MYCOTAXON
THE INTERNATIONAL JOURNAL OF FUNGAL TAXONOMY & NOMENCLATURE
VOLUME 124 APRIL-JUNE 2013
Chaetomium jatrophae sp. nov. (Sharma & al. — PLATE 2, p. 122) ROHIT SHARMA, artist
ISSN (PRINT) 0093-4666 http://dx.doi.org/10.5248/124 ISSN (ONLINE) 2154-8889 MYXNAE 124: 1-368 (2013)
EDITORIAL ADVISORY BOARD
WEN-YING ZHUANG (2003-2014), Chair Beijing, China HENNING KNUDSEN (2008-2013), Past Chair Copenhagen, Denmark Scott A. REDHEAD (2010-2015) Ottawa, Ontario, Canada SABINE HUHNDORE (2011-2016) Chicago, Illinois, U.S.A. PETER BUCHANAN (2011-2017) Auckland, New Zealand
KAREN HANSEN (2013-2018) Stockholm, Sweden
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MYCOTAXON
THE INTERNATIONAL JOURNAL OF FUNGAL TAXONOMY & NOMENCLATURE
VOLUME 124
APRIL-JUNE, 2013
EDITOR-IN-CHIEF
LORELEI L. NORVELL
editor@mycotaxon.com Pacific Northwest Mycology Service 6720 NW Skyline Boulevard Portland, Oregon 97229-1309 USA
NOMