepiota available in GenBank and Leucoagaricus leucothites (Vittad.) Wasser and

L. nympharum (Kalchbr.) Bon as outgroup. GenBank accession numbers and other

collection information for the sequences used in molecular analyses are given in Fic. 1.

Our alignment matrix, generated by MAFFT 7 (http://mafft.cbrc.jp/alignment/server/;

Katoh & Toh 2008) with auto strategy, was visually examined and manually corrected

using Geneious 7.0.6 (Kearse & al. 2012).

To identify the best nucleotide evolution model, we used the AlICc criterion -

Akaike Information Criterion corrected (Hurvich & Tsai 1991) in PartitionFinder

2.1.1 (Lanfear & al. 2017). After separating the ITS region into ITS1+ITS2 and

5.88, we estimated the evolution model for each partition. Maximum Likelihood

(ML) analysis was performed in RAxML v8.2.X (Stamatakis 2014) using the

GTR+G model with a rapid bootstrap analysis with 1000 replicates and search for

the best-scoring ML tree. The Bayesian inference (BI) analysis was performed with

MrBayes v3.2.6 (Ronquist & al. 2012) in the CIPRES Science Gateway 3.1 (Miller &

al. 2010) using two partitions with the TVM+G model for ITS1+ITS2 and the JC

model for 5.88, with two independent runs, four simultaneous independent chains,

and 10,000,000 generations with a sample frequency every 5000 generation. The

following abbreviations are used for statistical data: Bootstrap (BS) and Posterior

Probability (PP).

226 ... Freitas & Menolli

Results & discussion

We would prefer to sample additional collections with a broader

morphological variation to support our new taxa and to provide more

information about the ontogeny and distribution. Nonetheless, because only

five volvate Macrolepiota are currently known and because morphological

and molecular characters support our material as different taxa, we propose

new taxa here to advance Macrolepiota research: M. dunensis as a new

species and M. sabulosa var. velistellaris as a new variety that distinguished

from M. sabulosa var. sabulosa mainly by the presence of a distinct volva on

the stipe base.

Phylogeny

Both ML and BI analyses established the phylogenetic positions of the

three Brazilian volvate Macrolepiota taxa. The ML tree (Fic. 1) organized

the Macrolepiota species in three main clades corresponding to the sections

indicated by Ge & al. (2010)—M. sect. Macrolepiota, M. sect. Macrosporae

(Singer) Bon, and M. sect. Volvatae.

The clade corresponding to Macrolepiota sect. Volvatae includes

sequences from M. dunensis, M. eucharis, M. pulchella, M. velosa, and

other unidentified collections from Brazil and related to M. pulchella. All

identified specimens in this clade possess a distinct volva on the stipe base.

Macrolepiota pulchella clustered with other three sequences of unidentified

specimens from Brazil in a well-supported (100% BS, 1 PP) clade (Fazolino

Perez & al. 2018), with M. dunensis and M. eucharis on an external branch.

Although we still need to review the morphology of the unidentified

collections reported by Fazolino Perez & al (2018) as Macrolepiota sp. 3, our

molecular results indicate that they are most likely M. pulchella. The level of

sequence divergence among M. pulchella and Macrolepiota sp. 3 is up to 1%

and is represented by three different base pairs, with an additional different

base pair in Macrolepiota sp. 3 KY927723 and four ambiguous base pairs in

M. pulchella MC4411.

The other new volvate Macrolepiota (M. sabulosa var. velistellaris) clusters

in the M. sect. Macrolepiota clade with sequences of the non-volvate typical

variety of M. sabulosa Fazolino & R.M. Silveira from Brazil (Fazolino Perez

& al. 2018), and sequences of M. cyanolamellata Fazolino & al. from Brazil

(Fazolino Perez & al. 2018) and M. clelandii Grgur. from Australia and New

Zealand (Vellinga 2003, Vellinga & al. 2003). Although the sequence from

M. sabulosa var. velistellaris is 100% identical to that from the holotype

of M. sabulosa (and its autonymous variety M. sabulosa var. sabulosa)—

Macrolepiota dunensis sp. nov., M. sabulosa var. velistellaris var. nov. (Brazil) ... 227

HM246501 M. procera f fuliginosa*™

0.99/-] HM246503 M. procera f. permixta***

0.97/- HM246502 M. procera f. fuliginosa***

HM125511 M. procera

0.98/93) Aya edad M. procera

90 M. procera

Pevborsie M. procera

0.97/85 AY243588 M. procera

AY243591 M. aff. procera

1/96] HM125514 M. procera

HM125513 M. procera

1/100

0.98/-

HM246505 M. rhodosperma var. velicopia***

HM246507 M. rhodosperma***

HM246506 M. rhodosperma var. velicopia***

AY243598 M. rhodosperma**™*

1/98 | AY243597 M. thodosperma™**

0.98/- AF482841 M. rhodosperma**™*

0.98/75 AY243596 M. rhodosperma***

: U85311 M. colombiana

102 AY083196 M. clelandii

AY083198 M. clelandii

1/100] AY083195 M. clelandii

AY083202 M. clelandii

AF482838 M. clelandii

U85314 M. mastoidea***

U85313 M. mastoidea**™*

0.94/- | ay243599 M. mastoidea***

AF482844 M. mastoidea

HM125529 M. mastoidea

HM125531 M. mastoidea

HM125532 M. mastoidea

AY243603 M. konradii

AY243602 M. konradii

AY243601 M. konradii

0: 9304 AY243003 M. subsquarrosa

we, HM125524 M. orientiexcoriata

"4 HM125528 M. orientiexcoriata

y 1/100 Mic? AF482850 M. orientiexcoriata***

Pos AF482847 M. phaeodisca

182° agg | 8¥243607 M. excoriata

AF482840 M. excoriata

HM246504 M. excoriata

acrosporae

1/99

lacrolepiota sect.

1/100

0.98/90

0.92/71

AF482854 M. eucharis

1/100 0.97/100 NRE 1989 M. velosa

{82853 M. velosa

0.98/90, | Pears M. velosa

IN944095 M. velosa

HM125509 M. velosa

KJ524569 M. velosa

1/100 AF482865 Leucoagaricus nympharum

@ AF482868 Leucoagaricus leucothites

0.06

1/100] | M125518 M. dolichaula

HM125510 M. detersa

1100 - AY243586 M. detersa**

AF482851 M. detersa**

AF243587 M. detersa**

KY927721 Macrolepiota sp.

0.96/93] KY927722 Macrolepiota sp.

KY927723 Macrolepiota sp.

MC4411 M. pulchella

JJSO386 M. pulchella

NMJ161 M. dunensis

HM125521 M. dolichaula

HM125523 M. dolichaula

AF482839 M. dolichaula

AY083193 M. dolichaula

KY927715 M. sabulosa

1100] NMJ185 M. sabulosa var. velistellaris

1/100 KY927716 M. sabulosa

127714 M. cyanolamellata

1100 yoo M. cyanolameliata

Fig. 1. Bayesian Inference tree of Macrolepiota based on nrITS data and rooted with

Leucoagaricus leucothites and L. nympharum. The sequences generated in this work are in bold.

Support values (PP >0.90 and BS >70%) are shown above the node branches. *** indicates taxon

names modified according to Vizzini & al. (2011).

KY927715, we prefer to propose a new variety based on the morphological

differences discussed below.

Vizzini & al. (2011) described another volvate Macrolepiota taxon,

M. rhodosperma var. velicopia, which differs from the autonymous variety,

M. rhodosperma var. rhodosperma, primarily by the presence of a volva on

the stipe base. Macrolepiota rhodosperma var. velicopia, which clusters in

the M. sect. Macrolepiota clade, is not phylogenetically related to M. sect.

Volvatae. Vizzini & al. (2011) noted that the presence of a volva on the

stipe base is a character that developed independently over the evolution of

228 ... Freitas & Menolli

Macrolepiota species, suggesting that recognition of M. sect. Volvatae based

solely on the presence of volva is an artificial classification. Additionally, Ge

& al. (2010) highlighted the relatively small (usually <15 um) amygdaliform-

ellipsoid basidiospores and absence of clamp connections as additionally

diagnostic of M. sect. Volvatae. The position of M. sabulosa var. velistellaris

outside M. sect. Volvatae supports the view of Vizzini & al. (2011) regarding

the homoplasy of the volva within Macrolepiota.

Our sequence analyses also separate Macrolepiota dunensis and M. sabulosa

var. velistellaris from all known and sequenced volvate Macrolepiota taxa and

present for the first time molecular data for M. pulchella, the only volvate

Macrolepiota species previously recorded from Brazil. It is important to note

that our phylogeny was generated only from nrITS data and that the internal

position and the actual relationships of the sampled taxa may differ when

more markers are used.

Taxonomy

Macrolepiota dunensis D.S. Freitas & Menolli, sp. nov. FIGS 2A,D, 3

MB823088

Differs from Macrolepiota pulchella by its shorter stipe, its lack of triangular patches

or squamules on the pileus disc, its smooth and non-encrusted cheilocystidia, and its

shorter terminal elements in the pileus covering.

Type—Brazil. Rio Grande do Norte: Natal, Parque Estadual Dunas de Natal, Geology

trail, 12 July 2010, Menolli Jr. & al. NMJ161 (Holotype, SP466944; GenBank

MG136892).

ETYMOLOGY—dunensis is the Latinized form of dune, for the sand dune area in an

Atlantic Forest fragment where the holotype was collected.

Piteus 50 mm diam., plano-concave, with an obtuse umbo, dark brown

(N,,A,.M.,) at center, fully covered with appressed fibrils up to two-thirds of

the radius, slightly lighter (N,,A..M.,) elsewhere and with the innate fibrils

dissociating radially on cream background. LAMELLAE free, ventricose, crowded,

cream colored, with lamellulae. Stipe 55 x 3(apex)/7(base) mm, central, brown

(N.,A,.M.,) and slightly lighter at the apex, covered by inconspicuous zigzag

bands mainly on central part, tapering upwards and enlarged at base, fibrous

and slightly striate longitudinally. ANNULUs lost but with clear mark on the

central stipe. Votva limbate, membranous with a cottony base, dirty white to

light brown at the margin (N,,A,,M,,). CONTEXT, ODOR, TASTE, AND SPORE

PRINT COLOR not recorded.

Macrolepiota dunensis sp. nov., M. sabulosa var. velistellaris var. nov. (Brazil) ... 229

Fic. 2. A, D. Macrolepiota dunensis (holotype, SP466944). B, E. Macrolepiota pulchella

(B = SP466949; E = SP381606). C, E Macrolepiota sabulosa var. velistellaris (holotype,

SP466945). Scale bars = 1 cm. Photos: A, D = N. Menolli Jr.; B = J.J.S. Oliveira; C, F = I.G. Baseia;

E = M. Capelari.

BASIDIOSPORES [25/1/1] 9.0-12.0 x 6.0-8.0 um (Q = 1.4-1.8; Qm = 1.6;

Lm = 10.7 um; Wm = 6.8 um), ellipsoid to elongate, smooth, thick-walled,

hyaline, with an apical central truncate germ pore covered bya hyaline cap, some

with an inconspicuous apiculus, dextrinoid in Melzer’s and metachromatic

in Cresyl blue. Basrp1a 26-31 x 11.2-13.0 um, clavate, hyaline, thin-walled

to slightly thickened at the apex, 2-, 3-, and 4-spored in equal abundance.

CHEILOCYSTIDIA 32-47 x 7.5-11.2 um, cylindrical to narrowly clavate,

hyaline, thin-walled to slightly thickened towards the apex. HYMENOPHORAL

TRAMA irregular composed of thin-walled hyphae, 10.0-16.2 um diam.,

hyaline. PILEUS TRAMA hyphae thin-walled, 8.7-16.2 um diam., with light

brown content. PILEUS COVERING an intricate trichoderm, composed of

cylindrical elements with light brown content, thin-walled, terminal elements

77-171 x 5.0-7.5 um. STIPITIPELLIS hyphae thin-walled, 6.2-12.5 um diam.,

filled with light brown content. Votva hyphae undifferentiated, septate, loosely

interwoven, 2.5-10.0 um diam., thin-walled, colorless to slightly yellowish.

No CLAMP CONNECTIONS seen in all parts examined.

230 ... Freitas & Menolli

IO ae

Wi ed

SALT AG

‘. a WANK

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WN \

tN N\

Fic. 3. Macrolepiota dunensis (holotype, $P466944). A. Basidiospores; B. Basidia; C. Cheilocystidia;

D. Pileipellis elements. Scale bars = 10 um. Line drawings: D.S. Freitas.

EcoLtocy & DisTRIBUTION—Solitary on sandy soil. In a fragment of

shrubby-arboreal stratum on dunes, part of the Atlantic Forest domain in a

‘restinga (tropical coastal vegetation) area.

ComMMENTS—Macrolepiota dunensis is characterized by a dark brown pileus

fully covered with appressed fibrils up to two-thirds from the center and with

no velar remnants on the surface, a short stipe, cheilocystidia with slightly

thickened walls towards the apex, and short individual terminal elements in the

pileus covering. These and other additional morphological characters allied to

the phylogenetic position of the sampled taxa (Fic. 1), distinguish M. dunensis

from the other volvate Macrolepiota as discussed below.

Macrolepiota pulchella differs in its pileus with triangular brown-gray

squamules at center, longer (<130 mm) stipe, apical annulus, white membranous

volva, slightly shorter (18-40 um) cheilocystidia with verrucose apices, and

pileus covering composed of some pyriform and wider (23-40 um diam.)

hyphae or sometimes walls encrusted with brown pigments (Heinemann & de

Meijer 1996).

Macrolepiota eucharis differs in its pileus with black squamules on a brown-

gray background, longer stipe (<140 mm), slightly larger (10.8-15.5 x 7.1-9.1

um) basidiospores, and pileus surface with shorter and wider (20-80 x 6.0-14

um) terminal elements (Vellinga 2003).

Macrolepiota dunensis sp. nov., M. sabulosa var. velistellaris var. nov. (Brazil) ... 231

Macrolepiota brunnescens differs in its broader (<120 mm) pileus partly

covered by white woolly-membranous veil remnants, longer (115 mm) white

stipe, longer (35-44 um) basidia that have (sometimes) a brown resinous

encrustation or (rarely) brown contents, and an apparent absence of cystidia

(Singer 1959).

Macrolepiota velosa differs in its pileus surface with some purplish squamules

and white to dirty white universal veil patches, much longer (100-170 mm)

purplish brown tinged stipe, longer and narrower (44-68 x 4.5-7.5 um)

cheilocystidia with refractive apices and grayish-granular contents, and pileus

covering with wider (25 um diam.) terminal elements (Vellinga & Yang 2003).

Macrolepiota rhodosperma var. velicopia differs in its larger (<150 mm

diam.) pileus with white to dirty white membranous velar patches on the

surface; much longer (<180 mm) stipe; larger ((13—)13.5-15(-16.5) x 9-10.5(-

11.2) um) basidiospores; variously shaped cheilocystidia (narrowly clavate to

sub-fusiform, lageniform, utriform, or cylindrical) that are sometimes apically

capitulate, basally septate, or (rarely) with apical excrescences; and in the pileus

covering much longer (<250 um) individual terminal elements with several

secondary septa (pseudosepta) and some very thick-walled (1.5-2 um) hyphae

(Vizzini & al. 2011).

Finally, our other newly described taxon, M. sabulosa var. velistellaris, is

distinguished by its slightly rimose pileus surface cracking into very small

squamules towards the margin, slightly longer (95 mm) and distinctly

squamulose stipe, uniformly thin-walled cheilocystidia, and stipe covering with

distinct and ascendant terminal elements (see complete taxonomic description

below).

Macrolepiota pulchella de Meijer & Vellinga, Mycotaxon 85:184, 2003. FIGs 2B,E, 4

= Lepiotella brunnea Rick, Lilloa 2: 251. 1938.

= Volvolepiota brunnea (Rick) Singer, Bol. Soc. Arg. Bot. 8: 12. 1959.

“Chlorophyllum pulchellum” Gimenes & Capelari, [unpublished master’s thesis]. 2007.

Piteus 32-68 mm diam., conic to convex, light brown (A,,M_C,;

N,oAy™M,,.) to slightly greyish brown (N,,Y,,M,,), dark brown (A,.M,,C

60° 40 80° 50 90 “80?

N,,Y<)@,.) on the entire and prominent umbo, margin entire or sometimes

with inconspicuous velar remnants, surface completely scaly on fibrillose

background and covered with dark brown (A,,M.,C_,; N,,Y,,.M,,) scales (larger

at center, smaller toward the edge). Context white, 3.5 mm thick. LAMELLAE

free, remote, crowded, white to light cream (N,,Y,,M,,), with lamellulae. Stipe

232 ... Freitas & Menolli

Fic. 4. Macrolepiota pulchella (SP466949). A. Basidiospores; B. Basidia; C. Cheilocystidia;

D. Pileipellis elements. Scale bars = 10 um. Line drawings: D.S. Freitas.

75-135 x 3-3.5(apex)/5-10(base) mm, central, dark brown at base and slightly

lighter towards the apex, covered by inconspicuous zigzag bands mainly on

central part, almost cylindrical or slightly tapering upwards with a subbulbous

base, fibrous. ANNULUS apical, membranous, whitish upperside with brown

edge, margin slightly splitting. VoLvA saccate, membranous with a cottony

base, entire white. ODOR AND TASTE not recorded. SPORE PRINT white.

BASIDIOSPORES [187/9/9] (8.7—)9.0-13.7(-15.0) x (6.2-)7.5-10.0(-11.2)

um (Q = 1.1-2.0; Qm = 1.5; Lm = 12.2 um; Wm = 8.4 um), subglobose,

broadly ellipsoid, ellipsoid, elongate, smooth, thick-walled, hyaline, with a

slightly truncate germ pore covered by a hyaline cap, some with inconspicuous

apiculus, dextrinoid in Melzer’s and metachromatic in Cresyl blue. Basip1a

22-40 x 7.5-15.0 um clavate, hyaline, thin-walled or slightly thickened at apex,

2-3-4-spored. CHEILOCYSTIDIA 27-55 x 6.2-10.0 um, cylindrical to slightly

clavate, hyaline, thin-walled, some apically encrusted. HYMENOPHORAL TRAMA

irregular, hyphae thin-walled, 3.7-16.2 um diam., hyaline. PILEUs TRAMA

hyphae thin-walled, 3.7-18.7 um diam., hyaline to slightly light brown. PILEus

COVERING an intricate trichoderm, composed of thin-walled apically rounded

cylindrical elements with light brown contents, terminal elements 72-157 x

5.0-7.5 um; toward the center intermixed with clavate or pyriform elements.

STIPITIPELLIS hyphae thin-walled, 3.7-18.7 um diam., with brown contents.

Votva hyphae undifferentiated, 2.5-8.7 um diam., thin-walled, septate, loosely

interwoven, colorless. CLAMP CONNECTIONS not seen in all parts examined.

EcoLocy & DisTRIBUTION—Solitary on soil. In a ‘seasonal semi-deciduous

forest’ fragment, part of the Atlantic Forest domain.

SPECIMENS EXAMINED—BRAZIL. SAO PAULO STATE: Sao Paulo: Itapecerica da

Serra, 10 July 1997, A.M. Gugliotta AMG945 (SP466950); 24 July 1997, M. Capelari

Macrolepiota dunensis sp. nov., M. sabulosa var. velistellaris var. nov. (Brazil) ... 233

& A.M. Gugliotta MC3955 (SP466951); Parque Estadual das Fontes do Ipiranga,

9 October 2001, U.C. Peixoto PEFI13/2001 (SP381604); 21 September 2005,

C. Puccinelli & U.C. Peixoto CP 161 (SP466948); 3 October 2006, C. Puccinelli

CP193 (SP381606); 13 August 2006, L.J. Gimenes LJG130/06 (SP381605); 13

November 2007, Menolli Jr. & FE. Karstedt NMJ155 (SP466946); 19 October 2011,

J.J.S. Oliveira & P.O. Ventura JJSO386 (SP466949; GenBank MG136891); Parque

Estadual da Cantareira, 23 October 2008, M. Capelari & L.A.S. Ramos MC4411

(SP466947; GenBank MG136894).

CoMMENTS— The morphological features observed in new and old collections

confirm the occurrence of M. pulchella from Sao Paulo state, Southeast Brazil,

as first noted by Bononi & al. (1981, as V. brunnea). However, the collection

recorded by Bononi & al. (1981) does not represent M. pulchella; Macrolepiota

sp. (SP61013) has much larger basidiospores (13.5-16.5 x 9.0-12.0 um;

Q= 1.3-1.6 um; Qm = 1.4 um; Lm = 14.9 um; Wm = 10.6 um), a longer

(approx. 315 mm) stipe, and no evidence of a volva (or any velar remnants)

on the stipe base.

In addition to the Sao Paulo state record, Gimenes (2007) recorded

M. pulchella (as “Chlorophyllum pulchellum”) from the same area—Parque

Estadual das Fontes do Ipiranga—where we found most of our collections.

Gimenes (2007) justified the transfer to Chlorophyllum based on an unresolved

LSU sequence analysis and the morphology of the pileus covering (described

as hymenidermal) and basidiospores (described as lacking the characteristic

hyaline cap on the germ pore). However, our revision of the collections

studied by Gimenes (2007) plus our examination of new collections and

the molecular support confirms the identity of all materials as Macrolepiota

pulchella.

The most complete morphological study of M. pulchella was published

by Heinemann & de Meijer (1996) based on a unique basidioma (de

Meijer 2935) from Parana state in South Brazil. They also cited three other

collections, drawing attention to minor morphological variations such as

the pileus color, context, and squamules; stipe base shape; stability of the

annulus; and a volva that can be inconspicuous.

Compared to our M. pulchella collections, Heinemann & de Meijer

(1996) reported smaller basidiospores ((8.6-)9.41-10.3 x 5.8-6.19(-6.8)

um) and shorter basidia (24-26 um) and cheilocystidia (18-40 um), with

the cheilocystidia (4-8(-10) um diam.) sometimes narrowly lanceolate or

slightly verrucose at apex. They also described the pileus covering as:

“. in the center, palisadic, formed of erect hyphae, 5-10 um diam., with some

much larger, e.g. 23-40 um broad, pyriform elements which are seen at various

234 ... Freitas & Menolli

distance from surface; this structure is covered with a layer, 10-35 um thick,

made up of cylindrical, + radially oriented hyphae, 3-7 um diam., originating

probably from the underlying structure and constituting a general veil; in the

periphery, squamules made of cylindrical parallel hyphae, 6-11 um diam., wall

with brown incrusted pigment.’

We wish to highlight that among our specimens we observed the characteristic

trichodermal pileus covering with long, non-encrusted terminal elements

(72-157 x 5.0-7.5 um). We also noted i) only 2—spored basidia in NMJ155,

(23.7-27.5 x 10.0-12.5 um) similar in size to those described by Heinemann

& de Meijer (1996), and basidiospores ((9.5—)10.0-—11.5(-12.0) x 7.5-9.0 um)

slightly smaller than the other collections; ii) in JJSO386, the presence of

clavate to pyriform elements in the pileus disc covering; and iii) in JJSO386,

apically encrusted cheilocystidia possibly corresponding to what Heinemann

& de Meijer (1996) described as “slightly verrucose,’ which, however, does

not seem to be a constant character for the species.

Considering that Heinemann & de Meijer (1996) based their description

on a single collection, we feel we have accurately described additional

morphological characters for M. pulchella while providing the first molecular

data to contribute to a phylogeny of volvate Macrolepiota from Brazil.

Additional morphological examinations are needed to confirm the identity

of the specimens named by Fazolino Perez & al. (2018) as Macrolepiota sp. 3,

but our molecular analyses suggest they represent M. pulchella, which would

be a first record of M. pulchella from Rio Grande do Norte state in Northeast

Brazil.

Macrolepiota sabulosa var. velistellaris D.S. Freitas & Menolli, var. nov.

MB 823089 FIGS 2C,F, 5

Differs from the non-volvate autonymous variety Macrolepiota sabulosa var. sabulosa

by the presence of a distinct volva on the stipe base.

Type—Brazil. Rio Grande do Norte: Baia Formosa, Mata Estrela, Reserva Particular

de Patrimonio Natural Mata Estrela, 14 July 2010, Menolli Jr. & al. NMJ185 (Holotype,

SP466945; GenBank MG136893).

EryMoLocy—veli (L. for veil), referring to the presence of a volva and velar

remnants on the stipe base; stellaris (L. for star-shaped), referring to both the star-like

arrangement of the pileus squamules and Mata Estrela (Star Forest), the private area

where the holotype was collected.

PILEUS 62 mm, plano-convex, with an inconspicuous umbo, dark brown

(N,,A,.M,,) at center, covering slightly rimose and cracking into very small

squamules on one-third distant from the umbo before dissociating radially

Macrolepiota dunensis sp. nov., M. sabulosa var. velistellaris var. nov. (Brazil) ... 235

Fic. 5. Macrolepiota sabulosa var. velistellaris (holotype, SP466945). a. Basidiospores; b. Basidia;

c. Cheilocystidia; d. Pileipellis elements. Scale bars = 10 um. Line drawings: D.S. Freitas.

towards the margin into fibrils on cream background. LAMELLAE free,

crowded, white, with lamellulae. StipE 95 x 6(apex)/7(base) mm, central,

almost cylindrical, covered throughout with small scales forming distinct

zigzag bands that concolourous with the pileus disc, lighter at the apex.

ANNULUS apical, membranous, movable, whitish upperside with brown edge.

Votva limbate, membranous with a cottony base, dirty white. CONTEXT,

ODOR, TASTE AND SPORE PRINT COLOR not recorded.

BASIDIOSPORES [25/1/1] 9.5-12.0 x 6.0-8.0 um (Q = 1.3-1.8; Qm = 1.5;

Lm = 10.7 um; Wm = 6.9 um), ellipsoid to elongated, rarely broadly ellipsoid,

smooth, thick-walled, hyaline, with distinctive apiculus and an apical

central truncate germ pore covered by a hyaline cap, dextrinoid in Melzer’s

and metachromatic in Cresyl blue. Basrp1a 29-36 x 11.0-14.0 um, clavate,

hyaline, thin-walled, 2-3-4-spored. CHEILOCYSTIDIA 25-45 x 7.5-10.0 um,

cylindrical to narrowly clavate, hyaline, thin-walled, sometimes in clusters.

HYMENOPHORAL TRAMA irregular, hyphae thin-walled, 5.0-12.5 um diam.,

hyaline. PILEUS TRAMA hyphae thin-walled, 6.2-11.2 um diam., hyaline to

brownish. PILEUS COVERING an intricate trichoderm, composed of thin-

walled cylindrical elements with light brown content, terminal elements

measuring 118-193 x 5.0-8.7 um. SriprTrpELLis hyphae thin-walled,

5.0-11.2 umdiam., filled with light brown content; terminal elements elongated,

conspicuous, ascendant, 6.2-11.2 um diam. Votva hyphae undifferentiated,

septate, moderately branched, loosely interwoven, thin-walled, 3.7-10.0 um

diam., colorless to slightly yellowish. CLAMP CONNECTIONS not seen in all

parts examined.

236 ... Freitas & Menolli

EcoLocy & DIsTRIBUTION—Solitary on soil. In a ‘dense ombrophilous

forest’ fragment within the Atlantic Forest domain.

CoMMENTS—Macrolepiota sabulosa var. velistellaris is proposed as new based

on the presence of a distinct volva on the stipe base. Fazolino Perez & al.

(2018) described M. sabulosa based on two collections from the same state

in Northeast Brazil where we collected the new variety. They distinguished

M. sabulosa from M. pulchella due to the presence of a volva on M. pulchella,

a character neither shown in their illustrations nor mentioned in the

protologue of M. sabulosa. Additionally, M. sabulosa var. sabulosa has more

robust basidiomata (pileus 130-140 mm, stipe 165-175 x 11-12 mm), shorter

terminal elements in the pileus covering (28-63.5 x 5.5-8.5 um), and clamp

connections on hyphal septa in the stipe context and at the bases of basidia,

basidioles, and cheilocystidia (Fazolino Perez & al. 2018).

Macrolepiota sabulosa var. velistellaris morphologically differs from

M. dunensis as previously discussed in the notes under M. dunensis, as

well as from M. pulchella, M. eucharis, M. brunnescens, M. velosa, and

M. rhodosperma vax. velicopia.

Macrolepiota pulchella has differently arranged pileus squamules,

cheilocystidia with a slightly verrucose apex, and a stipe surface composed

of brown hyphae (3-9 um diam.) in a pseudoparenchyma under a palisade

structure (no mention of distinct or ascendant terminal elements; Heinemann

& de Meijer 1996).

Macrolepiota eucharis is distinguished by its brown-gray pileus

background, longer stipe (<140 mm), slightly larger (10.8-15.5 x 7.1-9.1 um)

basidiospores, mostly 4-spored basidia, slightly fusiform and slightly thick-

walled cheilocystidia, and wider (<14 um diam.) terminal pileipellis elements

(Vellinga 2003).

Macrolepiota brunnescens has a broader (<120 mm) pileus covered by a

woolly whitish membrane, undecorated and initially white stipe, encrusted

and pigmented basidia, and (apparently) no cystidia (Singer 1959).

Macrolepiota velosa differs in its purplish tinged pileus squamules, much

longer (100-170 mm) stipe, longer and narrower (44-68 x 4.5-7.5 um)

cheilocystidia with a distinct apex, and shorter (<100 um) pileipellis terminal

elements (Vellinga & Yang 2003).

Macrolepiota rhodosperma var. velicopia has a much broader (<150 mm

diam.) pileus covered with white universal veil remnants, muchlonger(<180mm)

stipe with a fringed annulus, larger ((13—)13.5-15(-16.5) x 9-10.5(-11.2) um)

Macrolepiota dunensis sp. nov., M. sabulosa var. velistellaris var. nov. (Brazil) ... 237

basidiospores, differently shaped cheilocystidia (varying among narrowly

clavate to sub-fusiform, lageniform, utriform or cylindrical), longer (<250

um) pileipellis elements that are normally thick-walled (1.5-2 tm thick) and

provided with internal pseudosepta (Vizzini & al. 2011).

Acknowledgments

The authors thank Dr. Alfredo Justo (New Brunswick Museum, Saint John

NB, Canada) and Dr. Ana Esperanza Franco-Molano (Universidad de Antioquia,

Medellin, Colombia) for critical review of the manuscript; Dr. Shaun R. Pennycook

for nomenclature review; Dr. Iuri Goulart Baseia for all the assistance during

the expeditions to the state of Rio Grande do Norte; Dr. Tarciso S. Filgueiras (1N

MEMORIAM) for help with the Latin names for the new taxa; and Alicia G. Knudson

for the English review. They also acknowledge financial support from FAPESP

(Fundagao de Amparo a Pesquisa do Estado de Sao Paulo-grant 04/04319-2), CAPES

(Coordenagao de Aperfeigoamento de Pessoal de Nivel Superior-PNADB grant

23038.000043/2010-43), CNPq (Conselho Nacional de Desenvolvimento Cientifico

e Tecnoldgico — PIBIC fellowship for D.S. Freitas).

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MY COTAXON

April-June 2019—Volume 134, pp. 241-251

https://doi.org/10.5248/134.241

Inocybe caroticolor from oak forests of Pakistan

A. NASEER’*, S. GHANT’, A. R. NiAzi’, A. N. KHALID?

"Centre for Undergraduate Studies & ’ Department of Botany,

University of the Punjab, Quaid-e-Azam Campus-54590, Lahore, Pakistan

* CORRESPONDENCE TO: arooj.hons@pu.edu.pk

ABSTRACT—Basidiomata of Inocybe caroticolor were collected from two different Quercus

forests in Swat, KP, Pakistan. Based on morphology and molecular phylogenetic analyses

of ITS and 28S rDNA regions, these basidiomata were found to represent I. caroticolor

and placed within the Inocybe viscata group. Our I. caroticolor specimens are molecularly

supported as conspecific with specimens from China and represent a first species record from

Pakistan.

Key worps—GenBank, Himalayan, Inocybaceae, LSU

Introduction

The genus Inocybe (Fr.) Fr. UInocybaceae) is distributed worldwide and

covers a wide geographical range from tropical to arctic-alpine regions (Favre

1955; Horak 1979, 1980, 1981, 1987; Kihner 1988; Buyck & Eyssartier 1999;

Watling 2001; Matheny & al. 2003; Jacobsson 2008). As one of the larger

ectomycorrhizal genera in Agaricales, Inocybe is an important component

of ectomycorrhizal temperate and boreal forest communities in alpine and

arctic habitats (Cripps & al. 2010, Kokkonen & Vauras 2012).

Between 70% and 80% of the species in Inocybaceae have been described

in associations primarily with ectomycorrhizal plant families Fagaceae,

Pinaceae, and Salicaceae (Kirk & al. 2008). Quercus (oak), an important

genus in the Fagaceae, is also known as the ‘king of trees’ due to its vital

ecological role. In Pakistan, oaks are among the dominant vascular plants

of the Himalayas, ranging from the subtropical to the sub-alpine zones

242 ... Naseer &al.

(Shrestha 2003). Himalayan oaks are evergreen, mostly gregarious, medium-

to large-sized trees distributed at elevations of 800-3800 m a.s.l. throughout

the Himalayan region. According to Alamgir (2004), established oak forests

cover approximately 16,800 ha, making Quercus an important genus in

temperate regions in Pakistan. Swat lies in Himalaya and its mycoflora is

poorly known.

More than 850 Inocybe species have been reported worldwide (Matheny

& al. 2009, 2012; Kobayashi & Onishi 2010; Horak & al. 2015; Jabeen & al.

2016). This number is increasing; during 2015-18 newly described species

of Inocybaceae included 100 from Australia (Matheny & Bougher 2017),

19 from New Zealand (Horak 2018), 17 from India (Latha & Manimohan

2017), and 13 from Thailand and Malaysia (Horak & al. 2015). Species

are also regularly being described in northern and southern Europe (e.g.,

Kokkonen & Vauras 2012; Esteve-Raventés & al. 2015, 2016, 2017; Bizio

& al. 2016; Franchi & al. 2016; Vauras & Larsson 2016). Inocybe has been

widely explored throughout Asia in China (Fan & Bau 2013, 2014), India

(Vrinda & al. 1996, 1997; Pradeep & Vrinda 2007, 2010; Latha & Manimohan

2015, 2016, 2017; Tibpromma & al. 2017), Japan (Kobayashi & Courtecuisse

1993, 2000; Kobayashi & Hongo 1993; Kobayashi 2009; Kobayashi & Onishi

2010), and the Himalayas (Horak 1981). Only a few studies of the genus,

however, have been undertaken in Pakistan. Since Ahmad & al. (1997)

published their first report of Inocybe from Pakistan, 29 Inocybe species have

been reported from different parts of the country (Ilyas & al. 2013, Saba &

al. 2015, Jabeen & al. 2016, Faroogi & al. 2017, Naseer & al. 2018, Ullah &

al. 2018), of which four—I. ahmadii Faroogi & al., I. kohistanensis Jabeen &

al., I. pakistanensis Z. Ullah & al., and I. shawarensis Naseer & Khalid—were

described as new to science (Jabeen & al. 2016, Farooqi & al. 2017, Naseer &

al. 2018, Ullah & al. 2018).

During 2014-16 we collected specimens on field research trips while

studying fungal communities associated with the oak forests of Pakistan. At

that time we encountered in Swat an Inocybe species previously unknown

from Pakistan. Its bright red carrot color easily distinguished this fungus

in the field, and in the laboratory, we identified it as Inocybe caroticolor.

Here we present a taxonomic description of our new Pakistani collections

with the results of our ITS1-5.8S-ITS2 and 28S rDNA sequence analyses

phylogenetically placing this species in the Inocybe viscata group of Inocybe

clade (Matheny & Bougher 2017).

Inocybe caroticolor from oak forests (Pakistan) ... 243

Material & methods

Collection & morphological examination

Specimens were collected from two different locations from the moist temperate

forest of Toa, Swat as described in Naseer & al. (2017) and Shawar Valley as described

in Naseer & al. (2018). These forests are dominated by Quercus incana Roxb. nom. illeg.

[= Q. oblongata D. Don] and to a lesser extent Q. dilatata Royle. Gross morphological

characters were recorded in the field and photographed using a Nikon D70S digital

camera. Basidiomata were wrapped in aluminium foil and kept separately in a

collection box to avoid mixing or crushing. Specimens were dried, sealed in plastic bag,

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

Lahore, Pakistan (LAH). Colors were designated by mColorMeter. For anatomical

examination, tissues from lamellae, pileipellis and stipitipellis were mounted on glass

slides and observed in Phloxine (1%), Melzer’s reagent, and KOH (5%) for pigmented

hyphae and walls of cystidia using a Meiji Techno MX4300H microscope. Dimensions

were measured for basidiospores, basidia and other elements from basidiomata under

the light microscope equipped with a camera lucida.

Molecular phylogenetic analyses

DNA was extracted from lamellae of basidiomata following a modified CTAB

method (Bruns 1995). The ITS region was amplified using primer pairs ITS1F/ITS4

(White & al. 1990) and the nrDNA 28S region using primer pairs LROR/LR5 (Vilgalys

& Hester 1990). Amplification parameters proceeded with an initial denaturation step

at 94°C for 4 min, followed by 34 cycles of 94°C for 40 sec, 55°C (ITS) or 52°C (28S)

for 40 sec, and 72°C for 1 min, and a final elongation step at 72°C for 8 min. The

PCR products were purified, and Macrogen (South Korea) conducted bidirectional

sequencing. The consensus sequences were subjected to BLAST, and compared with

available GenBank sequences. The newly generated sequences were aligned with the

closest matching NCBI sequences using MUSCLE v3.6 (Edgar 2004) and manually

adjusted where necessary. ML tree generation and bootstrap analyses were performed

for combined ITS and 28S data set on CIPRES Gateway (Miller & al. 2010). A maximum

likelihood (ML) tree was inferred using RAxML-HPC2 v8.1.11 (Stamatakis 2014) with

a GTR + gamma model of nucleotide substitution. One thousand bootstrap iterations

were performed with rapid bootstrapping. Significant support was considered to be

=70%. Sequences generated in this study were submitted to GenBank.

Taxonomy

Inocybe caroticolor T. Bau & Y.G. Fan, Mycotaxon 123: 170 (2013) Figs 1, 2

BASIDIOMATA small. PrLEus 12-32 mm diam., bright red (2.4YR 3.1/9.3)

in centre becoming yellowish (0.5Y 6.3/7) with patches of reddish orange;

conical when young, then plano-convex, with an obtuse umbo, slightly

rimulose, surface dry with appressed squamules over disc, radially fibrillose,

244 ... Naseer &al.

Fic. 1. Inocybe caroticolor. Basidiomata.

A, B. LAH35266; C, D. LAH35268; E-G. LAH35269.

Scale bars: A, B, E= 0.7 cm; C, D, F = 0.33.cm; G = 0.5 cm

rimose, margins inrolled (young) to incurved (mature). LAMELLAE light

yellow (4.6Y 8.9/4.2) when young to yellowish orange (2.6YR 2.5/5.4)

when mature, numbering 45-60, regularly arranged, adnexed, slightly

close to crowded, 2-3 mm deep, edges even, smooth; lamellulae %4 length

of lamellae, single-tiered and alternating with lamellae. Stipe 32 x 4 mm,

central, cylindrical, with subbulbous to non-marginate bulbous base,

fibrillose, pruinose entire length, densely so at apex, longitudinally striate;

yellow (4Y 6.2/6.5) in the middle region, and reddish orange (2.4YR 3.1/9.3)

at apex, basal tomentum white.

Inocybe caroticolor from oak forests (Pakistan) ... 245

Fic. 2. Inocybe caroticolor (LAH35266). A. Basidia; B. Basidiospores;

C. Cheilocystidia; D. Pleurocystidia; E. Caulocystidia with paracystidia; F. Pileipellis.

Scale bars: A = 7.90 um; B = 4.39 um; C = 9.54 um; D, E = 14 um; F = 6.7 um.

246 ... Naseer & al.

BasiprosporEs [n/b/p = 60/3/3], (6.3-)6.5-9.3(-9.5) x (4.5-)4.7-6.0

(-6.1) um, mean = 7.5 x 5.2 um, Q = (1.43-)1.47-1.58(-1.61), pale brown

in 5% KOH, weakly to strongly nodulose. Basrp1A 25-36 x 5-8 um, clavate,

4-spored, with yellowish pigments. PLEUROCysTIDIA 47-63 x 12-15 um,

fusiform, with crystalliferous apex, yellow in 5% KOH, thick walled, obtuse

base. CHEILOCYSTIDIA 48-62 x 12-15 um, fusiform, some cylindrical.

CAULOCYSTIDIA 49-75 x 9-13 um, variably shaped, abundant, thin-walled.

PILEIPELLIS 0.2-6.5 um, a cutis of cylindrical hyphae, smooth, pigmented,

thin-walled. STIPITIPELLIs 2—4.5 um, a cutis composed of regularly arranged

hyphae, filamentous, smooth, thin- to moderately thick-walled. CLamp

CONNECTIONS present.

MATERIAL EXAMINED: , Toa, 2100 m a.s.l, on soil under Quercus oblongata, 15 July

2015, Arooj Naseer, Abdul Rehman Niazi & Abdul Nasir Khalid AST47 (LAH35266;

GenBank MH473148, MH536985); , Shawar Valley, 2100 m a.s.l, solitary on soil

under Q. dilatata and Q. oblongata 14 July 2014, Arooj Naseer & Abdul Nasir Khalid

ASSW11 (LAH35268; GenBank MH473144, MH536984); 15 Aug 2015, Arooj

Naseer & Abdul Nasir Khalid ASS6 (LAH35267; GenBank MH473145, MH536986);

25 August 2014, Arooj Naseer & Abdul Nasir Khalid ASSW29 (LAH35269; GenBank

MH473146, MH536983).

Molecular phylogenetic characterization

Sequencing of the ITS PCR products from Inocybe caroticolor specimens

yielded 580-745 base pairs. NCBI BLAST searches of consensus sequences

from four collections showed these sequences as 99% identical with Inocybe

caroticolor (JX025772, JX025773 and JX025774) from China (100% query

coverage; 0.0 E value). These sequences also showed a 93% identity with

Inocybe sp. TR198-03 (KP636865) from Papua New Guinea.

Sequencing of the 28S PCR products yielded 867-980 base pairs.

Consensus sequences of the 835 base pairs BLAST searched at NCBI showed

97% identity with Inocybe sp. REH7418 (JN974931) from an oak forest in

Costa Rica and Inocybe sp.TR132-05 (KP171061) and Inocybe sp. TR88-06

(JN974992) from Castanopsis forests in Papua New Guinea with 100% query

cover and 0.0 E value.

Sequences representing the Inocybe viscata group (Inocybe clade; Matheny

& Bougher 2017), Indian species in a related clade (Latha & Manimohan

2017), and Auritella dolichocystis as outgroup were selected for preparation

of the ITS/28S-based phylogram. Our newly generated sequences clustered

with sequences from China with strong bootstrap value (Fic 3).

Inocybe caroticolor from oak forests (Pakistan) ... 247

Pakistan

Inocybe_caroticolor_ YGFan2011139

84|| Inocybe_caroticolor_YGFan2010216 China

Inocybe_caroticolor_YGFan2011123

Inocybe_sp_TR198_03

Inocybe_sp_CR1

Inocybe_sp_REH7418

Inocybe_sp_REH7965

Inocybe_sp_TR132-05

Inocybe_sp_TR88-06

Inocybe_kurkuriya_CAL1352

Inocybe_torresiae_PBM3722

Inocybe_torresiae_PBM2157

Inocybe_flavosquamulosa_CAL1355

Inocybe_floccosistipitata_CAL1256

Inocybe_sp_ZT10106

400) Inocybe_viscata_PBM3445

Inocybe_viscata_PBM3213

Inocybe_viscata_TENNO66

Inocybe_torresiae_PBM3722

Inocybe_insulana_CAL1258

Auritella_dolichocystis

0.5

Fic. 3. ML phylogram of Inocybe caroticolor in the I. viscata group based on combined ITS and

28S nrDNA as generated with RAxML with 1000 bootstrap iterations. The letters in bold refer to

sequences generated in this study, and sequences from Pakistan are highlighted in green.

Discussion

Inocybe caroticolor is characterized by the bright reddish carrot color

of the basidiomata, nodulose basidiospores, and thick-walled hymenial

cystidia. It can be easily recognized in the field by its bright red color,

distinct aromatic odor, and entirely pruinose stipe. ITS sequence analysis

clustered sequences from our Pakistani collections phylogenetically with

those reported from China by Fan & Bau (2013), while the 28S phylogeny

supported separation of the sequences from Pakistan in a separate clade

from the sequences from Inocybe sp. TR88.06, Inocybe sp. TR132.05, and

Inocybe sp. TR198.03 as observed in the ITS-based phylogram. However,

as no other 28S-based sequences were present in GenBank, they were not

included in a 28S-based phylogram.

The Pakistani collection of I. caroticolor differs somewhat morphologically

from Chinese collection, which Fan & Bau (2013) described with a smaller

248 ... Naseer & al.

pileus and brighter red carrot color. In China, I. caroticolor has been found

in association with Quercus variabilis Blume except for one collection found

under Pinus yunnanensis Franch. (Fan & Bau 2013). During this study,

all four collections from Pakistan were associated with Q. oblongata and

Q. dilatata forests, indicating a strong preference for Quercus. This hypothesis

is strengthened by the presence of other oak-specific species in the same

clade (Matheny & Bougher 2017).

Our collections represent first record of Inocybe caroticolor for Pakistan.

This species also exhibits a strong attraction for different oak species. In

Pakistan, oak forests are being cut extensively for burning fuel and thus

subject to decline. The presence of this unusual species may help develop an

awareness of the need to conserve, establish, and reclaim these oak forests.

Acknowledgements

We are sincerely grateful to Dr. Brandon Matheny (University of Tennessee,

Knoxville, USA) for his phylogenetic analyses and comments on an earlier version

of this paper. We are thankful to Dr. Ditte Bandini (Wiesenbach, Germany) and

Dr. Sana Jabeen (University of Education, Lahore, Pakistan) for their helpful

comments to improve this manuscript.

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315-322. https://doi.org/10.1016/b978-0-12-372180-8.50042-1

MY COTAXON

April-June 2019—Volume 134, pp. 253-256

https://doi.org/10.5248/134.253

Sporidesmium horizontale sp. nov. from China

ConG-Conc At’, JI-WEN XIA', XIU-GUO ZHANG’,

YUN GENG?“, L1-Guo Ma?®

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

College of Plant Protection, Shandong Agricultural University,

Taian, Shandong, 271018, China

’ Biotechnology Research Center, Shandong Academy of Agricultural Sciences,

Jinan, Shandong, 250100, China

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

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

“CORRESPONDENCE TO: “ tagy009@163.com, ® maliguo809@163.com

ABSTRACT—A new species, Sporidesmium horizontale, is described and illustrated from

specimens collected on dead branches in China. This fungus is characterized by horizontal,

obclavate, 4—5-septate, versicolored conidia with short rostra.

Key worps—anamorphic fungi, hyphomycete, Pleosporomycetidae, taxonomy

Introduction

Sporidesmium was erected by Link (1809) and typified with S. atrum Link.

Ellis (1971) defined the genus as having integrated, terminal, monoblastic

conidiogenous cells with acrogenous, solitary, transversely septate or

distoseptate conidia. Subramanian (1992) redefined the generic concept of

Sporidesmium in a stricter sense as simple conidiophores, determinate or

with irregular extending proliferations, and solitary, gangliar, acrogenous

and euseptate conidia. Wu & Zhuang (2005) expanded the generic concepts

to include fungi with typically lageniform, doliiform, or nodose, percurrently

extending conidiophores. Since then 31 new species have been described in the

genus, most of which were found on rotten wood, dead branches, or decaying

254 ... Ai & al.

leaves of various plants (e.g., Ma & Zhang 2007; Ma & al. 2012; Xia & al. 2017;

Yang & al. 2018; Zhang & al. 2008, 2017).

A new hyphomycete representing the genus of Sporidesmium was found

during research on saprobic fungi in China. Specimens are deposited in

the Herbarium of Department of Plant Pathology, Shandong Agricultural

University, Taian, Shandong, China (HSAUP) and the Mycological Herbarium,

Institute of Microbiology, Chinese Academy of Sciences, Beijing, China

(HMAS).

Taxonomy

Sporidesmium horizontale L.G. Ma, C.C. Ai & X.G. Zhang, sp. nov. FIG. 1

MB 830543

Differs from other Sporidesmium species by its horizontal, obclavate, 4—5-septate,

versicolored, and smooth conidia with a short, subhyaline rostrum.

Type: China, Yunnan Province, Xishuangbanna Tropical Botanical Garden, on dead

stems of an unidentified broadleaf tree, 2 Nov. 2011, L.G. Ma (Holotype, HSAUP

H2133; isotype, HMAS 146148).

Erymo coey: horizontale, referring to the conidia with horizontal growth.

COLONIES on natural substrate effuse, brown to dark brown, hairy. Mycelium

partly superficial, partly immersed in the substrate, composed of septate, pale

brown, smooth, 3.0-4.5 um diam. hyphae. ConIDIOPHORES macronematous,

mononematous, unbranched, erect, straight to flexuous, cylindrical, smooth,

thick-walled, brown to dark brown, 5-10-septate, 90-175 um long, 6-13.5 um

diam. at the base and 3-4.5 um diam. at the tapered apex. CONIDIOGENOUS

CELLS holoblastic, monoblastic, brown, cylindrical, smooth, thick-walled, 5-6.5

x 17-21.5 um. Conidial secession schizolytic. Conip14 solitary, horizontal on

the conidiophores, obclavate, thick-walled, brown, pale brown on the basal and

upper cells, smooth, 4—5-euseptate, 41-59 x 16-18 um, with a short, subhyaline

rostrum at the apex, sometimes embedded in a drop of mucilage.

CoMMENTS—Sporidesmium horizontale is unique in the genus by developing

conidia in a horizontal orientation, an unusual phenomenon that cannot be

explained at present. The species can also be separated from other Sporidesmium

species by its horizontal, obclavate, 4—5-septate, and versicolored conidia with

short, subhyaline rostra.

Acknowledgments

The authors express gratitude to Dr. Rafael EF Castafeda-Ruiz (Alejandro de

Humboldt, INIFAT, Cuba) and Dr. De-Wei Li (Connecticut Agricultural Experiment

Sporidesmium horizontale sp. nov. (China) ... 255

' cy

20um 20um

Fic. 1. Sporidesmium horizontale (holotype, HSAUP H2133).

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

Station Valley Laboratory, USA) for serving as pre-submission reviewers and for

their valuable comments and suggestions. This project was supported by National

Key R&D Program of China (2017YFD0201700), Young Talents Training Program

of Shandong Academy of Agricultural Sciences (CXGC2018E04), National Natural

Science Foundation of China (31400019, 31230001, 31093440), and the Ministry of

Science and Technology of the People’s Republic of China (2006FY 120100).

Literature cited

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

Surrey, England.

Link HE 1809. Observationes in ordines plantarum naturales. Dissertatio I. Magazin der

Gesellschaft Naturforschenden Freunde Berlin 3: 3-42.

Ma J, Zhang XG. 2007. Two new species of Sporidesmium from Yunnan, China. Mycotaxon 101:

73-76.

Ma J, Ma LG, Zhang YD, Xia JW, Zhang XG. 2012. New species and record of Sporidesmium

from southern China. Mycotaxon 119: 17-25. https://doi.org/10.5248/119.17

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

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

256 ... Ai & al.

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

Fungal Diversity Research Series 15. 351 p.

Xia JW, Li HH, Zhang XG. 2017. Two new species of hyphomycetes from southern China.

Mycosystema 36: 1483-1486. https://doi.org/10.13346/j.mycosystema.170140

Yang J, Maharachchikumbura SSN, Liu JK, Hyde KD, Jones EBG, Al-Sadi AM, Liu ZY. 2018.

Pseudostanjehughesia aquitropica gen. et sp. nov. and Sporidesmium sensu lato species from

freshwater habitats. Mycological Progress 17: 591-616.

https://doi.org/10.1007/s11557-017-1339-4

Zhang K, Ma J, Zhang XG. 2008. Taxonomic studies of Sporidesmium from Hainan, China.

Mycotaxon 104: 165-169.

Zhang H, Dong W, Hyde KD, Maharachchikumbura SSN, Hongsanan S, Bhat DJ, Al-Sadi

AM, Zhang D. 2017. Towards a natural classification of Annulatascaceae-like taxa:

introducing Atractosporales ord. nov. and six new families. Fungal Diversity 85: 75-110.

https://doi.org/10.1007/s13225-017-0387-z

MY COTAXON

April-June 2019—Volume 134, pp. 257-260

https://doi.org/10.5248/134.257

Sporidesmiella corniformis sp. nov. from China

ConG-Conc At’, JI-WEN XIA’, XIU-GUO ZHANG’,

YUN GENG?“, L1-Guo Ma?®

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

College of Plant Protection, Shandong Agricultural University,

Taian, Shandong, 271018, China

*Biotechnology Research Center, Shandong Academy of Agricultural Sciences,

Jinan, Shandong, 250100, China

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

Shandong Academy of Agricultural Sciences,

Jinan, Shandong, 250100, China

“CORRESPONDENCE TO: “tagy009@163.com, *maliguo809@163.com

ABSTRACT—A new species, Sporidesmiella corniformis, is described and illustrated from

specimens collected on decaying branches in China. This fungus is characterized by clavate

to cuneiform, 2-distoseptate, pale brown conidia with 4-8 short horns.

Key worps—anamorphic fungi, hyphomycete, Melanommataceae, taxonomy

Introduction

Numerous microfungi collected in tropical and subtropical forests in China

have recently been reported (Ma & al. 2016a, b). In our continual mycological

surveys on wood-inhabiting fungi in China, an anamorphic fungus with

morphological features typical of Sporidesmiella Kirk (1982) was collected.

However, this fungus is morphologically different from the currently described

Sporidesmiella species and is therefore proposed as new.

Specimens are deposited in the Herbarium of the Department of Plant

Pathology, Shandong Agricultural University, Taian, Shandong, China

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

Academy of Sciences, Beijing, China (HMAS).

258 ... Ai & al.

Materials & methods

Samples of partially decomposed woody debris collected were placed in separate

polyethylene bags for transport to the laboratory. Samples were incubated, examined,

and processed periodically following the methods described in Ma & al. (2012a).

Conidia and conidiophores were measured and photographed using a DP 27 camera

attached to a Olympus BX 53 microscope equipped with a 60x oil immersion objective.

Then photographic plate was prepared using Adobe Photoshop CSS.

Taxonomy

Sporidesmiella corniformis L.G. Ma, C.C. Ai & X.G. Zhang, sp. nov. FIG. 1

MB 830544

Differs from Sporidesmiella cornuta by its bigger, 2-distoseptate conidia with much

shorter, more numerous horns; and from S. coronata by its 2-distoseptate, wider conidia

with longer, more numerous horns.

Type: China, Yunnan Province, Xishuangbanna National Forest Park, on dead branches

of an unidentified broadleaf tree, 12 Oct. 2011, L.G. Ma (Holotype, HSAUP H2058;

isotype, HMAS 146151).

Erymo ocy: corniformis, referring to the conidial shape.

COLONIES on natural substrate effuse, brown to dark brown, hairy. Mycelium

partly superficial, partly immersed in the substrate, composed of septate, pale

brown, smooth, 1.5-3.0 um diam. hyphae. ConIDIOPHORES macronematous,

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

cylindrical, smooth, thick-walled, brown to dark brown, 4-11-septate,

55-175 x 3.5-5 um. CONIDIOGENOUS CELLS integrated, terminal, holoblastic,

monoblastic, brown, cylindrical, smooth, thick-walled, percurrently extended,

with 0-5 cylindrical extensions towards the apex. Conidial secession schizolytic.

ConIDIA acrogenous, solitary, clavate to cuneiform, thick-walled, pale brown,

smooth, 2-distoseptate, 21-35 um long, 11.5-17.0 um wide at the apex, 2.5-3.5

um wide at the truncate base, apex with 4-8 short horns 2.0-7.5 um long; cell

lumina reduced, less so in the basal cell, the basal cell darker and basal septum

convex and deeply pigmented.

ComMMENTS—Sporidesmiella was erected by Kirk (1982) with S. claviformis

P.M. Kirk as the type species. The genus is characterized by acrogenous,

solitary, distoseptate conidia seceding schizolytically from integrated, terminal,

monoblastic, percurrent, or rarely sympodially extended conidiogenous cells

on macronematous, mononematous, solitary conidiophores. Although it shares

some resemblance to Coronospora (Ellis 1971), the latter genus is distinguished

by polyblastic, sympodial, cicatrized conidiogenous cells bearing euseptate

conidia. Both Yanna & al. (2001) and Ma & al. (2012b) reviewed Sporidesmiella

Sporidesmiella corniformis sp. nov. (China) ... 259

Fic. 1. Sporidesmiella corniformis (holotype, HSAUP H2058). A. Conidiophores, conidiogenous

cells and conidia; B. Conidiophore with conidiogenous cell; C. Conidia.

and provided a key to the accepted species. Hitherto, 38 species have been

discovered, of which 12 were described from China.

Sporidesmiella corniformis is most similar to S. cornuta Kuthub. & Nawawi

and S. coronata (B. Sutton) P.M. Kirk. Sporidesmiella cornuta differs by its

conidia that are 2-3-distoseptate, cylindrical to cuneiform, 24-29 um long

and 8-13 um diam. towards the apex and 4-5 um diam. at the base and with

(2—)3(-4) acuminate horns that are 10-24 um long (Kirk 1982). Sporidesmiella

260 ... Ai & al.

coronata can be separated by its conidia, which are 2-3-distoseptate, 6-7.5

um diam. towards the apex and with 4 short (<4 um) horns (Kuthubutheen &

Nawawi 1993).

Acknowledgments

The authors express gratitude to Dr. Rafael E Castafeda-Ruiz (Alejandro de

Humboldt, INIFAT, Cuba) and Dr. De-Wei Li (Connecticut Agricultural Experiment

Station Valley Laboratory, USA) for serving as pre-submission reviewers and for their

valuable comments and suggestions. This project was supported by National Key R&D

Program of China (2016YFD0300700), Young Talents Training Program of Shandong

Academy of Agricultural Sciences (CXGC2018E04), National Natural Science

Foundation of China (31400019, 31230001, 31093440), the Ministry of Science and

Technology of the People’s Republic of China (2006FY120100), and Natural Science

Foundation of Shandong Province (BS2015SW020).

Literature cited

Ellis MB. 1971. Dematiaceous hyphomycetes. X. Mycological Papers 125. 30 p.

Kirk PM. 1982. New or interesting microfungi. VI. Sporidesmiella gen. nov. (hyphomycetes).

Transactions of the British Mycological Society 79: 479-489.

https://doi.org/10.1016/s0007-1536(82)80040-5

Kuthubutheen AJ, Nawawi A. 1993. Three new and several interesting species of Sporidesmiella

from submerged litter in Malaysia. Mycological Research 97: 1305-1314.

https://doi.org/10.1016/s0953-7562(09)80162-7

Ma LG, Ma J, Zhang YD, Castafieda-Ruiz RF, Zhang XG. 2012a. New species and records

of Heteroconium (anamorphic fungi) from southern China. Mycoscience 53: 466-470.

https://doi.org/10.1007/s10267-012-0190-3

MaJ, Zhang YD, Ma LG, Castaneda-Ruiz RF, Zhang XG. 2012b. Three new species of Sporidesmiella

from southern China. Mycoscience 53: 187-193. https://doi.org/10.1007/s10267-011-0152-1

Ma J, Zhang K, Zhang XG, Castafeda-Ruiz RF. 2016a. Three new species of Spadicoides from

Lushan Mountain, China. Mycological Progress 15: 43 [8 p.].

https://doi.org/10.1007/s11557-016-1185-9

Ma YR, XiaJW, Gao JM, Li XY, Castafieda-Ruiz RE, Zhang XG, Li Zhuang. 2016b. Atrokylindriopsis,

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

Mycological Progress 14: 77 [5 p.]. https://doi.org/10.1007/s11557-015-1071-x

Yanna, Ho WH, Hyde KD, McKenzie EHC. 2001. Sporidesmiella oraniopsis, a new species of

dematiaceous hyphomycete from North Queensland, Australia and synopsis of the genus.

Fungal Diversity 8: 183-190.

MY COTAXON

April-June 2019—Volume 134, pp. 261-269

https://doi.org/10.5248/134.261

New records of Amandinea and Buellia from China

ZUN-TIAN ZHAO™, XIAO ZHANG”, JIE-MENG Fu’, LING Hu”

'Key Laboratory of Plant Stress Research, College of Life Sciences &

? Institute of Environment and Ecology

Shandong Normal University, Jinan, 250014, PR. China

“CORRESPONDENCE TO: Hu_Ling_123@163.com

ABSTRACT—Otur research has revealed two species of Amandinea (A. occidentalis and

A. polyspora) and two species of Buellia (B. morsina and B. penichra), which we report

for the first time from China. Descriptions of morphological and chemical characters are

provided with the known distribution of each species.

Key worps—Caliciaceae, Caliciales, corticolous, lichen-forming fungi, taxonomy

Introduction

Amandinea M. Choisy ex Scheid. & M. Mayrhofer and Buellia De Not.

(Caliciaceae, Caliciales, Lecanoromycetes, Ascomycota) are closely related

genera characterized by a crustose thallus, apothecial ascomata, (4-)8

(—many)-spored asci, and brown 1-septate or (sub)muriform ascospores,

with or without oil droplets in the hymenium (Foucard & al. 2002, Coppins

& al. 2009). Amandinea can best be distinguished by the filiform conidia,

ascospores with or without inner wall thickenings and clearly ornamented

outer walls, and often lacking secondary metabolites detected by TLC,

while species of Buellia s. lat. have bacilliform conidia and usually contain

secondary compounds detectable by TLC (Coppins & al. 2009, Elix &

Kantvilas 2013a,b, Giralt & al. 2015). Amandinea is represented by 81

species and Buellia 454 species worldwide (Giralt & Nordin 2002; Kirk &

al. 2008; Senkardesler 2010; Giralt & al. 2011, 2015; Lumbsch & al. 2011;

*X1A0 ZHANG & ZUN-TIAN ZHAO contributed equally to this research.

262 ... Zhao, Zhang & al.

Giralt & van den Boom 2012; Fryday & Arcadia 2012; Tonsberg & al. 2012,

Elix & Kantvilas 2013a,b, 2016a,b; Kondratyuk & al. 2013, 2015, 2016; Elix &

al. 2015, 2017a,b, 2018; Blaha & al. 2016; Mayrhofer & al. 2016; Elix 2017a,b;

Elix & Mayrhofer 2017, 2018). In China, only one Amandinea species and

53 Buellia species have been reported (Wei 1991, Watanuki & al. 2017,

Wang & al. 2018).

This paper contributes to the knowledge of Amandinea and Buellia in

China and affords accurate data for the Lichen Flora of China project. In

this study we have identified four species new to the country: Amandinea

occidentalis, A. polyspora, Buellia morsina, and B. penichra.

Materials & methods

The specimens studied are preserved in the Lichen Section of Botanical Herbarium,

Shandong Normal University, Jinan, China (SDNU). The specimens were examined

morphologically and anatomically under a COIC XTL7045B2 stereo-microscope

and a Olympus CX41 polarizing microscope. We tested the thallus and medulla of

each specimen with K (10% aqueous KOH solution), C (aqueous CINaO solution),

and I (Lugol's iodine) for identification. Lichen substances were identified using

standardized thin-layer chromatography techniques (TLC) with system C (Orange &

al. 2010). The lichens were photographed using Olympus SZX16 and BX61 microscope

with a DP72 lens.

Taxonomy

Amandinea occidentalis Elix & Kantvilas,

Australas. Lichenol. 72: 9 (2013) FIG. 1A—D

Thallus crustose, smooth, thin, areolate, pale greyish to white, esorediate.

Prothallus present, but not apparent. Medulla white, I-. Apothecia black,

0.2-0.5 mm diam., sessile, epruinose, lecideine with a well developed

margin, weakly concave at first, then plane to convex, scattered or crowded.

Proper exciple dark brown, 50-70 um wide; epihymenium olive-brown,

K-; hymenium hyaline, inspersed with oil droplets; paraphyses simple;

hypothecium dark brown, K-. Asci (4—)8-spored, Bacidia-type. Ascospores

soon olive-brown, 1-septate, at first of the Physconia-type, then of the Buellia-

type, broadly fusiform to ellipsoid, 23-28 x 9-11 um, not constricted at the

septum. Pycnidia not observed.

Fic. 1 Amandinea occidentalis (Wang 20181068, SDNU). A: Thallus with apothecia;

B: Apothecium section; C: Ascus containing Physconia-type ascospores; D: Ascus containing

Buellia-type ascospores. Amandinea polyspora (Cheng 20120079, SDNU). E: Thallus with

apothecia; F: Apothecium section, without oil droplets in hymenium G: Ascus and ascospores;

H: Ascospores.

263

Amandinea & Buellia species new to China...

40um

pe N Ger

AAs GALES

Nee

Pret ae

264 ... Zhao, Zhang & al.

CHEMISTRY— Thallus K-, C-. No chemical substances detected by TLC.

SPECIMEN EXAMINED: CHINA. YUNNAN, Shangri-La, Mt. Tianbao, alt. 3700 m, on bark,

17 Aug. 2018, Chunxiao Wang 20181068 (SDNU).

DISTRIBUTION—Australia (Western Australia, New South Wales) (Elix &

Kantvilas 2013a; McCarthy 2018). New to China.

COMMENTS—Our specimens are similar to previously published

descriptions, except that the holotype has larger (0.2-1.2 mm) apothecia

and usually 4-spored asci (Elix & Kantvilas 2013a). Amandinea occidentalis

is characterized by the areolate thallus, Physconia-type to Buellia-type

ascospores, a hymenium inspersed with oil droplets, and the absence of lichen

substances. This species morphologically resembles A. lignicola var. australis

Elix & Kantvilas, which is distinguished by often having a subsquamulose

thallus, 8-spored asci with smaller ascospores (11-20 x 5-8 um), and a non-

inspersed hymenium (Elix & Kantvilas 2013a).

Amandinea polyspora (Willey) E. Lay & P.E. May,

Bryologist 100(2): 164 (1997) FIG. 1E-H

Thallus corticate, smooth, thin, pale brown to yellowish brown,

esorediate; medulla white, I-. Apothecia lecideine, black, 0.2-0.5 mm diam.,

sessile, epruinose, flat to convex, margin present at first but soon excluded.

Proper exciple narrow, dark brown, 20-30 um wide; epihymenium brown,

pigmentation continuous with the outer exciple; hymenium hyaline, not

inspersed with oil droplets; paraphyses simple to moderately branched,

swollen at upper part, with a brown pigmented cap; hypothecium reddish

brown. Asci Bacidia-type, mostly 16-spored, occasionally 32-spored.

Ascospores soon brown, 1-septate, Buellia-type, oblong to ellipsoid, not

constricted, 10-13(-15) x 4-5 um. Pycnidia not seen.

CHEMISTRY— Thallus K+ yellow, C-. Atranorin (trace) detected by TLC.

SPECIMEN EXAMINED: CHINA. JILIn, Helong, Mt. Zengfeng, alt. 1600 m, on bark, 19 Aug.

2011, Yuliang Cheng 20120079 (SDNU).

DIsTRIBUTION—Amandinea polyspora has been reported from Korea

(Kondratyuk & al. 2013) and from western, eastern, and southeastern USA

(Bungartz & al. 2007). New to China.

COMMENTS—Our specimens are similar to the protologue descriptions

(Bungartz & al. 2007). Amandinea polyspora is characterized by the small

ascospores and the polysporous asci. Buellia schaereri De Not. has similarly

small ascospores but differs in its consistently 8-spored asci and bacilliform

Amandinea & Buellia species new to China... 265

conidia, unlike the filiform conidia characteristic of A. polyspora (Bungartz

& al. 2007).

Buellia morsina A. Nordin, Symb. Bot. Upsal. 33(1): 74 (2000) Fig. 2A-C

Thallus crustose, smooth, rimose to areolate, brownish to grey, esorediate.

Prothallus absent. Medulla white, I-. Apothecia lecideine, 0.7-1.5 mm

diam.; sessile, disk black, epruinose, concave in young then flat; margin

black, obvious in young apothecia. Proper exciple dark brown, 50-100 um

wide; epihymenium dark brown, K-, C-. hymenium colorless, 80-125 um,

not inspersed with oil droplets. Hypothecium dark brown. Asci 8-spored;

ascospores soon brown, submuriform to muriform, 20-30 x 10-13 um.

Pycnidia not observed.

CHEMISTRY—Thallus K+ yellow turning red. Norstictic acid, atranorin

and connorstictic acid (trace) detected by TLC.

SPECIMEN EXAMINED: CHINA. YUNNAN, Luquan, Mt. Jiaozi, alt. 3600 m, on bark, 4 Nov.

2012, Hui Zhang 20129415 (SDNU).

DISTRIBUTION—Buellia morsina has previously been reported from Mexico

(Bungartz & al. 2007). New to China.

COMMENTS—Our specimens are similar to previously published

descriptions, except that the protologue describes smaller apothecia (0.2-1.2

mm) and the presence of a black prothallus (Bungartz & al. 2007). Buellia

morsina is characterized by its submuriform to muriform ascospores and

the presence of norstictic acid, atranorin, and connorstictic acid (trace). Its

thallus morphology, secondary chemistry, and exciple anatomy are similar

to B. lauri-cassiae (Fée) Mill. Arg., which is distinguished by its smaller,

3-septate spores (Nordin 2000, Giralt & Nordin 2002, Bungartz & al. 2007).

Buellia penichra (Tuck.) Hasse, Contr. U.S. Natl. Herb. 17: 122 (1913) Fig. 2D-F

Thallus corticate, smooth, greyish white, rimose to dispersed areolate.

Prothallus black. Medulla white, I-. Apothecia lecideine, black, round, 0.4-

0.8 mm diam., sessile, dispersed, flat at first soon convex, disc epruinose;

margin black, distinct in young apothecia, later excluded. Proper exciple

brown, pigmentation continuous with the outer exciple; epihymenium

brown to dark green; hymenium hyaline, inspersed with oil droplets,

paraphyses simple with distinct apical cap; hypothecium dark brown to

dark green. Asci 8-spored, ascospores submuriform, pale green then brown,

broadly fusiform to ellipsoid, 25-28 x 10-13 um. Pycnidia not observed.

CHEMISTRY—Thallus K+ yellow, C-. Atranorin detected by TLC.

266 ... Zhao, Zhang & al.

20m

Fic. 2 Buellia morsina (Zhang 20129415, SDNU). A: Thallus with apothecia; B: Apothecium

section; C: Ascus and ascospores. Buellia penichra (Wang 20181108, SDNU). D: Thallus with

apothecia; E: Apothecium section; F: Ascospores, showing stages of development.

SPECIMENS EXAMINED: CHINA. YUNNAN, Shangri-La, Mt. Tianbao, alt. 3790 m, on

bark, 17 Aug. 2018, Chunxiao Wang 20181131 (SDNU)); alt. 3845 m, on bark, 17 Aug.

2018, Chunxiao Wang 20181108 (SDNU).

Amandinea & Buellia species new to China ... 267

DIsTRIBUTION— Western North America (Rocky Mountains) (Bungartz &

al. 2007). New to China.

CoMMENTS—The protologue describes Buellia penichra as containing

atranorin and sometimes placodiolic acid (Bungartz & al. 2007), but our

specimens contain only atranorin. This species is characterized by the

submuriform spores lacking a perispore and presence of atranorin. Buellia

penichra might be confused with B. muriformis A. Nordin & Tonsberg on

the basis of its ascospores lacking a perispore, but B. muriformis differs in its

secondary chemistry (containing placodiolic acid, atranorin, isousnic acid,

brialmontin 1 and brialmontin 2) and larger (25-39 x 13-18 um) ascospores

(Nordin 1999).

Acknowledgements

The authors thank Dr. John A. Elix (Australian National University, Canberra) and

Dr. Shou-Yu Guo (State Key Laboratory of Mycology, Chinese Academy of Sciences,

Beijing) for guidance in specimen identification and presubmission reviews. We also

thank Shu-Kun Yan and other laboratory members for assistance during this study.

This investigation was supported by the Emergency Management Project of National

Natural Science Foundation of China (31750001), the National Natural Science

Foundation of China (31600100) and the Scientific Research Foundation of Graduate

School of Shandong Normal University (SCX201945).

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Elix JA, Kantvilas G. 2013a. New taxa and new records of Amandinea (Physciaceae, Ascomycota) in

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Elix JA, Kantvilas G. 2016b. New species and new records of buellioid lichens (Ascomycota,

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Ascomycota) from New Zealand. Telopea 20: 75-84. https://doi.org/10.7751/telopea1 1334.

Elix JA, Mayrhofer H. 2018. Three new species and ten new records of buellioid lichens (Ascomycota,

Caliciaceae) from New Zealand. Australasian Lichenology 82: 68-79.

Elix JA, Malcolm WM, Knight A. 2015. New records and new combinations of buellioid lichens

(Physciaceae, Ascomycota) from New Zealand. Australasian Lichenology 77: 36-41.

Elix JA, Kantvilas G, McCarthy PM. 2017a. Thirteen new species and a key to buellioid lichens

(Caliciaceae, Ascomycota) in Australia. Australasian Lichenology 81: 26-67.

Elix JA, Mayrhofer H, McCarthy PM. 2017b. New species and a new record of buellioid lichens

(Physciaceae, Ascomycota) from Australia. Australasian Lichenology 80: 28-37.

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Physciaceae) from New Zealand. Australasian Lichenology 78: 11-12.

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America. Bryologist 102(2): 249-264. https://doi.org/10.2307/3244364

Amandinea & Buellia species new to China ... 269

Nordin A. 2000. Taxonomy and phylogeny of Buellia species with pluriseptate spores

(Lecanorales, Ascomycotina). Symbolae Botanicae Upsalienses 33(1). 117 p.

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Senkardesler A. 2010. Additions and corrections of types in the genus Buellia s. lat. (Physciaceae)

described by J. Steiner. Lichenologist 42(4): 439-448.

https://doi.org/10.1017/S0024282910000241

Tonsberg T, Nordin A, Tibell L. 2012. Amandinea lignicola, a new species from the Pacific coast

of North America. Graphis Scripta 24(2): 60-64.

Wang QD, Liu FY, Wu XH, Zhao X, Jia ZF. 2018. Buellia taishanensis sp. nov. and new Buellia

records from Mt. Tai, China. Mycosystema 133: 165-172. https://doi.org/10.5248/133.165

Watanuki O, Hara K, Harada H, Komine M, Fuji S. 2017. Buellia numerosa and B. subnumerosa,

two new species of the lichen genus Buellia (Caliciaceae) from Japan. Bryologist 120(1):

25-36. https://doi.org/10.1639/0007-2745-120.1.025

Wei JC. 1991. An enumeration of lichens in China. Beijing, International Academic Publishers.

278 p.

MY COTAXON

April-June 2019—Volume 134, pp. 271-273

https://doi.org/10.5248/134.271

Introduction of subfamily names for four clades in

Cladoniaceae and Peltigeraceae (Lecanoromycetes)

H. THORSTEN LUMBSCH! & STEVEN D. LEAVITT?

' Science & Education, The Field Museum,

1400 S Lakeshore Dr, Chicago, IL 60605-2496

? Department of Biology and M.L. Bean Life Science Museum, Brigham Young University,

Provo, UT 84602, USA

* CORRESPONDENCE TO: tlumbsch@fieldmuseum.org

ABSTRACT—In a recent proposal for a classification of orders and families in

Lecanoromycetidae and Ostropomycetidae based on a temporal approach, Squamarinaceae and

Stereocaulaceae were synonymized with Cladoniaceae, and Lobariaceae and Nephromataceae

with Peltigeraceae. Since these four synonymized families represent easily recognized,

monophyletic lineages and are well-established names, we here propose to accept them at

subfamilial rank, as Lobarioideae, Nephromatoideae, Squamarinoideae, and Stereocauloideae.

Key worps—classification, Lecanorales, Peltigerales, phylogeny, taxonomy

Introduction

Recently we proposed a revised classification of orders and families in the two

major subclasses of Lecanoromycetes, Lecanoromycetidae and Ostropomycetidae

(Kraichak & al. 2018) based on a temporal banding approach (Avise & Johns

1999, Kraichak & al. 2017). This classification proposal differed at the family

level from other recently proposed classifications (Jaklitsch & al. 2016; Lucking

& al. 2016, 2017; Lumbsch & Huhndorf 2010; Wijayawardene & al. 2018) in

accepting some clades at family level (such as Diploschistaceae, Thelotremataceae,

and Varicellariaceae) while synonymizing other clades previously accepted

as families within older families. The latter clades represented monophyletic

lineages but were shown to have diverged more recently than 111-235 Mya,

272 ... Lumbsch & Leavitt

which was the temporal band identified for the family rank. Four of the

families synonymized in Kraichak & al. (2018) have been widely used names,

and, hence, we suggest they should be accepted at subfamily rank. This allows

recognition of these well-established and monophyletic clades while avoiding

confusion due to incomparable ranks of clades.

Below we formally propose new subfamilies for the previously accepted

families Lobariaceae, Nephromataceae, Squamarinaceae, and Stereocaulaceae.

With the description of these subfamilies, Cladonioideae will become an

available name for Cladoniaceae s.str. (i.e., excluding Squamarinoideae and

Stereocauloideae) and Peltigeroideae for Peltigeraceae s.str. (i.e., excluding

Lobarioideae and Nephromatoideae).

Taxonomy

Cladoniaceae Zenker, in Goebel & Kunze, Pharmac. Waarenk. 1(3): 124 (1827).

Squamarinoideae Lumbsch & S.D. Leav., stat. nov.

MB 830561

BASIONYM: Squamarinaceae Hafellner, Beih. Nova Hedwigia 79: 342 (1984).

Stereocauloideae Lumbsch & S.D. Leav., stat. nov.

MB 830562

BASIONYM: Stereocaulaceae Chevall., Fl. Gén. Env. Paris 1: 596 (1826).

Peltigeraceae Dumort., Comment. Bot.: 68 (1822).

Lobarioideae Lumbsch & S.D. Leav., stat. nov.

MB 830563

BastonyM: Lobariaceae Chevall., Fl. Gén. Env. Paris 1: 609 (1826).

Nephromatoideae Lumbsch & S.D. Leav., stat. nov.

MB 830564

BastonyM: Nephromataceae Wetmore ex J.C. David & D. Hawksw.,

Syst. Ascom. 10: 15 (1991).

Acknowledgements

We wish to thank Dr. Brendan Hodkinson (Janssen Research & Development,

Spring House, PA USA) and Dr. Christian Printzen (Senckenberg Research Institute,

Frankfurt/Main, Germany) for reviewing this manuscript.

Literature cited

Avise JC, Johns GC. 1999. Proposal for a standardized temporal scheme of biological classification

for extant species. Proceedings of the National Academy of Science of the United States of

America 96: 7358-7363. https://doi.org/10.1073/pnas.96.13.7358

Cladoniaceae & Peltigeraceae: new subfamilies ... 273

Jaklitsch WM, Baral HO, Liicking R, Lumbsch HT. 2016. Syllabus of plant families —

A. Engler’s Syllabus der Pflanzenfamilien, Part 1/2 Ascomycota. Gebr. Borntraeger

Verlagsbuchhandlung. 322 p.

Kraichak E, Crespo A, Divakar PK, Leavitt SD, Lumbsch HT. 2017. A temporal banding

approach for consistent taxonomic ranking above the species level. Scientific Reports

7:2297 [7 p.]. https://doi.org/10.1038/s41598-017-02477-7

Kraichak E, Huang JP, Nelsen M, Leavitt SD, Lumbsch HT. 2018. A revised classification

of orders and families in the two major subclasses of Lecanoromycetes (Ascomycota)

based on a temporal approach. Botanical Journal of the Linnean Society 188: 233-249.

https://doi.org/10.1093/botlinnean/boy060

Liicking R, Hodkinson BP, Leavitt SD. 2016. The 2016 classification of lichenized fungi in

the Ascomycota and Basidiomycota - approaching one thousand genera. Bryologist 119:

361-416. https://doi.org/10.1639/0007-2745-119.4.361

Liicking R, Hodkinson BP, Leavitt SD. 2017. Corrections and amendments to the 2016

classification of lichenized fungi in the Ascomycota and Basidiomycota. Bryologist 120:

58-69. https://doi.org/10.1639/0007-2745-120.1.058

Lumbsch HT, Huhndorf SM. 2010. Myconet volume 14. Part one. Outline of Ascomycota—2009.

Fieldiana Life and Earth Sciences 1: 1-42. https://doi.org/10.3158/1557.1

Wijayawardene NN, Hyde KD, Lumbsch HT, Liu JK, Maharachchikumbura SSN, Ekanayaka

AH, Tian Q, Phookamsak R. 2018. Outline of Ascomycota: 2017. Fungal Diversity 88:

167-263. https://doi.org/10.1007/s13225-018-0394-8

MY COTAXON

April-June 2019—Volume 134, pp. 275-279

https://doi.org/10.5248/134.275

Cacumisporium fusiforme sp. nov.

from Jiangxi, China

ZHAO-HUAN XU’, KAI ZHANG}, XIU-GUO ZHANG‘,

RAFAEL FE, CASTANEDA-RUuiIZz, JIAN Ma’?*

' College of Agronomy & Jiangxi Key Laboratory for Conservation & Utilization of Fungal Resources,

Jiangxi Agricultural University, Nanchang, Jiangxi 330045, China

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

‘Department of Plant Pathology, Shandong Agricultural University,

Taian, Shandong 271018, China

° 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: majian821210@163.com; jxaumj@126.com

ABSTRACT—A new species, Cacumisporium fusiforme, is described and illustrated from a

specimen collected on dead branches of an unidentified broadleaf tree in Jiangxi, China. The

fungus differs from other described Cacumisporium species in its conidial shape and size.

A key to Cacumisporium species is provided.

KEY worDs—asexual fungi, hyphomycetes, saprobes, taxonomy

Introduction

Cacumisporium Preuss, was established by Preuss (1851) forasingle species

C. tenebrosum Preuss. The genus is diagnosed by differentiated, unbranched,

septate conidiophores and solitary, acropleurogenous, euseptate, pigmented

conidia that secede schizolytically from polyblastic, integrated, terminal,

flattened or inconspicuous denticulate conidiogenous cells after several

holoblastic sympodial elongations on enteroblastic percurrent regenerated

portions of the conidiophores (Goos 1969, Tsui & al. 2001, Castafeda-

Ruiz & al. 2007a,b, Wongsawas & al. 2009, Seifert & al. 2011). Saccardo

276 ... Xu & al.

(1877) recombined C. tenebrosum as Acrothecium tenebrosum (Preuss)

Sacc., but Acrothecium is now treated as an unaccepted genus based on an

unidentifiable type (Hughes 1958, Seifert & al. 2011). In modern literature,

Cacumisporium tenebrosum is treated as a junior synonym of C. capitulatum

(Corda) S. Hughes [= Helminthosporium capitulatum Corda; = Acrothecium

capitulatum (Corda) Ferraris] (Mangenot 1953, Hughes 1958, Sutton 1973,

Seifert & al. 2011). Eight other species have been published in Cacumisporium

(Mercado-Sierra & Castaneda-Ruiz 1987, Castaneda-Ruiz & Kendrick 1991,

Kirk 1992, Tsui & al. 2001, Castafteda-Ruiz & al. 2007a,b, Wongsawas &

al. 2009, Rambelli & Raimondo 2014). However, “C. oceanicum” Rambelli

is invalid (Index Fungorum 2018), and C. curvularioides R.F. Castaneda

& W.B. Kendr. has been synonymised with C. pleuroconidiophorum

(Davydkina & Melnik) R.F. Castafeda & al. (Castafteda-Ruiz & al. 2007a),

leaving Cacumisporium currently with seven valid taxa. The criteria used to

delimit species are based primarily on conidial shape, size, septation, and

pigmentation (Kirk 1992; Tsui & al. 2001; Castafteda-Ruiz & al. 2007a,b;

Wongsawas & al. 2009). A key to Cacumisporium species was provided by

Castafieda-Ruiz & al. (2007b).

The forests of Jiangxi have a rich fungal diversity, and many anamorphic

fungi have been discovered there (Ma & Zhang 2015, Ma & al. 2016, Xu & al.

2017). During an ongoing mycological survey in these forests, an interesting

hyphomycete with morphological features of Cacumisporium was collected

on dead branches. It was found to be distinct from previously described taxa

and is therefore proposed as new to science. The specimens are conserved in

the Herbarium of the Department of Plant Pathology, Jiangxi Agricultural

University, Nanchang, China (HJAUP).

Cacumisporium fusiforme Z.H. Xu, Jian Ma, X.G. Zhang &

R.F. Castaneda, sp. nov. Fic. 1

MB 829269

Differs from Cacumisporium sigmoideum by its smaller, 3(-4)-euseptate, fusiform

conidia with a rounded apex.

Type: China, Jiangxi Province, Jiulianshan National Nature Reserve, on dead branches

of an unidentified broadleaf tree, 3 November 2014, J. Ma (Holotype, HJAUP M0307).

EryMo .oey: refers to the fusiform conidial shape.

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

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

smooth, subhyaline to pale brown hyphae. CoONIDIOPHORES macronematous,

Cacumisporium fusiforme sp. nov. (China) ... 277

TT)

TTT

cand a Nh O00

Fic. 1. Cacumisporium fusiforme (holotype, HJAUP M0307).

A-C. Conidiophores, conidiogenous cells, and conidia; D. Conidia.

mononematous, simple, erect, straight or flexuous, smooth, septate, cylindrical,

often with 1-2 enteroblastic percurrent extensions, dark brown at the base,

pale brown or subhyaline at the apex, 73-185 x 4-7 um. CONIDIOGENOUS

CELLS polyblastic, integrated, terminal, sympodial elongated, with flattened

or inconspicuous denticles at the conidiogenous loci, brown, pale brown to

subhyaline, 35-61.5 x 5.5-6.5 um. Conidial secession schizolytic. CONIDIA

solitary, acrogenous, fusiform, occasionally curved, 3(-4)-euseptate, end

cells subhyaline or pale brown to pale yellow, middle cells yellow-brown,

smooth, 23.5-33.5 x 7.5-9 um, apex rounded, base truncate, 2-2.5 um wide,

accumulating in brown, somewhat mucilaginous masses.

COMMENTS —Among Cacumisporium species, C. fusiforme is most similar to

C. sigmoideum Mercado & R.F. Castafieda, which is distinguished by its falcate,

mostly sigmoid, 3-euseptate, larger (26-41 x 7.6-10.2 um) conidia with an

acute apex (Mercado-Sierra & Castaneda-Ruiz 1987).

278 ... Xu & al.

Key to Cacumisporium species

1. Conidia 1-euseptate, versicolorous, ovoid,

S268 1416 MIN tes eo es ead eee eine a eines che a ete ret C. uniseptatum

1. Conidia 3(—4)- or 7-euseptate, botuliform, fusiform, obturbinate,

sigmoid, obovoid to broadly ellipsoid... 2.1 ..s. x eeays vehex seer se ages ees 2

2. Conidia 7-euseptate,

narrowly ellipsoid to somewhat fusiform, 26-32 x (3-)4(-5) um ... C. spooneri

DEA OMICER SS (Se AC UISCDLAUE! <5 We ren tha tl run i, bes Seed a Wo sed We bracdg te See Ma ts erent =a Sree aba drser ath Pee 3

3. Conidia rugose to verrucose,

obovoid to broadly ellipsoid, 28-35 x 14-20 um ................. C. rugosum

Du Ae UNA CIO OMI Tereyors 5S." aan, sual a ge esl gp Seo ok ye em Agee oad gh SOLE sy OL ee SL Lg ae 4

4, Conidia botuliform, 16-22 x 4.5-5um ........ eee eee eee eee C. capitulatum

4. Conidia obturbinate, sigmoid or fusiform, at least 6 um diam. .................. 5

5. Conidia concolorous, broad fusiform, sometimes navicular or

somewhat ellipsoid, 12-17 x 6-8 UM ........ eee eee ee eee C. tropicale

Se GONidiasVersiCOlOe wy oh. 8. 2c bon we ely een Ee eth Reig ee 2 eee ee Ree A 5 8 6

6. Conidia with one swollen cell strongly pigmented,

1s PA Se Gf osates 1G egCbect Sc PAE CRORE ae AOR RN C. pleuroconidiophorum

6. Conidia with two cells strongly pigmented ............. 00. cee eee ee eee eee eee 7

7. Conidia fusiform, 23.5-33.5 x 7.5-9 um, 3(-4)-euseptate,

CUMS REE OLB 0G Cot lez) BT 4g SMe cosine Fae Ral eh pe PE RE Ra ete Ae a C. fusiforme

7. Conidia falcate, mostly sigmoid, 26-41 x 7.6-10.2 tum, 3-euseptate,

MiCLinweloty: (e101 02270) < en Me ee, Ineeeal. eee ceeeeets Sen eR eine C. sigmoideum

Acknowledgments

The authors express gratitude to Dr. D.W. Li (The Connecticut Agricultural

Experiment Station Valley Laboratory, Windsor CT, USA) and Dr. J.S Monteiro (Museu

Paraense Emilio Goeldi, Belém, Brazil) for serving as pre-submission reviewers and to

Dr. Shaun Pennycook for nomenclatural review and Dr. Lorelei L. Norvell for editorial

review. This project was supported by the National Natural Science Foundation of

China (Nos. 31360011, 31870016), and the Education Department of Jiangxi Province

of China (No. GJJ160357).

Literature cited

Castafieda-Ruiz RE, Kendrick B. 1991. Ninety-nine conidial fungi from Cuba and three from

Canada. University of Waterloo Biology Series 35. 132 p.

Castaneda-Ruiz RF, Heredia-Abarca G, Arias RM, Saikawa M, Minter DW, Stadler M. 2007a.

Anamorphic fungi from submerged plant material: Phaeomonilia pleiomorpha, P. corticola and

Cacumisporium pleuroconidiophorum. Mycotaxon 100: 327-336.

Castaneda-Ruiz RF, Gusmao LFP, Guarro J, Stchigel AM. 2007b. Two new anamorphic fungi from

Brazil, Cacumisporium tropicale and Acrodictys irregularis. Mycotaxon 102: 91-99.

Cacumisporium fusiforme sp. nov. (China) ... 279

Hughes SJ. 1958. Revisiones hyphomycetum aliquot cum appendice de nominibus rejiciendis.

Canadian Journal of Botany 36(6): 727-836. https://doi.org/10.1139/b58-067

Index Fungorum. 2018. Fungal names search [accessed 25 December 2018].

http://www.indexfungorum.org/names/Names.asp

Kirk PM. 1992. New or interesting microfungi XVI. Hyphomycetes from the British Isles.

Mycotaxon 43: 231-236.

MaJ, Zhang XG. 2015. A preliminary report of dematiaceous hyphomycetes from dead branches in

Jiangxi Province. Biological Disaster Science 38(4): 290-293.

Ma J, Xia JW, Zhang XG. 2016. Three new species of Hemicorynespora and Solicorynespora from

southern China. Mycotaxon 131: 263-268. https://doi.org/10.5248/131.263

Mangenot F. 1953. Sur quelques hyphales dematiées lignicoles. Revue de Mycologie 18: 133-148.

Mercado-Sierra A, Castafteda-Ruiz RF. 1987. Nuevos o raros Hifomicetes de Cuba. I. Especies de

Cacumisporium, Gueda, Rhinocladium, y Veronaea. Acta Botanica Cubana 50: 1-7.

Preuss CGT. 1851. Uebersicht untersuchter Pilze, besonders aus der Umgegend von Hoyerswerda.

Linnaea 24: 99-153.

Rambelli A, Raimondo FM. 2014. Interesting dematiaceous hyphomycetes on Lodoicea

maldivica dead fragments. Plant Biosystems 148(3): 467-472.

https://doi.org/10.1080/11263504.2013.785449

Saccardo PA. 1877. Fungi italici autographice delineati. Michelia 1(1): 73-100.

Seifert K, Morgan-Jones G, Gams W, Kendrick B. 2011. The genera of hyphomycetes. CBS

Biodiversity Series 9. 997 p.

Sutton B. 1973. Hyphomycetes from Manitoba and Saskatchewan, Canada. Mycological Paper

132. 143 p.

Tsui CKM, Goh TK, Hyde KD, Hodgkiss IJ. 2001. New species or records of Cacumisporium,

Helicosporium, Monotosporella and Bahusutrabeeja on submerged wood in Hong Kong

streams. Mycologia 93(2): 389-397. https://doi.org/10.2307/3761660

Wongsawas M, Wang HK, Hyde KD, Lin FC. 2009. Two new hyphomycetes from submerged

wood collected in China. Sydowia 61(2): 345-351.

Xu ZH, Hu DM, Luo YQ, Ma J. 2017. Three species of Linkosia and Spadicoides new to China.

Mycotaxon 132: 243-250. https://doi.org/10.5248/132.243

MY COTAXON

April-June 2019—Volume 134, pp. 281-287

https://doi.org/10.5248/134.281

Acarospora squamulosa, the correct name for A. peliocypha

KERRY KNUDSEN’ , LINDA IN ARCADIA’, 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

’*Kastri, 22013, Arkadias, Greece

* CORRESPONDENCE TO: knudsen@fzp.czu.cz

ABSTRACT—The epithet of the species widely known as Acarospora “peliscypha” should be

restored to its original orthography, A. peliocypha. The oldest name for this species is Lichen

squamulosus, which has priority. Lichen squamulosus is lectotypified. Acarospora squamulosa

replaces the later name A. peliocypha. Acarospora squamulosa is reported for continental

North America from Baffin Island, Nunavut Territory, Canada.

KEY worpDs—Acarospora rugulosa, nomenclature, priority, taxonomy

Introduction

Although stability in nomenclature is desirable, it is not always achievable.

Improvements in the understanding of taxonomy often necessitate changes in

names, and the molecular revolution has caused, and will continue to cause,

massive changes to the taxonomy of lichenized fungi, with corresponding

changes in names. Another source of instability can be the need to achieve

consensus about names. Consensus is achieved by applying the rules of the

International Code of Nomenclature, as those rules are accepted throughout

the botanical and mycological community, but “consensus” and “stability” come

into conflict in cases where the rules imply that a commonly used name is not

the correct one. In such cases, a judgment must be made about whether to accept

a change in name or to invoke the cumbersome procedure of conservation

to preserve the more familiar usage. In this paper we resurrect the name

Acarospora squamulosa for the species currently known as A. peliocypha, which

282 ... Knudsen, Arcadia, Kocourkova

frequently occurs in the literature as A. “peliscypha’, a misspelling published by

Theodore Fries (1860) based on the orthographic variant Parmelia pelioscypha

listed by Wallroth (1831: 474). The orthographic variants A. peliocypha and

A. pelioscypha are essentially synonyms: the first element from Greek pelios

(meAtoc) means “black; dark,’ and the alternative second elements are either

from scyphos (oxv@oc) meaning “cup” or from cyphos (kv@dc) meaning

“curved; the round of a cup.” Since both variants are authentic, the original

version peliocypha is not a correctable error and must be accepted.

The oldest epithet for this species is Schrader’s squamulosus, which Acharius

and other early 19" century authors accepted as having priority. It is unfortunate

that the later epithet “peliscypha” was taken up following Fries’s work. We do

not consider that conservation would be appropriate in this case. We prefer to

follow the Code of Nomenclature (which seems to us a matter of justice, too)

and resurrect the older epithet.

Material & methods

Specimens from GZU, H, M and the private herbaria of Kocourkova & Knudsen

(hb. K&K) and Jiti Malicek (hb. Mali¢ek) were studied using standard microscopy

and spot tests. Specimens were sectioned by hand and measurements were made

in water. Amyloid reactions of the hymenium were studied in fresh undiluted

Merck Lugol’s solution (IKI) following Knudsen & Kocourkova (2018). The

excellent description by A.H. Magnusson (1929) was revised. Macrophotographs

were taken using an Olympus DP74 digital camera mounted on Olympus SZX

7 stereomicroscope equipped with PRO-SZM1-Focus Drive Motorization for

stacking pictures and stacked using the Olympus DeepFocus 3.4 module. The figure

plates were processed with QuickPhoto Camera 3.2 software fitted with Promicra

Publisher Module and eventually refined with Adobe Photoshop CS4 Extended ver.

Dlg.

Taxonomy

Acarospora squamulosa (Schrad.) Trevis., Rivista Period. Lav. Regia Accad.

Sci. Lett. Arti Padova 1: 263. 1853. Fic. 1

= Lichen squamulosus Schrad., Ann. Bot. (Usteri) 22: 84. 1797.

TYPE: Systematische Sammlung Cryptogamischer Gewachse, Fascicle I, no.

153, Germany. Niedersachsen, Harz, on rocks (lectotype designated here,

M! MBT385224; isolectotypes, GOTE! L!).

= Parmelia squamulosa (Schrad.) Ach., Methodus, sect. post.: 181. 1803.

= Parmelia peliocypha Wahlenb., in Acharius, Methodus, suppl.: 40. 1803.

Type: Norway. Lappland, Nord Cap, on rocks, Wahlenberg s.n. [type not

seen or located].

= Acarospora peliocypha (Wahlenb.) ‘Th. Fr. [as “peliscypha”], Lich. Arct.: 89. 1860.

Acarospora peliocypha a synonym of Acarospora squamulosa ... 283

Fic. 1. Acarospora squamulosa (lectotype, Schrader No. 15, M): A. Subsquamulose imbricate

thallus with apothecia; B. Crustose areolate thallus; C. Detail of apothecia with thalline margin and

rugulose disc. Scale bars: A, B = 1 mm; C = 500 um.

THALLUS of areoles quite variable in size, as small as 0.5 mm and up to 3 mm

diam., 100-500 um thick, broadly attached, becoming sometimes lobulate

and subsquamulose and sometimes imbricate, replicating by division. Upper

284 ... Knudsen, Arcadia, Kocourkova

surface dark to light brown or reddish brown, shiny or matte, epruinose. Lower

surface brown. Epicortex lacking or thin. Cortex 30-50 um thick, upper layer

brown, lower layer hyaline, cells round, 3-5 um diam. Algal layer continuous

to thin and scattered, 50-200 um thick, algal cells 6-15 um diam. Medulla

thickening corresponding with areole size, hyphae thin-walled, 2-3 um diam.

APOTHECIA usually 1 per areole, beginning immersed and (usually)

becoming elevated in a thalline margin or expanding and thus reducing

the areole to a thalline margin (pseudo-lecanorine), <1 mm diam., the disc

epruinose, usually rugulose, usually round. Parathecium expanding around disc

10-30 um without excluding thalline margin, 30-60 um diam. Epihymenium

15-30 um thick, coherent, brown to yellowish brown. Hymenium (including

epihymenium but not accretions on disc surface) 100-120(-130) um tall,

hymenial gel IKI+ blue to red (hemiamyloid). Paraphyses 1.5-2 um diam. at

mid-level, apices unexpanded or expanded <3 um. Asci 70-90 x 16-20 um,

100-200 ascospores per ascus. Ascospores 3-6 x 1-2 um. Subhymenium

25-50 um thick, IKI+ blue (euamyloid). Hypothecium 10-30 um tall. Pycnidia

inconspicuous, ca. 100 x 60 um, ampuliform conidiogenous cells 10-15 um

tall, conidia mostly 2 x 1 um. Gyrophoric acid in cortex, C+, KC+ pinkish red

in thin section, rarely cortical parts lacking a distinct reaction.

SELECTED SPECIMENS EXAMINED: CANADA. Nunavut. Baffin Island, Head of Clyde

Fiord, on gneiss rock below a gyrfalcon nest, July 1950, M.E. Hale Jr. 372 (H). CZECH

REPUBLIC, CENTRAL BOHEMIA, Distr. Rakovnik, Krivoklatsko Protected Landscape

Area, Hracholusky, Certova skdéla Nature Reserve, 49°59’52.535”N, 13°47’27.885’E,

345 m, on top of high outcrop above Berounka River, 14 July 1997, J. Kocourkova 3727

(hb. K&K), Nezabudické skakly nature reserve, 50°1’10.02”N 13°51’2.34’E, 255 m, on

spilite, 11 April 1996, J. Kocourkova 9855 (hb. K&K), Roztoky, Na Babé Nature Reserve,

50°1'50.676”N, 13°52’18.069”E, 290 m, on steep rocky slope above Berounka River, on

rhyolite, 17 June 1997, J. Kocourkova 8471 (hb K&K), Tytovice, Tytov National Nature

Reserve, Tytovické skaly, w-facing rocks above Berounka River, 49°58’59”N 13°47'39’E,

330-420 m, on volcanic rock, 11 Jan. 2018, J. Mali¢ek 12585 (hb Malicek). WESTERN

BoHEMIA. Distr. Plzem-sever, Nectiny, Preitenstein castle ruins, s- to sw-facing

rocks below the castle, 49°57’37”N, 13°09’41”E., 550-580 m, on on spilite basalt, 13

April 2014, J. Malicek 7697 (hb Malicek). FINLAND. JAmsA, Seppola, 21 April 1934,

A. Horkiverr (H); Turku, Ruissalo Kalliolla, on rock, 19 July 1922, V. Rasanen (H);

Kavipja, on rock, 3 June 1908, K. Linkola (H). NORWAY. OppLanp, Ringebu, near

the village, on boulders in stone fence, 28 June 1925, A.H. Magnusson, Lichenes selecti

scandinavici No 67 (H, GZU). SWEDEN. SGDERMANLAND, Flemingsberg parish,

ca. 800 meters S of Masmo and just E of Lake Albysjon, 40 m, on S-facing exposed

siliceous rock, 59°14’30”N 17°52’49”E, 10 April 2009, M. Westberg TU-3 (hb. K&K, 2

specimens). RUSSIAN FEDERATION. KurkyokI! [KuRKIYEK1/Kypxuexu], on schist, 8

August 1935, V. Rasanen (H). ORENBURG REGION, Mednogorsk district, 14 km NE of

Mednogorsk, vicinity of Yumaguzino the 2"¢ village, 51°32’41”N 58°38’50’E, 357 m, on

basalt outcrops, 30 May 2013, A. Paukov (hb. K & K).

Acarospora peliocypha a synonym of Acarospora squamulosa ... 285

ECOLOGY & DISTRIBUTION. Asia, Europe, North America, on various siliceous

rock, on gneiss, rhyolite and basalt including spilite (Magnusson 1929,

Golubkova 1988).

Discussion. Schrader’s exsiccatae Systematische Sammlung Cryptogamischer

Gewachse were published in two fascicles in 1796 and 1797. According to

Sayre (1969), they did not include any descriptive text, although schedae for

each fascicle were published in volumes 20 (1796) and 22 (1797) of Usteri'’s

Annalen der Botanik. According to Stafleu & Cowan (1985), the schedae

were also published in 1796 and 1797 as independent publications. It is not

known therefore, whether the journal contains preprints or reprints, but we

have assumed the former. We have not been able to consult the independent

publications but assume that their text is word-for-word identical with the

journal.

The name Lichen squamulosus was validly published in the journal adjacent

to the number “153”, an unambiguous reference to number 153 in the exsiccata.

The name must therefore be typified on number 153 in the exsiccata. Because

the exsiccata was issued in multiple copies, and we cannot be sure that each

number represents a single gathering, a lectotype should be designated. We

were able to locate only three specimens of number 153, one in the herbarium

of Munich (M), one in Leiden (L), and one in Gottingen (GOET). The

specimen in M is 7 cm in diameter and is designated here as the lectotype, as

it can be easily studied without any damage. The specimen in L is too scanty

for non-destructive microscopic and chemical examination, but does appear to

belong to the same species, and is acceptable as an isolectotype. The specimen

in GOET, which is smaller than the specimen in M and identified by F. Arnold

who also did spot tests, is also an excellent isolectotype.

The lectotype and isolectotypes belong to the species widely known as

either Acarospora “peliscypha” or A. peliocypha. Theodore Fries cited several

names in synonymy, but as they are not validly published, proposed at an infra-

specific rank, or cited with “p.p.” or “?’, his name is legitimate (Fries 1860).

The correct spelling of Fries’s epithet should be peliocypha, because he based

his combination on Parmelia peliocypha Wahlenb., which he miscited as P

“peliscypha.” Acharius (1803) himself considered Parmelia peliocypha to be

merely a synonym of Parmelia squamulosa; but Wahlenberg, who named P.

peliocypha, regarded it as referring to a good species. ICN (Shenzhen) Art. 36.1,

Ex 3 (Turland & al. 2018) supports the name as validly published. We could

not locate Wahlenberg’s collection at BM, H, UPS, or S or in the literature.

Magnusson (1929) did not state that he actually saw Wahlenberg’s specimens,

286 ... Knudsen, Arcadia, Kocourkova

only that Wahlenberg eventually incorporated P. peliocypha “for the most part”

into his concept of Lichen cervinus Wahlenb., which may explain the lack of

annotated peliocypha type material. We choose not to neotypify the name

P. peliocypha, because we believe a type may well be found among Wahlenberg

specimens of L. cervinus. It is also possible the type actually represents the

morphologically similar species currently known as Acarospora rugulosa Korb.

We base our synonymy on the consensus of Acharius (1803), Arnold (1880),

and Magnusson (1929) that Wahlenberg applied P peliocypha to the species

known as L. squamulosus.

As often happened in 19" century, names were often based on one-

line descriptions without examining the types. In this case the name Lichen

squamulosus was misapplied to the species A. macrospora A. Massal. ex Bagl.

(Magnusson 1929). In his discussion of the nomenclature of A. macrospora,

Magnusson stated that L. squamulosus in Schrader’s type was “for the most

part” the species later called Parmelia peliocypha, but he wrote “to not cause

perpetual confusion it is best to let this name sink into oblivion” (Magnusson

1929). It seems fairer that we not propose to conserve the name P. peliocypha

but recognize the nomenclatural priority of Lichen squamulosus and not let it

sink into oblivion. We oppose conservation because E. Acharius, F. Arnold,

and A.H. Magnusson all recognized L. squamulosus as the same species as

P. peliocypha (Acharius 1803, Arnold 1880, Magnusson 1929). Magnusson

arbitrarily decided to accept the name P peliocypha even though he recognized

that the taxon was originally called L. squamulosus.

Acarospora squamulosa is an arctic and montane species in Europe, although

it has been collected at elevations as low as 255 m in the Czech Republic

(Magnusson 1929). Collections have been identified by A.H. Magnusson

from Fennoscandia to Russia in Europe (Magnusson 1929). Golubkova (1988)

reported A. squamulosa in eastern Siberia in Asia, and Magnusson (1929)

reported the species from Greenland in North America. We identified a

specimen from Canada during this study but have seen no other specimens

of A. squamulosa from continental North America. The taxon reported as A.

“peliscypha” from California and the Sonoran region is rare and represents an

undescribed species that needs further study (Knudsen 2007; for picture see

Knudsen & al. 2017).

Acarospora squamulosa can be confused with A. rugulosa [= A. montana

H. Magn.]. Acarospora rugulosa has a higher hymenium (110-)125-140(-175)

uum vs. 100-120(-130) um in A. squamulosa, and phylogenetic analysis supports

A. squamulosa and A. rugulosa as distinct species (Westberg & al. 2015). The

Acarospora peliocypha a synonym of Acarospora squamulosa ... 287

two species can be identified accurately only by hymenium height, as they share

overlapping morphological characters. Acarospora rugulosa is on the North

American lichen checklist (Esslinger 2018) but we have yet to confirm this report.

Acknowledgements

We thank our reviewers, James Lendemer (The New York Botanical Garden,

Bronx, USA) and Andrei Tsurykau (F Skorina Gomel State University, Belarus),

and the curators of GOTE, GZU H, L, and M. We also thank T. Ahti, M. Appelhans,

R. Bijmoer, A. Nordin, C. Schollaard, and H.J.M. Sipman for assistance in locating

isolectotypes. We thank J. McNeill, J. Mali¢ek and A. Tsurykau for their assistance and

the late J.R. Laundon for bringing this problem to our attention. The work of Kerry

Knudsen and Jana Kocourkova was financially supported by the grant Environmental

Aspects of Sustainable Development of Society 42900/1312/3166 from the Faculty of

Environmental Sciences, Czech University of Life Sciences, Prague.

Literature cited

Acharius E. 1803. Methodus qua omnes detectos lichenes. Supplementum. Leipzig. 52 p.

Arnold F, 1880. Lichenologische Fragmente. XXII. Flora (Regensburg) 63: 371-385.

Esslinger TL. 2018. A cumulative checklist for the lichen-forming, lichenicolous and allied fungi

of the continental United States and Canada. Version#22. Opuscula Philolichenum 17: 6-268.

Fries TM. 1860. Lichenes arctoi. Uppsala. 298 p.

Golubkova N. 1988. The lichen family Acarosporaceae Zahlbr. in the U.S.S.R. Leningrad: Nauka.

(In Russian). 136 p.

Knudsen K. 2007. Acarospora. 1-38, in: TH Nash III & al. (eds). Lichen Flora of the Greater

Sonoran Desert Region. Volume 3. Lichens Unlimited, Arizona State University, Tempe.

Knudsen K, Kocourkova J. 2018. Two new calciphytes from Western North America, Acarospora

brucei and Acarospora erratica (Acarosporaceae). Opuscula Philolichenum 17: 342-350.

Knudsen K, Lendemer JC, Schultz M, Kocourkova J, Sheard JW, Pigniola A, Wheeler T. 2017.

Lichen biodiversity and ecology in the San Bernardino and San Jacinto Mountains in southern

California (U.S.A.). Opuscula Philolichenum 16: 15-138.

Magnusson AH. 1929. A monograph of the genus Acarospora. Kungliga Svenska Vetenskaps-

Akademiens Handlingar 7(4). 400 p.

Sayre G. 1969. Cryptogamae exsiccatae: an annotated bibliography of published exsiccatae of Algae,

Lichenes, Hepaticae, and Musci. Memoirs of the New York Botanical Garden 19. 174 p.

Stafleu FA, Cowan RS. 1985. Taxonomic literature, 2nd edition, vol. 5: Sal-Ste. The Hague &

Boston. 1066 p.

Turland NJ, Wiersema JH, Barrie FR, Greuter W, Hawksworth DL, Herendeen PS, Knapp S,

Kusber WH, Li DZ, Marhold K, May TW, McNeill J, Monro AM, Prado J, Price MJ, Smith GF

(eds). 2018. International Code of Nomenclature for algae, fungi, and plants (Shenzhen Code)

adopted by the Nineteenth International Botanical Congress Shenzhen, China, July 2017.

Regnum Vegetabile 159. Glashiitten: Koeltz Botanical Books. 254 p.

Wallroth FG. 1831. Flora cryptogamica germaniae, vol. 1. Filices, lichenastra, muscos et lichenes.

Nuremberg. 654 p.

Westberg, M, Millanes AM, Knudsen K, Wedin M. 2015. Phylogeny of the Acarosporaceae

(Lecanoromycetes, Ascomycota, Fungi) and the evolution of carbonized ascomata. Fungal

Diversity 70: 145-158. https://doi.org/10.1007/s13225-015-0325-x

MY COTAXON

April-June 2019—Volume 134, pp. 289-294

https://doi.org/10.5248/134.289

Megalaria yunnanensis sp. nov. from Yunnan, China

CHUN-XIAO WANG’, XIAO ZHANG’, CHUAN-FENG ZHENG’, LING Hu?*

'Key Laboratory of Plant Stress Research, College of Life Sciences &

?Institute of Environment and Ecology,

Shandong Normal University, Jinan, 250014, P. R. China

* CORRESPONDENCE TO: Hu_Ling_123@163.com

ABSTRACT—Megalaria yunnanensis from southern China is described as a new species.

The lichen is diagnosed by its bi-layered exciple, presence of atranorin, zeorin, and

fumarprotocetraric acid in the thallus, ascospore size (20-25 x 5—7.5 um), and the lack of

indentations from the cell lumina into the center of the spore septum.

Key worps —East Asia, Lecanorales, lichenized fungi, Ramalinaceae, taxonomy

Introduction

Megalaria (Ramalinaceae) was erected for the single species M. grossa

(Pers. ex Nyl.) Hafellner (Hafellner 1984). The genus is recognized by its

crustose thallus, biatorine apothecia, greenish to dark purple epihymenium,

(2—)8-spored ascus, Lecanora-, Bacidia-, or Biatora-type asci, and 1-septate,

ellipsoidal to subglobose, colourless, thick-walled ascospores (Ekman &

Tonsberg 1996; Brodo & al. 2001: 428-429; Galloway 2004; Lendemer 2007;

Sanderson 2009; Fryday & Lendemer 2010; McCarthy & Elix 2016; McMullin

& Lendemer 2016).

Megalaria contains about 48 species worldwide (Lendemer & al. 2016,

McMullin & Lendemer 2016, McCarthy & Elix 2016, Su & Ren 2017, Elix &

McCarthy 2018). In China, only two species, M. hainanensis Q. Ren and M.

laureri (Hepp ex Th. Fr.) Hafellner have been reported (Su & Ren 2017). During

our study of the lichen flora of Mt. Wuliang in Jingdong county, Yunnan

province, China, we collected a species of Megalaria new to science.

290 ... Wang & al.

Materials & methods

The specimens were collected in Yunnan Province, China, and are preserved in

Lichen Section of the Botanical Herbarium, Shandong Normal University, Jinan,

China (SDNU). The specimens were examined morphologically using a COIC

XTL7045B2 stereo-microscope and anatomically with an Olympus CX41 polarizing

microscope and photographed under Olympus SZX16 and BX61 with DP72. Thallus

and medulla were tested with K (a 10% aqueous solution of potassium hydroxide),

C (a saturated solution of aqueous sodium hypochlorite) for identification. Lichen

substances were identified using standardized thin layer chromatography techniques

(TLC) with system C (Orange & al. 2001).

Total genomic DNA was extracted with DNeasy Plant Mini Kit according to

the manufacturer’s instructions and purified with PCR quick-spin™ PCR Product

Purification Kit. The ITS1-5.8S-ITS2 regions were amplified in a C1000TM automatic

thermocycler using primers ITS1IF (Gardes & Bruns 1993) and ITS4 (White & al.

1990) in a 50 uL volume containing 5 units of 1 ul Taq DNA Polymerase (Sangon

Biotech), 5 uL ITS buffer, 37.2u] ddH20, 0.8 uL 10 uM F primer solution, 0.8 uL

10 uM R primer solution, 3.2 uL 2.5 mM per dNTP solution, and 2 uL genomic

DNA. PCR thermal cycling parameters were initial denaturation at 98°C for 2 min,

followed by 35 3-step cycles (98°C for 10 s, 54°C for 10 s, 72°C for 1 min), and

a final 5 min extension at 72°C. PCR products were sanger-sequenced by Sangon

Biotechnology Company.

All raw sequences were assembled and edited using SeqMan 7.0. The ITS data

set comprised two newly generated sequences from the holotype and a paratype of

our new species and eight relevant ITS sequences of Megalaria (plus two outgroup

sequences) downloaded from GenBank. The sequences were aligned in MAFFT

version 7 (Katoh & al. 2005) with the parameters set to default values. Ambiguous

regions were excluded using Gblocks (Talavera & Castresana 2007) with default

settings. There were 479 positions total in the final dataset. Maximum likelihood

(ML) was used to reconstruct the phylogenetic tree (Fic. 2). Biatora subduplex

(Nyl.) Printzen and B. meiocarpa (Nyl.) Arnold were used as outgroup (McMullin

& Lendemer 2016). All characters were equally weighted and treated as unordered.

Taxonomy

Megalaria yunnanensis C.X. Wang & L. Hu, sp. nov. Fie. 1

MB 829259

Differs from Megalaria albocincta by its longer ascospores and its lack of indentations

from the cell lumina into the center of the spore septum.

Type: China. Yunnan province, Jingdong county, Mt. Wuliang, 24°24’02”N 100°45’44”E,

alt. 2200 m, on bark, 7 Aug. 2017, R. Tang 20170963 (Holotype, SDNU; GenBank

MK348528).

ErymMo.oey: The specific epithet yunnanensis refers to Yunnan province, where this

species was found.

Megalaria yunnanensis sp. nov. (China) ... 291

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Fic. 1. Megalaria yunnanensis. (Holotype, SDNU (Tang 20170963)). A. Thallus and apothecia;

B. Apothecial section; C. Asci and ascospores; D. Amyloid reaction of ascus, Biatora-type; E.

Ascospores; F. TLC chromatograms (R = Lethariella cladonioides).

THALLUS crustose, verrucose protuberance, corticolous, white, without

soredia. APOTHECIA biatorine, adnate, circular in outline, 0.38-1 mm

diam., margins pale, waxy white, becoming excluded with age; piscs black,

flat, not pruinose, occasionally with few crystals. Epihymenium dark green

to black, 22.5—52.5 um tall, K-, N+ red; hymenium hyaline, 50—67.5 um tall,

K-, N-; subhymenium colourless to red-brown or black, 37.5—50 um tall,

K+ red, N+ red-brown to red; hypothecium black, 25-65 um tall, K-,

N+ red; paraphysis hyaline, simple, easily free, I+ blue. Ascr 8-spored,

45—-62.5 x 11.25-15 um, Biatora-type. Ascosporgs ellipsoid, colourless,

thick walled, 1-septate, not halonate, 20—25 x 5—7.5 um (n = 32). PYCNIDIA

not seen.

CHEMISTRY—Cortex and medulla K+ yellow, C-, KC-; thallus UV-;

atranorin, zeorin, and fumarprotocetraric acid detected by TLC.

ADDITIONAL SPECIMENS EXAMINED: CHINA. YUNNAN, Jingdong, Mt. Wuliang,

24°24’02”N 100°45’44”E, alt. 2200 m, on bark, 7 Aug. 2017, R. Tang 20170805,

292 ... Wang & al.

GenBank MK348527; 20170806; 20170820; 20170838; 20170856; 20170870;

20170896; 20170904; 20170918; 20170923; 20170926; 20170956; 20170962;

20170966 (SDNU).

DIsTRIBUTION—The type specimen was collected from Mt. Wuliang,

Jingdong, Yunnan, which has a subtropical monsoon climate. The average

annual temperature, relative humidity, and rainfall are 18.3°C, 77%, and

108.7 cm. This species grows on bark and is known only from the type

locality.

Discussion

Megalaria yunnanensis is similar to M. albocincta (Degel.) Tonsberg,

M. alligatorensis Lendemer, M. anaglyptica (Kremp.) Fryday & Lendemer, and

M. pulverea (Borrer) Hafellner & E. Schreiner, all of which are characterized

by a bi-layered exciple with a prosoplectenchymatous outer layer and an

inner layer composed of a textura intricata with large intercellular spaces

and a thalline chemistry of atranorin, zeorin, and fumarprotocetraric acid.

However, the other four species are separated from M. yunnanensis by

their shorter or smaller ascospores—M. albocincta: 13-17 x 6.5—8.5 um,

M. alligatorensis: 12-14 x 3.8-5.5 um, M. anaglyptica: 17-22 x 4-6 um,

and M. pulverea: 10—16(—19) x 4.5—6.5 um (Ekman & Tonsberg 1996, Kalb

2007, Sanderson 2009, Lendemer & al. 2016).

Furthermore, M. albocincta has two-sided indentations from the cell

lumina into the center of the spore septum (Ekman & Tonsberg 1996);

M. alligatorensis has a Bacidia-type ascus and greenish-blue thallus that

forms small circular patches (Lendemer & al. 2016); M. anaglyptica has a

thick granular thallus and dirty greyish brown subhymenium (Kalb 2007);

and M. pulverea has a sorediate granular thallus and bigger apothecia

(0.5—2.2 mm diam.; Sanderson 2009).

As shown in Fic. 2, M. yunnanensis is sister to M. pulverea, but the

genetic distance (Kimura 2-parameter model) between these two species is

quite large (0.198—0.207). Interestingly, a sterile sorediate species M. alleniae

Lendemer & McMullin from southeastern North America (McMullin

& Lendemer 2016) is chemically similar to M. yunnanensis, but our new

species obviously differs from M. alleniae morphologically and molecularly

(with a 0.212 genetic distance separating M. alleniae and M. yunnanensis).

Acknowledgements

The authors thank Dr. J.C. Lendemer (Institute of Systematic Botany, New York

Botanical Garden, Bronx, New York, USA) and Dr. Shou- Yu Guo (State Key Laboratory

Megalaria yunnanensis sp. nov. (China) ... 293

MK348527 Megalaria yunnanensis

MK348528 Megalaria yunnanensis [T]

KX660735.1 Megalaria pulverea

FR799227.1 Megalaria pulverea

KX132969.1 Megalaria pulverea

KX660734.1 Megalaria alleniae

AF282074.1 Megalaria grossa

FR799225.1 Megalaria grossa

FR799224.1 Megalaria grossa

FR799226.1 Megalaria grossa

KP314369.1 Biatora subduplex

AM292667.1 Biatora meiocarpa

0. 050

Fic. 2. Molecular phylogenetic tree illustrating the position of Megalaria yunnanensis in

Megalaria s.l. based on ITS sequences. Biatora species were selected as the outgroup. Maximum

likelihood bootstrap support values >50% are shown on nodes.

of Mycology, Institute of Microbiology, CAS, Beijing) for presubmission reviews and

providing great help during the study. We also thank Dr. Alan M. Fryday (Michigan

State University, USA) for providing suggestions and useful literature during the study.

This work was supported by Emergency Management Project of National Natural

Science Foundation of China (31750001), the National Natural Science Foundation of

China (31400015), the National Natural Science Foundation of China Youth Science

Foundation (31600100), and the Scientific Research Foundation of the Graduate

School of Shandong Normal University (SCX201945) .

294 ... Wang & al.

Literature cited

Brodo IM, Sharnoff SD, Sharnoff S. 2001. Lichens of North America. Yale University Press, New

Haven & London. 795 p.

Ekman S, Tonsberg T. 1996. A new species of Megalaria from the North American

West Coast, and notes on the generic circumscription. Bryologist 99: 34-42.

https://doi.org/10.2307/3244435

Elix A, McCarthy M. 2018. Ten new lichen species (Ascomycota) from Australia. Australasian

Lichenology 82: 20-59.

Fryday AM, Lendemer JC. 2010. Reassessment of the genus Catillochroma (lichenized

Ascomycota, Ramalinaceae). Lichenologist 42: 587-600.

https://doi.org/10.1017/S0024282910000320

Gardes M, Bruns TD. 1993. ITS primers with enhanced specificity for basidiomycetes—

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

Galloway DJ. 2004. New lichen taxa and names in the New Zealand mycobiota. New Zealand

Journal of Botany 42: 105-120. https://doi.org/10.1080/0028825X.2004.95 12893

Hafellner J. 1984. Studien in Richtung einer natiirlicheren Gliederung der Sammelfamilien

Lecanoraceae und Lecideaceae. Beihefte zur Nova Hedwigia 79. 241-371.

Kalb K. 2007. New or otherwise interesting lichens. III. Bibliotheca Lichenologica 95: 297-316.

Katoh K, Kuma K, Toh H, Miyata T. 2005. MAFFT version 5: improvement in accuracy of multiple

sequence alignment. Nucleic Acids Research 33: 511-518. https://doi.org/10.1093/nar/gkil98

Lendemer JC. 2007. Megalaria beechingii (lichenized Ascomycota), a new species from eastern

North America. Opuscula Philolichenum 4: 39-44.

Lendemer JC, Harris RC, Ruiz AM. 2016. A review of the lichens of the Dare Regional

Biodiversity Hotspot in the Mid-Atlantic Coastal Plain of North Carolina, eastern North

America. Castanea 81: 1-77. https://doi.org/10.2179/15-073R2

McCarthy M, Elix A. 2016. A new species of Megalaria (lichenized Ascomycota, Ramalinaceae)

from north-eastern Queensland, Australia. Australasian Lichenology 79: 20-25.

McMullin RT, Lendemer JC. 2016. Megalaria allenae [sic] (Ramalinaceae), a new sorediate

species from southeastern North America previously confused with M. pulverea. Bryologist

119(3): 290-297. https://doi.org/10.1639/0007-2745-119.3.290

Orange A, James PW, White FJ. 2001. Microchemical methods for the identification of lichens.

British Lichen Society, London. 101 p.

Sanderson NA. 2009. Megalaria. 565-567, in: CW Smith & al. (eds). The Lichens of Great

Britain and Ireland. British Lichen Society, London.

Su QX, Ren Q. 2017. A new species of Megalaria (Ascomycota, Ramalinaceae) and M. laurerinew

to mainland China Phytotaxa 313(1): 147-150. https://doi.org/10.11646/phytotaxa.313.1.13

Talavera G, Castresana J. 2007. Improvement of phylogenies after removing divergent and

ambiguously aligned blocks from protein sequence alignments. Systematic Biology 56:

564-577. https://doi.org/10.1080/10635150701472164

White TJ, Bruns T, Lee S, Taylor JW. 1990. 38-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. San Diego, CA: Academic Press.

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

MY COTAXON

April-June 2019—Volume 134, pp. 295-300

https://doi.org/10.5248/134.295

The conserved type of Lichen fuscatus [= Acarospora fuscata]

KERRY KNUDSEN’, JIRi MALICEK?, 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

? The Czech Academy of Sciences, Institute of Botany,

Zamek 1, CZ-252 43 Pruhonice, Czech Republic

*CORRESPONDENCE TO: knudsen@fzp.czu.cz

ABSTRACT—The conserved type of Lichen fuscatus, the basionym of Acarospora fuscata, is

described, and two genes, ITS and mrSSU, are made available through GenBank for further

phylogenetic research.

Key worps—Acarospora gallica, A. variegata, nomenclature, taxonomy

Introduction

Acarospora fuscata is the most common species of the genus in Europe

(Magnusson 1929). In crustose lichen communities on siliceous rock, its pale

brown color is distinctive, and it is often collected. But despite A. fuscata being

common, it did not have a designated type or legitimate name (Arcadia & al.

2015). Because the taxon is common and the name A. fuscata is in wide use,

a formal proposal was made to conserve the name Lichen fuscatus Schrad.

(Arcadia & al. 2015) and has been accepted (May 2017). A modern conserved

type that could be sequenced was designated that matched the species concept

of A.H. Magnusson, who did the most extensive study of the species in Europe

(Magnusson 1929). The taxon’s most typical phenotype was collected in central

Europe to serve as conserved type. In this paper, we present a description of

the conserved holotype and isotypes and designate GenBank sequences to aid

in the identification and phylogenetic study of A. fuscata and other Acarospora

taxa containing gyrophoric acid that have been confused with it.

296 ... Knudsen, Mali¢ek, Kocourkova

Material & methods

The description is based only on the conserved holotype and isotypes. Specimens

were studied with standard microscopy. Measurements were made in water. Undiluted

fresh Merck Lugol’s (IKI) was used to test amyloid reactions of thin squashed sections

of apothecia following Knudsen & Kocourkova (2018). Ascus structure was studied

with IKI with or without pretreatment with KOH (Hafellner 1993). Thin-layer

chromatography (TLC) was used to verify the presence of secondary metabolites

(Orange & al. 2001).

DNA was extracted from the holotype (Kocourkova 8499, S) using the Invisorb

Spin Plant Mini Kit. The fungal ITS rDNA and mitochondrial SSU were amplified with

the following primers: ITS1F (Gardes & Bruns 1993) and ITS4 (White & al. 1990), and

mrSSU1 and mrSSU3R (Zoller & al. 1999). PCR reactions of nrITS and mtSSU were

prepared for a 20 ul final volume containing 14 ul double-distilled water, 4 ul MyTaq

polymerase reaction buffer, 0.2 ul MyTaq DNA polymerase, 0.4 tl of each of the 25 mM

primers, and 1 ul of the sample. Amplifications of both loci comprised an initial 1 min

denaturation at 95 °C; 35 cycles of 1 min at 95 °C, 1 min at 56 °C, 1 min at 72 °C; anda

final 7 min extension at 72 °C. The PCR products were visualized on a 0.8% agarose gel

and cleaned with ExoSAP-IT, according to Thermo Fisher Scientific protocols. Both

newly produced sequences, ITS (MK178225) and mrSSU (MK178225), were checked

in BioEdit 7.2.5 (Hall 1999).

Macrophotographs were taken with an Olympus DP74 camera mounted on

an Olympus SZX 7 stereomicroscope equipped with PRO-SZM1-Focus Drive

Motorization for stacking pictures and stacked using the Olympus DeepFocus 3.4

module. The figure plates were processed with QuickPhoto Camera 3.2 software fitted

with Promicra Publisher Module and eventually refined with Adobe Photoshop CS4

Extended ver. 11.0. The type locality picture was taken with Panasonic DMC LX5.

Taxonomy

Acarospora fuscata (Schrad.) Arnold,

Flora 53 (30-31): 469. 1871 [“1870"]. Fic. 1

= Lichen fuscatus Schrad., Spic. Fl. Germ.: 83. 16 Mai-5 Jun 1794, nom. cons. prop.

Type: Czech Republic. West Bohemia, Distr. Tachov, between Mezholezy and Racov,

Prirodni park Sedmihofi (Natural Park), 49°37’53”N 12°51’41”E, 524 m, granite

outcrops in Pinus sylvestris forest, 1 Oct. 2014, J. Kocourkova 8499 & K. Knudsen (S,

conserved holotype, designated in Arcadia & al. 2015; B, FH, KRAM, GZU, H, NY,

PRA, UCR, UPS, conserved isotypes).

THALLUS indeterminate of areoles, often contiguous, occasionally imbricate,

covering areas sometimes more than a meter wide, or dispersed, sometimes

forming lines along fractures in the rock. Areoles irregular in shape, often

lobulate with edges free and often with a black margin, sometimes reduced

to lecanorine-like apothecia, broadly attached to the substrate, replicating

by division, 0.1-3.0 mm wide, up to 1 mm thick. Upper surface pale brown,

Lichen fuscatus conserved ... 297

Fic. 1. Lichen fuscatus (conserved holotype, J. Kocourkova 8499, S): A. Habit of the areolate thallus

with apothecia; B. Marginal sublobate areole with sunken apothecia; C. Detail of areoles with

apothecia. Scale bars: A = 2 mm; B, C = 500 um.

epruinose, matte, smooth. Lower surface light or dark brown, corticate under

the margins, becoming black through melanization. Epicortex thin (<10 um

298 ... Knudsen, Maliéek, Kocourkova

thick) or absent. Cortex 25-50 um thick, upper layer yellow brown, lower layer

hyaline, paraplectenchymatous, with cells 3-5 um diam., perpendicularly

arranged and usually distinct, containing crystals visible in polarized light.

Algal layer 50-100 um thick, uninterrupted by hyphal bundles, with cells

5-12 um diam. Medulla forming a thick mycelial base (gomphate), 0.1-0.8

mm thick, not distinctly stipitate, the outer surface usually carbonized,

medullary hyphae intricate, thin-walled, septate, 3-4 um thick.

APOTHECIA solitary or up to 10 per areole, immersed, punctiform

0.1-0.3 um or expanding <0.6 mm wide, disc round or irregular, reddish-

brown, darker than the thallus, epruinose, smooth or rough. Parathecium

10-20 um diam., not expanding around the disc. Epihymenium 10-15 um

thick, reddish-yellow, coherent. Hymenium (80-—)100-—120(-140) um high,

paraphyses (1.0-)1.5-2.0 um diam. at mid-level, not branching, septate,

often with oil drops, with the apices barely expanded or capitate, 3-5 um

diam., hymenial gel IKI+ red or blue turning red (hemiamyloid). Asci 60-90

x 15-18 um, ascus stain Acarospora-type, ascospores 4.0—5.0 x 1.5-2.0 um,

narrowly ellipsoid to ellipsoid. Subhymenium 30-60 um thick, IKI+ blue

(euamyloid). Hypothecium 10-20 um thick. Pycnidia 100-110 x 60-70 um,

conidiogenous cells ampulliform, conidia 1.0-2.0 x 0.5-1.0 um. Secondary

metabolite: gyrophoric acid in cortex, C+, KC+ pinkish red in thin section.

Sequences available in GenBank: ITS (MK178225) and mrSSU (MK178225).

ECOLOGY & SUBSTRATE— The conserved type was collected at a low (310 m)

elevation in full sunlight in Pinus sylvestris forest, where it covered the tops

of granite outcrops without other associated lichens (Fic. 2)

COMMENTS— Using our description and simple spot tests many specimens

of A. fuscata can now be easily identified. Our description based on the

conserved type does not differ significantly from the descriptions of

Magnusson (1929) or Fletcher & al. (2009), except with respect to hymenium

height. In the type the hymenium measures (80-)100-120(-140) um,

varying 60 um in height depending on apothecial size. Magnusson (1929)

gives a height of (70—)85-100 um, a range of 30 um. The British flora cites

(70—)80-120 um, a range of 50 um (Fletcher & al. 2009).

Nonetheless, Acarospora fuscata exhibits a wide phenotypic variability

in Europe, which led to the description of nine forms and one variety

(Magnusson 1929). Now that we have gene sequences available from the

conserved type of A. fuscata, unusual specimens can be verified with

molecular analyses. Additional sequences can also be made from the

conserved type or matching specimens, so that researchers can finally

Lichen fuscatus conserved ... 299

Fic. 2. Acarospora fuscata type locality in West Bohemia, Czech Republic

establish if A. fuscata is polymorphic or a complex of several semi-cryptic

or cryptic taxa. Candidates for a full genomic study can be selected that

might reveal whether there is horizontal gene transfer with other Acarospora

species. In Europe, the current taxonomic circumscriptions of A. gallica

H. Magn. and A. variegata H. Magn. can also be revised. The British

submontane taxon “A. peliscypha” sensu auct. Brit., included in A. fuscata,

can be analyzed as well as pruinose specimens (Fletcher & al. 2009). The

name A. fuscata, like other old European names, was applied to brown C+

red Acarospora taxa around the world. Additionally, collections from North

America, Australia, or Asia identified as A. fuscata can be revised (Knudsen

2007, McCarthy & Elix 2017, Ohmura & Kashiwadani 2018).

Acknowledgements

We thank our reviewers, Gintaras Kantvilas (Tasmanian Herbarium, Tasmanian

Museum and Art Gallery, Australia) and Andrei Tsurykau (F. Skorina Gomel State

University, Belarus). The work of Kerry Knudsen and Jana Kocourkova was financially

supported by the grant Environmental Aspects of Sustainable Development of Society

(42900/1312/3166) from the Faculty of Environmental Sciences, Czech University of

Life Sciences, Prague. The contribution by Jifi Mali¢ek was supported by the Long-

Term Research Development Project RVO 67985939.

300 ... Knudsen, Malicek, Kocourkova

Literature cited

Arcadia L, Knudsen K, Westberg M. 2015. (2341) Proposal to conserve the name Lichen fuscatus

Schrad. (Acarospora fuscata) against L. fuscatus Lam. with a conserved type (lichenised

Ascomycota: Acarosporaceae). Taxon 64: 168-169. https://doi.org/10.12705/641.7

Fletcher A, James PW, Purvis, OW. 2009. Acarospora A. Massal. (1852). 125-132, in: CW Smith

& al. (eds.) The lichens of Great Britain and Ireland. London: British Lichen Society.

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

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

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

Hafellner J. 1993. Acarospora und Pleopsidium - zwei lichenisierte Ascomycetengattungen

(Lecanorales) mit zahlreichen Konvergenzen. Nova Hedwigia 56: 281-305.

Hall TA. 1999. BioEdit: a user-friendly biological sequence alignment editor and analysis

program for Windows 95/98/NT. Nucleic Acids Symposium Series 41: 95-98.

Knudsen K. 2007 [“2008”]. Acarospora. 1-38, in: TH Nash III & al. (eds). Lichen Flora of the

Greater Sonoran Desert Region. Volume 3. Lichens Unlimited, Arizona State University,

Tempe, Arizona.

Knudsen K, Kocourkova J. 2018. Two new calciphytes from western North America, Acarospora

brucei and Acarospora erratica (Acarosporaceae). Opuscula Philolichenum 17: 342-350.

Magnusson AH. 1929. A monograph of the genus Acarospora. Kungliga Svenska Vetenskaps-

Akademiens Handlingar, ser. 3, 7. 400 p.

May TW. 2017. Report of the Nomenclature Committee for Fungi: 20. Taxon 66: 483-495.

https://doi.org/10.12705/662.15

McCarthy P, Elix JA. 2017. Two new species and a new record of Acarosporaceae (lichenized

Ascomycota) from eastern Australia. Australasian Lichenology 80: 16-27.

Ohmura Y, Kashiwadani H. 2018. Checklist of lichens and allied fungi of Japan. National

Museum of Nature and Science Monographs 49. 140 p.

Orange A, James PW, White FJ. 2001. Microchemical methods for the identification of lichens.

London: British Lichen Society. 101 p.

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

fungal ribosomal RNA genes for phylogenetics. 315-322, in: PCR Protocols: a Guide

to Methods and Applications. MA Innes & al. (eds): Academic Press, New York.

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

Zoller S, Scheidegger C, Sperisen C. 1999. PCR primers for the amplifications of mitochondrial

small subunit ribosomal DNA of lichen-forming Ascomycetes. Lichenologist 31: 511-516.

https://doi.org/10.1006/lich.1999.0220

MY COTAXON

April-June 2019—Volume 134, pp. 301-305

https://doi.org/10.5248/134.301

Beltraniopsis cyclobalanopsidis sp. nov.

from Guizhou, China

ZHONG-J1U XIAO? , XIAO-XIA Li}, CHANG-HONG CHU °,

TrnG Liu*?, ZHENG-ZHENG Lu’, PEI- YONG SONG’?

"College of Biological & Agricultural Science & Technology &

* College of Resources & Environment,

Zunyi Normal University, Zunyi, Guizhou 563002, China

* Key Laboratory of Regional Characteristics for Conservation & Utilization

of Plant Resource in Chishui River Basin, Zunyi, Guizhou, 563002, China

* CORRESPONDENCE TO: xzj198099@163.com

ABSTRACT—A new species, Beltraniopsis cyclobalanopsidis, is described and illustrated from

a specimen collected on dead twigs of Cyclobalanopsis stewardiana |= Quercus stewardiana]

in Guizhou Province, China. The fungus is characterized primarily by its longer, setiform

conidiophores with inflated apexes. A dichotomous key to Beltraniopsis species is provided.

Key worps—anamorphic fungi, lignicolous fungi, taxonomy

Introduction

Batista & Bezerra (1960) erected Beltraniopsis based on the type species,

Beltraniopsis esenbeckiae Bat. & J.L. Bezerra. The genus is characterized by

macronematous, simple or branched setiform conidiophores and polyblastic,

discrete, sympodial extended, denticulate conidiogenous cells with separating

cells that produce solitary, acropleurogenous, aseptate, biconic, smooth, brown

conidia.

Beltraniopsis currently contains ten valid species found as saprobes on rotten

leaves (Matsushima 1971, 1993; Pirozynski 1972; Castafieda-Ruiz & Arnold

1985; Rambelli & Ciccarone 1985; Gusmao & al. 2000; Castafieda Ruiz & al.

2006; Crous & al. 2014; Lin & al. 2017). A survey of saprobic microfungi on

302 ... Xiao & al.

dead twigs from a subtropical forest in Guizhou Province, China, has revealed a

previously undescribed Beltraniopsis species. The specimen is deposited in the

Mycological Herbarium of Zunyi Normal College, Zunyi, China (HMZNC).

Beltraniopsis cyclobalanopsidis Z.J. Xiao & Xiao X. Li, sp. nov. FIG. 1

FN570598

Differs from all other Beltraniopsis species by its longer setiform conidiophores inflated

rather abruptly at the apical cell.

Type: China, Guizhou Province: Xishui National Nature Reserve, saprobic on dead twigs

of Cyclobalanopsis stewardiana (A. Camus) Y.C. Hsu & H.W. Jen [= Quercus stewardiana

A. Camus], 6 Jun. 2018, X.X. Li (Holotype, HMZNC 0450).

ErymMo_oey: Latin, referring to the substrate genus Cyclobalanopsis.

CoLonigs on dead twigs effuse, velutinous, brown to dark brown. Mycelium

partly superficial and immersed in the substrate, composed of branched, septate,

pale brown to brown, cylindrical, smooth-walled hyphae. CoNIDIOPHORES

macronematous, mononematous, setiform, sometimes becoming fertile at the

apex, erect, slightly flexuous, unbranched or mostly branched in the lower-

middle part, thick-walled, cylindrical, 13-25-septate, smooth or sometimes

the lower-middle part with verrucose, arising from flat, radially lobed basal

cells, brown to dark brown at the lower part, pale brown to hyaline at the

upper part, 420-1110 um long, 5.5-7.5 um diam. at the base, tapering to

the upper part, but apical cell inflated rather abruptly, rounded, thin-walled,

hyaline, 2.2-4.5 um diam. CONIDIOGENOUS CELLS polyblastic, mostly discrete

intercalary, sometimes terminal, integrated, located in the lower-middle

part of conidiophores, sympodial extended, 1-3-denticulate apex, smooth,

ampulliform, flask-shaped, pale brown, 9.5-12.5 um long, 5.5-7 um diam. in

the broadest part. SEPARATING CELLS ellipsoidal to ovoid, thin-walled, smooth,

hyaline, denticulate apex, 7-10.5 um long, 4.5-5.5 um diam. in the broadest

part. Conrp1A solitary, biconic to broad navicular, smooth, light brown,

constricted at the supra-equatorial zone with subhyaline transverse band,

29-35 x 5.5-7.5 um, apex rostrate, 0.5-1.5 um wide, base obtuse.

COMMENTS—Based on the synopsis and key to Beltraniopsis species, B. cyclo-

balanopsidis is superficially similar to B. longiconidiophora C.G. Lin & K.D.

Hyde and B. miconiae Gusmao & Grandi. Beltraniopsis longiconidiophora

Fic. 1. Beltraniopsis cyclobalanopsidis (holotype, HMZNC 0450). A. Conidiophores arising

from twig surface; B, C. Conidiophores, conidiogenous cells, separating cells, and conidia;

D. Conidiophore with broken upper part; E. Conidiogenous cells, separating cells, and conidia;

EF. Conidia arising from conidiogenous cells; G. Inflated apex of the conidiophores; H. Conidia.

Beltraniopsis cyclobalanopsidis sp. nov. (China) ... 303

304 ... Xiao & al.

differs from B. cyclobalanopsidis by its shorter setiform conidiophores

(100-680 um) tapering to a pointed apex and by its shorter conidia (18-29 um;

Lin & al. 2017); and B. miconiae differs by its much shorter setiform

conidiophores (77-147 um) with a slightly inflated apical cell (2-3.5 um)

and by its smaller separating cells (6-8 x 3-4.6 um; Gusmao & al. 2000).

Key to Beltraniopsis species

b-Seuuorin coMmdigphoréacuteat the apen s 03! chr oa Macnee s heb wrdldeg parti ath gra Hutt ye dle 2

1. Setiform conidiophore rounded or inflated at the apex ...................0006. 6

Zesetiiorn: conidiophorde-verrucosé: .. 2545.02 44 5a soa sees spare ke B. asperisetifera

De SO LIGEM COMI ATO PMOL STOO HS a: Fe gk accel £2. veal Ep. cect Op incest Ep tace epee EP aoe Es 3

3; Setiform branched-conidiophore apex not fertile ss 24.28 foe ft Bite Binds £08 4

3. setiform unbranchéd:conidiophoreapexfertile os os eae ee ihe 5

AxGonidians=8, piigiiaiit, whan, cet etal ae et ae Bala otto ate B. longiconidiophora

4 Gohidia4-5unr diat. «2% < yee et eget ered cig e ee loce 8 ey nae wy Ig, CRS B. ramosa

Se Gonidia 20=25 KAS60M » «obs. dat nnn eaeatnee gente ened B. esenbeckiae

psConidia 22-28: X 6, 54805 (iy hack tat Aeeiak Malek Mae nt Male coe let ey B. tanzaniensis

6. Setiform conidiophore rounded at the apex ........ 0... cece cece eee eee eee di

6. Setitom conidiophore inflated atthe apexes. lates, satan wee nate + tees oe 9

7. Conidia without rostrum and transverse band ................-..0-- B. aquatica

7. Conidia with rostrum and transverse band ............ 0... cece eee eee eens 8

8. Conidia 30-37 x 9 um, separating cells 4-4.5x 4m ................ B. fabularis

8. Conidia 30-33 x 7-8 um, separating cells 7-10 x 4-6 um _............ B. neolitseae

9. Conidia without rostrum and transverse band ................... B. rhombispora

SsConidia with rostrum and transverse band, « ..k,..12hc Fads cee Soselers Lodesere Peon ee Bakes 10

10. Setiform conidiophore 77-147 um long ............ 0. cece eee B. miconiae

10. Setiform conidiophore 420-1110 um long ................... B. cyclobalanopsidis

Acknowledgments

The authors express gratitude to Dr. R.E Castafeda Ruiz (Alejandro de

Humboldt—INIFAT, Havana, Cuba) and Dr. Jian Ma (Jiangxi Agricultural

University, Nanchang, China) for serving as pre-submission reviewers and for their

valuable comments and suggestions. Dr. Lorelei L. Norvell’s editorial review and

Dr. Shaun Pennycook’s nomenclature review are greatly appreciated. This project

was supported by the National Natural Science Foundation of China (Ne.31600030),

Science and Technology Project of Guizhou Province (NeLKZS[2014]20 and

Ne[2017]1206), Guizhou Provincial Department of Education Foundation

(NEQJHKY[2018]032, NeQJGF[2017]158, Ne[2010]087, NeQSZHZ[2012]146

and NZYRCPYJH[2016]9, Guizhou Science and Technology Cooperation Plan

(No. LH[2015]7031).

Beltraniopsis cyclobalanopsidis sp. nov. (China) ... 305

Literature cited

Batista AC, Bezerra JL. 1960. Beltraniopsis - novo género de fungos dematiaceae. Publicacdes do

Instituto de Micologia da Universidade do Recife 296: 1-13.

Castaneda RF, Arnold GRW. 1985. Deuteromycotina de Cuba. I. Hyphomycetes. Revista del Jardin

Botanico Nacional Universidad de la Habana 6: 47-67.

Castaneda Ruiz RF, Minter DW, Stadler M, Saikawa M, Camino-Vilaré M. 2006. Anamorphic

fungi from submerged leaves in Cuba: Brevicatenospora enteroproliferata gen. et sp. nov. and

Beltraniopsis aquatica sp. nov. Mycotaxon 96: 151-158.

Crous PW, Shivas RG, Quaedvlieg W, et al. 2014. Fungal Planet description sheets: 214-280.

Persoonia 32: 184-306. https://doi.org/10.3767/003158514X682395

Gusmao LFP, Grandi RAP, Milanez AI. 2000. A new species of Beltraniopsis from Brazil, with a key

to the known species. Mycological Research 104: 251-253.

https://doi.org/10.1017/S0953756299001239

Lin CG, Hyde KD, Lumyong S, McKenzie EHC. 2017. Beltrania-like taxa from Thailand.

Cryptogamie, Mycologie 38: 301-319. https://doi.org/10.7872/crym/v38.iss3.2017.301

Matsushima T. 1971. Microfungi of the Solomon Islands and Papua-New Guinea. Published by the

author: Kobe, Japan.

Matsushima T. 1993. Matsushima mycological memoirs no. 7. Published by the author: Kobe,

Japan.

Pirozynski KA. 1972. Microfungi of Tanzania. I. Miscellaneous fungi on oil palm. I. New

hyphomycetes. Mycological Papers 129. 64 p.

Rambelli A, Ciccarone C. 1985. Two new dematiaceous hyphomycetes from humid tropic forest

litter. Giornale Botanico Italiano 119: 291-294. https://doi.org/10.1080/11263508509426684

MY COTAXON

April-June 2019—Volume 134, pp. 307-312

https://doi.org/10.5248/134.307

Perichaena acetabulifera sp. nov. from Juarez City, Mexico

Marcos LIzARRAGA!', GABRIEL MORENO? , IRAIS FLORES-ROMERO!

" Dpto. Ciencias Quimico Bioldgicas, ICB, Univ. Auténoma de Ciudad Juarez,

Anillo Envolvente Pronaf y Estocolmo s/n, Ciudad Juarez, Chihuahua, México

? Dpto. Ciencias de la Vida (Botanica), Facultad de Ciencias, Univ. de Alcala,

Alcala de Henares, 28805 Madrid, Spain

" CORRESPONDENCE TO: gabriel.moreno@uah.es

AsBstTRAcT—Perichaena acetabulifera is described as a new species of myxomycete that

fruited profusely in moist chamber cultures on bark samples collected from Populus deltoides

and Ulmus pumila growing in urban areas of Juarez City, Chihuahua, Mexico. Photographs of

its diagnostic macroscopic and microscopic features are provided.

Key worps—Amoebozoa, myxobiota, SEM, taxonomy, Trichiales, Trichiaceae

Introduction

The city of Juarez is located in the northern part of the Mexican state of

Chihuahua, adjacent to El Paso (Texas) and Las Cruces (New Mexico), both in

the United States. The city has an extreme climate, with temperatures ranging

from 25 to 40°C in the summer and -4 to 13°C in the winter. In the spring, it

is frequently lashed by strong winds that reach speeds up to 100 km/h. The

dominant vegetation surrounding the city consists of xerophytic shrubs, with

species of Larrea (Zygophyllaceae), Yucca (Agavaceae), Opuntia (Cactaceae), and

Prosopis (Mimosaceae) as the prominent taxa, while the city interior contains a

large number of ornamental plants in several recreational parks and gardens.

These include species of Cupressus, Thuja, Pinus, Morus, Populus, Fraxinus,

Salix, Melia, Nerium, Vitex, Platanus, Tamarix, Chilopsis, and Quercus. Several

studies are being carried out to determine which myxomycetes develop on the

bark of these urban plants, and the first results were published by Lizarraga &

al. (2015, 2018).

308 ... Lizarraga, Moreno, Flores-Romero

Perichaena Fr. is characterized by its fructifications: sessile, stipitate or

short plasmodiocarps. Generic characters include a simple peridium with

a membranous inner surface and a granular and fragile outer surface that

sometimes is crusted with calcium oxalate. The spores are yellow or reddish

brown. The capillitium, which comprises tubular filaments with irregular or

regular constrictions, is superficially ornamented with warts or spines and

lacks spirals and birefringence (Nannenga-Bremekamp 1991). Although the

absence of spiral-like bands in the capillitium is the most distinctive character

separating Perichaena from Trichia Haller and Oligonema Rostaf., some species

lacking a capillitium are placed within Perichaena, including P. heterospinispora

Novozh. & al. (Novozhilov & al. 2008), P. taimyriensis Novozh. & Schnittler

(Novozhilov & Schnittler 2001), B pachyderma D.W. Mitch. & al. (Mitchell &

al. 2011), and Perichaena nigra D. Wrigley & al. (Lado & al. 2014).

According to Lado (2018), 34 species are currently recognized worldwide, of

which 13 have been recorded for Mexico (Moreno & al. 2007, Estrada-Torres &

al. 2009, Rojas & al. 2010, Mitchell & al. 2011, Lizarraga & al. 2016).

Materials & methods

Samples for light microscopy were mounted in Hoyer’s medium. Macroscopic

photographs were taken with a Nikon SM2800 microscope, micrographs were

obtained with a Nikon Eclipse 2000 microscope, and SEM micrographs were taken

with a Zeiss DSM-950 microscope after critical point drying and sputtering, following

Moreno & al. (2015). The specimens are stored in the Herbarium, Universidad de

Alcala, Madrid, Spain (AH) and the Herbarium, Instituto de Ciencias Biomédicas,

Universidad Autonoma de Ciudad Juarez, Chihuahua, México (UACJ). Nomenclature

follows Lado (2018).

Taxonomy

Perichaena acetabulifera Lizarraga, G. Moreno & Flores-Rom., sp.nov. — PLATE 1

MB 829449

Differs from Perichaena vermicularis by its peridial inner surface and capillitium

producing ocelli and craters.

Type. Mexico: Chihuahua, Juarez City, Biomedic Science Institute (Spanish acronym:

ICB), on bark of Ulmus pumila L., placed into moist chamber 19-VIII-2015, obtained

22-IX-2015, leg. I. Flores-Romero & M. Lizarraga (Holotype, AH 49096; isotype, UACJ

2789).

Erymo oey: acetabulifera from the Latin acetabulum, referring to a shallow concave

vessel used as a cruet (and therefore the name given by Plinio to the suckers of an

octopus) and fer/fero meaning to carry. The name draws attention to the resemblance of

the capillitium and internally ornamented peridium to the suckers of octopus tentacles.

Perichaena acetabulifera sp. nov. (Mexico) ... 309

PLATE 1 Perichaena acetabulifera (holotype, AH 49096): 1. Fructifications; 2. Detail of fructifications;

3-5. Capillitium by LM; 6. Spores; 7. Detail of the inner surfaces of peridium by SEM; 8-10. Detail

of capillitium with craters by SEM; 11-12. Spores by SEM; 13. Detail of spore ornamentation by

SEM. Scale bars: 1, 2 = 1 mm; 3-6 = 10 um; 7-12 = 2 um; 13 = 1 um.

310 ... Lizarraga, Moreno, Flores-Romero

FRUCTIFICATIONS scattered to gregarious, sessile, occasionally substipitate,

globose, subglobose to short plasmodiocarpic, in this case somewhat

sinuous, occasionally centrally depressed, to 0.8-1.5 x 0.2-0.6 mm. Stipe

absent, occasionally with a blackish-brown small base up to 0.1 mm total

height. PERrp1uM simple, membranous, olivaceous yellow, inner part with

papillae and small crests and craters that project over the surface. The

outer surface granulose with deposits of amorphous material, brownish to

olivaceous brown. HyPOTHALLUS absent or barely noticeable. CAPILLITIUM

2-6 um in diam., somewhat abundant in the sample studied, formed by

filaments with irregular edges, branched, with ocellus-like structures and

craters, giving it a pitted appearance under OM. By SEM a capillitium

lacking spines and reticulum can be observed, with rare and minute

warts, and isolated small ocelli and craters of variable size, <2.5 um in

diam., resembling the suckers of an octopus. Spores 11-14(-16) um in

diam., ochraceous in mass, yellowish ochre by OM, globose, subglobose to

oval, with a paler area, with sparse and irregular warts. By SEM the spore

ornamentation is formed by warts with a flattened apex or bacula, unevenly

distributed over the surface.

OTHER MATERIAL EXAMINED: MEXICO. CHIHUAHUA, Juarez City: Federal Public

Park El Chamizal, on the bark of Ulmus pumila, placed into moist chamber 18-VIII-

2015, obtained 29-IX-2015, leg. I. Flores-Romero & M. Lizarraga (UAC] 2827); on

bark of Populus deltoides Marshall, placedinto moist chamber 16—VIII-2016, obtained

30-VIII-2016 (UACJ 3317); obtained 20-IX-—2016 (UAC]J 3318, AH 49095); obtained

27-IX-2016 (UACJ 3319); obtained 17—X—2016 (UACJ 3320, AH 49100; UAC] 3321)

obtained 18—X-2016 (UACJ 3322); obtained 31—X-—2016 (UACJ 3323); obtained

10—XI-2016( UAC] 3324); obtained 1 1—XI-2016 (UAC] 3325); obtained 10—XII-2016

(UAC] 3326). Institutes of Engineering, Technology, Architecture, Art and Design

(IIT-IADA), on bark of Ulmus pumila, placed into moist chamber 19-VIII-2016,

obtained 14—X-2016, leg. I. Flores-Romero & M. Lizarraga (UAC] 3327; UACJ 3328).

Institute of Biomedical Sciences, on bark of Ulmus pumila, placed into moist chamber

19-VIII-2015, obtained 14-IX-2015, leg. I. Flores-Romero & M. Lizarraga (UACJ

2717); obtained 21-IX-2015 (UACJ 2773); obtained 22-IX-2015 (UACJ 2790, AH

49097); Populus deltoides, obtained 22—XI-2016, leg. I. Flores-Romero & M. Lizarraga

(UAC) 3329, AH 49099); obtained 6—-IX-2017 (UACJ 3307); obtained 13-IX-2017

(UACJ 3330); placed into moist chamber 9-IV-2018, obtaining fructifications

23-IV-2018 (UACJ 3063, AH 49098).

OBSERVATIONS—Perichaena acetabulifera is characterized by its sporocarpic

to plasmodiocarpic fructifications and especially by its papillate peridium

with isolated ocelli and craters, and a capillitium also with ocelli and craters,

with rare warts visible only by SEM. The morphologically closest species are

described below.

Perichaena acetabulifera sp. nov. (Mexico) ... 311

Perichaena vermicularis (Schwein.) Rostaf. is similar to the new species in

its plasmodiocarpic fructifications, spore measurements, and the presence

of papillae on the inner peridial and capillitial surfaces (Rammeloo 1981),

but it lacks the ocelli and craters found in the new species.

Perichaena areolata Rammeloo is a foliicolous species with short

sporothecae and craters and ocelli in the capillitium, similar to the new

species. However, it can be separated its spherical areolated sporothecae,

inner peridial surface lacking craters and ocelli (Rammeloo 1981), and its

capillitium ornamented by spines or small appendixes.

Perichaena poronema Yu Li & H.Z. Li is distinguished by its globose

sporothecae that are sometimes pulvinate, its circumscissile dehiscence by

a definite preformed lid, its minutely warted and incompletely reticulated

capillitium that is occasionally covered with holes, and small (8.75-9.5 um

in diam.) spores (Li & al. 1990).

Perichaena stipitata Lado & al. is a succulenticolous species that shares

with P. acetabulifera the presence of craters or ocelli on the inner peridium,

but which can be distinguished by its stalked fructifications and an inner

peridium lacking papillae and crests but with dense ocelli and craters.

Moreover, the capillitium has holes but lacks craters and ocelli.

Acknowledgements

We wish to express our gratitude to A. Priego and J.A. Pérez (Electron Microscopy

Service, University of Alcala) for their invaluable help in producing the SEM

images. We also thank Luis Monje and A. Pueblas (Department of Drawing and

Scientific Photography, Alcala University) for their help in the digital preparation

of the photographs, and we are grateful to J. Rejos, curator of the AH herbarium,

for his assistance with the examined specimens in the present study. We want to

express our gratitude to S.L. Stephenson (University of Arkansas, Fayetteville, USA)

and M.M. Dios (Universidad Nacional de Catamarca, Argentina) for reviewing

this manuscript. We also thank Shaun Pennycook and Luis Parra for their help

in choosing the specific epithet and D. Lépez-Pefia and H. Singer for their help

with the manuscript translation. M. Lizarraga extends his thanks to Antonio de la

Mora Covarrubias and Alberto Constance (DCQB-ICB, Universidad Autonoma of

Ciudad Juarez) for their economic support. I. Flores-Romero thanks CONACYT

for the scholarship given to carry out her master’s studies.

Literature cited

Estrada-Torres A, Wrigley de Basanta D, Conde E, Lado C. 2009. Myxomycetes associated with

dry land ecosystems of the Tehuacan-Cuicatlan Valley Biosphere Reserve, Mexico. Fungal

Diversity 36: 17-56.

312 ... Lizdrraga, Moreno, Flores-Romero

Lado C. (2018) An online nomenclatural information system of Eumycetozoa. Real Jardin

Botanico, CSIC. Madrid, Spain. Available from: http://www.nomen.eumycetozoa.com

(accessed 14 June 2018)

Lado C, Wrigley de Basanta D, Estrada-Torres A, Garcia-Carvajal E. 2014. Myxomycete diversity

of the Patagonian Steppe and bordering areas in Argentina. Anales del Jardin Botanico

de Madrid 71(1): e006 [35 p.]. https://doi.org/10.3989/ajbm.2394

Li Y, Li HZ, Wang Q, Chen S. 1990. Myxomycetes from China VII: new species and new records

of Trichiaceae. Mycosystema 3: 93-98.

Lizarraga M, Moreno G, Escobar-Zapata S. 2015. Myxomycetes del area urbana de Ciudad

Juarez, Chihuahua, México. Boletin de la Sociedad Micolégica de Madrid 39: 83-92.

Lizarraga M, Moreno G, Esqueda M. 2016. New records of myxomycetes from México.

Mycotaxon 131: 511-520. https://doi-org/10.5248/131.511

Lizarraga M, Moreno G, Flores-Romero I, Jiménez-Vega FE. 2018. A new species of Physarum

(Myxomycetes) from Chihuahua, Mexico. Boletin de la Sociedad Micoldgica de Madrid

42: 21-26.

Mitchell DW, Moreno G, Lizarraga M. 2011. A new species of coprophilous Perichaena from

New Mexico. Boletin de la Sociedad Micoldgica de Madrid 35: 103-108.

Moreno G, Lizarraga M, Illana C. 2007. Catalogo de los myxomycetes de México. Boletin de la

Sociedad Micoldgica de Madrid 31: 187-229.

Moreno G, Castillo A, Deschamps JR, Giménez G, Hladki A, Lopez- Villalba A. 2015. Critical

revision of some myxomycetes deposited in the Buenos Aires Herbaria BAFC, BA and the

Tucuman Herbarium LIL. IV. Boletin de la Sociedad Micoldgica de Madrid 39: 129-140.

Nannenga-Bremekamp NE. 1991. A guide to temperate myxomycetes. Biopress Limited, Bristol,

England. 409 p.

Novozhilov YK, Schnittler M. 2001. A new coprophilous species of Perichaena (Myxomycetes)

from the Russian Arctic (the Taimyr Peninsula and the Chukchi Peninsula). Karstenia 40:

117-122. https://doi.org/10.29203/ka.2000.360

Novozhilov YK, Zemlyanskaya IV, Schnittler M, Stephenson SL. 2008. Two new species of

Perichaena (Myxomycetes) from arid areas of Russia and Kazakhstan. Mycologia 100(5):

816-822. https://doi.org/10.3852/08-047

Rammeloo J. 1981. Flore illustrée des champignon d’Afrique centrale. Trichiales (Myxomycetes).

Fascs. 8-9. Ministére de Agriculture, Meise; Jardin Botanique National de Belgique.

Rojas C, Stephenson SL, Estrada-Torres A, Valverde R, Morales O. 2010. New records of

myxomycetes from high-elevation areas of Mexico and Guatemala. Mycosphere 1: 73-82.

MY COTAXON

April-June 2019—Volume 134, pp. 313-319

https://doi.org/10.5248/134.313

Gliocladiopsis wuhanensis sp. nov. from China

N1-PING ZHAI, ZI-QUAN SUN, YA-LONG ZHANG, RUI ZANG,

CHAO Xu, YUE-Hua GENG’, MENG ZHANG’

College of Plant Protection, Henan Agricultural University,

95 Wenhua Street, Zhengzhou, Henan 450000, China

* CORRESPONDENCE TO: gengyuehua@163.com, zm2006@126.com

ABSTRACT—A new species, Gliocladiopsis wuhanensis, was isolated from soil collected

in Hubei Province, China. A combination of ITS, HIS3, and TUB2 DNA sequences were

analyzed to evaluate its phylogenetic status. Phylogenetically G. wuhanensis is closely related

to G. guangdongensis. Morphologically, G. wuhanensis can be distinguished from related

species by its tertiary conidiophore branching.

KEY worps—asexual fungi, hyphomycetes, multiple gene, subtropics, taxonomy

Introduction

Gliocladiopsis S.B. Saksena was established with the type G. sagariensis

S.B. Saksena, characterized by densely penicillate and branched conidiophores

(Saksena 1954, Lombard & al. 2015). Crous & Wingfield (1993) synonymized

the type species under G. tenuis (Bugnic.) Crous & M.J. Wingf. based on

morphological studies; however, accurate identification of the Gliocladiopsis

species based on morphology alone is difficult (Liu & Cai 2013). Through

analyzing the ITS, TUB2, His3, 28S LSU, and TEFl-a DNA loci, Lombard

& Crous (2012) found G. sagariensis and G. tenuis phylogenetically distinct

and described five new species. Although its teleomorph was described as

Glionectria Crous & C.L. Schoch (Schoch & al. 2000), the “anamorphic” name

Gliocladiopsis has nomenclatural priority and is now the accepted generic name

under “One Fungus: One Name”. Gliocladiopsis currently comprises 14 species

(Index Fungorum 2018) mostly soilborne (Lombard & al. 2015) except for

314 ... Zhai & al.

G. guangdongensis F. Liu & L. Cai (Liu & Cai 2013) and G. aquaticus Y.Z. Lu &

al. (Hyde & al. 2018) isolated from decaying wood in freshwater.

Materials & methods

Isolation

Soil samples from different regions were investigated using a dilution plate

method: sterilized water agar medium cooled to 45 °C was mixed with 100

uL ampicillin sodium (100 ng/uL), and poured into a sterile plate with sterile

wheat-straw.; for each soil sample, three plates were evenly coated with a 200 uL

soil suspension diluted 100 times, incubated ca. 3 wks at 25 °C in a biochemical

incubator, and observed under stereomicroscope. Single spores were selected and

transferred to PDA and synthetic nutrient-poor agar (SNA) (Nirenburg 1981) to

obtain pure cultures. The fungi were examined morphologically on slides prepared

with lactic acid glycerin. Measurements and descriptions of microscopic structures

were made using a Nikon Eclipse 80i light microscope. One isolate was identified

as representing Gliocladiopsis, from which more than 30 conidiogenous cells and

30 conidia were randomly selected for measurement. A dried culture specimen and

living cultures were deposited in the Herbarium of Henan Agricultural University,

Zhengzhou, China (HEAC).

DNA extraction, PCR amplification & sequencing

Total genomic DNA was extracted from fresh mycelia using the CTAB method

(Doyle & Doyle 1990). PCR amplification primers used were: ITS region—universal

primers ITS1 and ITS4 (White & al. 1990); partial 6-tubulin gene—T1 (O’Donnell

& Cigelnik 1997) and CYLTUBIR (Crous & al. 2004); partial Histone H3 gene—

CYLH3F and CYLH3R (Crous & al. 2004). The DNA was amplified using 20 uL

volumes containing 1 wL 50 ng/uL genomic DNA, 1 uL each of 10 uM primers, 10

uL Premix Ex Taq (Version 2.0, TaKaRa, containing 0.625 U DNA polymerase, 200

mM dNTP, and 1.5 mM Mg”*), and 7 uL ddH,O. The amplification protocol in an

Eppendorf Mastercycler gradient thermal cycler followed an initial denaturation

step of 5 min at 94 °C with 35 cycles of 45 s at 94 °C, 30 s at 58 °C, and 1min at

72 °C and a final elongation step of 10 min at 72 °C concluding with incubation at

4 °C. PCR products were examined on 1% agarose gel and sequenced on an ABI

3730XL automated DNA Analyzer by Sangon Biotech (Shanghai) Co, Ltd.

Sequence alignment & phylogenetic analyses

DNA sequences generated by each primer set were emended using SeqMan

v. 7.1.0 in the DNASTAR Lasergene core suite software. Sequences were aligned

and edited manually using MEGA v6.0 (Tamura & al. 2013) and then blasted in

GenBank. Gene sequences of 39 strains were downloaded from NCBI, including the

outgroup Calonectria brachiatica (CBS 123700) and C. brassicae (CBS 111869). The

Bayesian Inference (BI) phylogenetic tree generated with MrBayes v.3.1.2 with the

Gliocladiopsis wuhanensis sp. nov. (China) ... 315

Markov Chain Monte Carlo (MCMC) consisted of 4 runs (Ronquist & Huelsenbeck

2003), with each run analyzing 4 chains analyzed with 10 million generations (trees

sampled every 1000 generations, a sample frequency of 100, stopping when average

standard deviation of split frequencies reached below <0.01, and a burn-in of 25%

(Huelsenbeck & Ronquist 2001; Ronquist & Huelsenbeck 2003). Trees were viewed

in FigTree v. 4.0, then modified using Adobe Illustrator CS6 and saved in Adobe

Photoshop in JPG format. Newly generated sequences were deposited in GenBank.

g710.78f0- curvata CBS 111194 (JQ666122/ JQ666012 / JQ666050)

eolieG. curvata CBS 112935 (JQ666127 / JQ666017 / JQ666051)

-10.8-—G. curvata CBS 112365 (JQ666126/ JQ666016 / JQ666050)

G. curvata CBS 110840 (JQ666121 / JQ666011 / JQ666045)

10.98) G. forsbergii BRIP 60984 (KX274036/KX274053 / KX274070)

69/0.9814G. forsbergii BRIP 61349a (KX274037 / KX274054 | KX274071)

62/0.80,¢G. whileyi BRIP 64430 (KX274052/ KX274069 / KX274086)

ovyG. guangdongensis LC1340 (KC7776124/ KC776120/ KC776122)

seit} 'G. guangdongensis LC1349 (KC7776125/ KC776121 | KC776123)

| LG. wuhanensis HEAC17307 (MH169602 / MH255786 / MH024520)

G. pseudotenuis CBS 114763 (JQ666139 /JQ666029 / JQ666062)

9aLG. pseudotenuis CBS 116074 (JQ666140 / JQ666030 / AF220981)

. peggii BRIP 54049 (JN243766/ JN243767 / JN243765)

10.60 G. peggii BRIP 60983 (KX274038 / KX274065 / KX274083)

a G. peggii BRIP 63709b (KX274046 / KX274056 / KX274073)

1)4/0.97,G. peggii BRIP 60987 (KX274040 / KX274062 / KX274074)

40.6G. peggii BRIP 60988 (KX274043 / KX274063 / KX274082)

G. mexicana CBS 110938 (JQ666137 / JQ666027 / JQ666060)

100nlG. mexicana CBS 111131 (JQ666138/ JQ666028 / JQ666061)

9010-75] 1G. irregularis CBS 111142 (0666124 / JQ666024 / JQ666057)

gaiif O- irregularis CBS 111176 (JQ666135 / JQ666025 / JQ666058)

G. irregularis CBS 114667 (JQ666136 / JQ666026 / JQ666059)

a G. irregularis CBS 755.97 (JQ666133 / JQ666023 / AF220977)

7110.85 ] ,G. sumatrensis CBS 111198 (10666143 / JQ666033 / JQ666065)

gti [4 | G. sumatrensis CBS 111213 (JQ666144 / JQ666034 / JQ666066)

9710.73} go] FG. Sumatrensis CBS 111368 (JQ666145 / JQ666035 / AF220978)

G. sumatrensis CBS 754.97 (JQ666142/ JQ666032 / JQ666064)

G. indonesiensis CBS 116090 (JQ666132/ JQ666022 / JQ666056)

G. tenuis CBS 111961 (JQ666146 / JQ666036 / JQ666067)

G. tenuis CBS 111964 (JQ666147 / JQ666037 / JQ666068)

100 1G. tenuis CBS 114147 (J666148 / 0666038 / JQ666069)

G. tenuis CBS 114148 (JQ666149/ JQ666039 / JQ666070)

7110.9 G. tenuis IMI 68205 (JQ666150/ JQ666040 / AF220979)

Gliocladiopsis sp. 1 CBS 111038 (JQ666151 / JQ666041 / JQ666071)

sor S: elghollii CBS 116104 (JQ666131 / JQ666021 / JQ666055)

se] 1G. elghollii CBS 206 94 (JQ666130 / JQ666020 / JQ666054)

100! G. sagariensis CBS 199 55 (JQ666141 / JQ666031 / JQ666063)

G. aquaticus MFLUCC 17-1841 (MG574421 | MG734182/ MG543924)

gaitG. aquaticus MFLUCC 17-2028 (MG574422 / MG734183 / MG543925)

1001 Calonectria brassicaeCBS 111869 (AF232857 / DQ190720 / GQ280576)

Calonectria brachiaticaCBS 123700 (FJ696388 / FJ696396 (GQ280555)

8710.97

0.04

Fic. 1. Maximum-likelihood (ML) Gliocladiopsis tree obtained from the combined DNA sequence

data from three loci (internal transcribed spacer region, histone H3, and f-tubulin). Calonectria

brachiatica (CBS 123700) and C. brassicae (CBS 111869) were included as the outgroup. Bootstrap

support values >50 %, and Bayesian posterior probability values >0.95, are shown at the nodes

(BS/PP). Branches supported by BS = 100 % and PP = 1 are depicted as black thickened lines.

Ex-type strains are in bold.

316... Zhai & al.

Phylogenetic results

Amplicons of approximately 400-600 base pairs were determined for

TUB2 (489 bp), HIS3 (477 bp), and ITS (461 bp). The phylogenetic analysis

comprised 15 ingroup and two outgroup [Calonectria brachiatica (CBS

123700), C. brassicae (CBS 111869)] taxa. The combined data set comprised

1427 characters, including 35 gaps and 775 constant characters. Analysis

of the 305 parsimony-informative characters yielded one tree (Fic. 1).

GTR+I+G was the model selected for Bayesian analysis and Maximum

Likelihood (ML). The tree (Fic. 1) divides Gliocladiopsis isolates into two

main clades, the first (PP = 1) containing the extype G. sagariensis strain

(CBS 199.55), two G. elghollii strains (CBS 206.94, CBS 116104), and two

G. aquaticus strains (MFLUCC 17-1811, MFLUCC 17-2028), and the second

(PP = 0.73) containing the other 12 Gliocladiopsis species: (G. curvata,

G. forsbergii, G. guangdongensis, G. indonesiensis, G. irregularis, G. mexicana,

G. peggii, G. pseudotenuis, G. sumatrensis, G. tenuis, G. whileyi, and

G. wuhanensis.) on the main branch.

Taxonomy

Gliocladiopsis wuhanensis Meng Zhang, N.P. Zhai & Y.H. Geng, sp. nov. FIG. 2

MB 825569

Differs from Gliocladiopsis guangdongensis, G. curvata, G. forsbergii, and G. whileyi by its

tertiary conidiophore branching and its longer conidia.

Type: China, Hubei Province, Wuhan, from potato farm soil, 15 Jul. 2017, Niping Zhai

(Holotype, HEAC17056 [dried culture]; ex-type living culture, HEAC17307; GenBank

MH024520, MH169602, MH255786).

EryMoLoey: wuhanensis, referring to the city where the type was collected.

CoLonlgs on the SNA effuse, velvet or powdery, whitish to yellow, reverse

green, reaching 6 cm diam. in two wks at 25 °C; mycelium mostly superficial

or immersed, hyphae smooth, septate, branched, hyaline, 1.0-5 um.

CONIDIOPHORES penicillate, without stipe extensions and terminal vesicles,

pale brown, smooth, primary branches aseptate or 1-septate, 13.0-38.5 x

2.0-4.5 um; secondary branches aseptate, 9.5-30.5 x 2.0-4.5 um; tertiary

branchesaseptate, 2.5-13.0 x 1.0-2.5um. CONIDIOGENOUS CELLS polyblastic,

hyaline to light brown, 13.0-37.5 x 2.0-3.5 um. ConipIA single, smooth,

hyaline, cylindrical, straight or slightly curved, 0-1-septate, 13.5-18.5 x

2.0-2.5 um. TELEOMORPH unknown.

Gliocladiopsis wuhanensis sp. nov. (China) ... 317

Fic. 2. Gliocladiopsis wuhanensis (ex-type culture). A, B. Sporulation phenotype on PDA after 10 d;

C, D. Colonies on SNA after 10 d (C = top view, D = reverse view); E-I, K. Penicillate conidiophores;

J, L, M. Conidia. Scale bars = 10 um.

Discussion

Gliocladiopsis wuhanensis is morphologically similar to G. guangdongensis,

G. curvata L. Lombard & Crous, G. forsbergii L.E. Parkinson & al., and

G. whileyi L.E. Parkinson & al. However, G. guangdongensis differs by its

binary conidiophore branching and its shorter conidia (13.5-16 um; Liu &

Cai 2013); G. curvata differs by its quaternary conidiophore branching and

its shorter, wider conidia (17-21 x 3-5 um; Lombard & Crous 2012), and

G. forsbergii and G. whileyi differ by their quaternary conidiophore branching

and their shorter conidia (15.5-19 um and 17-21 um; Parkinson & al. 2017).

Phylogenetically, Gliocladiopsis wuhanensis is closely related to G. guang-

dongensis. However, for the HIS3 gene, G. wuhanensis is separated by 12

bp from G. guangdongensis, greater than that from G. forsbergii (10 bp) and

G. whileyi (9 bp), suggesting that they are phylogenetically more distant.

318 ... Zhai & al.

The known Gliocladiopsis species were isolated from different habitats

(roots of diseased plants, plant litter, water, or soil) (Lombard & Crous 2012).

Gliocladiopsis species occur quite often in soil associated with plant roots.

Dann & al. (2012) conducted tests on plant roots, but the pathogenicity was

not clear, and possibly they were mainly saprobic.

Acknowledgements

The author would like to thank the reviewers, De-Wei Li (The Connecticut

Agricultural Experiment Station Valley Laboratory, CT USA) and Ying Zhang

(Institute of Microbiology, Beijing Forestry University, P.R. China), for their valuable

comments. We also thank the curator of CABI Bioscience for making the ex-type

strain of G. tenuis available for study.

This study was financially supported by NSFC 31770029, 31400129 &

18IRTSTHN021. The authors declare no potential conflict of interest for this study.

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MY COTAXON

April-June 2019—Volume 134, pp. 321-328

https://doi.org/10.5248/134.321

Four new records of Haematomma

from Southern China

RONG TANG *, XIAO ZHANG *, CHUN-XIAO WANG, LU-LU ZHANG *

Key Laboratory of Plant Stress Research, College of Life Sciences

Shandong Normal University, Jinan, 250014, P. R. China

* CORRESPONDENCE TO: 675359138@qq.com

ABSTRACT—Haematomma accolens, H. collatum, H. fenzlianum, and H. flexuosum are

reported as new for China; and H. fenzlianum is also new for Asia. Detailed taxonomic

descriptions with photos are provided for the four species.

Key worps—East Asia, Haematommataceae, Lecanorales, lichenized Ascomycota, taxonomy

Introduction

Haematomma A. Massal., introduced by Massalongo in 1852, is a genus of

crustose lichens with blood-red apothecia having thallus-coloured margins

and transversely septate to muriform ascospores. For the most part, species

of Haematomma are found in the tropics and subtropics (Staiger & Kalb

1995, Brodo & al. 2008), so it is not surprising that in China Haematomma

is mainly distributed in the southern provinces, where the terrain is diverse

with plains, basins, and plateaus, dominated by tropical subtropical monsoon

climate and with high temperatures and abundant rain in summer and mild

temperatures with less rain in winter. With its diverse climate types (tropical,

subtropical, temperate, and cold temperate) and plant ecosystems (tropical rain

forest, evergreen broadleaf forest, deciduous broadleaf forest, coniferous forest,

meadow, desert, etc.), and altitudes of 72-6740 m, Yunnan Province has an

especially rich range of lichen species.

* RONG TANG & XIAO ZHANG contributed equally to this research.

322 ... Tang, Zhang & al.

In the world monograph of Haematomma by Staiger & Kalb (1995), only

one species is mentioned as occurring in China: H. rufidulum (Fée) A. Massal.

Previously, H. fauriei Zahlbr. and H. puniceum (Ach.) A. Massal. had been

recorded from China (Zahlbruckner 1930, Xu 1989), but as we have been unable

to find any specimens of these two species to verify the reports, we cannot

confirm their Chinese occurrence. Since 1995, four other species have been

added to the Chinese list: H. africanum (J. Steiner) C.W. Dodge, H. caperaticum

Brodo & al., H. persoonii (Fée) A. Massal., and H. wattii (Stirt.) Zahlbr. (Wei

1991, Aptroot & Sparrius 2003, Obermayer 2004, Tang & al. 2018), to which

we here add another four species: H. accolens, H. collatum, H. fenzlianum, and

H. flexuosum based primarily on recently collected material from Southern

China. Hopefully, this paper will provide the basis for further research.

Materials & methods

The specimens studied are housed in: Lichen Section of the Botanical

Herbarium, Shandong Normal University (SDNU); Kunming Institute of Botany,

Chinese Academy of Sciences (KUN-L); Sectio Lichenum, Herbarium Mycologici

Academiae Sinicae (HMAS-L); and Lichen Herbarium of the College of Life Sciences,

Liaocheng University (LCU). The macro- and micromorphological characters of the

specimens were examined under an Olympus SZ51 stereomicroscope and Olympus

CX21 polarizing microscope. Both thallus and medulla were tested with K (a 10%

aqueous solution of potassium hydroxide) and C (a saturated solution of aqueous

sodium hypochlorite). The lichen substances were identified using standardized

thin layer chromatography techniques (TLC) with solvent system C (Orange & al.

2010). Photos of the voucher specimens were taken under Olympus SZX16 and

BX61 with DP72.

Taxonomic descriptions

Haematomma accolens (Stirt.) Hillmann,

Repert. Spec. Nov. Regni Veg. 49: 34 (1940) Fic. 1A-E

Thallus crustose, smooth or roughened, slightly cracked, without isidia or

soredia, pale yellowish white; prothallus present, white. Apothecia red, sessile

or constricted at base, dispersed or crowded, smooth, epruinose, 0.4-1.0

mm diam., margin distinct, smooth; epihymenium orange-red, K+ purple,

dissipating; hymenium hyaline, 75-90 um tall; hypothecium hyaline; asci

clavate, 8-spored; ascospores persistently colorless, parallel in ascus, several-

septate, fusiform, with 7-12 cells in optical view, 38-60 x 3-4(-5) um. Pycnidia:

not observed.

CHEMISTRY—Cortex and medulla K+ yellow, C-, KC-, IKI-. Atranorin,

placodiolic acid, and haematommone detected by TLC.

Haematomma spp. new to China... 323

Hasitat—Growing on the bark of shrubs trees (e.g., Betula and Michelia)

at elevations of 400-1230 m and in China usually growing together with

Phaeographis spp..

DiIsTRIBUTION—North, Central, and South America, Africa, Australia, Asia

(Staiger & Kalb 1995, Brodo & al. 2008). New to China.

SPECIMENS EXAMINED: CHINA. GUANGDONG, Shixing, Lulang, Che eight Ling National

Nature Reserve, alt. 400 m, on bark, 14 May 2015, Zefeng Jia GD15156 (LCU); YUNNAN,

Dali Co., Nanjian, Gonglang Village, Yangjiehe, alt. 1230 m, on bark, 15 Jun. 2012, Wang

Li-song, Wang Xin-yu & Liu Dong 12-34410A, 12-34410C (KUN-L).

COMMENTS—Haematomma accolens is characterized by the 7-12-celled

ascospores and K+ purple epihymenium. The description of the Chinese

material is similar to previously published descriptions of H. accolens,

although Brodo & al. (2008) reported the spores with (5-)6-8(-12) cells.

Haematomma accolens is morphologically similar to H. flexuosum, but they

have a different chemistry: H. accolens contains atranorin, placodiolic acid, and

haematommone, whereas H. flexuosum contains atranorin, isoplacodiolic acid,

isopseudoplacodiolic acid, and haematommone.

Haematomma collatum (Stirt.) C.W. Dodge,

Beih. Nova Hedwigia 38: 41 (1971) Fic. 1F-J

Thallus crustose, continuous, rugose or rimose, without isidia or soredia,

yellowish white; prothallus usually present, white. Apothecia rounded, orange-

red, sessile or constricted at base, dispersed, smooth, (0.5-)0.8-1.5 mm diam.,

margin distinct, crenulate, usually prominent above disk; epihymenium orange

to orange-red, K+ red; hymenium hyaline, 80-95 um tall; hypothecium hyaline

or slightly yellowish, K-; paraphyses branched and anastomosing; asci clavate,

8-spored; ascospores persistently colorless, twisted in ascus, multiseptate,

fusiform, curved, with 10-14 cells in optical view, locules sometimes not equal

in size, the tail cell longer than middle cells, (53-)65-75 x 4-5 um. Pycnidia:

not observed.

CHEMISTRY—Cortex and medulla K+ yellow, C-, KC-, IKI-. Atranorin,

sphaerophorin, isosphaeric acid (minor), and russulone detected by TLC.

Hasitat—Growing on bark at elevations of 670-2500 m. The specimen

from Yunnan was found in a moist evergreen broadleaf forest; compared with

other Haematomma spp. in this area, H. collatum grows very well. We also find

that, in China, H. collatum likes to grow in the same environment as lichens in

the Graphidaceae.

DISTRIBUTION—Central and South America, Africa, Australia, Asia Minor

(Staiger & Kalb 1995; Brodo & al. 2008). New to China.

324 ... Tang, Zhang & al.

Haematomma spp. new to China... 325

SPECIMENS EXAMINED: CHINA. YUNNAN, Jingdong, Mt. Ailao, 24°32’29”N

101°01’37”E, alt. 2500 m, on bark, 5 Aug. 2017, R. Tang & al. 20170319 (SDNU);

HAINAN, Mt. Wuzhi, 18°46’N 109°31’E, alt. 670 m, on bark, 28 Jun. 2008, Jia Zefeng.

119392 (HMAS-L).

CoMMENTS—Haematomma collatum can be characterized by its crenulate

apothecial margins, 10-14-celled ascospores, medullary sphaerophorin,

and epithecial russulone. The Chinese material closely matches previously

published descriptions (Staiger & Kalb 1995). This species and H. persoonii

contain the same chemical substances, but H. persoonii has ascospores with

fewer (6-8) cells (Brodo & al. 2008).

Haematomma fenzlianum A. Massal.,

Mem. Reale Ist. Veneto Sci. 10: 58 (1861) Fic. 2A-E

Thallus thick, crustose, rugose or rimose, areolate, without isidia or soredia,

dull yellowish white; prothallus not seen. Apothecia rounded, deep reddish,

sessile or constricted at base, dispersed, smooth, epruinose, 0.5(-0.8)—2.0

mm diam., margin distinct, crenulate or flexuose, usually prominent above

disk; epihymenium orange-red, K+ red; hymenium hyaline, 70-75 um tall;

hypothecium hyaline or pale yellowish; asci clavate, 8-spored; ascospores

persistently colorless, multiseptate, fusiform, straight or slightly curved, with

(3-)4-7 cells in optical view, (30-)33-43(-48) x 4.5-5 um. Pycnidia: not

observed.

CHEMISTRY—Cortex and medulla K+ yellow, C-, KC-. Atranorin,

sphaerophorin, isosphaeric acid, and russulone detected by TLC.

DISTRIBUTION—Europe, Africa, Australasia, America (Staiger & Kalb 1995,

Nash & al. 2004, Brodo & al. 2008). New to China, and new to Asia.

HasitatT—Growing on non-calcareous rock. Haematomma fenzlianum has

rarely been collected in China, probably because it likes to grow in dry exposed

areas, such as those occurring in North America and Argentina; comparable

habitats are rare in southern China.

SPECIMEN EXAMINED: CHINA. YUNNAN, Lijiang, Yupang, alt. 2600 m, on rock, 15 Aug.

1982, Wang Li-song 82-1081 (KUN-L).

COMMENTS—Haematomma fenzlianum is characterized by its thick thallus

and 4-7-celled ascospores, and by its substrate. The Chinese material

Fic. 1. Haematomma accolens (LCU-L Jia GD15156). A. Thallus; B. Apothecium; C. Apothecium

section; D. Ascus; E. Ascospores. Haematomma collatum (HMAS-L Jia 119392). F. Thallus;

G. Apothecium; H. Apothecium section; I. Ascus; J. Ascospores.

326 ... Tang, Zhang & al.

closely matches previously published descriptions (Staiger & Kalb 1995).

Haematomma fenzlianum is the only saxicolous species of Haematomma in

China. Haematomma ochroleucum (Neck.) J.R. Laundon is also saxicolous but

differs in containing zeorin and being abundantly sorediate (vs. H. fenzlianum

which lacks zeorin and (usually) soredia).

Haematomma flexuosum Hillmann,

Repert. Spec. Nov. Regni Veg. 49: 35 (1940) Fic. 2F-J

Thallus crustose, continuous, rugose or rimose, roughened, without isidia

or soredia, yellowish white; prothallus present, white. Apothecia rounded, deep

reddish, sessile or constricted at base, dispersed, smooth, 0.4-0.8 mm diam.,

margin distinct, even with disk; epihymenium red, K+ purple, dissipating;

hymenium hyaline, 63-70 um tall; hypothecium hyaline or pale yellowish,

K-; paraphyses branched and anastomosing; asci clavate, 8-spored; ascospores

persistently colorless, multiseptate, fusiform, curved or straight, with 5-7 cells

in optical view, (35-)50-63 x 4-5 um. Pycnidia: not observed.

CHEMISTRY—Cortex and medulla K+ yellow, C-, KC-. Atranorin,

isoplacodiolic acid, isopseudoplacodiolic acid, and haematommone detected

by TLC.

DISTRIBUTION—Central and South America, Africa, Australia, West Indies

(Staiger & Kalb 1995; Brodo & al. 2008). New to China.

Hasitat—Growing on bark around 1000 m above sea level. We found this

species growing together with H. rufidulum.

SPECIMENS EXAMINED: CHINA. YUNNAN, Najian, Mt. Manhai, 24°42’57”N

100°25’17’E, alt. 1266 m, on bark, 30 Jun. 2015, X. Ye & W.C. Wang 15-47956(A), 15-

47956(D) (KUN-L). YUNNAN, Luxi, Mt. Santai, 24°26’N 98°35’E, alt. 1340 m, on bark,

28 Jun. 1980, Jiang Yu-mei 051540 (HMAS-L).

COMMENTS—Haematomma flexuosum is characterized by 5-7-celled

ascospores and by its special chemistry. The Chinese material closely matches

the description by Staiger & Kalb (1995). Haematomma matogrossense

Kalb & Staiger is similar but differs by having 8-14-celled spores and by

containing methylisopseudoplacodiolic acid and isopseudoplacodiolic acid

(rather than 5-7 cells and isoplacodiolic acid and isopseudoplacodiolic acid

in H. flexuosum).

Fic. 2. Haematomma fenzlianum (KUN-L Wang 82-1081). A. Thallus; B. Apothecium;

C. Apothecium section; D. Ascus; E. Ascospores. Haematomma flexuosum (KUN-L Ye

15-47956(D)). F. Thallus; G. Apothecium; H. Apothecium section; I. Ascus; J. Ascospores.

Scale bars: A= 5 mm;B=1 mm.

O20

Haematomma spp. new to China...

328 ... Tang, Zhang & al.

Acknowledgements

We thank Dr. Irwin Brodo (Canadian Museum of Nature, Ottawa, Canada),

Dr. S. Y. Guo (State Key Laboratory of Mycology, Chinese Academy of Sciences,

Beijing, China) and Dr. John Elix (Australian National University, Canberra,

Australia) for providing great guidance during the study. Thank Dr. Li-song Wang

(Kunming Institute of Botany, China), Dr. Ze-feng Jia (Liaocheng University, College

of Life Sciences, China) and Dr. Hong Deng (State Key Laboratory of Mycology,

Institute of Microbiology, Chinese Academy of Sciences, Beijing, China) for

sending specimens to us. This work was supported by the National Natural Science

Foundation of China Youth Science Foundation (31600100), and Emergency

management project of National Natural Science Foundation of China (31750001).

Literature cited

Aptroot A, Sparrius LB. 2003. New microlichens from Taiwan. Fungal Diversity 14: 1-50.

Brodo IM, Culberson, WL, Culberson, CE. 2008. Haematomma (Lecanoraceae) in North and

Central America, including the West Indies. Bryologist 111: 363-423.

https://doi.org/10.1639/0007-2745(2008)111[363:hlinac]2.0.co;2

Nash TH, Ryan BD, Diederich P, Gries C, Bungartz F. 2004. Lichen flora of the Greater Sonoran

Desert Region, vol. 2. Tempe, Lichens Unlimited.

Obermayer W. 2004. Additions to the lichen flora of the Tibetan region. Bibliotheca

Lichenologica 88: 479-526.

Orange A, James PW, White FJ. 2010. Microchemical methods for the identification of lichens.

2nd edition. London, British Lichen Society.

Staiger B, Kalb K. 1995. Haematomma-Studien: I. Die Flechtengattung Haematomma.

Bibliotheca Lichenologica 59. 198 p.

Tang R, Yan SK, Sun MJ, Zhang LL. 2018. New records of Haematomma and Ophioparma from

China. Mycotaxon 133: 175-181. https://doi.org/10.5248/133.175

Wei JC. 1991. An enumeration of lichens in China. Beijing, China. International Academic

Publishers.

Xu BS. 1989. Cryptogamic flora of the Yangtze Delta and adjacent regions. Shanghai Scientific

and Technical Publishers: 158-266.

Zahlbruckner A. 1930. Lichenes. Symbolae Sinicae, vol. 3. 254 p.

MY COTAXON

April-June 2019—Volume 134, pp. 329-334

https://doi.org/10.5248/134.329

Cordana meilingensis and C. lushanensis spp. nov.

from Jiangxi, China

ConG-Conc AI’, JIAN Ma*3, KAI ZHANG‘,

RAFAEL FE. CASTANEDA-RUIZ*, XIU-GUO ZHANG’*

"Department of Plant Pathology, Shandong Agricultural University,

Taian, Shandong 271018, China

College of Agronomy, Jiangxi Agricultural University, Nanchang, Jiangxi 330045, China

"Jiangxi Key Laboratory for Conservation and Utilization of Fungal Resources,

Jiangxi Agricultural University, Nanchang, Jiangxi 330045, China

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

‘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: zhxg@sdau.edu.cn

ABSTRACT—Two new anamorphic fungi, Cordana meilingensis and C. lushanensis,

were collected from dead branches in Jiangxi Province, China. Cordana meilingensis is

characterized by its oblong or cylindrical, medially 1-septate, brown, smooth conidia with

a prominent hilum. Cordana lushanensis is distinguished by its ellipsoidal to obovoid, pale

brown, aseptate, smooth conidia with a prominent basal scar. They are described, illustrated,

and compared with similar taxa.

KEY worDs—asexual fungi, Cordanaceae, Cordanales, hyphomycetes, taxonomy

Introduction

Preuss (1851) established Cordana with three species, without designating

a type species, and subsequently added a fourth species (Preuss 1853).

However, Saccardo (1886) redescribed Cordana, retaining only one species,

C. pauciseptata Preuss, which thus effectively became the type species

(Hughes 1958, Seifert & al. 2011; epitypified by Hernandez-Restrepo &

330... Ai & al.

al. 2014). Cordana is mainly characterized by distinct, simple or sparely

branched conidiophores, and solitary, acropleurogenous, 0-1-septate conidia

seceding schizolytically from polyblastic, integrated, terminal and becoming

also intercalary, sympodial conidiogenous cells with small denticles, the

conidia are ellipsoidal, ovoid, obovoid, pyriform or cylindrical, often with

a prominent hilum (Ellis 1971, Markovskaja 2003). Castafieda-Ruiz & al.

(1999) reviewed 17 species of Cordana and provided a comparative table that

distinguished 11 accepted Cordana species. However, Markovskaja (2003)

regarded C. miniumbonata R.F. Castaheda & al. as a problematic species

due to its conidial shape and septation mode. Cordana currently contains

19 recognized species (Hernandez-Restrepo & al. 2014), distinguished

primarily by conidial features including shape, size, septation, pigmentation,

ornamentation, and presence or absence of a synanamorph (Castafeda-Ruiz

& al. 1999, Markovskaja 2003, Hernandez-Restrepo & al. 2014).

During our continuing survey (2005-18) of microfungi from plant debris

in the forests of southern China, two species referable to the genus Cordana

were collected on dead branches in Jiangxi Province. A close examination of

the two fungi showed that they have significant differences from previously

described Cordana species and are therefore proposed as new to science.

Cordana meilingensis C.C. Ai, Jian Ma, X.G. Zhang & R.F. Castafieda, sp.nov. Fic. 1

IF 555813

Differs from Cordana johnstonii, C. uniseptata, and C. versicolor by its smaller oblong

or cylindrical, medially 1-septate, concolorous conidia; and from C. mercadoana by its

larger, medially 1-septate, brown conidia.

Type: China, Jiangxi Province: Meiling National Park, on dead branches of an

unidentified broadleaf tree, 8 October 2013, J. Ma (Holotype, HJAUP M0144).

EryMoLoGey: refers to the locality where the type specimen was found.

CoLonligs on the natural substratum effuse, brown to dark brown. Mycelium

partly superficial, partly immersed, composed of branched, septate, smooth,

subhyaline to pale brown hyphae. CONIDIOPHORES macronematous,

mononematous, unbranched, erect, straight or flexuous, cylindrical, smooth,

septate, brown to dark brown, paler towards the apex, 73-185 x 4-5 um,

occasionally swollen at the base, 5.5-9 um diam. CONIDIOGENOUS CELLS

polyblastic, integrated, terminal and intercalary, with subhyaline and slightly

prominent scars, cylindrical. Conidial secession schizolytic. CoNrp1A

solitary, acropleurogenous, dry, medially 1-septate, often slightly constricted

at the septa, oblong or cylindrical, brown, smooth, 10-13 x 5.5-7 um, with a

prominent hilum, 0.5-1 x 0.5 um.

Cordana meilingensis & C. lushanensis spp. nov. (China) ... 331

204m

20um

4 i

Fic. 1. Cordana meilingensis (holotype, HHAUP M0144).

A. Conidia; B-F. Conidiophore with conidia.

20nm

COMMENTS—Cordana meilingensis resembles C. johnstonii M.B. Ellis,

C. uniseptata L. Cai & al., C. versicolor D.J. Soares & R.W. Barreto, and

C. mercadoana Hern.-Restr. & al. in conidial shape (TaBLE 1). However,

C. johnstonii has larger broadly ellipsoidal conidia and grows only on

plants in the genus Musa (Ellis 1971); C. uniseptata produces larger broadly

ellipsoidal, asymmetrically 1-septate, versicolored conidia (Cai & al. 2004);

C. versicolor differs by its larger broadly ellipsoidal conidia with a paler basal

cell and is parasitic on Canna denudata Roscoe [= C. paniculata Ruiz & Pav.]

332 ... Ai &al.

(Soares & al. 2005); and C. mercadoana differs by its smaller, versicolored,

0-1-septate conidia (Hernandez-Restrepo & al. 2014).

TABLE 1. Comparisons of conidia and substrates of Cordana meilingensis

and similar species

CONIDIA

SPECIES SS = Lk SUBSTRATE,

SHAPE SIZE(um) COLOR SEPTATION

C. johnstonii! Broadly 20-30 x Uniform Medially Parasitic on

ellipsoidal 12-18 1-septate Musa spp.

a ae Chonan eS a Ee Gia Mane? Oe ee

cylindrical 5.5-7 1-septate dead branches

“Co mercadoana® Oblong, 6-10x Variable 0-I-septate Saprobicon

obovoid, or 3-4 dead twig

cylindrical

ane? Saye ee nT ee aa ASG eae

ellipsoidal 8.5-11.5 1-septate on bamboo

Beye ave Pr epee Bees sparrata cotta eaaees Mai ee pesca eile

ellipsoidal 10-15 1-septate Canna

denudata

Data from ' Ellis (1971); ? Hernandez-Restrepo & al. (2014); * Cai & al. (2004); *Soares & al. (2005)

Cordana lushanensis C.C. Ai, Jian Ma, X.G. Zhang & R.F. Castafieda, sp. nov. Fic. 2

IF 555814

Differs from Cordana verruculosa by its larger smooth conidia; from C. semaniae and

C. solitaria by its smaller ellipsoidal to obovoid, pale brown conidia with a prominent

basal scar; and further from C. solitaria by lacking a Bispora-like synanamorph.

Type: China, Jiangxi Province: Lushan (Mount Lu), on dead branches of an

unidentified broadleaf tree, 8 November 2017, J. Ma (Holotype, HHAUP M5406).

EryMo_oey: refers to the locality where the type specimen was found.

Co.onigs on the natural substratum effuse, brown to dark brown. Mycelium

partly superficial, partly immersed, composed of branched, septate, smooth,

subhyaline to pale brown hyphae. CoNIDIOPHORES macronematous,

mononematous, simple or branched, erect, straight to flexuous, cylindrical,

with intercalary nodes, 7-14 um diam, brown, paler toward the apex, smooth,

<240 x 6.5-7.5 um. CONIDIOGENOUS CELLS integrated, polyblastic, terminal

and intercalary, with subhyaline small denticles, proliferations percurrent,

cylindrical to lageniform. Conidial secession schizolytic. Conip1a solitary,

acropleurogenous, dry, 0-septate, ellipsoidal to obovoid, pale brown, smooth,

5.5-8 x 2.5-4 um, with a prominent basal scar, 0.3-0.5 um diam.

Cordana meilingensis & C. lushanensis spp. nov. (China) ... 333

000

10nm

Fic. 2. Cordana lushanensis (holotype, HHAUP M5406).

A. Colonies on natural substratum; B—D. Conidiophores and conidia; E. Conidia.

ComMENTs - Among the known species, only C. semaniae Davydkina & al.,

C. solitaria V. Rao & de Hoog, and C. verruculosa Hern.-Restr. & al. resemble

C. lushanensis in producing aseptate conidia. However, C. verruculosa differs

by its smaller (3-5.5 x 2-3.5 um) verruculose conidia (Hernandez-Restrepo

& al. 2014); C. semaniae differs by its larger (21-27 x 9-15 um) obovoid black

conidia with an acute basal cell (Davydkina & Melnik 1989); C. solitaria differs

by its broader (4.5-6.5 um diam) obovoid conidia with a slightly papillate base,

and the presence of a Bispora-like synanamorph (Rao & de Hoog 1986).

Acknowledgments

The authors express gratitude to Dr. De-Wei Li (The Connecticut Agricultural

Experiment Station Valley Laboratory, Windsor CT, USA) and Dr. Li-Guo Ma

(Institute of Plant Protection, Shandong Academy of Agricultural Sciences, Jinan,

China) for serving as pre-submission reviewers and to Dr. Shaun Pennycook for

nomenclatural review. This project was supported by the National Natural Science

Foundation of China (No. 31360011, 31870016), and the Jiangxi Province Department

of Education, China (No. GJJ160357).

Literature cited

Cai L, McKenzie EHC, Hyde KD. 2004. New species of Cordana and Spadicoides from decaying

bamboo culms in China. Sydowia 56(2): 222-228.

334 ... Ai &al.

Castafeda-Ruiz RF, Iturriaga T, Guarro J. 1999. A new species of Cordana from Venezuela.

Mycotaxon 73: 1-8.

Davydkina TA, Mel'nik VA. 1989. Two new hyphomycetes from the genera Cordana and

Pyriculariopsis. Mikologiya i Fitopatologiya 23(2): 110-113.

Ellis MB. 1971. Dematiaceous hyphomycetes. X. Mycological Papers 125. 30 p.

Hernandez-Restrepo M, Gené J, Mena-Portales J, Cano J, Madrid H, Castaneda-Ruiz RF, Guarro

J. 2014. New species of Cordana and epitypification of the genus. Mycologia 106(4): 723-734.

https://doi.org/10.3852/13-122

Hughes SJ. 1958. Revisiones hyphomycetum aliquot cum appendice de nominibus rejiciendis.

Canadian Journal of Botany 36: 727-836.

Markovskaja S. 2003. A new species of Cordana from Lithuania. Mycotaxon 87: 179-185.

Preuss CGT. 1851. Ubersicht untersuchter Pilze, besonders aus der Umgegend von Hoyerswerda.

Linnaea 24: 99-153.

Preuss CGT. 1853. Ubersicht untersuchter Pilze, besonders aus der Umgegend von Hoyerswerda.

Linnaea 25: 723-742.

Rao VG, de Hoog GS. 1986. New or critical hyphomycetes from India. Studies in Mycology 28.

84 p.

Saccardo PA. 1886. Sylloge hyphomycetum. Sylloge Fungorum 4. 807 p.

Seifert K, Morgan-Jones G, Gams W, Kendrick B. 2011. The genera of hyphomycetes. CBS

Biodiversity Series 9. 997 p.

Soares DJ, Nechet KL, Barreto RW. 2005. Cordana versicolor sp. nov. (dematiaceous hyphomycete)

causing leaf-spot on Canna denudata (Cannaceae) in Brazil, with observations on Cordana

musae. Fungal Diversity 18: 147-155.

MY COTAXON

April-June 2019—Volume 134, pp. 335-352

https://doi.org/10.5248/134.335

Coprophilous fungi from Brazil:

new records for the Neotropics

ROGER ER. MELoO”™, DANIEL B.P. DO MONTE’, NICOLE H.B. GONDIM’,

LEONOR C. MAtA', ANDREW N. MILLER?

' Universidade Federal de Pernambuco, Departamento de Micologia,

Av. da Engenharia, s/n, 50740-600, Recife, Brazil

? University of Illinois at Urbana-Champaign, Illinois Natural History Survey,

1816 S. Oak St., Champaign, IL 61820, USA

* CORRESPONDENCE TO: rogerfrmelo@gmail.com

ABSTRACT— Twelve ascomycetes associated with herbivore dung, collected throughout an

edaphic and climatic gradient from the Atlantic Forest complex to the semi-arid Caatinga

in Pernambuco, Northeastern Brazil, represent new records: Ascobolus castaneus, Arnium

hirtum, Cercophora anisura, Chaetomium citrinum, C. spirale, Coniochaeta philocoproides,

Coprotus albidus, and Delitschia vulgaris are new records for the Neotropics; Cercophora

sordarioides, Chaetomium cochliodes, Sordaria lappae, and Thelebolus microsporus are new

records for Brazil. Ascobolus castaneus is recorded for the second time worldwide, revealing

a much wider geographical distribution for a species previously reported only from China.

Descriptions and updated geographic distributions are presented, and information regarding

the records is discussed.

KEY worps—Ascomycota, dung fungi, taxonomy

Introduction

Coprophilous fungi constitute a rich and diverse ecological group of

saprobes especially adapted to withstand conditions and use resources available

in herbivore dung (Kirschner & al. 2015). These fungi can be found in dung

of many herbivores, domesticated or wild, all over the world and throughout

the year (Richardson 2001). Fungi on dung are represented mainly by

members of Ascomycota (e.g., Ascobolaceae, Lasiosphaeriaceae), Mucoromycota

336 ... Melo & al.

(e.g., Pilobolaceae, Mucoraceae), and Basidiomycota (e.g., Coprinaceae,

Psathyrellaceae). As in other ecological groups, a gradient ranging from

specialized species to opportunistic saprobes can be found in coprophilous

fungal communities (Newcombe & al. 2016). Despite this, the coprophilous

mycobiota retains some similarity and shows somewhat regular patterns of

occurrence and abundance throughout continents and time (Basumatary &

McDonald 2017). The Brazilian coprophilous mycobiota is poorly known,

and recent works have focused on the taxonomy and distribution of the most

common genera and in establishing regional and national checklists (Calaca

& al. 2014; Melo & al. 2012, 2017; Souza & al. 2017). As most coprophilous

species are restricted to their substrates, their geographical distribution is only

known where such inventories have been conducted, and some species thought

to be restricted to temperate regions are being reported in the tropics as well.

This work presents the first report of twelve records of Ascomycota associated

with herbivore dung, all new records either for the Neotropics, South America,

or Brazil.

Materials & methods

A total of 270 dung samples from goat, cattle, and horse were sampled between

2011 and 2013 from three animal precincts: (1) Universidade Federal Rural de

Pernambuco, Veterinary Medicine Department, Recife (8°00°54”S 34°56’59”W),

(2) Instituto Agronémico de Pernambuco (IPA), Caruaru (8°01’59”S 36°06’59”W),

and (3) Instituto Agronémico de Pernambuco (IPA), Serra Talhada (7°54’59”S

38°17'00”W). These areas are located along a latitudinal transect of ~420 km in the

state of Pernambuco, Northeastern Brazil and encompass two biomes: the Atlantic

Rainforest to the east and the semiarid Caatinga to the west. Additional material,

from Rio Grande do Sul state, were also received. Fresh dung samples were collected

in clean plastic bags, taken to the laboratory, and incubated in moist chambers at

room temperature (28 + 2 °C) for at least 75 days under alternating natural light

and dark periods. The material was observed directly on the substrata with the aid

of a stereomicroscope, and ascomata (with or without their asexual morphs) were

mounted in water, lactic acid, or lactophenol with cotton blue for identification

under light microscopy.

Species were identified according to Ames (1961), Brummelen (1967),

Kimbrough & al. (1972), von Arx & al. (1986), Bell (1983, 2005), Doveri (2004),

Lundqvist (1972), and Richardson & Watling (1997). Careful review of the literature

and national database searches were conducted to access information regarding

previous records of coprophilous ascomycetes in Brazil. All voucher material was

deposited at the Herbarium Padre Camille Torrend, Departamento de Micologia,

Universidade Federal de Pernambuco, Recife, Brazil (URM). Additional information

regarding all records and deposited specimens, along with high quality micrographs,

Coprophils new for Brazil ... 337

are available at the INCT - Herbario Virtual da Flora e dos Fungos database website

(http://inct.florabrasil.net).

Taxonomy

Dothideomycetes, Pleosporales, Delitschiaceae

Delitschia vulgaris Griffiths, Mem. Torrey Bot. Club 11: 104 (1901) FIG. 1A-C

PsEUDOTHECIA immersed, scattered, obpyriform, dark brown to black,

1275-1450 x 350-460 um, with body adorned with light brown to golden,

thin, flexuous, simple or branched hyphoid hairs with a slightly thickened wall,

septate, about 1.5-2 um diam., <150 um long; NEcK long, cylindrical, black,

with papilliform rounded apex, 690-725 x 165-210 um, tomentose; Asci

8-spored, cylindrical, fissitunicate, uniseriate, 275-300 x 20-25.5 um, with

rounded apices and short stipe; AscosporEs 2-celled, with a medial septum,

oblong-ellipsoid to fusoid, weakly constricted, with slightly narrowed ends,

light brown when young, becoming dark brown at maturity, smooth, 32.5-37.5

x 13-15 um; germ slits longitudinal, spanning the entire length of each cell.

SPECIMENS EXAMINED—BRAZIL. PERNAMBCUO: Recife, Universidade Federal Rural de

Pernambuco (UFRPE), on horse dung, 13.11.2012, R-ER. Melo s.n. (URM86772a, b, c).

DIsTRIBUTION—Europe (Sweden), North America (Canada, USA), Oceania

(Australia). This is the first record for the Neotropics.

ComMENtTs—The Delitschia vulgaris material from Brazil presents larger

ascospores compared to the material cited by Luck-Allen & Cain (1975).

Considering Delitschia species with tomentose pseudothecia and ascospores

with a transverse septum, D. araneosa Cain is similar to D. vulgaris, but differs

by its slightly narrower ascospores (10-11 um; Luck-Allen & Cain 1975).

Leotiomycetes, Thelebolales, Thelebolaceae

Coprotus albidus (Boud.) Kimbr., Amer. J. Bot. 54: 22 (1967) FIG. 1D, E

APOTHECIA scattered, globose to lenticular, subhyaline to creamy-yellow,

translucent, 300-600 um diam., sessile, glabrous; MARGIN rounded, entire and

smooth; Disk concolorous with the receptacle, scurfy to slightly villose due

to the ascal protrusions; MEDULLARY EXCIPULUM with intricate cells (textura

intricata); ECTAL EXCIPULUM composed of elongated cells, 7.5-10 um diam.;

PARAPHYSES cylindrical, becoming clavate towards the apex, enlarged to

5-6.5 tum wide, unbranched, septate, without visible refractive globules or

pigments; Asci 32-spored, clavate, with ascospores irregularly disposed,

80-110 x 22.5-30 um, with a rounded apex; Ascospores 1-celled, ellipsoid

338 ... Melo & al.

to elongated-ellipsoid, with tapered ends, hyaline, smooth, (7.5-)10-12.5 x

5-7.5(-10) um, mostly with a de Bary bubble.

SPECIMEN EXAMINED—BRAZIL. R10 GRANDE Dv SOL: Santana do Livramento, Area

de Protegao Ambiental do Ibirapuita, on horse dung, 13.VI.2015, Xavier de Lima s.n.

(URM92164).

DiIsTRIBUTION—Europe (Denmark, England, Finland, France). This is the first

record for the Neotropics.

ComMMENTS—The material described here matches closely C. albidus as

described by Kimbrough & al. (1972). Among Coprotus species with whitish to

translucent apothecia bearing 32-spored asci and growing on dung, C. albidus

can be confused with C. rhyparobioides (Heimerl) Kimbr., which presents larger

asci (120-175 x 50-75 um) and longer ascospores (13.5-17.5 x 7-8 um), as

well as paraphyses frequently branched toward apices (Kimbrough & al. 1972).

Thelebolus microsporus(Berk. & Broome) Kimbr.,

Ann. Rep, Inst. Ferment. Res. Osaka: 50 (1967) FIG. 15, G

APOTHECIA scattered to gregarious, with cleistohymenial development

pattern, initially globose to obpyriform, becoming cupulate to lenticular,

exposing the disk later in the telohymenial phase, white to pale yellow,

becoming yellowish to pale luteous, 95-250 um diam., sessile, glabrous;

MARGIN rounded, entire and smooth; pisk concolorous with the receptacle,

almost smooth; HYMENIUM ~45-100 um thick; MEDULLARY EXCIPULUM and

hypothecium undifferentiated; ectal excipulum composed of intertwined thick

hyphae (textura epidermoidea), 7.5-12.5 um diam.; PARAPHYSES cylindrical,

branched, septate, 2-3 um wide at the base, without visible refractive globules

or pigments, branched close to its base and conspicuously broadened towards

apices, enlarged up to 6 um diam.; Asci 8-spored, cylindric-clavate, irregularly

biseriate or uniseriate, 80-95 x 10-25 um, with a short stipe, rounded above;

Ascosporss 1-celled, ellipsoid, with rounded ends, hyaline, smooth, 7.5-10

x 3-5 um, occasionally with a de Bary bubble close to one end (absent when

mounted in water).

SPECIMENS EXAMINED—BRAZIL. PERNAMBUCO: Caruaru, Instituto Agrondmico de

Pernambuco (IPA), on cattle dung, 16.X.2011, R-ER. Melo s.n. (URM86783); on horse

dung, 13.V1I.2012, R.ER. Melo s.n. (URM86784); Serra Talhada, Instituto Agronémico

de Pernambuco (IPA), on goat dung, 21.IX.2011, R.ER. Melo s.n. (URM86785).

DISTRIBUTION—Antarctica, Europe (Austria, Denmark, England, Estonia,

Faroe Islands, France, Germany, Ireland, Netherlands, Norway, Sweden,

Switzerland, Ukraine), North America (Canada USA), Oceania (Australia,

New Zealand), South America (Argentina). This is the first record for Brazil.

Coprophils new for Brazil ... 339

COMMENTS—Some measurements of the Brazilian material of Thelebolus

microsporus, such as ascospore length and width, showed slight variation

compared with material cultivated by Hoog & al. (2005) from Antarctica.

This species differs from other 8-spored members of Thelebolus mainly by its

developmental pattern: the apothecia expose their asci late in the mesohymenial

or telohymenial phase, resulting in a typical apothecioid morphology, in

contrast with species such as T: globosus Brumm. & de Hoog, which forms

globose apothecia.

Pezizomycetes, Pezizales, Ascobolaceae

Ascobolus castaneus Teng, Sinensia 11: 109 (1940) FIG. 1H-J

APOTHECIA usually gregarious, with eugymnohymenial development

pattern, initially globose to subglobose, becoming scutellate to discoid when

mature, at first yellowish-brown, later chestnut brown, drying rust brown to

finally blackish, 0.8-2 mm diam., sessile, almost glabrous; margin rounded,

smooth, bending outwards with age; Disk at first concave and yellow in

maturation, finally convex, concolorous with the receptacle, smooth to slightly

villose, not furfuraceous; HYPOTHECIUM 30-47.5 um thick, mostly with

strongly compact, isodiametric cells (textura angularis); ECTAL EXCIPULUM

composed of irregular ellipsoid cells, becoming more globose towards the

margins (textura globosa near the margin, textura angularis near the base),

30-50 um wide; PARAPHYSES cylindrical, branched, septate, 2-3 um diam. at

the base, with brownish intercellular pigment, slightly to not enlarged at the

apex; AscI 8-spored, cylindric-clavate, irregularly biseriate or uniseriate, 120-

175 x 15-17.5 um, with a short stipe, rounded above; Ascosporgs 1-celled,

ellipsoid, with rounded ends, at first hyaline, then brown to reddish brown,

smooth, 21.5-26.5 x 11.5-12.5 um.

SPECIMEN EXAMINED—BRAZIL. PERNAMBUCO: Recife, Universidade Federal de

Pernambuco, on horse dung, 06.V1.2017, D.B.P. Monte s.n. (URM92165).

DISTRIBUTION— Asia (China). This is the first record for the Neotropics.

CoMMENTS—Ascobolus castaneus is distinguished by its chestnut brown

mature apothecia and completely smooth ascospores. It can be confused

with other common members of A. sect. Gymnascobolus Brumm., such as A.

scatigenus (Berk. & M.A. Curtis) Brumm., when young—both present similar

apothecia up to the mesohymenial phase but can be immediately distinguished

when mature. Ascobolus scatigenus has relatively large receptacles, white to

pale luteous, furfuraceous disks composed of protruding asci with ellipsoid

340 ... Melo & al.

ascospores (22-24.5 x 10-15 um according Melo & al. 2014), usually with a

single to few oblique epispore fissures. The material was collected in situ on

horse dung and not observed in moist chamber incubation like the others

mentioned in this work. The ascospores are smaller than those in the Chinese

material described by Van Brummelen (1972; 15-19.5 x 7.5-9 um).

Sordariomycetes, Coniochaetales, Coniochaetaceae

Coniochaeta philocoproides (Griffiths) Cain,

Univ. Toronto Stud., Biol. Ser. 38: 65 (1934) FIG. 1K-O

PERITHECIA Scattered to gregarious, partially immersed, subglobose to

obpyriform, brown to dark brown, 325-410 x 310-390 um; NECK short,

papilliform, occasionally indistinct; PER1IDrIUM membranaceous, opaque,

dark brown, becoming dark brown to finally black towards the neck; Hairs

dark brown to black, rigid, usually straight, simple, thick-walled, non-septate,

becoming thinner at the apex, abundant at the neck; Asci 32-spored, cylindric

to cylindric-clavate, irregularly triseriate to tetraseriate, 112-120 x 14-20 um,

with a short stipe, rounded above; Ascosporss 1-celled, variable in length,

ellipsoid to oblong in frontal view, conical in lateral view, light brown to golden

when young, becoming dark brown at maturity, smooth, 7.5-12(-22) x 57.5 um,

with a lateral straight germ slit through the entire length of the spore, without

conspicuous gelatinous sheath; CoNrDIoPpHORES hyaline, simple or branched,

septate; CONIDIOGENOUS CELLS enteroblastic, hyaline, consisting of discrete

phialides, ampulliform to lageniform, 10-15(-22.5) x 2.5-3 um, with

cylindrical collarettes, 1-2 um long; Conip1a not observed.

SPECIMEN EXAMINED—BRAZIL. PERNAMBUCO: Caruaru, Instituto Agrondmico de

Pernambuco (IPA), on horse dung, 28.11.2013, R-ER. Melo s.n. (URM86634).

DISTRIBUTION—Europe (Norway), North America (USA), and Oceania (New

Zealand). This is the first record for the Neotropics.

ComMENTS— The specimen from Brazil matches closely the material described

by Asgari & al. (2007). Coniochaeta philocoproides, a rare species reported on

herbivore dung, is one of the few documented multispored species in this genus.

Asci of C. polymegasperma M.J. Richardson are 64-spored, and its ascospores

are longer (13-16.5 um) and larger (9.5-13.5 um) than C. philocoproides.

Figure 1. Brazilian dung fungi: Delitschia vulgaris. A. pseudothecium on dung; B. ascus; C. mature

ascospore, with two visible germ slits, once at each cell. Coprotus albidus. D. apothecia on dung;

E. mounted apothecium, with asci in different developmental stages. Thelebolus microsporus.

E apothecia on dung; G. ascospores. Ascobolus castaneus. H. apothecia on dung; I. hymenium,

Coprophils new for Brazil... 341

with asci in different developmental stages; J. ascospores. Coniochaeta philocoproides.

K. perithecium on dung; L, M, N. ascospores from the same ascus, considerably different in length;

O. phialide. Arnium hirtum. P. perithecia on dung; Q. perithecial hairs; R. mature ascospores.

Scale bars: A = 750 um; B, E = 20 um; C, J = 10 um; D = 500 um; F = 100 um; G = 7.5 um; H =

mm; I = 25 um; K = 200 um; L-O = 5 um; P = 400 um; Q = 2.5 um; R= 15 um.

342 ... Melo & al.

Coniochaeta hansenii (Oudem.) Cain is reported with 64-128-spored asci and

slightly different ascospore measurements (6-9 x 5-9 um) (Asgari & al. 2007).

Sordariomycetes, Sordariales, Lasiosphaeriaceae

Arnium hirtum (E.C. Hansen) N. Lundgq. & J.C. Krug,

Symb. Bot. Upsal. 20(1):218 (1972) FIG. 1P—-R

PERITHECIA scattered to isolated, semi-immersed to superficial, obpyriform,

690-930 x 480-540 um, reddish brown to dark brown, translucent, covered

by hyphoid, slender, flexuous hairs, septate, 2-2.5 um thick; NECK cylindrical

to conical, opaque, 150-165 x 130-140 um, tomentose, neck hairs more

rigid and setose, <7 um long, becoming thinner towards the apex; PERIDIUM

membranaceous, subopaque to olivaceous brown, darkening towards the neck

to finally black, 3-layered—outer peridial layer composed of large, inflated,

angular cells, thick-walled, 5-10 um diam., forming a short palisade that

resembles inflated perithecial hairs; PARAPHYSES interspersed with the asci,

ventricose, persistent or collapsing; Asc1 8-spored, cylindric to cylindric-

clavate, 350-400 x 25.5-35.5 um, usually persistent, with rounded apex and

a long stipe; Ascosporgs at first fusoid to ellipsoid, hyaline, later becoming

ochraceous to olivaceous brown, with several oil droplets, and finally dark

brown, bicaudate; 35-47.5 x 18.5-22.5 um, with an eccentric germ pore; APICAL

CAuDA single, 7.5-9 um diam., becoming thinner towards its apex, up to 42.5

um long, usually persistent in mountings, longitudinally striped, covering the

germ pore; BASAL CAUDA similar in morphology, slightly thinner and longer,

5-6 um wide at the base when not inflated, up to 50 um long, persistent or

collapsing when mounted.

SPECIMENS EXAMINED— BRAZIL. PERNAMBUCO: Serra Talhada, Instituto Agronémico

de Pernambuco (IPA), on goat dung, 24.X.2011, R-ER. Melo s.n. (URM86719); ,

Universidade Federal Rural de Pernambuco (UFRPE), on horse dung, 23.11.2012, R-ER.

Melo s.n. (URM86720).

DiIsTRIBUTION—Africa (Morocco), Europe (Belgium, England, Denmark,

France, Germany, Ireland, Norway, Scotland, Sweden) and North America

(USA). This is the first record for the Neotropics.

COMMENTS—Arnium species are distinguished from other dark-coloured

perithecial dung ascomycetes by their unicellular ascospores with gelatinous

caudae at both ends. Arnium hirtum presents a wide array of morphological

variation, having as its main characteristics the substrate (herbivore dung) and

the ascospore morphology: equilateral, non-septate and bicaudate (35-47.5

x 18.5-22.5 um). The present material fits well with the description of the

Coprophils new for Brazil ... 343

European specimens (Lundqvist 1972), with similar measurements and general

morphology. It is a common species in Europe, but rare outside that continent,

although more surveys will probably reveal new records for tropical areas.

Cercophora anisura N. Lundq., Symb. Bot. Upsal. 20(1): 91 (1972) FIG. 24, B

PERITHECIA isolated, semi-immersed, obpyriform, subglobose or conical,

480-515 x 350-400 um, light brown to golden, translucent, covered by hyphoid,

flexuous, occasionally branched hairs, about 2 um thick; NECK cylindrical to

conical, opaque, carbonaceous, dark brown to finally black close to the apex,

100-110 x 80-100 um, adorned with tufts of short, inflated and agglutinated

hairs, 2-2.5 um thick, septate, concolorous with the peridium in the lower

part of the perithecium; PERIDIuM pseudo-parenchymatous, membranaceous,

subopaque, darkening towards the neck to finally black, 3-layered—outer

peridial layer composed of angular to rounded cells, thin-walled, 4-9 um

diam.; PARAPHYSES interspersed with the asci, ventricose, persistent or

collapsing; Asci 8-spored, cylindric-clavate, 175-225 x 17.5-25 um, with a

long stipe and tapered apex, thickened, simple apical ring present; subapical

globule subglobose to ellipsoid, smooth to slightly verrucose, 4-5 um diam.;

Ascosportss at first cylindrical vermiform to slightly sigmoid, hyaline, with

several oil droplets, occasionally geniculate at the base, non-septate, smooth,

bicaudate, later becoming swollen in the upper part and finally 2-celled after

the formation of a transverse septum, delimiting the region of the spore that

will become the pigmented head cell above, and another part for the hyaline

pedicel below, irregularly biseriate to triseriate; HEAD CELL ellipsoid, truncated

at the base, non-septate, brown to dark brown, smooth, 15-19 x 7.5-8 um with

an eccentric germ pore; PEDICEL cylindrical, hyaline, occasionally geniculate

near the base, usually collapsing, 30-35 x 4-5 tum; APICAL CAUDA single,

cylindrical to conical, tapering towards the apex, 10-20 um long, 2-2.5 um

diam. at the base, usually collapsing; BASAL CAUDA similar in morphology,

thicker and longer, 20-28.5 x 3-3.5, usually collapsing.

SPECIMENS EXAMINED—BRAZIL. PERNAMBUCO: Recife, Universidade Federal Rural de

Pernambuco (UFRPE), on horse dung, 11.XIJ.2012, R.ER. Melo s.n. (URM86721a, b, c).

DISTRIBUTION—Europe (Sweden). This is the first record for the Neotropics.

ComMENTS— The Brazilian material was sparse and little information regarding

its morphological variation is presented here. Cercophora anisura was proposed

by Lundqvist (1972) in his monographic treatment of the Nordic Sordariaceae

from material on cattle dung in Sweden. The species can be confused with

C. mirabilis Fuckel, one of the most common Cercophora on herbivore dung,

344 ... Melo &al.

which differs by its larger perithecia (650790 x 340475 um), longer inflated

hairs (2-3.5 um), and ascospores with even caudae (Lundqvist 1972). In our

material of C. anisura, mature ascospores with multiseptate pedicels, ascospores

with both cells pigmented, and ascospores without the transverse septum were

commonly observed.

Cercophora sordarioides (Speg.) N. Lundg.,

Symb. Bot. Upsal. 20(1): 112 (1972) FIG. 2C-E

PERITHECIA scattered to isolated, semi-immersed to superficial, obpyriform,

subglobose or conical, 650-850 x 550-600 um, olivaceous to golden, translucent,

glabrous; NECK conical to slightly cylindrical, opaque, carbonaceous, dark

brown to finally black close to the apex, 200-350 x 190-200 um; PERIDIUM

pseudo-parenchymatous, membranaceous, subopaque, darkening towards the

neck to finally black, 3-layered—outer peridial layer with angular to rounded

cells, thin-walled, 7-10(-12.5) um diam.; PARAPHYSES interspersed with the

asci, filiform, persistent or collapsing; Asc 8-spored, clavate, 155.5-185 x

17.5-25 um, with a long stipe and tapered apex, thickened, simple apical ring

present; subapical globule subglobose to globose, smooth to slightly verrucose,

4-5.5 um diam.; Ascosporgs at first cylindrical to sigmoid, hyaline, with

several oil droplets, occasionally geniculate at the base, non-septate, smooth,

bicaudate, later becoming swollen in the upper part and finally 2-celled after

the formation of a transverse septum, delimiting the region of the spore that

will become the pigmented head cell above, and another part for the hyaline

pedicel below, irregularly biseriate to triseriate; HEAD CELL ellipsoid, truncated

at the base, usually with an eccentric transverse septum, brown to dark

brown, smooth, 17.5-22.5 x 7.5-11 um with a apical or subapical germ pore;

PEDICEL cylindrical, hyaline, occasionally geniculate near the base, persistent

or collapsing, 45.5-49 x 29.5-31 um; APICAL CAUDA single, cylindrical,

tapering towards the apex, 27.5-55 um long, 2.5-3 um wide at the base, usually

collapsing; BASAL CAUDA similar in morphology.

SPECIMEN EXAMINED—BRAZIL. PERNAMBUCO: Caruaru, Instituto Agrondmico de

Pernambuco (IPA), Caruaru, on horse dung, 23.X1.2012, R-ER. Melo. s.n. (URM86728).

DISTRIBUTION—Europe (Sweden). This is the first record for the Neotropics.

COMMENTS—Cercophora sordarioides is regarded as a rare species. It was

reported from Argentina by Spegazzini. The key characteristics are the

glabrous, olivaceous to dark-yellow perithecia with thin-walled peridial

cells and ascospores with septate head cells. The Brazilian material presents

longer pedicels in relation to Argentine material (37-38 um), as described in

Lundqvist (1972).

Coprophils new for Brazil ... 345

Sordariomycetes, Sordariales, Chaetomiaceae

Chaetomium citrinum Udagawa & T. Muroi,

Trans. Mycol. Soc. Japan 22: 15 (1981) FIG. 2F-H

PERITHECIA scattered to isolated, superficial, subglobose to obovoid,

yellowish to greenish in mass, dark brown when isolated, 250-300 x 240-280 um,

attached to the substrate by weakly pigmented rhizoids, light brown to brown,

thin-walled, dichotomously branched, non-septate, flexuous, usually smooth,

2.5-2.8 um diam.; TERMINAL HAIRS of one type — numerous, septate, wavy

to loosely coiled, forming distant and weakly organized coils along its length,

making 2-3 convolutions, 2.4-3 um diam., with almost constant diameter in

each turn towards the apex, light brown to golden, becoming less pigmented

close to the apex, extending up to 400(-450) um, markedly punctate in all its

extension, clustered above to form a strongly compact sporiferous head above

the perithecium, ellipsoid to columnar; LATERAL Hairs slightly similar in

morphology — numerous, long, filiform, tortuous, smooth, sparsely septate,

thin-walled, light brown to golden, slightly less pigmented than the terminal

hairs, 1.9-2.4 um, extending up to 350 um; PERIDIUM pseudo-parenchymatous,

membranaceous, fragile, with flattened, angular to elongate, slightly thick-

walled cells; Asci 8-spored, clavate, spore-bearing part 9-12.5 um diam., with a

short stipe, evanescent; ASCosPorREs variable in shape, ellipsoid, long-ellipsoid,

ovoid, fusoid to limoniform, biapiculate or not, dark brown when mature,

smooth or with superficial cracklings, 7.4-10.6 x 4.8-6.5 um.

SPECIMENS EXAMINED—BRAZIL. PERNAMBUCO: Caruaru, Instituto de Pesquisas

Agronémicas, on goat dung: 27.11.2012, R.ER. Melo s.n. (URM86708), 25.V1.2012, R.ER.

Melo s.n. (URM86709a, 86709b), Recife, Universidade Federal Rural de Pernambuco

(UFRPE), on goat dung, 16.1V.2012, R-ER. Melo s.n. (URM86710).

DISTRIBUTION— Asia (Japan), North America (Canada). This is the first record

for the Neotropics.

COMMENTS—Chaetomium citrinum presents a striking diagnostic feature of

greenish-yellow perithecia, especially when occurring in groups. Ascospore

shapes are quite variable in this species, even irregular or with protuberances. It

is a rarely encountered species of Chaetomium. The present material has larger

perithecia than the Japanese strain (160-250 um), as described by von Arx &

al. (1986).

Chaetomium cochliodes Palliser, N.Amer. Fl. 3(1): 61 (1910) FIG. 21-K

PERITHECIA scattered to isolated, superficial, subglobose to doliform,

grayish in stereomicroscopy, dark brown to black when mounted, 310-340

346 ... Melo & al.

x 280-295 um, attached to the substrate by hyaline, thin-walled rhizoids,

1.5-2.5 um diam.; TERMINAL HAIRS of two types — (1) initially straight,

from the proximal end until up to half of its length, becoming loosely spaced

and coiled toward the apex, making 3-4 convolutions and progressively

smaller in diameter (“corkscrew” type), brown to golden, becoming less

pigmented close to the apex, extending up to 300 um above the sporiferous

head, 5-7.5 um diam., smooth to roughened, without conspicuous septa,

loosely clustered to form a weakly compact sporiferous head above the

ascoma; (2) wavy, thinner, flexuous, septate, not forming coils, projecting

above the sporiferous head, usually intertwined with the coiled hairs,

markedly roughened, olivaceous brown to pale brown, up to 3 um diam.;

LATERAL HAIRS numerous, long, filiform, tortuous, smooth, septate, thin-

walled, light brown to golden, slightly less pigmented than the terminal hairs,

3.5-5.5 um, extending <200 um, slightly bulbous at the base; PERrpIUM

pseudo-parenchymatous, membranaceous, fragile, with flattened, angular

to slightly elongate, slightly thick-walled cells, 7.5-15 um diam.; Asc1

8-spored, clavate, spore-bearing part 30-40 x 15-22.5 um, with a short stipe,

evanescent; AscosporEs limoniform, biapiculate, light brown to olivaceous

brown when mature, smooth, 8.5-10 x 7-8 um.

SPECIMENS EXAMINED—BRAZIL. PERNAMBUCO: Serra Talhada, Instituto Agronémico

de Pernambuco (IPA), on goat dung, 16.V.2012, R-ER. Melo s.n. (URM8671 1a, b, c).

DIsTRIBUTION—Africa (South Africa), Asia (India, Japan, Nepal),

Europe (Belgium, England, Netherlands, Norway), North America

(Canada, USA), and South America (Argentina). This is the first record

for Brazil.

COMMENTS—Chaetomium cochliodes is a common species in indoor

air samples, occasionally reported in herbivore dung. The material

examined in Brazil was consistent with the protologue description:

terminal hairs of two types: one long, simple, becoming coiled above the

sporiferous head, and the other type septate, wavy, usually rough-walled.

It can be confused with C. globosum Kunze, a common species with wide

circumscription, but differing by the thinner terminal hairs (3.5-4.5 um)

and larger ascospores (9-12 x 7.5-8 um) (Ames 1961).

FiGuRE 2. Brazilian dung fungi: Cercophora anisura. A. perithecium on dung; B. ascospore.

Cercophora sordarioides. C. perithecia on dung; D. ascal apical globule; E. mature ascospore.

Chaetomium citrinum. F. perithecia on dung; G. mounted perithecium; H. ascospores.

Chaetomium cochliodes. 1. mounted perithecium; J. perithecial rhizoids; K. ascospores.

Coprophils new for Brazil ... 347

Chaetomium spirale. L. perithecium on dung; M. coiled terminal hair; N. ascospores. Sordaria

lappae. O. perithecia on dung; P. mounted perithecium; Q. ascospore. Scale bars: A, F = 250 um;

B= 7.5 um; C = 750 um; D, K, Q=5 m; E, M = 15 um; G = 100 um; H, J, N = 10 um; I = 150 um;

L, P = 50 um; O = 300 um.

348 ... Melo & al.

Chaetomium spirale Zopf,Nova Acta Acad. Caes. Leop.-Carol.

German. Nat. Cur. 42(5): 275 (1881) FIG. 2L-N

PERITHECIA Scattered to isolated, superficial, globose to ovoid, grayish

in stereomicroscopy, dark brown when mounted, 150-175 x 150-165 um,

attached to the substrate by weakly pigmented, light brown to brown, thin-

walled rhizoids, 2.5-3 um diam.; TERMINAL HAIRS of one type—initially

curved from the proximal end until up to half of its length, becoming

strongly coiled toward the apex, making 7-12 (usually nine) convolutions,

<30 um diam., progressively smaller in diameter towards the apex

(“corkscrew” type), olivaceous brown to golden, usually not becoming less

pigmented close to the apex, extending up to 350 um above the sporiferous

head, 2.5—-5 um diam.., finely punctate to verrucose, loosely clustered to form

a weakly compact sporiferous head above the perithecium; LATERAL HAIRS

scarce, filiform, tortuous, smooth to slightly punctate, usually setose, light

brown to golden, slightly less pigmented than the terminal hairs, 3.5-4 um,

extending up to 150 um, slightly bulbous at the base; PERIDIUM pseudo-

parenchymatous, membranaceous, fragile, with flattened, angular, slightly

thick-walled cells, 4.5-12 um diam.; Asci 8-spored, clavate, spore-bearing

part 32.5-37 x 7.5-12.5 um, with a short stipe, evanescent; ASCOSPORES

subglobose to limoniform, weakly biapiculate, light brown to olivaceous

brown when mature, smooth, 8.5-9 x 5.5—7.5 um.

SPECIMEN EXAMINED—BRAZIL. PERNAMBUCO: Caruarru, Instituto Agronoémico

de Pernambuco (IPA), Caruaru, on goat dung, 7.X.2011, R.ER. Melo s.n.

(URM867 16).

DISTRIBUTION—Africa (Egypt, Iraq, Kenya, Kuwait, Libya, Swaziland,

Sudan, Tanzania, Zambia), Asia (India, Japan, Malaysia), Europe (Czech

Republic, England), North America (USA) and Oceania (Australia, New

Zealand, Papua New Guinea). Probably worldwide. This is the first record

for the Neotropics.

CoMMENTS—Among similar Chaetomium species that present coiled

“corkscrew” terminal hairs with convolutions progressively decreasing in

diameter toward the apex, C. spirale differs from C. convolutum Chivers by

its smaller, more globose perithecia and by the terminal hairs with more

convolutions (7-12). It also differs from C. cochliodes by lacking a second

type of terminal hair besides the coiled ones, and from C. aterrimum Ellis

& Everh. ex Palliser in which the terminal hairs are uniform in diameter.

The material presented here fits well the description provided by Ames

(1961).

Coprophils new for Brazil ... 349

Sordariomycetes, Sordariales, Sordariaceae

Sordaria lappae Potebnia, Ann. Mycol. 5: 13 (1907) FIG. 20-Q

PERITHECIA usually scattered, rarely isolated, semi-immersed to

superficial, obpyriform to slightly obovoid, brown to golden, 475-550 x

300-315 um, translucent, glabrous; neck cylindrical to papilliform, opaque,

carbonaceous, dark brown to finally black close to the apex, 150-170

x 140-150 um; PERIDIUM pseudo-parenchymatous, membranaceous,

subopaque, darkening towards the neck to finally black—outer peridial

layer with angular cells, thick-walled, 7.5-17.5 um long, occasionally

inflated; PARAPHYSES not observed; Ascr 8-spored, cylindrical, 160-190

x 16-17.5 um, with a short stipe and tapered apex; AscosporeEs obovoid

to subglobose, light brown to golden when young, dark brown when

mature, smooth, 18-22.5 x 15-17.5 um, guttulate, with a basal germ pore,

surrounded by a thin, hyaline gelatinous sheath except at the germ pore.

SPECIMENS EXAMINED—BRAZIL. PERNAMBUCO: Recife, Universidade Federal

Rural de Pernambuco (UFRPE), on horse dung, 04.IV.2012, R.ER. Melo s.n.

(URM86770a, b).

DIsTRIBUTION—Africa (Kenya, Tanzania, Uganda, Zaire), Europe (Czech

Republic, England, Finland, France, Italy, Russia, Sweden), North America

(Canada, USA) and South America (Argentina, Colombia). Possibly

worldwide. This is the first record for Brazil.

CoMMENTS—‘Sordaria lappae, although recorded in many countries, is of

difficult determination. Sordaria fimicola (Roberge ex Desm.) Ces. & De

Not. differs from S. lappae by its less brightly coloured peridium and less

inflated cells and by its smaller and less apically rounded ascospores (15-

22.5 x 10-12.5 um) (Lundqvist 1972). The material from Brazil presents

slightly wider ascospores when compared to the material described by

Lundqvist (13-15 um; 1972).

Discussion

Recent surveys and inventories examining understudied substrates in

tropical areas has led to novel information on the geographical distribution

of fungal species. Coprophilous fungi as an ecological group are considered

to occur worldwide, but their occurrence is closely tied to herbivore

occurrence, so the factors that determine their geographical range are still

subject to discussion. Considering that the discovery of new fungal records

is quite frequent, this group may serve as an indicator of the advances

in taxonomical and ecological knowledge in a region. In the present

350 ... Melo & al.

study, representatives of three major ascomycete groups were recorded.

Sordariomycetes represents a large clade, including non-lichenized

ascomycetes with perithecial ascomata (Zhang & al. 2006). Arnium hirtum

is known from Africa, Europe, and North America. The new record for

South America suggests that this species may have a worldwide distribution.

Members of Chaetomium, recently studied by Wang & al. (2016), were

reported for the first time in South America. Although not specialized

on dung, these fungi can be common in cellulose-rich substrates, and

incubation in moist chambers could subvert limiting factors that would

otherwise prevent its sampling and identification. Leotiomycetes contains

mostly forms formerly known as inoperculate discomycetes: apothecial

ascomycetes with asci with a non-operculum apical apparatus (Wang & al.

2006). Thelebolales is the main group of this class that occurs on herbivore

dung. The discovery of Coprotus albidus updates the Coprotus species from

Brazil recently studied by Melo & al. (2015a). Among the Pezizomycetes,

usually regarded as “operculate discomycetes,’ Ascobolaceae species are the

most well known on herbivore dung. The Ascobolus castaneus record is the

most significant for the group, considering that A. castaneus was previously

recorded only from China. Similarly, Syncephalis Tiegh. & G. Le Monn., a

zygosporic mycoparasite of coprophilous ascomycetes previously recorded

only in Taiwan, produced in Brazil two second worldwide records: S. clavata

(Melo & al. 2011) and S. obliqua (Melo & al. 2015b). The coprophilous

mycobiota of Brazil is obviously rich and diverse, and further studies will

increase our understanding of their patterns of occurrence, distribution,

and substrate not only in South America but on other continents as well.

Acknowledgments

The authors thank Laise de Holanda Cavalcanti Andrade (Universidade Federal

de Pernambuco, Brazil) and Nadja Santos Vitéria (Universidade do Estado da Bahia,

Brazil) for helpful suggestions and pre-submission review of the manuscript. The

authors would also like to thank the Coordenacao de Aperfeigoamento de Pessoal de

Nivel Superior (CAPES) and the Conselho Nacional de Desenvolvimento Cientifico

e Tecnolégico (CNPq-Ciéncia sem Fronteiras; INCT-Herbario Virtual da Flora e

dos Fungos). L.C. Maia acknowledges the research fellowship and grants provided

by CNPq (INCT-HVFFE, Protax, Sisbiota).

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MY COTAXON

April-June 2019—Volume 134, pp. 353-357

https://doi.org/10.5248/134.353

Two Heterostelium species newly recorded from China

Pu Liu, SHUNHANG ZHANG, XIAOYAN ZHOU, JIANJUN ZHAO, Yu LI*

Engineering Research Center of Chinese Ministry of Education for Edible and

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

* CORRESPONDENCE TO: fungi966@126.com

ABSTRACT—Two species of dictyostelid cellular slime molds new to China (Heterostelium

anisocaule and H. arachnoideum) were isolated from samples of soil/humus collected in East

China. Descriptions and illustrations based on these isolates are provided.

Key worps—Amoebozoa, taxonomy, forest soils

Introduction

Dictyostelid cellular slime molds (dictyostelids) are key components of the

soil microbiota and play a role in maintaining the balance that exists between

bacteria and other organisms (Raper 1984). Dictyostelids have been reported

from several localities in China (He & Li 2010; Liu & Li 2012a,b, 2014, 2017;

Liu & al. 2019), but there are relatively few reports of this group from East

China (Yeh & Chien 1983, Hagiwara & al. 1985, Zhao & al. 2017). East China

includes Shandong Province, Jiangsu Province, Anhui Province, Zhejiang

Province, Jiangxi Province, Fujian Province, Shanghai, and Taiwan. Two species

of dictyostelids, reported originally from the Southern Hemisphere, were

isolated from samples of soil/humus collected from this region. Descriptions

and illustrations based on these isolates are provided herein.

Materials & methods

Samples were collected during 2012 from Jiangsu Province, Fujian Province, and

Taiwan. Each sample consisted of 30-50 g of soil/humus and was placed in a sterile

whirl-pack plastic bag. Each sample bag was numbered and the sample itself preserved

354 ... Liu & al.

at 4 °C in the herbarium of the Mycological Institute of Jilin Agricultural University,

Changchun, China (HMJAU). Isolation methods followed Cavender & Raper (1965),

with some minor modifications outlined by Liu & al. (2019). Isolates were identified

with reference to Raper (1984) and following the new classification system of Sheikh

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

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

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

sorocarps were prepared with water as the mounting medium. Features of spores,

sorophores, and sorocarps were observed and measured using a Zeiss Axio Imager A2

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

were taken with Zeiss Axiocam 506 color microscope camera. Spores from these plates

were frozen in HL 5 media (Cocucci and Sussman 1970) and stored at — 80 °C in

HMJAU.

Taxonomy

Heterostelium anisocaule (Cavender &al.) S. Baldauf, S. Sheikh & Thulin,

Protist 169: 14. 2018. PLATE 1a-e

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

sorocarps white, erect to prone, solitary to clustered, prostrate, phototropic,

normally 3-9 mm. Sorophores asymmetrical, sinuous, with 1-7 whorls, each

whorl comprising 1-4 branches. Sorophore tips and the upper portion of the

sorophore consist of one tier of cells, bases round. Terminal sori white, globose,

commonly 60-160 um in diam. Lateral sori white, globose, 15-85 um in diam.

Spores elliptical 5-9.5x3-5.5 um, with unconsolidated granules. Aggregations

of the “violaceum” type.

SPECIMEN EXAMINED— CHINA, JIANGSU PROVINCE, Jintan, Mao Mountains, soil

(S2593) collected in broadleaf forest, 19 Aug. 2012, Liu & al.; isolated in 2014 (HMJAU

MRI174).

ComMMENTS—Heterostelium anisocaule was first isolated from a sample

collected in a podocarp/broadleaf forest in Northland, New Zealand. This

species has a wide range of temperature adaptation (18-30 °C). Variations in

temperature and light exposure result in a considerable variation in sorocarp

size. The optimum temperature is 22-23 °C (Cavender & al. 2002).

Heterostelium arachnoideum (Vadell & Cavender) S. Baldauf, S. Sheikh &

Thulin, Protist 169: 14. 2018. PLATE 1f-n

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

white, erect to prone, sometimes prostrate, solitary to clustered, with slightly

phototropic growth, 5-14 mm or more. Sorophores asymmetrical, with 3-20

whorls, each whorl with 2-5 uneven branches. The lower portion of the

Two Heterostelium spp. new for China... 355

PLATE 1. Heterostelium anisocaule: a, b. Sorocarps; c. Aggregation; d. Sorophores and branches;

e. Spores. Heterostelium arachnoideum: f, g, i. Sorocarps; h. Aggregation; j, k. Sorophore tip;

1. Sorophores and branches; m. Sorophore base; n. Spores. Scale bars: a, b, f, g = 1 mm; c,h = 100 um;

d, j, k= 20 um; e, n= 5 um; i = 0.5 mm; 1 = 50 um; m = 30 um.

sorophore sometimes with secondary whorls. Sorophore tips consist of one

tier of cells, sinuous, becoming tangled together to form a delicate spider-

web-like structure, especially sometimes without sori present. Bases clavate,

356 ... Liu & al.

irregular. Terminal sori white, globose, commonly 30-135 um in diam. Lateral

sori white, globose, commonly 20-50 um in diam. Spores elliptical or oblong,

mostly 5.5-8x3-4.5, with unconsolidated polar granules. Aggregations of the

“violaceum” type. Late sorogens sometimes curved.

SPECIMEN EXAMINED- SPECIMEN EXAMINED: CHINA, JIANGSU PROVINCE, Yixing, soil

(S2570) collected in broadleaf forest, 22 Aug. 2012, Liu & al.; isolated in 2014 (HMJAU

MRI175). FUJIAN PROVINCE, Fuzhou, Gu Mountain, soil (S2031) collected in mixed

forest, 11 Mar. 2012, Liu & al.; isolated in 2014 (HMJAU MR209). Tatwan, Yilan,

Lotung Sports Park, soil (S2936) collected in humus, 11 Sep. 2012, Liu & al.; isolated in

2014 (HMJAU MR210).

ComMMENTS—Heterostelium arachnoideum was originally isolated from soil litter

collected from three sites within Iguazu National Park, Misiones, Argentina.

This species is characterized by its spider web-like terminal sorophores (Vadell

& Cavender 2007).

Acknowledgments

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

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

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

study was supported by the National Natural Science Foundation of China (No.

31870015, 31300016), Science and Technology Development Program of Jilin Province

(No. 20180101273JC), 111 Project (No. D17014), and the Science and Technology

Research Programs of the Education Department of Jilin Province in the Thirteenth

Five-Year Plan (No. JJKH20180671KJ).

Literature cited

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52: 294-296.

Cavender JC, Stephenson SL, Landolt JC, Vadell EM. 2002. Dictyostelid cellular slime moulds

in the forests of New Zealand. New Zealand Journal of Botany 40: 235-264.

https://doi.org/10.1080/0028825X.2002.9512786

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

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

Hagiwara H, Yeh ZY, Chien CY. 1985. Dictyostelium macrocephalum, a new dictyostelid cellular

slime mold from Taiwan. Bulletin of the National Science Museum, Series B, 11: 103-108.

He XL, Li Y. 2010. A new species of Dictyostelium from Tibet, China. Mycotaxon 111:

287-290. https://doi.org/10.5248/111.287

Liu P, Li Y. 2012a. Dictyostelids from Heilongjiang Province, China. Nova Hedwigia 94(1-2):

265-270. https://doi.org/10.1127/0029-5035/2012/0094-0265

Liu P, Li Y. 2012b. New records of dictyostelids from China. Nova Hedwigia 94(3-4): 429-436.

https://doi.org/10.1127/0029-5035/2012/0010

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

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

Two Heterostelium spp. new for China ... 357

Liu P, Li Y. 2017. Dictyostelids from Jilin Province, China. II. Phytotaxa 323(1): 077-082.

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

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

molds in high-elevation habitats on the Qinghai-Tibet Plateau, China. Scientific Reports

9:5 [13 p.]. https://doi.org/:10.1038/s41598-018-37896-7

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

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

Nanjundiah V, Queller DC, Strassmann JE, Spiegel FW, Stephenson SL, Vadell

EW, Baldauf SL. 2018. A new classification of the dictyostelids. Protist 169: 1-28.

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

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

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

https://doi.org/10.1080/15572536.2007.11832606

Yeh ZY, Chien CY. 1983. Cellular slime molds in Taiwan I: Four newly recorded species.

Biological Bulletin of National Taiwan Normal University 18: 69-86.

Zhao M, Liu P, An Y, Yao Y, Li Y. 2017. Dictyostelium annularibasimum (Dictyosteliaceae,

Dictyostelida), a new purple species from China. Nova Hedwigia 104(1-3): 351-358.

https://doi.org/info:doi/10.1127/nova_hedwigia/2016/0381

MY COTAXON

April-June 2019—Volume 134, pp. 359-367

https://doi.org/10.5248/134.359

Pseudocercospora meliosmicola sp. nov. and

three new Pseudocercospora records from China

QIAN ZHAO’, BAo-Ju Li’, YAN-XIA SHIT’,

XUE-WEN XIE’, A-LI CHAI’, YING-LAN GUO?*

"Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences,

Beijing 100081, PR. China.

? Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, PR. China

* CORRESPONDENCE TO: guoyl@im.ac.cn

ABSTRACT—A new species, Pseudocercospora meliosmicola, and three new records, P. deglupta,

P. eucalyptigena, and P. infuscans, are reported from China. Descriptions, illustrations, and

discussions of the species concerned are provided. The examined specimens are deposited

in HMAS.

Key worps—biodiversity, cercosporoid fungi, Meliosma parviflora, Mycosphaerellaceae,

taxonomic novelty

Introduction

Pseudocercospora Speg. (Spegazzini 1910), with more than 1600 names

recorded, represents the largest genus of cercosporoid hyphomycetes. The genus

comprises plant pathogens either causing distinct necrotic spots or forming

effuse fungal colonies without any definite spots, on leaves, stems, flowers, and

fruits. Some species are known to cause severe diseases on important crops,

which is also reflected in plant quarantine regulations. On the other hand,

some have been used as biological control agents of weeds (Den Breeyen &

al. 2006). More than 400 Pseudocercospora species have been reported from

China (Liu & Guo 1998). Here we propose Pseudocercospora meliosmicola on

Meliosma parviflora as a new species, and report P. deglupta, P. eucalyptigena,

and P infuscans as new records for China.

360 ... Zhao & al.

Taxonomy

Pseudocercospora deglupta Crous, Mycol. Mem. 21: 127, 1998. FIG. 1

Leaf spots amphigenous, subcircular to angular, without definite margin,

1-5 mm wide, confluent, pale gray to gray to brown, with a pale gray to

pale olivaceous brown halo on the upper surface, pale grayish brown to

pale brown on the lower surface. Fruiting hypogenous. Mycelium internal.

Stromata absent or well-developed, substomatal, subglobose, dark brown,

20-40 um diam. Conidiophores few, emerging through stomata, or in dense

fasciculate, olivaceous brown to dark olivaceous brown, paler towards

the apex, regular in width, cylindrical, unbranched or branched, smooth

<20in een Sen BES SEE VEL oe,

ok ee ae Oe

Fic. 1 Pseudocercospora deglupta (HMAS 247119).

a. Conidia; b. Conidiophores. Scale bar = 25 um.

Pseudocercospora meliosmicola sp. nov. (China) ... 361

to finely verruculose, straight to curved, obtuse to conical at the apex,

1-7-septate, 20-60(-80) x 3.2-5.4 um. Conidiogenous loci inconspicuous,

unthickened and not darkened. Conidia solitary, obclavate-cylindrical to

cylindrical, olivaceous to moderately olivaceous brown, straight to cured,

finely verruculose, obtuse at the apex, obconically truncate at the base, hila

neither thickened not darkened, 3-10-septate, 32-90 x 3-4.5 um.

SPECIMEN EXAMINED: On living leaves of Eucalyptus globulus Labill. (Myrtaceae):

CHINA, YUNNAN PROVINCE, Fenglu, 27.IX.1987, coll. Guo Y.L. no. 1141 (HMAS

247119).

Notes: Pseudocercospora basitruncata Crous on Eucalyptus sp. is similar

to P. deglupta, but differs in having amphigenous fruiting, sympodially

or percurrently proliferating conidiogenous cells, and paler (hyaline to

olivaceous) and narrower conidia (25—70(-90) x 2.5-3(-3.5) um; Crous

1998).

Pseudocercospora eucalyptigena U. Braun, Fungal Diversity 8: 42, 2001. FIG. 2

Leaf spots amphigenous, irregularly angular, without definite margin,

1-4.5 mm wide, vein-limited, often confluent, yellowish brown to brown

on the upper surface, pale brown to graying brown on the lower surface.

Fruiting hypogenous. Mycelium internal and external; hyphae pale

olivaceous to olivaceous, branched, smooth, septate, 2-3 um wide. Stromata

absent or small, substomatal, subglobose, olivaceous brown, 10-30 um diam.

Conidiophores few, emerging through stomatal, loose fasciculate or arising

singly as lateral branches from external mycelial hyphae, pale olivaceous

to pale olivaceous brown, paler towards the apex, irregular in width, not

branched, straight to curved, 0-1-geniculate, denticulate, obtuse to conical

at the apex, 0-1(-2)-septate, primary conidiophores 8-35 x 2.5-3.5 um,

secondary conidiophores (arising from superficial hyphae) 5-25 x 2.5-3.5 um.

Conidiogenous loci inconspicuous, unthickened and not darkened. Conidia

solitary, cylindrical to cylindrical-obclavate, pale olivaceous, straight to

strongly curved, acute to obtuse at the apex, subtruncate to obconically

truncate at the base, hila neither thickened nor darkened, indistinctly

3-12-septate, 35-80(-100) x 2-3 um.

SPECIMEN EXAMINED: On living leaves of Eucalyptus sp. (Myrtaceae): CHINA,

TAIWAN, Taibei, 20.VII.1926, coll. Sawada K., no. 9196 (HMAS 05196).

Notes: Pseudocercospora cubae Crous, P. deglupta Crous, and P. denticulata

Crous on Eucalyptus sp. all differ from P eucalypticola in having finely

verruculose conidiophoresand conidia. Additionally, P cubae differsin having

362 ... Zhao & al.

Fic. 2 Pseudocercospora eucalyptigena (HMAS 05196).

a. Conidia; b. Conidiophores; c. Stroma. Scale bar = 25 um.

sympodially or percurrently proliferating conidiogenous cells (Crous 1998);

P. deglupta possesses longer and wider conidiophores (60-120 x 4-6 um)

and wider conidia (3.0-4.0(-4.5 um); Crous 1998); and P denticulata has

amphigenous fruiting, larger stromata (60 um wide, 30 um high), and darker

(moderately brown), wider conidiophores (3-5 um; Crous 1998).

Pseudocercospora meliosmicola sp. nov. (China) ... 363

Pseudocercospora infuscans (Ellis & Everh.) U. Braun,

Monogr. Cercosp. Ramul. Allied Gen. 2: 402, 1998. FIG. 3

MISAPPLIED: Pseudocercospora toxicodendri sensu Liu & Guo (1989)

Leaf spots amphigenous, subcircular to angular, 1-4 mm wide, grayish

white to red-brown on the upper surface, yellowish brown to pale red-

brown on the lower surface. Fruiting hypogenous. Mycelium internal and

40 um

Fic. 3 Pseudocercospora infuscans (HMAS 60028).

a. Conidia; b. Conidiophores; c. Stroma. Scale bar = 40 um.

364 ... Zhao & al.

external, subhyaline, branched, smooth, septate, 2-3 um diam. Stromata

absent. Conidiophores 2-5, emerging through stomata or arising singly as

lateral branches from external mycelial hyphae, olivaceous to pale olivaceous

brown, uniform in color, irregular in width, branched, geniculate, denticulate,

straight to curved, rounded to conical at the apex, 1-7-septate, sometimes

constricted, 6.5-125 x 3-5.5 wm. Conidiogenous loci inconspicuous,

unthickened and not darkened. Conidia solitary, cylindrical to cylindrical-

obclavate, pale olivaceous, straight to curved, obtuse at the apex, obconically

truncate at the base, hila neither thickened not darkened, 2-11-septate,

30-120 x 3-6.5 um.

SPECIMENS EXAMINED: On living leaves of Rhus chinensis Mill. (Anacardiaceae):

CHINA, HUNAN PRovINcE, Longshan, 28.VIII.1958, coll. Chen Q.T., Liang LS.

no. 135 (HMAS 55957); no. 143 (HMAS 55958). SICHUAN PROVINCE, Emeishan,

17.X.1958, coll. Song M.H. no. 235 (HMAS 60028).

Notes: Guo & Liu (1989) introduced the combination Pseudocercospora

toxicodendri (Ellis) X.J. Liu & Y.L. Guo (= Cercospora toxicodendri Ellis)

and erroneously applied this name to Pseudocercospora collections on Rhus

chinensis Mill. in China. Based on examination of the type material, Braun

(1998) revealed that C. toxicodendri represents a species of Cercosporella,

reallocated Cercospora infuscans to Pseudocercospora, and emphasized that

the Chinese collections, erroneously assigned to P. toxicodendri, agree with

P. infuscans, although the conidia are somewhat narrower (3-5.5 um), and

the conidia longer (to 120 um) compared to North American samples, which

are characterized by broader (3-8 um) conidiophores and shorter (30-70)

conidia (Braun 1998). The re-examination of the Chinese collections on

R. chinensis cited under “specimens examined” showed that they should be

properly referred to as Pseudocercospora infuscans as emphasized by Braun

(1998) and Braun & al. (2016).

Pseudocercospora meliosmicola Y.L. Guo, Qian Zhao & Y.X. Shi, sp. nov. FIG. 4

MB 823728

Differs from Pseudocercospora sabiae by its longer, denticulate conidiophores and

its paler, longer conidia.

Type: China, Zhejiang Province, Hangzhou, on living leaves of Meliosma parviflora

Lecomte (Sabiaceae), 27.1X.1961, coll. Ma Qi-ming, Liu Xi-jin 581 (Holotype,

HMAS 168350).

EryMo.ocy: Derived from the host genus Meliosma.

Leaf spots amphigenous, circular to angular, 1-6 mm wide, often confluent,

red-brown, dark brown to almost black, with a pale olivaceous brown halo

Pseudocercospora meliosmicola sp. nov. (China) ... 365

SES ye

Soa

7 VE EER? Ringe

re, Ee Pes eae

Fic. 4 Pseudocercospora meliosmicola (holotype, HMAS 168350).

a. Conidia; b. Conidiophores; c. Stroma. Scale bar = 25 um.

on the upper surface, olivaceous brown to pale brown, with an olivaceous

brown halo on the lower surface. Fruiting amphigenous. Mycelium

internal and external; hyphae olivaceous, branched, smooth, septate,

2-2.5 um wide. Stromata substomatal, only a few brown globose cells to

well-developed, subglobose, dark brown, 20-40 um diam. Conidiophores

few, emerging through stomata, loose to dense fasciculate or arising

singly as lateral branches from external mycelial hyphae, olivaceous to

pale olivaceous brown, uniform in color, irregular in width, straight to

geniculate-curved, branched, smooth, rounded to conical at the apex,

0-1-septate, 8-35 x 3-3.5(-4) um. Conidiogenous cells cylindrical, 8-24

366 ... Zhao & al.

um long, conidiogenous loci inconspicuous, unthickened, not darkened.

Conidia solitary, cylindrical to narrowly obclavate-cylindrical, olivaceous

to very pale olivaceous brown, smooth, straight to curved, obtuse at the

apex, obconically truncate at the base, hila neither thickened nor darkened,

indistinctly 3-7(-10)-septate, 25-70(-85) x 2.5-3.5 um.

Notes: Cercosporoid fungi previously reported on hosts in Sabiaceae

include Pseudocercospora sabiae Y.L. Guo & W.X. Zhao on Sabia sp. (Guo

& Zhao 1989) and Cercospora meliosmae Y.L. Guo & Li Xu on Meliosma

sp. (Guo & Xu 2004). Pseudocercospora sabiae is distinguished from

P. meliosmicola by its epigenous fruiting habit; conidiophores that are

darker (olivaceous to pale brown), denticulate, and longer (15-162 um);

and conidia that are obclavate to cylindrical, paler (pale olivaceous), and

longer and slightly wider (40-112.5 x 3-4.3 um; Guo & Zhao 1989). Its

placement in another genus clearly separates Cercospora meliosmae from

Pseudocercospora meliosmicola.

Acknowledgments

This work was supported by the Project for Fundamental Research on Science

and Technology, Ministry of Science and Technology of China (No. 2013FY110400);

Science and Technology Innovation Program, Chinese Academy of Agricultural

Sciences (CAAS-ASTIP-IVFCAAS); Key Laboratory of Horticultural Crops

Genetic Improvement, Ministry of Agriculture in China (IVF2017ZF01). We

express our deep appreciation to Dr. U. Braun (Martin Luther Universitat, Halle,

Germany) and Prof. S.Y. Guo (Institute of Microbiology, Chinese Academy of

Sciences, Beijing) for their valuable suggestions, kind help, and assistance in the

course of submission of this manuscript. We are grateful to Ms. X.F. Zhu for inking

line drawing.

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hyphomycetes). Eching: Ihw- Verlag 2: 402 p.

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https://doi.org/10.5598/imafungus.2016.07.01.10

Crous PW. 1998. Mycosphaerella spp. and their anamorphs associated with leaf spot diseases of

Eucalyptus. Mycologia Memoirs 21: 170 p.

Den Breeyen A, Groenewald JZ, Verkley GJM, Crous PW. 2006. Morphological and molecular

characterisation of Mycosphaerellaceae associated with the invasive weed, Chromolaena

odorata. Fungal Diversity 23: 89-110.

Guo YL, Liu XJ. 1989. Studies on the genus Pseudocercospora in China I. Mycosystema 2:

225-240.

Pseudocercospora meliosmicola sp. nov. (China) ... 367

Guo YL, Xu L. 2004. A new species of Cercospora causing leaf spot of Meliosma sp.

Mycosystema 23: 18-19.

Guo YL, Zhao WX. 1989. Studies on hyphomycetes of Zhangjiajie in Hunan I. Pseudocercospora.

Acta Mycologica Sinica 8: 118-122.

Liu XJ, Guo YL. 1998. Flora fungorum sinicorum vol. 9: Pseudocercospora. Beijing: Science

Press.

Spegazzini CL. 1910. Mycetes argentinenses (Series V). Anales del Museo Nacional de Historia

Natural de Buenos Aires 20: 329-467.

MY COTAXON

April-June 2019—Volume 134, pp. 369-376

https://doi.org/10.5248/134.369

Heteroplacidium compactum

reported as a genus new to China

XIANG-MIN CHENG", DA-LE Lru*3, XIN-L1I WEI*’?, JIANG-CHUN WEI’?

"State Key Laboratory of Mycology, Institute of Microbiology,

Chinese Academy of Sciences, Beijing 100101, China

? University of Chinese Academy of Sciences, Beijing 100049, China

> College of Life Sciences, Southwest Forestry University, Kunming 650224, China

* CORRESPONDENCE TO: weixl@im.ac.cn

ABSTRACT—Three specimens of Heteroplacidium compactum (Verrucariaceae), isolated

from arid areas in Northwest China, were identified by morphological and phylogenetic

comparisons with other species, including species of Endocarpon and Placidium (common

verrucariaceous genera in the same area). This is the first report of Heteroplacidium from

China.

Key worps—desert, ITS, lichen, morphology, taxonomy

Introduction

Breuss (1996) separated four genera—Anthracocarpon, Clavascidium,

Heteroplacidium, and Involucropyrenium—from Catapyrenium Flot. s. lat.

(Verrucariaceae) based on morphological characters. Neocatapyrenium

H. Harada, Placidium A. Massal., and Scleropyrenium H. Harada are

also morphologically related (Prieto & al. 2010b) but are independent

monophyletic lineages within Verrucariaceae, according to phylogenetic

analyses (Gueidan & al. 2007, 2009; Prieto & al. 2010b).

Endocarpon Hedw. (Verrucariaceae) was relatively common during

our field survey in the arid region of Northwest China (Yang & Wei 2008,

Zhang & al. 2017a), but no species of Heteroplacidium Breuss was recorded.

Heteroplacidium, which grows primarily on soil and rock in warm temperate

370 ... Cheng & al.

Inner Mongolia

@ Sampling

Fic. 1. Heteroplacidium compactum collection sites in China.

regions, is characterized by a crustose-areolate to squamulose thallus, a

paraplectenchymatous or subparaplectenchymatous cortex, and clavate

asci with biseriate ascospores (Prieto & al. 2010a); twelve species have been

accepted (Knudsen & al. 2014). We report Heteroplacidium in China for the

first time, based on three specimens of H. compactum.

Material & methods

Phenotypic analysis

All Heteroplacidium samples were collected from Ningxia Hui Autonomous

Region and Inner Mongolia (Fic. 1) and are preserved in the Lichen Section of

Herbarium Mycologicum Academiae Sinicae, Beijing, China (HMAS-L). Zeiss

Stemi SV11 dissecting and Zeiss Axioskop 2 plus compound microscopes were

used for morphological examinations.

DNA extraction, amplification, sequencing

DNA was extracted from our three fresh specimens of Heteroplacidium (TABLE 1)

following the modified CTAB method (Rogers & Bendich 1988). The internal

transcribed spacer of nuclear ribosome DNA (nrDNA ITS), the standard DNA

barcoding marker (Schoch & al. 2012), was chosen as the genetic marker. Primers

LR1 (Vilgalys & Hester 1990) and ITS1 (White & al. 1990) were used. Reactions

were carried out in 50 uL reaction volume comprising 3 uL total DNA, 1 uL each

primer (10 uM), 25 uL 2xTaq MasterMix (CWBIO) and 20 uL ddH,O. The DNA

was amplified in a Biometra T-Gradient thermal cycler as follows: initial heating for

5 min at 95 °C; 35 cycles of 30s at 94 °C, 30s at 56 °C, and 90 s at 72 °C; ending witha

final extension of 8 min at 72 °C, after which the samples were kept at 4 °C. Negative

controls were prepared for each amplification series. PCR products were purified

using a CWBIO gel extraction kit following the manufacturer's instructions.

Heteroplacidium new to China... 371

Sequence alignment and phylogenetic tree construction

PCR products from the three putative Heteroplacidium specimens were

sequenced using ABI 3730 XL Sequencer. Additional sequences of Endocarpon

(3 spp.), Heteroplacidium (4 spp.), and Placidium (3 spp.) were downloaded from

GenBank (TaBLE 1) with the Endocarpon sequences selected as outgroup. The

sequences were aligned using ClustalW Multiple Alignment (Thompson & al.

1994) in BioEdit 7.2.5 (Hall 1999). The program Gblocks v0.91b (Castresana

2000, Talavera & Castresana 2007) was used to delimit and remove regions

of alignment uncertainty, using options for a “less stringent” selection on

the Gblocks web server (http://molevol.cmima.csic.es/castresana/Gblocks_

server.html). We confirmed the phylogenetic position of Heteroplacidium

sp. represented by our three specimens by subjecting the alignment to a

maximum likelihood (RAxML) analysis; nodal support was assessed using

1000 bootstrapping pseudoreplicates with RAxML-HPC v. 8.2.6 (Stamatakis

2014) and MrBayes v.3.2.6 (Huelsenbeck & Ronquist 2001, Ronquist &

Huelsenbeck 2003) on the CIPRES Science Gateway (http://www.phylo.org).

In the ML and Bayesian analyses, substitution models for ITS were estimated

using jModelTest-2.1.9 (Guindon & Gascuel 2003, Darriba & al. 2012). Based

TABLE 1. Specimens and sequences of Endocarpon, Heteroplacidium, and Placidium

used in the phylogenetic analysis

SPECIES VOUCHER ORIGIN GENBANK No.

E. adscendens CG671 Switzerland KF959777

E. pusillum CG470 _ JQ927447

E. tenuissimum Lendemer 29447 USA KM371592

Lendemer 27013 USA KM371593

H. acervatum LI 271015 Spain GU228954

M. Prieto 399 Spain GU228955

H. compactum M. Prieto 1701 Spain GU228952

M. Prieto 1607 Spain GU228949

XL2017117 China MH930459*

ALS2018002 China MH930460*

ALS2018036 China MH930461*

H. congestum LI 552268 USA GU228950

LI 297536 USA GU228951

H. divisum LI 218428 Italy GU228953

P pilosellum M. Prieto 3 Spain GU228993

VEL_PLG Slovakia KY981585

P. podolepis LI 297365 Argentina GU228956

P. squamulosum M. Prieto 336 Spain GU228994

* = sequences newly generated for this study by the authors

372 ... Cheng & al.

on these results, we used the TrN+I+G model with 1000 pseudoreplicates in the

ML analysis, and two parallel Markov chain Monte Carlo (MCMC) runs were

performed in MrBayes, each using 8 million generations and sampling every

1000 steps. A 50% majority-rule consensus tree was generated from the combined

sampled trees of both runs after discarding the first 25% as burn-in. Tree files

were visualized with FigTree v.1.4.2 (http://tree.bio.ed.ac.uk/software/figtree/).

Endocarpon adscendens KF959777

100/1.00 Endocarpon tenuissimum KM371592

100/1.00

80y-- Endocarpon tenuissimum KM371593

Endocarpon pusillum JQ927447

Heteroplacidium acervatum GU228955

100/1.00

85/0.97 Heteroplacidium acervatum GU228954

Heteroplacidium divisum GU228953

94/0.99 95/1.b0 Heteroplacidium congestum GU228950

Heteroplacidium congestum GU228951

88/0 9a7— @ Heteroplacidium compactum MH930461

Heteroplacidium compactum GU228952

0D/4|.00

00/1.00

--/D195 - Heteroplacidium compactum GU228949

@ Heteroplacidium compactum MH930460

93/0|97

@Heteroplacidium compactum MH930459

Placidium podolepis GU228956

100/1.00

Placidium squamulosum GU228994

100/1.00

Placidium pilosellum KY981585

D9/0.99

Placidium pilosellum GU228993

0.07

Fic. 2. The maximum likelihood tree of Endocarpon, Heteroplacidium, and Placidium

species based on ITS sequences. The numbers in each node represent bootstrap support

(BS) and posterior probability (PP) values. Bootstrap values 275 and posterior probability

values 20.95 were plotted on the branches of the RAxML tree. Except for the three samples

of Heteroplacidium compactum marked by the solid circle “@®% all other sequences were

downloaded from GenBank. Scale bar = 0.07 substitution per site.

Heteroplacidium new to China... 373

Phylogenetic and species delimitation analyses

The aligned matrix contained 455 unambiguous ITS nucleotide

positions. The phylogenetic tree (Fic. 2) included 10 taxa representing three

genera within Verrucariaceae. Heteroplacidium formed a well-supported

monophyletic clade (BS = 94, PP = 0.99) with the distinct species clearly

separated. Our three Chinese specimens all grouped with Spanish sequences

of H. compactum.

Taxonomy

Heteroplacidium compactum (A. Massal.) Gueidan & Cl. Roux,

Bull. Inf. Ass. Franc. Lichén. 33(1): 25, 2008. Fic. 3

THALLUS areolate, areoles 0.6-1.8 mm diam., angular to rounded, flat

to slightly convex; upper surface dark brown to black, dull to somewhat

shiny.

THALLUS LAYER $300 um thick; upper cortex paraplectenchymous,

12.5-37.5 um thick; algal layer 62.5-150 um thick, cells 12.5-17.5 um diam;

medulla 112.5-187.5 um thick; lower cortex paraplectenchymous, 10-12.5

um thick or not clearly delimited. RHIZOHYPHAE simple, intersecting,

Fic. 3. Heteroplacidium compactum (HMAS-L 140900): A. Habit of thallus; B. Anatomical

structure of thallus; C. Perithecia in thallus; D. Ascospores; E. Separate areole with abundant

rhizines tangling with soil particles. Scale bars: A = 1 mm; B = 20 um; C = 100 um; D = 50 um;

E=0.5 mm.

374 ... Cheng & al.

4.5-7.5 um diam. PERITHECIA nearly spherical, 300-500 um diam, exciple,

pale darkening with age. Ascr clavate, 52.5-62.5 x 15-17.5 um. ASCOSPORES

simple, ellipsoid, 12.5-17.5 x 10-12.5 um. PYCNIDIA not seen.

SPECIMENS EXAMINED. CHINA. INNER MONGOLIA: 39°28’22”N 101°04’04”E,

1563 m alt., on sand, 5 June 2018, D.L. Liu & al. ALS2018002 (HMAS-L 0141672;

GenBank MH930460); 39°32’31”N 101°06’34’”E, 1478 m alt., on sand, 5 June

2018, D.L. Liu et al. ALS2018036 (HMAS-L 0141671; GenBank MH930461).

NinGxia: Zhongwei City, Cui Liu Gou. 37°24’35”N 104°35’09”E, 1577 m

alt., on sand, 15 July 2017, D.L. Liu & R.D. Liu XL2017117 (HMAS-L 140900;

GenBank MH930459).

DISTRIBUTION: Heteroplacidium compactum has previously been reported

from Asia (Mongolian People's Republic; Pakistan), Europe, and northern

Africa (Breuss 1994; Prieto & al. 2010a). In China, it is now known from

arid desert areas in Inner Mongolia and Ningxia.

Discussion

The characters of Chinese H. compactum are essentially similar to the

previous descriptions (Breuss 1994; Prieto & al. 2010a) in the areolate

thallus, dark brown upper surface, paraplectenchymous upper and

lower cortex, and the shape and size of the perithecia and asci. Among

the differences noted are a thinner thallus (300 um versus 600 um) and

broader ascospores (10-12.5 um versus 8-10 um). Furthermore, pycnidia

were not seen in the Chinese materials.

In the arid desert area of China, Endocarpon is quite common and

comprises a key component of biological soil crust communities (BSC).

The biodiversity reported for these Chinese BSCs includes several newly

described Endocarpon species (Yang & Wei 2008, Zhang & al. 2017a) with

the predominant species, E. pusillum, studied physiologically (Zhang &

Wei 2011), ecologically (Ding & al. 2013), and genomically (Wang & al.

2014, 2015, Li & Wei 2016, Zhang & al. 2017b). Heteroplacidium (with

Dermatocarpon-type pycnidia) is clearly different from Endocarpon (with

Endocarpon-type pycnidia).

The most similar genus to Heteroplacidium is Placidium A. Massal.,

which is also characterized by Dermatocarpon-type pycnidia, but the

two genera have differently sized thallus squamules (Heteroplacidium

0.3-3 mm diam; Placidium 2-8(-15) mm diam). In the phylogenetic tree

(Fic. 2), Heteroplacidium is most closely related to Placidium, clustering

in a well-supported clade (BS = 100, PP = 1) and relatively distant from

Endocarpon.

Heteroplacidium new to China... 375

Acknowledgments

This research was funded by one of National Key Research and Development

Program of China (2016YFE0203400) and the National Natural Science

Foundation of China (31770022, 31470149). The authors sincerely thank the two

peer reviewers, Dr. Steve Leavitt (Brigham Young University, USA) and Dr. Ze-

Feng Jia (Liaocheng University, China), for giving constructive suggestions and

kindly help in making careful modifications. Ms. H. Deng gave considerable

assistance during our studies in HMAS-L.

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M, Lutzoni F. 2009. Generic classification of the Verrucariaceae (Ascomycota) based on

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MY COTAXON

April-June 2019—Volume 134, pp. 377-389

https://doi.org/10.5248/134.377

Geopora ahmadii sp. nov. from Pakistan

M. SaBA"?”3, D,. HAELEWATERS?*, T. ITURRIAGA?>>,

T. ASHRAF’, A.N. KHALID’, D.H. PFISTER?

“Department of Plant Sciences, Quaid-i-Azam University,

Islamabad, 45320, Pakistan

?Department of Botany, University of the Punjab,

Quaid-e-Azam Campus, Lahore 54590, Pakistan

*Farlow Reference Library and Herbarium of Cryptogamic Botany, Harvard University,

Cambridge, MA 02138, USA

*Faculty of Science, University of South Bohemia, 37005 Ceské Budéjovice, Czech Republic

°*Plant Pathology and Plant-Microbe Biology Section, School of Integrative Plant Science,

College of Agriculture & Life Sciences, Cornell University, Ithaca, NY 14853, USA

*CORRESPONDENCE TO: rustflora@gmail.com

ABSTRACT—A new species, Geopora ahmadii, is described and illustrated based on material

from Punjab, Pakistan. This species is characterized by sessile, cup- to saucer-shaped, partly

immersed apothecia with whitish to grayish hymenial surfaces; broad ellipsoid, mostly

uniguttulate ascospores; and brown excipular hairs. Phylogenetic analyses of the nrDNA ITS

region with maximum parsimony, maximum likelihood, and Bayesian inference methods

reveal that G. ahmadii is distinct from other described Geopora species. A collection

previously identified as Geopora arenosa from Rawalakot, Pakistan, likely represents a second

locality of G. ahmadii.

KEY worps—Ascomycota, hypogeous fungi, Pezizales, Pyronemataceae, taxonomy

Introduction

Geopora Harkn. (Pyronemataceae, Pezizales) is characterized by (1)

ascomata that occur entirely or partially below ground and are covered

with brown, septate excipular hairs; (2) a whitish, grayish, or yellowish-

gray hymenium; and (3) smooth, mostly uniguttulate ascospores. Because

ascomata appear infrequently and are hypogeous at some developmental

378 ... Saba & al.

stage, Geopora specimens are infrequently collected. Species delimitation

is additionally challenging due to the scarcity of distinctive morphological

characters, with measurement ranges overlapping among species (Tamm &

al. 2010, Guevara-Guerrero & al. 2012, Flores-Renteria & al. 2014).

Identification of Geopora species has relied primarily on ascospore

shape and size, position of apothecia in the ground, and the length of

excipular hairs (Burdsall 1965, 1968; Tamm & al. 2010; Flores-Renteria

& al. 2014). Molecular analyses by Tamm & al. (2010) showed that well

supported clades are not congruent with morphological species concepts.

The combination of molecular and morphological data is considered the

most reliable approach to define species in this genus (Southworth & Frank

2011, Guevara-Guerrero & al. 2012, Flores-Renteria & al. 2014).

Perry & al. (2007) studied the phylogenetic relationships of

Pyronemataceae. Using LSU ribosomal DNA (rDNA) sequence data, they

suggested that Geopora is monophyletic. Hansen & al. (2013) determined

that Geopora was sister to Tricharina in the larger Scutellinia—Trichophaea

lineage. Neither study included species representing Phaeangium Pat. or

Picoa Vittad. Stielow & al. (2013) suggested that Geopora was paraphyletic

including species of both Phaeangium and Picoa. They also found that

Geopora pellita (Sacc.) T. Schumach. was phylogenetically isolated from

other Geopora species and created the new genus Hoffmannoscypha Stielow

& al. to accommodate this taxon.

During our studies of ectomycorrhizal fungi, we found Geopora

specimens growing in groups on damp soil in Punjab, Pakistan. Molecular

phylogenetic analysis ofthe ITS rDNA region combined with morphological

evaluation support the recognition of our collections as a new species. ‘This

species is described, illustrated, and compared with other Geopora species.

Materials & methods

Morphological studies

Ascomata were collected and dried in a food dehydrator at 39 °C for 7-9 hours.

Shape, texture, and dimensions of important characters were recorded from fresh

ascomata. Colors were compared to the Munsell Soil Color Charts (1975). Dried

voucher specimens are deposited at the University of the Punjab Herbarium, Lahore,

Pakistan (LAH) and the Farlow Herbarium, Harvard University, Cambridge, MA,

USA (FH).

Sections of specimens were mounted in water and Congo red in ammonia (0.3%

in commercial ammonia cleaner) to increase contrast for microscopic observations.

Micromorphological analysis, photographs, and measurements were made

Geopora ahmadii sp. nov. (Pakistan) ... 379

using an Olympus Bx40 light microscope with Olympus XC50 digital camera

and Microsuite Special Edition software 3.1. Sections were made using a freezing

microtome. Measurements include the typical range with extremes given in

parentheses. Q values (length/width ratios) are given for ascospores.

DNA extraction, PCR amplification, DNA sequencing

Genomic DNA was extracted from a small piece of an ascoma by a modified

CTAB method (Gardes & Bruns 1993). The internal transcribed spacer region

(ITS1+5.8S+ITS2) of the nuclear ribosomal RNA gene was amplified using the

primer pair ITSIF and ITS4 (White & al. 1990, Gardes & Bruns 1993) and the

RED Extract-N-Amp PCR ReadyMix. PCR cycling parameters comprised initial

denaturation (94° C for 1 min), 35 cycles (94 °C for 1 min, 53 °C for 1 min, and

72 °C for 1 min), and final extension 72 °C (8 min). Amplified PCR products

were outsourced to Macrogen, (Seoul, Republic of Korea) for purification and

bidirectional sequencing.

Sequence alignment & phylogenetic analyses

Geopora sequences downloaded from GenBank included those studied by

Tamm & al. (2010) and sequences representing recently described species—

G. cercocarpi D. Southw. & J.L. Frank, G. gilkeyae (Burds.) G. Guevara & al.,

G. pinyonis Flores-Rent. & Gehring, and G. tolucana G. Guevara & al. (Flores-

Renteria & al. 2014, Guevara-Guerrero & al. 2012, Southworth & Frank 2011).

Tarzetta catinus (Holmsk.) Korf & J.K. Rogers and Trichophaea hybrida (Sowerby)

T. Schumach. (Pyronemataceae, Pezizales) were selected as the outgroup because

they are closely related to Geopora (Perry & al. 2007).

Manually edited sequences were assembled in BioEdit v7.2.6 (www.mbio.

ncsu.edu/bioedit/bioedit.html). All sequences were trimmed with the conserved

motifs 5’-(...GAT)CATTA- and —-GACccT(CAAA...)-3" (Dentinger & al. 2011), and

the alignment portions between them were included in the analysis. Sequences

retrieved from NCBI GenBank were aligned by Muscle v3.7 (Edgar 2004) with

default parameters using Molecular Evolutionary Genetics Analysis (MEGA)

software (Tamura & al. 2011).

Maximum parsimony (MP) analysis was performed with PAUP 4.0b on

XSEDE (Swofford 1991), available on the Cipres Gateway v3.3 (Miller & al. 2010).

All characters were equally weighted and gaps were treated as missing data. The

heuristic search option with tree-bisection-reconnection (TBR) branch swapping

and 1000 random sequence additions were used to infer trees. Clade robustness

was assessed using a bootstrap analysis with 500 replicates (Felsenstein 1985).

A maximum likelihood (ML) analysis was carried out with RAxML XSEDE on

the Cipres Gateway, using the general time-reversible (GTR) model of nucleotide

substitution (Stamatakis & al. 2008). Nodal support was determined from 1000

bootstrap replicates.

Bayesian analysis was done with a Markov chain Monte Carlo (MCMC)

coalescent approach implemented in Beast v1.8.2 (Drummond & Rambaut 2007),

380 ... Saba & al.

TABLE 1. Geopora isolates and outgroup included in phylogenetic analyses

[Clade designations sensu Tamm & al. 2010]

ORIGINAL ID

Tarzetta catinus

Trichophaea

hybrida

G. cercocarpi

G. pinyonis

G, tolucana

G. tolucana

G. ahmadii

G. arenicola

G. sp.

G. arenicola

G. foliacea

Ectomycorrhizal

uncultured

G. cf. sepulta

G. cooperi

G. gilkeyae

G. arenicola

G. sepulta

CLADE

wm Ms RM OM

Vill

VIII

VII

COUNTRY

Estonia

USA, OR

USA, AR

Mexico

Pakistan

Estonia

France

Spain

Estonia

USA, CA

Estonia

VOUCHER

TAAM 192291

TAAM 192334

SOC 1590

DGB 27586

ITCV 1081

MSM#0091 [T]

MSM#00163

TAAM 192329

TAAM 192324

TAAM 192330

TAAM 192323

ECM 2

ECM 95

Riv-4

TAAM 113526

101GA

108GC

109GC

src515

TAAM 188666

TAAM 188339

TAAM 117708

TAAM 188293

TAAM 188292

TAAM 135060

TAAM 116784

TAAM 192311

GENBANK

FM206478

FM206477

HQ283090

NR121491

KF768653

KF768652

HQ184961

HQ184960

KY805995

KY805996

FM206473

FM206471

FM206472

FM206470

AJ410862

AJ410865

EF484934

FM206476

AF387651

AF387649

AF387650

DQ974731

FM206449

FM206446

FM206460

FM206440

FM206439

FM206433

FM206462

FM206432

REFERENCE

Tamm & al. 2010

Southworth & Frank 2011

Flores-Renteria & al. 2014

Guevara-Guerrero & al. 2012

This paper

Tamm & al. 2010

El Karkouri & al. 2004

Rincon & al. 2007

Tamm & al. 2010

Gutierrez & al. (unpubl.)

Smith & al. 2007

Tamm & al. 2010

Geopora ahmadii sp. nov. (Pakistan) ... 381

ORIGINAL ID CLADE COUNTRY VOUCHER GENBANK REFERENCE

G. sepulta Vil Estonia TAAM 192333 FM206431 Tamm &al. 2010

G. foliacea Vv Finland H RS-34685 FM206428 Tamm &al. 2010

G. foliacea Vv Finland H RS-29584 FM206424 Tamm &al. 2010

G. cervina V Finland H RS-17984 FM206426 ‘Tamm & al. 2010

G. cervina VI Estonia TAAM 192232 FM206420 ‘Tamm &al. 2010

G. tenuis VI Estonia TAAM 192302 FM206429 ‘Tamm & al. 2010

G. sp. VI Tajikistan TAAM 116668 FM206475 Tamm &al. 2010

G. tenuis IV Estonia TAAM 188326 FM206397. Tamm & al. 2010

G. tenuis IV Finland H RS-09584 FM206402. Tamm &al. 2010

G. tenuis IV Estonia TAAM 188331 FM206396 ‘Tamm &al. 2010

G. cervina IV Estonia TAAM 192293 FM206401 ‘Tamm &al. 2010

G. cervina Ill Estonia TAAM 117479 FM206413 ‘Tamm &al. 2010

G. cervina Ill Finland H RS-07186 FM206406 ‘Tamm &al. 2010

G. cervina Il Estonia TAAM 117884 FM206409 ‘Tamm &al. 2010

G. cervina Il Estonia TAAM 192321 FM206410 ‘Tamm &al. 2010

G. arenicola Ill Estonia TAAM 117952 FM206412 ‘Tamm &al. 2010

G. cervina II Estonia TAAM 188304 FM206417 ‘Tamm &al. 2010

G. cervina II Estonia TAAM 117898 FM206419 ‘Tamm &al. 2010

G. arenicola II Estonia TAAM 188517 FM206418 ‘Tamm &al. 2010

G. cervina I Finland H RS-06986 FM206387 ‘Tamm &al. 2010

G. cervina I Estonia TAAM 188655 FM206390 ‘Tamm &al. 2010

G. cervina I Estonia TAAM 117854 FM206391 ‘Tamm &al. 2010

G. cervina I Estonia TAAM 117659 FM206389 ‘Tamm & al. 2010

with an uncorrelated lognormal relaxed clock for rate variation across the tree.

A Bayesian skyride coalescent tree GMRF prior with the GITR+I+G model of

nucleotide substitution was used in all simulations, with a randomly generated

starting tree. Four independent runs of 10 million generations were undertaken.

Tracer v1.6.0 (Drummond & Rambaut 2007) was used to check the effective

sample size (ESS), and burn-in values were adjusted to achieve a net ESS of at least

200. Upon removal of a portion of each run as burn-in, log files and trees files

were combined in LogCombiner v.1.8.2. Finally, a consensus tree (0% burn-in) was

generated using TreeAnnotator v1.8.2 and visualized in FigTree v1.4.2.

Sequences of Geopora ahmadii generated during this study were submitted to

GenBank. Accession numbers for the sequences downloaded from GenBank and

those sequenced during this study are given in TABLE 1.

382 ... Saba & al.

Taxonomy

Geopora ahmadii Saba, T. Ashraf, Khalid & Pfister, sp. nov. Fics 1, 2

MB 822666

Differs from Geopora arenicola by its partly immersed apothecia that are larger in

diameter, cupulate when young but saucer-shaped when older, by its broadly ellipsoid

ascospores, and by its ITS sequence with 49-52 autapomorphies.

Type: Pakistan, Punjab, Lahore, University of the Punjab, Department of Botany,

Botanical Garden, 31°29’56”N 74°17'57’E, 6 March 2009, leg. M. Saba, T. Ashraf &

A.N. Khalid, MSM#0091 (Holotype, LAH 310019; GenBank KY805995).

EryMo.Locy: Named in honor of Dr. Sultan Ahmad (1910-1983), eminent pioneering

mycologist in Pakistan.

APOTHECIA partly immersed in soil, sessile, fleshy, cup-shaped at early stages

and saucer-shaped at older stages when becoming thin, flat-discoid, <15-25

mm in diameter and 4 mm deep when fresh; when dry shrinking to 6-10 mm

diam; disc grey and smooth when fresh, whitish beige to whitish grey when

Fic. 1. Geopora ahmadii (holotype, LAH310019). a. Section of ascoma showing hymenium,

subhymenium, excipulum, and excipular hairs; b. Excipular cells with single excipular hair; c. Asci

and paraphyses; d. Asci and paraphyses, with detail of an ascospore with a single guttule (insert).

Scale bars: a = 200 um; b, d = 50 um; c = 100 um.

Geopora ahmadii sp. nov. (Pakistan) ... 383

}

Fic. 2. Geopora ahmadii (holotype, LAH310019). Ascospores, each with a single guttule:

a. Mounted in Congo red in ammonia; b. Mounted in water. Scale bars = 10 um.

rehydrated; receptacle dark brown, warted and hairy. Margin wavy at young

stages, splitting into 5-6 lobes at older stages.

HyMENIUM 180-220 um thick. Ascr cylindrical, |e (175-)210-250(-340)

x 15-21 um, with 8 ascospores. SUBHYMENIUM dense textura intricata,

dark brown, compact. Ascospores uniseriate, broadly ellipsoid, with a

single guttule, and smaller guttules at the poles, 19-26.0 x (11-)12-15 um,

Q = 1.68-1.94; wall 1.0-1.5 um thick, hyaline. ParApuysss slender, hyaline,

broadly clavate, 7-10 um diam. at the tip, 5-6 um diam. in the middle, septate.

ECTAL EXCIPULUM 75-87(-115) um thick, dark brown of textura globulosa to

textura angularis, cells round to irregular to polygonal, 16-30 x 13-16 um,

walls brown, 1-2 um thick (especially thicker in the outermost cells), outer

cells aggregated to form warts. MEDULLARY EXCIPULUM (40-)50-70(-80) um

thick; dense textura intricata, cells appearing angular to irregular to roundish,

becoming smaller towards the subhymenium, cells 8.5-20.5 x 5-12 um. Hairs

arising from globose ectal excipular cells, septate and forming a mat of brown

hyphae holding soil particles. When young thin, straight, light brown, smooth,

(6-)7.5-10.0(-16) um diam.; with age branched, twisted and curved, dark

brown, <(40-)50-70(-80) um diam., with dark granular walls and granular

content; hair walls 0.8-1.5(—2) um thick.

384 ... Saba & al.

ADDITIONAL MATERIAL STUDIED: PAKISTAN, Punyjas, Lahore, University of the

Punjab, Botanical Garden, 31°29’56”N 74°17'57’E, 16 June 2011, MSM#00160 (LAH

310099!); 5 May 2013, leg. M. Saba, T. Ashraf & A.N. Khalid, MSM#00163 (FH

01142414; GenBank KY805996).

Molecular analyses of the genus Geopora

Initial BLAST analysis of the G. ahmadii ITS sequence showed a

maximum identity of 84% with collection TAAM 192330 (GenBank

accession number FM206472). This collection belongs to clade IX (sensu

Tamm & al. 2010) but was identified based on morphology as G. arenicola

(Lév.) Kers. Our data matrix included 53 isolates, all representing identified

Geopora species except for three unidentified ectomycorrhizal isolates (El

Karkouri & al. 2004, Rincén & al. 2007) and the two outgroup sequences

(TaBLE 1). The final aligned data matrix included 722 characters, of which

272 were constant and 351 were parsimony-informative.

Our MP and ML analyses (Fic. 3) largely agree. One exception is the

placement of G. gilkeyae (Burds.) Guevara & al. (as “Geopora cooperii var.

gilkeyi” in Smith & al. 2007) and G. tolucana Guevara & al. In the MP

topology (not shown), these species are sister taxa and form a branch basal

to all other Geopora species (sensu Tamm & al. 2010). However, there is no

bootstrap support for this placement. Also, the placement of G. gilkeyae

in the ML analysis is unresolved (no bootstrap support). In the three

analyses (MP, ML, Bayesian), a number of the basal branches lack support,

and phylogenetic reconstructions based on ITS alone cannot resolve

relationships of Geopora at deeper nodes.

Clades I through X (sensu Tamm & al. 2010) are recognized here with

high support from MP, ML, and Bayesian analyses (Fics 3, 4). Geopora

ahmadii is inferred in Geopora clade IX from Tamm & al. (2010) with

maximum support. The morphological characters of G. ahmadii (fruit-

body features, ascospore dimensions, Q ascospore values) are consistent

with those of clade IX as given by Tamm & al. (2010). The species is

recovered as sister to “Geopora sp. b” (sensu Schumacher 1979; defined in

the study of Tamm & al. 2010) in clade IX. Geopora sp. b is known only

from northern Europe, with records from Estonia (Tamm & al. 2010) and

Norway (Schumacher 1979).

Discussion

Tamm & al. (2010) reviewed the history of the genus Geopora and

confirmed that delimitations of species within Geopora are difficult. All

Geopora ahmadii sp. nov. (Pakistan) ... 385

e Tarzetta catinus

100 Trichophaea hybrida

Geopora cercocarpi NR_121491

Geopora cercocarpi HQ283090

Geopora pinyonis KF768653

Geopora pinyonis KF768652

Geopora tolucana HQ184961

Geopora tolucana HQ184960

Geopora ahmadii KY805995

Geopora ahmadii KY805996

= Geopora arenicola FM206472

Geopora sp. FM206471

Geopora foliacea FM206470

Pe Geopora arenicola FM206473

99 - ECM 2 AJ410862

00 ECM 95 AJ410865

99 ECM Riv-4 EF484934

$8 Geopora cf. sepulta FM206476

75| Geopora cooperi AF387651

Geopora cooperi AF387649

Geopora cooperi AF387650

Geopora gilkeyae DQ974731

Geopora arenicola FM206449 |

100} Geopora arenicola FM206446

74 |- Geopora arenicola FM206460

82] Geopora arenicola FM206440

se Geopora arenicola FM206439

Geopora arenicola FM206433

Geopora arenicola FM206462

Geopora sepulta FM206432

Geopora sepulta FM206431

Geopora foliacea FM206428

Geopora cervina FM206426

Geopora foliacea FM206424

Geopora cervina FM206420

95 l96-— Geopora tenuis FM206429

90 95L_— Geopora sp. FM206475

93 Geopora tenuis FM206397

93 Geopora tenuis FM206402

Geopora tenuis FM206396

80 Geopora cervina FM206401

100 B Geopora arenicola FM206412

100 Geopora cervina FM206413

99 Geopora cervina FM206406

96 pa Geopora cervina FM206409

97 aal| 99 'Geopora cervina FM206410

93 Geopora arenicola FM206418 |

I+I+II1

9g|} Geopora cervina FM206417

1004 Geopora cervina FM206419

Geopora cervina FM206387

Geopora cervina FM206390 |

gs| Geopora cervina FM306391

97 'Geopora cervina FM206389

Fic. 3. Phylogeny of Geopora species produced from maximum likelihood (ML) analysis of the ITS

rDNA dataset. The best-scoring ML tree (log likelihood of -6541.843463) is shown. Only bootstrap

values >70% are given. Maximum parsimony (MP) bootstrap values are given above the branches;

ML bootstrap values are below the branches. Branches in bold have maximum support.

known taxa are ectomycorrhizal (Tedersoo & al. 2006) but we have not

been able to determine the mycorrhizal associate(s) for our new species.

The ectomycorrhizal host could be Pinus roxburghii, which grows near the

site where this species has been repeatedly collected in the Botanical Garden

386 ... Saba & al.

Tarzetta catinus

Trichophaea_hybrida_TAA192334

Geopora ahmadii KY805995

Geopora ahmadii KY805996

Geopora arenicola FM206473 IX

Geopora foliacea FM206470

Geopora arenicola FM206472

0.7 Geopora sp. FM206471

7 ECM 2 AJ410862

ECM 95 AJ410865

0.9 ECM Riv-4 EF484934

09 Geopora cf. sepulta FM206476 X

Geopora cooperi AF387650

Geopora cooperi AF387651

Geopora cooperi AF387649

Geopora cercocarpi NR_121491

Geopora cercocarpi HQ283090

Geopora pinyonis KF768653

0.8 Geopora pinyonis KF768652

Geopora tolucana HQ184961

Geopora tolucana HQ184960

Geopora gilkeyae DQ974731

Geopora sepulta FM206432

Geopora sepulta FM206431

Geopora arenicola FM206449

Geopora arenicola FM206446

Geopora arenicola FM206462

Geopora arenicola FM206460

Geopora arenicola FM206433

Geopora arenicola FM206440

Geopora arenicola FM206439

Geopora foliacea FM206428

0.9 - Geopora foliacea FM206424

Geopora cervina FM206426 |

Geopora cervina FM206420 |

0.7

Geopora sp. FM206475

Geopora tenuis FM206429

Geopora tenuis FM206402

Geopora tenuis FM206397

Geopora tenuis FM206396

Geopora cervina FM206401

Geopora cervina FM206413

Geopora arenicola FM206412

Geopora cervina FM206406

Geopora cervina FM206410

Geopora cervina FM206409

Geopora arenicola FM206418

Geopora cervina FM206419

Geopora cervina FM206417

0.7 - Geopora cervina FM206390

Geopora cervina FM206387

Geopora cervina FM206389

Geopora cervina FM306391

I+11+lll

Fic. 4. Phylogeny of Geopora species produced from Bayesian inference of the ITS rDNA dataset.

Only posterior probabilities 20.9 are shown. Branches in bold have maximum support.

of the Department of Botany, University of the Punjab, Lahore, Pakistan.

Associations of the new species should be evaluated with sequences from

root tip samples.

Geopora ahmadii sp. nov. (Pakistan) ... 387

Geopora ahmadii is distinct from other Geopora species based on

apothecial shape, partial immersion of the apothecium in the soil, hymenial

color, and ascospore size. Other partly immersed species are G. tenuis

(Fuckel) T. Schumach. and G. cervina (Velen.) T. Schumach. (Yao & Spooner

1996), which are distinguished by different spore sizes ((20.3—)21.4(—23.0) x

(10.8—)11.6(—12.1) for G. tenuis; (20.8—)23.8(—26.2) x (10.8—)12.1(-14.2) um

for G. cervina; Tamm & al. 2010) and placement in different clades in our

analyses. The hymenial color in G. ahmadii is similar to G. sepulta (Fr.) Korf

& Burds., which differs by complete immersion in the soil and placement

in another clade.

Our molecular analyses place G. ahmadii in clade IX sensu Tamm &

al. (2010). Its morphological characters—including the position and shape

of the ascomata and ascospore dimensions—are consistent with placement

among the other clade IX species. This clade includes specimens initially

identified as G. arenicola and G. foliacea (Schaeff.) S. Ahmad. Geopora

arenicola, as defined by Tamm & al. (2010), has a completely immersed

ascoma. The name G. foliacea has been variously applied.

Once again, this study underscores the difficulties of using morphology

for species delimitation in this group. Southworth & al. (2011) also

noted the difficulty in using morphology to describe their new species,

G. cercocarpi, citing its exclusive association with Cercocarpus ledifolius and

that other Geopora specimens showed similar host fidelity. Unfortunately

we do not know the associate of G. ahmadii but we plan to collect root tip

samples under trees of its most likely candidate ectomycorrhizal associate,

Pinus roxburghii.

One comment regarding G. arenosa (Fuckel) S. Ahmad: Ahmad (1978)

transferred Peziza arenosa Fuckel [= Humaria arenosa (Fuckel) Fuckel]

(Fuckel 1864, 1866, 1870) to Geopora based on a collection he studied from

Rawalakot, Pakistan. According to his description, G. arenosa has globose

to subglobose apothecia, a whitish gray hymenial surface, and ellipsoid

ascospores measuring 20-24 x 13-14.5 um. Yao & Spooner (1996) studied

the type material of G. arenosa (Fuckel’s Fungi Rhenani exsiccati, No. 1212,

K—K(M) 69362, designated as lectotype by Yao & Spooner 2003), and

their description contrasts with the material studied by Ahmad (1978) in

ascospore shape and size (ellipsoid to fusoid, 27-30 x 13.5-15 um in the

G. arenosa type vs. ellipsoid, 20-24 x 13-14.5 um in Ahmad’s material).

Because of these discrepancies, we believe that Ahmad’s collection from

Rawalakot represents a previous collection of our new species. We have not

388 ... Saba & al.

been able to locate Ahmad’s specimen and it is likely lost. It is not present

in LAH and no sequence data are available for the Rawalakot material.

Further collecting in Rawalakot may help to resolve the identity of Ahmad’s

species and to determine if it is indeed conspecific with G. ahmadii.

Acknowledgments

We are highly indebted to Higher Education Commission (HEC), Islamabad,

Pakistan, for funding this project under Phase II, Batch I, Indigenous PhD fellowships

program for 5000 scholars and through International Research Support Initiative

Program (IRSIP). Finally, we thank Dr. Rosanne Healy and Dr. Matthew E. Smith (both

of Dept. Plant Pathology, University of Florida, Gainesville, USA), and Nomenclature

Editor Dr. Shaun B. Pennycook for critically reviewing the manuscript, which would

have been a lot meagerer if it weren't for their valuable comments for improvement.

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Bulletin 58(1): 247-252. https://doi.org/10.2307/4119373

MYCOTAXON

April-June 2019—Volume 134, pp. 391-398

https://doi.org/10.5248/134.391

Seychellomyces sinensis sp. nov. from China

Min Q1A0%, HuA ZHENG**S, ZHE ZHANG’, ZE-FEN YU”*

‘Laboratory for Conservation and Utilization of Bio-resources,

Key Laboratory for Microbial Resources of the Ministry of Education,

Yunnan University, Kunming, Yunnan, 650091, P. R. China

’ School of Life Sciences, Yunnan University,

Kunming, Yunnan, 650091, P. R. China

* CORRESPONDENCE TO: zfyugm@hotmail.com

ABSTRACT —An aquatic hyphomycete, Seychellomyces sinensis, isolated from China, is

proposed as a new species. Phylogenetic analysis of combined ITS, LSU, 60S, and MCM7

sequences place Seychellomyces in Ceratocystidaceae. The new species, characterized by

septate conidia and elliptical to cylindrical (sub)hyaline endoconidia, is distinguished from

S. hexagonus by its larger, 1-3-septate conidia.

KEY worps—asexual fungi, Microascales, Sordariomycetes, taxonomy

Introduction

Aquatic hyphomycetes are fungi that most commonly occur on dead

leaves in streams (Barlocher 1992). These fungi are a polyphyletic group

that play a key role in processing organic matter in freshwater (Belliveau &

Barlocher 2005). Traditionally, species have been identified through their

conidial morphology, but phylogenetic placements of many species have

not yet been determined. The introduction of molecular analysis to fungal

taxonomy has recently verified the phylogenetic placement of some species

(Duarte & al. 2015).

Seychellomyces Matsush. was proposed for a single species, S. hexagonus

Matsush. (Matsushima 1981) and remains monotypic. It is characterized by

‘These authors contributed equally to this research.

392 ... Qiao, Zheng & al.

producing septate brown to pale brown conidia that are hexagonal in cross-

section and aseptate hyaline endoconidia (Matsushima 1981, Seifert & al.

2011).

Several new hyphomycetes species have been reported from southwest

China (Qiao & al. 2017a,b; 2018a,b, 2019; Yu & al. 2019). During further

study of the aquatic hyphomycetes, a specimen with two types of conidia

was found growing on submerged dicotyledonous leaves. The specimen was

quite close to Seychellomyces hexagonus in its conidiogenous features but

differed in possessing larger and more septate conidia. Here we describe it

as a new species and present the phylogenetic position of Seychellomyces as

determined by combined sequences of internal transcribed spacer (ITS),

large subunit nuclear ribosomal RNA (LSU), 60S ribosomal protein gene

RPL10 (60S), and the minichromosome maintenance complex component

7 (MCM7).

Materials & methods

Collection, fungal isolation, and morphological characterization

Submerged dicotyledonous leaves were collected from a stream in Sangsi,

Guangxi Province, China. Samples were preserved in zip-lock plastic bags,

labelled, and transported to the laboratory. Each rotted leaf was cut into several

3-4 x 4-5 cm fragments that were incubated on CMA (20 g cornmeal, 18 g agar,

40 mg streptomycin, 30 mg ampicillin, 1000 ml distilled water) for 5 days at room

temperature. Individual conidia were isolated using a sterilized toothpick under

a BX51 microscope and cultivated on CMA plates. Morphological observations

were conducted on cultures growing on CMA, V-8 juice agar (200 ml Campbell

V-8 juice, 3.0 g CaCO,, 18 g agar, 800 ml distilled H,O), and PDA (200 g potato,

20 g dextrose, 18 g agar, 1000 ml distilled H,O) after incubation at 25°C for one

week.

Pure cultures were deposited in the Herbarium of the Laboratory for

Conservation and Utilization of Bio-resources, Yunnan University, Kunming,

Yunnan, P.R. China (YMF; formerly Key Laboratory of Industrial Microbiology

and Fermentation Technology of Yunnan).

DNA extraction, PCR amplification, and sequencing

Genomic DNA was extracted from fresh mycelia grown on potato dextrose agar

(PDA) at 25°C as described by Turner & al. (1997). Four gene regions were amplified:

ITS with primer pair ITS5/ITS4 (White & al. 1990); LSU with LROR/LR7 (Vilgalys

& Hester 1990); 60S ribosomal protein RPL10 (60S) with 60S-506F/60S-908R

(Nel & al. 2018); and MCM7 with MCM7-for/MCM7-rev (Nel & al. 2018). The

PCR thermal cycle programs for the amplifications of these four DNA fragments

followed those described in de Beer & al. (2014). PCR products were then purified

Seychellomyces sinensis sp. nov. (China) ... 393

TABLE 1. Strains and GenBank accession numbers of sequences used in molecular

phylogenetic analyses

GENBANK NUMBER

SPECIES STRAIN

ITS LSU 60S MCM7

Ambrosiella xylebori CMW110.61 NR_144921 KM495318 KM495495 KM495407

Berkeleyomyces basicola CMW6714 MF952423 MF948658 =MF967072 MF967079

CMW49351 MF952428 MF948659 MF967075 MF967102

CMW25439 MF952427 — _ MF967099

Berkeleyomyces rouxiae CMW5472 MF952406 MF948657. + =MF967074 = MF967080

CMW 14219 MF952402 MF948660 MF967076 MF967086

Ceratocystis diversiconidia CMW22445 FJ151440 KM495334. =KM495511 KM495423

Ceratocystis fimbriata CMW 15049 AY953378 KM495343, =KM495520 KM495432

Ceratocystis pirilliformis CMW6579 AF427105 KM495365 KM495542 KM495453

Ceratocystis platani CMW 14802 DQ520630 KM495366 KM495543 KM495454

Chalaropsis ovoidea CMW22733 FJ411343 KM495400 =KM495577. =. KM495487

Chalaropsis thielavioides CMW 22736 FJ411342 KM495402, =KM495579 =KM495489

Davidsoniella australis CMW2333 FJ411325 KM495396 KM495573 KM495483

Davidsoniella eucalypti CMW3254 FJ411327 KM495338 KM495515 KM495427

ame note CMW26365 -FJ411322. = KM495329 KM495506 KM495418

Endoconidiophora douglasii §CMW26367 NR_120295 KM495335 KM495512 KM495424

Graphium pseudormiticum CMW503 AY 148186 KM495390 KM495567. =KM495477

Huntiella chinaeucensis CMW24658 JQ862729 KM495327 KM495504 KM495416

Huntiella moniliformis CMW 10134 FJ151422 KM495355 = KM495532. =KM495443

Huntiella oblonga CMW23803 EU245019 KM495359 =9.KM495536 =9§ KM495447

Seychellomyces sinensis YMF1.04178 KU549178 MG830704 MK554714. MK554715

Thielaviopsis cerberus CMW36668 NR_145225 KM495326 KM495503 KM495415

Thielaviopsis musarum CMW 1546 JX518325 KM495357. = KM495534. KM 495445

Thielaviopsis paradoxa CMW36689 JX518342 KM495363, =KM495540 KM495451

Thielaviopsis radicicola CMW 1032 KF612023 KM495371 KM495548 KM495459

using a Bioteke DNA Gel/PCR Purification Kit and forward and reverse sequenced

with a LI-COR 4000L automatic sequencer, using a Thermo Sequenase-kit as

described by Kindermann & al. (1998).

394 ... Qiao, Zheng & al.

4007 CMW11536 C. eucalypticola

62|' CMW15049 C. fimbriata

100|_ CMW14802 C. platani Ceratocystis

top CMW22445 C. diversiconidia

98 CMW6579 C. pirilliformis

CMW22733 Ch. ovoidea

CMW22736 Ch. thielavioides

He. CMW49352 B. basicola

100 CMW6714 B. basicola

100|‘ CMW25439 B. basicola Berkeleyomyces

CMW14219 B. rouxiae

°°! CMW5472 B. rouxiae

YMF1.04178 Seychellomyces sinensis Seychellomyces

CMW1032 T. radicicola

106 CMW36668 T. cerberus Thie laviopsis

CMW1546 T. musarum

'°l__ CMW36689 T. paradoxa

“ soo CMW2333 D. australis

Ul ina CMW3254 D. eucalypti

CMW26365 E. coerulescens

CMW26367 E. douglasii

so07- CMW23803 H. oblonga

100 | CMW24658 H. chinaeucensis Huntiella

ree CMW10134 H. moniliformis

CBS110.61 A. xylebori Ambrosiella

ie Chalaropsis

Davidsoniella

100 Endoconidiophora

CMW503 G. pseudormiticum Graphium

0.07

aeoepiskoojelay

PLATE 1. MrBayes tree generated from a combined alignment of ITS, LSU, 60S, and MCM7

sequences of Ceratocystidaceae, with Graphium pseudormiticum (Microascaceae) as outgroup.

Sequence alignment and phylogenetic analysis

The sequence dataset contained the ITS, LSU, 60S and MCM7 sequence data for 21

species representing eight genera in the Ceratocystidaceae (Microascales). Graphium

pseudormiticum M. Mouton & M.J. Wingf. (Microascaceae) was chosen as outgroup

based on Nel & al’s (2018) phylogenetic study, and the sequence data were downloaded

from the NCBI GenBank database (TABLE 1).

Seychellomyces sinensis sp. nov. (China) ... 395

DNA sequence data were aligned using ClustalX 1.83 (Higgins 1994) with default

parameters, and the consensus sequences were adjusted manually and linked in

BioEdit v.7.0 (Hall 1999). MrBayes (Ronquist & Huelsenbeck 2003) was used to

calculate the concatenated sequence-based tree, with following parameters: ngen =

1,000,000; samplefr = 1000; printfr = 1000.

Phylogenetic results

The final alignment included a total of 2022 base pairs. The phylogenetic

tree was generated by combining the ITS, LSU, 60S, and MCM7 sequences and

using Bayesian analysis. The topology of the tree is shown with the bootstrap

support value for each node (PLATE 1). The tree supports Seychellomyces

within the Ceratocystidaceae; the genus does not group with any other genera

but is associated with the lineage (94% support) containing Berkeleyomyces,

Chalaropsis, and Ceratocystis in a basal position.

Taxonomy

Seychellomyces sinensis Z.F. Yu & Hua Zheng, PLATE 2

MB 829084

Differs from Seychellomyces hexagonus by its larger 1-3-septate conidia.

Type: China, Guangxi province, Shangsi, Shiwan Mountain, 21°34’49”N 109°14’20’E,

alt. 23 m, on decaying leaf of an unidentified broadleaf tree, 7 July 2013, Z.F. Yu.

(Holotype, YMFT 1.04178 [permanent slide]; ex-type living culture, YMF 1.04178;

GenBank KU549178, MG830704, MK554714, MK554715).

EryMo.oey: From the Latin, sinensis, for the country of origin, China.

COLONIES attaining 3 cm diam. after 7 days on CMA, effuse, pale brown

to brown. Mycelium sparse, hyphae branched, septate, hyaline or pale

brown. CONIDIOPHORES arising from the creeping hyphae, cylindrical,

2(-5)-septate, erect, unbranched, smooth, 12-40 x 5-7 um (narrowing to

3.5-5 um at the apex). CONIDIOGENOUS CELLS monoblastic, integrated,

terminal, determinate. Conip1A ellipsoidal, brown to pale brown, smooth or

sometimes with a warty surface, growing singly or 2-3 in chains, 1-3-septate

(mainly 2-septate), with short (4-13 um long) cylindrical conidiophore

remnants at both ends, in cross-section hexagonal (occasionally pentagonal

or quadrangular). ENDOCONIDIA originating endogenously within phialides,

catenulate, variable in length, hyaline or subhyaline, elliptical to cylindrical,

truncate at ends, occasionally repetitiously germinating by producing a

short conidiophore from which a phialide is formed to produce a chain of

endoconidia. CHLAMYDOSPORES ochraceous, globose, smooth, 5-10 um in

diam.

396 ... Qiao, Zheng & al.

CONIDIAL SIZES ON DIFFERENT MEDIA—ON V-8 JUICE AGAR: 1-septate,

20-23 x 12-16 um, total length (including short cylindrical conidiophore

remnants) 34-39 tm; 2-septate, 29-39 x 16-21 um, total length 40-56 um;

3-septate 35-49 x 11-15 um, total length 55-35 um; endoconidia 5-21 x

3=5;/-pams

On PDA: 1-septate, 20-27 x 13-21 um, total length 31-45 um; 2-septate,

38-50 x 19-30 um, total length 42-69 um; 3-septate 38-56 x 19-27 um, total

length 53-73 um; endoconidia 7-39 x 3-5 um.

On CMA: 1-septate, 23-30 x 15-21 um, total length 34-47 um; 2-septate,

31-51 x 19-31 um, total length 42-70 um; 3-septate, 42 x 20 um, total length

58 um; endoconidia 6-42 x 3-5 um.

Discussion

Seychellomyces has been treated as monotypic since the genus was

established in 1981. Based on characteristic of conidia and endoconidia,

our strain was easily identified to Seychellomyces. However, S. sinensis is

distinguished from the type species, S. hexagonus, by having 1-3-septate

conidia, while conidia of S. hexagonus are always 2-septate. In

addition, conidia of S. hexagonus are always hexagonal in cross-section

(Matsushima 1981), whereas those of S. sinensis are mostly hexagonal,

but occasionally pentagonal or quadrangular. The size of conidia of

S. sinensis differs based on growth medium and conidial septation; but

on V-8 juice agar medium the 2-septate conidia of S. sinensis are larger

than those of S. hexagonus (25-33 x 14-18 um; Matsushima 1981).

Globose chlamydospores were observed in culture of S. sinensis, but not

mentioned in S. hexagonus.

Among Ceratocystidaceae, only Thielaviopsis sensu lato has a

superficial resemblance to Seychellomyces in conidial shape (Seifert & al.

2011). However, Thielaviopsis, Chalaropsis, and Ceratocystis significantly

differ from S. sinensis phylogenetically.

Acknowledgments

This work was financed by the National Natural Science Foundation Program

of the PR China (31770026, 31570023). We are very grateful to Dr. Rafael F.

Castafieda-Ruiz (INIFAT Alejandro de Humboldt, Havana, Cuba) and Prof. Jun-

Zhi Qiu (Fujian Agriculture & Forestry University, Fuzhou, China) for critically

reviewing the manuscript and providing helpful suggestions to improve this paper,

and to Dr. Shaun Pennycook and Dr. Lorelei Norvell for their critical review and

suggestions.

Seychellomyces sinensis sp. nov. (China) ... 397

PLATE 2. Seychellomyces sinensis (holotype, YMF 1.04178): A-C. Conidia; D, E. Conidia with

conidiophores; F. Chlamydospores; G. Cross-section of conidia. H. Endoconidia. I. Conidiogenous

cells of endoconidia. Scale bars: A-C, F, H, I = 10 um; D, E = 20 um; G = 100 um.

398 ... Qiao, Zheng & al.

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Mycotaxon. 134:177-181. https://doi.org/10.5248/134.177

MY COTAXON

April-June 2019—Volume 134, pp. 399-406

https://doi.org/10.5248/134.399

Ramaria flavescentoides sp. nov. with clamped basidia

from Pakistan

MUHAMMAD HAnIkF’*, ABDUL NASIR KHALID’, RONALD L. EXETER?

' Department of Botany, Government College University,

Lahore, 54600, Pakistan

? Department of Botany, University of the Punjab,

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

* Dallas, Oregon, 97338, USA

* CORRESPONDENCE TO: dr.mhanif @gcu.edu.pk

AsBsTRACT—During an investigation of macrofungi from Pakistan, a new species with

clamped basidia within Ramaria subg. Laeticolora and associated with Abies pindrow was

found, which is proposed here as Ramaria flavescentoides. The new fungus morphologically

resembles R. flavescens and clusters phylogenetically within the same clade. Ramaria

flavescentoides is characterized by an obovate to obconic basidioma with pale orange to light

orange branches and apices that are tinged light greenish grey, a monomitic hyphal system,

ampulliform clamp connections, stellate crystals distributed within the upper branches,

multiguttulate rhizomorphic strands, and oblong to ellipsoid verrucose basidiospores.

Key worps—Himalayan moist temperate forest, ITS region, molecular identification,

polychotomous branching

Introduction

Ramaria Fr. ex Bonord. (Ramariaceae) is a widespread non-gilled

basidiomycete genus comprising nearly 220 species worldwide (Kirk & al.

2008). The basidiomata, which generally repeatedly branch dichotomously,

are usually divided into stipe, branches, and apices. Important diagnostic

characters include color, stipe consistency (fibrous vs gelatinous), odor,

staining reactions to reagents, presence or absence of clamp connections,

and spore size and ornamentation (Exeter & al. 2006). Villegas & al. (1999)

AOO ... Hanif, Khalid, Exeter

placed Ramariaceae in Gomphales, a placement also supported by molecular

phylogenetic studies (Hibbett & al. 1997, Bruns & al. 1998, Humpert & al.

2001, Giachini 2004). Ramaria, which includes both saprobic and

ectomycorrhizal species, is currently divided into four subgenera: R. subg.

Echinoramaria Corner, R. subg. Lentoramaria Corner, R. subg. Ramaria,

and R. subg. Laeticolora Marr & D.E. Stuntz (Marr & Stuntz 1973, Exeter

& al. 2006). Previously these divisions were based on spore ornamentation,

monomitic vs. dimitic rhizomorphic strands, humicolous vs. lignicolous

substrate, and presence or absence of clamp connections.

Ramaria flavescentoides is described here based on morphological and

molecular data as a new species in R. subg. Laeticolora. Both R. subg. Laeticolora

and R. subg. Ramaria tend to develop larger fruiting bodies than R. subg.

Echinoramaria and R. subg. Lentoramaria. Ramaria subg. Laeticolora is further

characterized by non-striate basidiospores that are almost smooth to distinctly

ornamented at 1000x (vs. striate in R. subg. Ramaria), tissues that are primarily

non-amyloid (vs. amyloid in R. subg. Ramaria), single to fasciculate stipe bases,

usually brightly colored branches and apices, and a terrestrial habit (Exeter &

al. 2006).

Materials and methods

Collections

The sampling site was selected based on the richness and abundance of

aphyllophoralean fungi in an area of probable high basidiomycete biodiversity in

a moist temperate coniferous forest in Khyber Pakhtunkhwa, Pakistan where air

temperatures range from -4 to 25°C, and the soil is loamy with gravel and variably

sized stones. Field notes and photographs were made of all collections, and the

specimen was morphologically characterized following Corner (1950). Features

(stipe, branches, rhizomorphic hyphae) were measured microscopically at 1000x

using a compound microscope equipped with an ocular micrometer. Measurements

of basidiospores, basidia, cystidia, and rhizomorphic hyphae were determined from

at least 20 measurements per character. Small (~1 cm) portions of the hymenium

were stored in 2% CTAB buffer in 1.5 ml Eppendorf vials and stored at —20°C for

molecular analysis; the remaining material was dried using a fan heater and stored

in sealed plastic bags. The collections are curated at the mycological herbarium at

the University of the Punjab, Lahore, Pakistan (LAHMH).

DNA extraction, PCR amplification & sequencing

Hymenial tissue was removed with sterile forceps for DNA extraction by a CTAB

method following Gardes & Bruns (1993), modified for Gene-Clean silica emulsion

binding and purification. The polymerase chain reaction (PCR) followed Gardes &

Ramaria flavescentoides sp. nov. (Pakistan) ... 401

Mi Ramaria flavescentoides MG760617

@ Ramaria flavescentoides MG760618

@ Ramaria flavescentoides MG760616 [T]

@ Ramaria flavescentoides MG760615

@ Ramaria flavescentoides MG760614

@ Ramaria flavescentoides MG760613

24 @ Ramaria flavescentoides MG760612

Ramaria sp. DQ974713 Clade | [Clamped Basidia]

94 , Ramaria flavescens AJ408366

77 Ramaria flavescens AJ408365

58 Ramaria rasilisporoides JQ408242

99 - Ramaria flavoides AJ408381

Ramaria flavoides AJ408380 Subgenus Laeticolora

99 Ramaria largentii EU652344

82 Ramaria maculatipes EU669390

92 Ramaria ignicolor AJ408386

Ramaria conjunctipes EU846301

Ramaria mediterranea AJ408370

85 , Ramaria araiospora EU837208

99 [TL Ramaria sp. DQ365620 Clade II [Non-Clamped Basidia]

Ramaria araiospora EU852809

30 Ramaria sp. DQ365603

Ramaria cystidiophora EU597077

93 |Ramaria cystidiophora DQ384590

Ramaria formosa DQ365648

99 Ramaria flaccida AJ408390

89 Ramaria flaccida AJ408371 peo Echinoramaria

34 Ramaria curta AJ408358

Ramaria stricta EU819419

Ramaria stricta DQ367910 Clade Ill

Ramaria rubella AY854078

Ramaria abietina FJ627035

51 Ramaria gracilis AJ408378

g9 + Ramaria gracilis EU258553

Ramaria rubripermanens AJ408368

Ramaria botrytis AF377055

Ramaria concolor f. marrii AF213110

Ramaria rainierensis AY574694

Ramaria eumorpha AF139973

Ramaria botrytis EU652385

Ramaria rubrievanescens AF213061

93 | Ramaria rubrievanescens AF213060

Clade IV

0.1

Fic. 1. Maximum Likelihood phylotree of the evolutionary history of ITS sequences of Ramaria

species, including R. flavescentoides. The tree with the highest log likelihood (-10063.0715)

is shown, drawn to scale with branch lengths indicating the number of substitutions per site.

Bootstrap support is shown next to the branches.

Bruns (1993), using the fungus-specific ITS1F primer (CTTGGTCATTTAGAGGAAGT)

and the eukaryotic ITS4 primer (TCCTCCGCTTATTGATATGC) to amplify the nuclear

rDNA-ITS region. The hot-start enzyme JumpStart catalyzed the PCR with 2 min

at 94°C; followed by 30 cycles of 30 s at 94°C, 30 s at 53°C, and 40s + 5s per cycle

at 72°C; and finishing with 5 min at 72°C. The PCR products were purified with

QIAquick, sequenced bi-directionally using the reverse and forward primers and

BigDye 3.1 on an ABI 3730 DNA sequencer, and edited in Sequencher 4.5 by Jodrell

Laboratory, Royal Botanical Gardens, Kew, UK. The DNA sequences were BLAST

searched at NCBI (https://www.ncbi.nlm.nih.gov/) and submitted to GenBank.

Sequences were aligned and corrected manually for phylogenetic analysis with

MacClade 4.08. Percentage identities were calculated using DNAStar software.

402 ... Hanif, Khalid, Exeter

Phylogenetic analysis

The rDNA ITS sequences were aligned with sequences of other Ramaria species

selected from GenBank. The original dataset comprising 1108 characters including

gaps was trimmed by 252 characters excluded from 3’ and 5’ ends of the sequences. All

gaps were treated as missing data. Of the 856 characters (of ‘unord’ type and equally

weighted) analysed, 202 characters were constant, 616 were parsimony-uninformative,

and 507 were parsimony-informative. The Branch-and-Bound search method was

used for bootstrapping with 1000 bootstrap replicates in MEGA5 (Tamura & al. 2011).

Results

ITS rDNA-based identification and phylogenetic analysis

The amplification of the rDNA ITS from Ramaria flavescentoides produced

630-665 bp fragments. The consensus sequence of R. flavescentoides showed

91% identity and 65% query coverage with an undescribed Ramaria sp.

(DQ974713.1) and 88% identity and 75% query coverage with R. flavescens

(AJ408365).

Species included in this study were selected from all four Ramaria

subgenera based on the diagnostic characters of each subgenus. The

maximum likelihood phylogram represents four clades (Fic. 1). Clade I

represents species belonging to R. subg. Laeticolora with clamped basidia,

Clade II represents species belonging to R. subg. Laeticolora with non-

clamped basidia, and Clades III and IV represent species belonging to

R. subg. Echinoramaria, R. subg. Lentoramaria, and R. subg. Ramaria.

The Clade I sequences from R. flavescentoides clustered with those from

R. flavescens, R. flavoides, R. largentii, R. maculatipes, R. rasilisporoides, and

Ramaria sp. (bootstrap = 77%). Within the clade, R. flavescentoides was

sister to R. flavescens, R. rasilisporoides, and Ramaria sp. (bootstrap = 94%).

Percentages of genetic characters shared by R. flavescentoides with other

species were: Ramaria sp.—83%, R. flavescens—82%, R. largentii—81%,

R. rasilisporoides—79%, R. maculatipes—78%, and R. flavoides—77%.

Taxonomy

Ramaria flavescentoides Hanif & Khalid, sp. nov. Fic. 2

MB 823965

Differs from Ramaria flavescens by its obconic to obovate basidioma, its non-bulbous

fasciculate stipe, and the presence of stellate crystals in the upper branches.

Type: Pakistan. Khyber Pakhtunkhwa province, Hazara Division, Ayubia—Khanspur,

34°01’24”’N 73°24’14’E, 2159 m asl, under Abies pindrow, 8 August 2009, M. Hanif

MH88 (Holotype, LAHMH0889; GenBank MG760616).

EryMoLoGy—Named for its macroscopic resemblance with R. flavescens.

Ramaria flavescentoides sp. nov. (Pakistan) ... 403

Fic. 2. Ramaria flavescentoides (holotype, LAHMH0889). A. Basidiocarps; B. Basidiospores;

C. Basidia and cystidia; D. Stipe tramal hyphae; E. Overlapped cellular hyphae of upper

branches; F. Ampulliform tramal hyphae of upper branches; G. Stellate crystals among stipe

hyphal mass; H. Cellular hyphae of upper branches; I. Tramal elements of upper branches;

J. Dimitic hyphae of rhizomorph. Scale bars: A = 2 cm; B, C = 3 um; D, G = 15 um; E, H = 17 um;

F = 13.5 um; I = 6 um; J = 37.5 um.

BASIDIOMA 13-15 cm high, <10 cm broad, repeatedly branched, usually

obovate to obconic in outline; orange to light orange on drying. STIPE <3(-

404 ... Hanif, Khalid, Exeter

4) cm high, compound to fasciculate in groups of 2-5, orange white at the

base, upward pale orange, tapering gradually downward into a basal mycelial

mat or white tomentum covering the stipe base. RHIZOMORPHS white,

involving significant amounts of substrata, drying light orange. FLESH white,

solid, gelatinous or slippery, dried flesh firm but easily penetrated. Mayor

BRANCHES 4-6, 1.5-2 cm thick, stout, more or less terete, ascending, orange

white below, upward pale. BRANCHES 4—7 ranks, ascending, polychotomous,

smooth and gelatinous from exterior, pale orange; axils more or less rounded.

ApicEs 2—4 mm long, acute to blunt, light orange up to the tips with light

greenish grey tint. TASTE, ODOR, and MACROCHEMICAL REACTIONS not

recorded.

STIPE TRAMAL HYPHAE 6-11 um wide, hyaline, thin-walled, parallel,

slightly ampulliform, occasionally clamped, grainy encrusted with crystalline

material of various shapes; gloeoplerous hyphae not observed. TRAMAL

HYPHAE OF UPPER BRANCHES 6-8 um diam., hyaline, thin-walled, parallel,

ampulliform; skeletal generative hyphae present, clamps occasional, stellate

crystalline material 32 um diam. HYPHAE OF BASAL MAT monomitic, 3-4 um

diam., thin-walled, branched, clamped, 6 um thick at clamps, grainy

encrusted with crystalline material of various shapes; generative hyphae

7-10 um diam. RHIZOMORPHIC HYPHAE smooth to encrusted, diameters

vary. SUBHYMENIUM rudimentary, hyphal structure is monomitic with thin-

walled and thick-walled hyphae of various diameters, overlapped, septate;

frequent ampulliform hyphae (27 x 15 um). Hymenium thick during

maturation. BAsIDIA 32—50(—70) x 9-12 um, clavate, clamped; contents

homogeneous when young, slightly grainy, hyaline or slightly yellowish,

slightly cyanophilous, 4-spored; sterigmata 6-8 um high. BASIDIOSPORES

(10.7-)11—13(-15) x (4.8-)5—6(—7) um, verrucose, multiguttulate when

mature, oblong to ellipsoid, slightly curved at one end, contents weakly

cyanophilous; wall moderately thick; ornamentation strongly cyanophilous,

roughened outline of low coarse warts and delicate meandering ridges

covering significant areas of wall.

HABITAT & DISTRIBUTION—Terrestrial, solitary, under Abies pindrow.

Known only from the type area, Ayubia-Khanspur, fruiting in July-August

at 1900-2200 m asl.

ADDITIONAL SPECIMENS EXAMINED—PAKISTAN. KHYBER PAKHTUNKHWA

PROVINCE, Hazara Division, Ayubia~Khanspur, 34°01’24”N 73°24’14”E, 2159 m asl,

17 Aug 2009, M. Hanif MH136 (LAHMH1789.1; GenBank MG760618); Helipad,

34°01’31”N 73°25'19”E, 1974 m asl, 25 Jul 2008, M. Hanif MH253 (LAHMH2578.2;

GenBank MG760612); M. Hanif MH254 (LAHMH2578.3; GenBank MG760617);

Ramaria flavescentoides sp. nov. (Pakistan) ... 405

M. Hanif MH96 (LAHMH2578.5; GenBank MG760615); M. Hanif MHI101

(LAHMH2578.1; GenBank MG760613); 16 Aug 2009, M. Hanif MH97 (LAHMH 1689;

GenBank MG760614).

CoMMENTS—Ramaria flavescentoides is characterized by medium sized

obovate to obconic basidiomata with pale orange to light orange colored

branches and lightly tinged greenish grey apices and a compound to

fasciculate stipe that narrows gradually downward into a mat of white

mycelia and rhizomorphic strands.

The new species resembles R. flavescens (Schaeff.) R.H. Petersen in

shape, branching pattern, and basidioma color, also sharing a similar spore

shape and ornamentation and the presence of basidial clamp connections.

However, our specimens differ morphologically from R. flavescens, which

produces smaller (10 x 7 cm) obpyriform profusely branched basidiomata

with a rather abrupt ball surmounting the stipe and smaller basidiospores

(11-12.5 x 4.8-5.6 um; Petersen 1974). The presence of stellate crystalline

material in the hyphae of upper branches is another important character of

R. flavescentoides. Similar crystals have been reported by Zhang & al (2005)

in the stipe and upper branches of Ramaria luteoaeruginea P. Zhang & Zhu

L. Yang in R. subg. Echinoramaria. Phylogenetically, R. flavescentoides nests

adjacent to R. flavescens; and R. flavescentoides and R. flavescens cluster

together in clade I with other species having clamped basidia (Fic. 1).

Ramaria flavescentoides shares similar branching pattern, clamped

basidia, warted (verrucose) spores with morphological similarities with

its phylogenetic sister species, R. rasilisporoides Exeter, which clusters in

the same clade with strong (94%) bootstrap support (Fic. 1). Ramaria

rasilisporoides differs from R. flavescentoides in its pale yellow basidiomata,

amyloid stipe tissues, deeply rooting stipe with numerous abortive

branchlets, and smaller basidiospores (Exeter & al. 2006).

Ramaria mairei Donk, the only species of subg. Laeticolora previously

reported from Pakistan (Ahmad 1972; Ahmad & al. 1997), is distinguished

by larger (18 cm) basidiomata, a 8 cm high stipe, numerous pale or deep

lemon to pale ochraceous and rather lax or fairly crowded branches, and

slightly longer (10-15-um) oblong to ellipsoid roughened pale ochraceous

basidiospores (Ahmad 1972).

Phylogenetic analysis also supports the morphological identification.

Ramaria flavescentoides clusters with the clamped species of subg.

Laeticolora. This clade seems to be paraphyletic and originated with non-

clamped species of this subgenus with unsupported bootstrap frequency.

A406 ... Hanif, Khalid, Exeter

Humpert & al. (2001) reported the genus Ramaria as polyphyletic, which

our analyses support.

Acknowledgments

The authors would like to thank Dr. Ping Zhang (Hunan Normal University,

China) and Dr. Sana Jabeen (University of Education, Lahore, Pakistan) for their

helpful comments and critical review of this manuscript.

Literature cited

Ahmad S. 1972. Basidiomycetes of Pakistan. Biological Society of Pakistan. Monograph 6, Lahore.

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

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

Bruns TD, Szaro TM, Gardes M, Cullings KW, Pan JJ, Taylor DL, Horton TR, Kretzer A, Garbelotto

M, Li Y. 1998. A sequence database for the identification of ectomycorrhizal basidiomycetes by

phylogenetic analysis. Molecular Ecology 7: 257-272.

https://doi.org/10.1046/j.1365-294X.1998.00337.x

Corner EJH. 1950. A monograph of Clavaria and allied genera. Annals of Botany Memoirs.

1. 740 p.

Exeter RL, Norvell L, Cazares E. 2006. Ramaria of the Pacific Northwestern United States.

United States Department of Interior, Bureau of Land Management: Salem, Oregon USA.

156 p.

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

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

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

Giachini AJ. 2004. Systematics of the Gomphales: the genus Gomphus Pers. sensu lato. PhD

Dissertation, Oregon State University, Corvallis, OR, USA

Hibbett DS, Pine EM, Langer E, Langer G, Donoghue MJ. 1997. Evolution of gilled

mushrooms and puffballs inferred from ribosomal DNA sequences. Proceedings of

the National Academy of Sciences of the United States of America 94: 12002-12006.

https://doi.org/10.1073/pnas.94.22.12002

Humpert AJ, Muench EL, Giachini AJ, Castellano MA, Spatafora JW. 2001. Molecular phylogenetics

of Ramaria and related genera: evidence from nuclear large subunit and mitochondrial small

subunit rDNA sequences. Mycologia 93: 465-477. https://doi.org/10.2307/3761733

Kirk PM, Cannon PF, Minter DW, Stalpers JA. 2008. Ainsworth & Bisby’s dictionary of the fungi.

10th edn. CAB International, U.K. https://doi.org/10.1079/9780851998268.0000

Marr CD, Stuntz DE. 1973. Ramaria of western Washington. Bibliotheca Mycologia 38. 232 p.

Petersen RH. 1974. Contribution toward.a monograph of Ramaria. 1.Some classic species redescribed.

American Journal of Botany 61: 739-748. https://doi.org/10.1002/j.1537-2197.1974.tb12296.x

Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S. 2011. MEGAS5: molecular

evolutionary genetics analysis using maximum likelihood, evolutionary distance, and

maximum parsimony methods. Molecular Biology and Evolution 28(10): 2731-2739.

https://doi.org/10.1093/molbev/msr121

Villegas M, de Luna E, Cifuentes J, Torres AE. 1999. Phylogenetic studies in Gomphaceae sensu

lato (Basidiomycetes). Mycotaxon 70: 127-147.

Zhang P, Yang ZL, Ge ZW. 2005. Two new species of Ramaria from southwestern China.

Mycotaxon 94: 235-240.

MYCOTAXON

April-June 2019—Volume 134, p. 407

https://doi.org/10.5248/134.407

Regional annotated mycobiota new to the Mycotaxon website

ABSTRACT—In May, Mycotaxon added the following new annotated species distribution

list to our 132 previously posted free-access fungae. The 24-page “Aphyllophoroid fungi in

Teide National Park (Tenerife, Canary Islands)” by Esperanza Beltran-Tejera, Jesus Laura

Rodriguez-Arma, Miguel Jonathan Diaz Armas, and Luis Quijada may be downloaded

from our website via http://www.mycotaxon.com/mycobiota/index.html

EUROPE

Portugal & Spain (Iberian Peninsula)

ESPERANZA BELTRAN-TEJERA, JESUS LAURA RODRIGUEZ-ARMA, MIGUEL

JONATHAN Diaz Armas, Luis QuyapA. Aphyllophoroid fungi in Teide

National Park (Tenerife, Canary Islands). 24 p.

ABSsTRACT—Data on aphyllophoroid fungi in Teide National Park Tenerife are

summarized, and 102 species are recorded. Twenty eight species are new to Tenerife,

out of which 17 are also new records for the Canary Islands (Athelia pyriformis,

Cartilosoma ramentaceum, Ceriporia excelsa, Dendrocorticium lilacinoroseum,

Hyphoderma sibiricum, Hyphodermella rosae, Hyphodontiella multiseptata,

Melzericium bourdotii, Phanerochaete avellanea, Phanerochaete cremeo-ochracea,

Phlebia griseoflavescens, Phlebia lacteola, Phlebia lilascens, Phlebia_tuberculata,

Sistotrema pistilliferum, Skeletocutis amorpha, and Tubulicrinis angustus). The list

is supplemented with 7 species of Peniophora recorded for this area, published

previously (Diaz Armas et al. 2019).

Mycobiota of the two highest mountain areas of the Canary Islands (Teide

National Park in Tenerife and Taburiente National Park in La Palma) are compared

regarding their richness of genera, species, abundance and substrates. Aphyllophoroid

fungi were found on 16 endemic vascular species, the majority were on Spartocytisus

supranubius, which is dominant in both abundance and distribution in the study area.

MYCOTAXON

April-June 2019—Volume 134, pp. 409-412

https://doi.org/10.5248/134.409

BOOK REVIEWS AND NOTICES:

ELSE C. VELLINGA

861 Keeler Avenue, Berkeley CA 94708 USA

ABSTRACT—Books reviewed include Syllabus of Plant Families—A. Engler’s

Syllabus der Pflanzenfamilien. 1/3 Basidiomycota and Entorrhizomycota by Begerow,

McTaggart, and Agerer (W. Lang ed.) and Fungi lepiotoidei (Agaricaceae) edited by

Zhu-Liang Yang (2019).

GENERAL

Syllabus of Plant Families—A. Engler’s Syllabus der Pflanzenfamilien. 1/3

Basidiomycota and Entorrhizomycota. 13" ed. By Wolfgang Frey (ed.)

2018. Borntraeger Science Publishers, Johannesstr. 3A, 70176, Stuttgart, Germany.

xiit+471p. 43 figs. ISBN 978-3-443-01098-0. Price (excl. postage): € 139. www.

borntraeger-cramer.com/9783443010980

Two years after the volume on Ascomycota (Jaklitsch & al. 2016), the

current work was published, giving an overview of the Basidiomycota and

the Entorrhizomycota.

Only three authors wrote the text: Dominik Begerow and Alistair

McTaggart took care of the Pucciniomycotina, Ustilagomycotina, and

Entorrhizomycota, whereas Reinhard Agerer is the author for the

Agaricomycotina.

This is a systematic overview, incorporating the newest insights and

giving standardized descriptions for all genera in these groups. As such it

replaces long out-of-date works such as Singer’s 1986 monumental “The

* Book reviews or books for consideration for coverage in this column should be sent to the

EDITOR-IN-CHIEF <editor@mycotaxon.com> 6720 NW Skyline, Portland OR 97229 USA.

A410 ... MycoTaxon 134

. . b>]

Wottgang Frey cate) “84ricales in modern taxonomy,’ although

the present work lacks keys and infrageneric

Syl labus a "* classifications and is much wider in scope.

of Plant Families

J, Feeder’ Syleinac der Plenzenieniien I am impressed with the morphological

descriptions provided, which present a

1/3 Basidiomycota and Entorrhizomycota wealth of knowledge.

an? i ;

Ms have not been used in decades, but this does

It is always a bit of a balancing act which

genera to include, as modern data are not

available for all genera. There are names that

not mean that they do not represent good

solid genera. In some genera, most species—

but not all—have been transferred to a

more fitting genus, such as is the case for Torrendia. But as one Australian

Torrendia species has not yet been combined in Amanita, the genus is still

treated here separately.

Unfortunately, there are quite a number of strange mistakes, such

as the statement that Saproamanita species are ectomycorrhizal. This

genus was explicitly erected to accommodate the non-ectomycorrhizal

species in Amanita, as is also reflected in the genus name (Redhead &

al. 2016). Another example is Notholepiota, which here is still included

in the Agaricaceae, although it was shown to be based on a secotioid

bolete species, now called Boletus semigastroideus (Nuhn & al. 2013).

Some genera, such as Bonomyces and Cruentomycena are not included, for

unknown reasons.

Therearealso differences between the treatments of the Pucciniomycotina

and Ustilagomycotina on the one hand, and the Agaricomycotina on the

other. For instance, in Agaricomycotina the type species of a genus is only

given for monotypic genera but is always cited for genera in the other two

subphyla. Discussion on genus borders etc. are also given for the genera in

the first two subphyla, and absent in the Agaricomycotina.

Of course, the description of new families and genera did not grind

to a halt when this work was sent to the publisher. Mythicomycetaceae

(Vizzini et al. 2019) and Sarcoporiaceae (Audet 2018) have recently been

distinguished at the family level, and Zhulangiomyces (Redhead 2019)

and Clitopaxillus (Alvarado & al. 2018) are just two examples of recently

published genera in the Agaricales.

Book Reviews ... 411

This overview is an especially useful and thorough piece of work that

hopefully will be the standard for years to come. As such the SYLLABUS 1/3

deserves a place on many mycologists’ bookshelves.

Alvarado P, Moreau P-A, Dima B, Vizzini A, Consiglio G, Moreno G, Setti L, Kekki T,

Huhtinen S., Liimatainen K, Niskanen T. 2018. Pseudoclitocybaceae fam. nov.

(Agaricales, Tricholomatineae), a new arrangement at family, genus and species level.

Fungal Diversity 90: 109-133. https://doi.org/10.1007/s13225-018-0400-1

Audet S, 2018. Sarcoporiaceae. Mushrooms nomenclatural novelties no. 18: 1-6.

Jaklitsch W, Baral H.-O, Licking R, Lumbsch HT, Frey W (eds). 2016. Syllabus of Plant

Families. A. Engler’s Syllabus der Pflanzenfamilien Part 1/2: Ascomycota.

Nuhn ME, Binder M, Taylor AFS, Halling RE, Hibbett DS, 2013. Phylogenetic overview

of the Boletineae. Fungal Biology 117: 479-511. https://doi-org/10.1016/j.funbio

Redhead SA, 2019. Nomenclatural novelties. Index Fungorum 385: 1.

Redhead SA, Vizzini A, Drehmel DC, Contu M. 2016. Saproamanita,a new name for both

Lepidella E.-J. Gilbert and Aspidella E.-J. Gilbert (Amaniteae, Amanitaceae). IMA

Fungus 7: 119-129. https://doi.org/10.5598/imafungus.2016.07.01.07

Singer, R. The Agaricales in modern taxonomy. Ed. 4. Koeltz Scientific Books, Koenigstein,

Federal Republic of Germany, 908 p., 88 pl.

Vizzini A., Consiglio G., Marchetti M., 2019. Mythicomycetaceae fam. nov. (Agaricineae,

Agaricales) for accommodating the genera Mythicomyces and Stagnicola, and

Simocybe parvispora reconsidered. FUSE 3: 41-56. https://doi.org/10.3114/

fuse.2019.03.05

BASIDIOMYCETES

Fungi lepiotoidei (Agaricaceae). By Zhu-Liang Yang (ed.). 2019. Flora

FUNGORUM SINICORUM VOL. 52. http://www.sciencep.com Science Press,

Beijing, China. 228 p. 2 colour plates, 112 black & white figures, Hardcover.

ISBN 978-7-03-060680-8. Price (excl. postage): 198.00 ¥; cited at $76 by

www.chinascientificbooks.com

This 52nd volume in the series that aims

at providing keys and descriptions of all

fungi in China focuses on the lepiotoid

fungi, to be more precise, all agaricoid

genera in the Agaricaceae except Agaricus

and Micropsalliota, a total of 14 genera.

Most of these are white-spored, but some

species in Chlorophyllum and the genera

Heinemannomyces, Hymenagaricus, and

Melanophyllum have coloured basidiospores.

412 ... MycoTAXON 134

An introduction to the group gives an overview of the recent literature,

classification and phylogeny of the group (which represents only part of a

monophyletic clade), and the morphology.

After the key to the genera, the genera are treated in alphabetical order,

with keys to species, descriptions, and high-quality line drawings of all

species that have been confirmed for the country. All collections on which

the descriptions are based are listed. Those species listed in earlier works

whose presence is questionable are briefly discussed. For 15 species, colour

photos are provided. An extensive list of references and an index complete

the work.

This book is the result of 15 years of work by Prof. Zhu-Liang Yang and

his students (now professors themselves) Zai- Wei Ge and Jun-Feng Liang.

Lepiota has the highest number of taxa (48)—it interesting is to see

how some species have a distribution area spanning from western Europe

throughout the Northern Hemisphere into China, whereas others,

especially those from southern, tropical China, have a much more limited

distribution. Thanks to DNA comparisons of collections from Europe,

North America and China, real distribution patterns emerge. In the past,

when neither these techniques nor literature were available, names were

adopted indiscriminately from other regions.

Some species listed under Lepiota (L. shixingensis and L. furfuraceipes)

belong to Leucoagaricus. Hymenagaricus taiwanensis is now considered in

the genus Xanthagaricus.

The last word on the Chinese lepiotoids has not been said yet—I am

convinced that there are more species in Leucoagaricus than presented

here, but that does not diminish the value of the present work, a value that

extends far beyond China. The only drawback, of course, is the language,

as the book is written in Chinese, but fortunately the names are presented

in Latin.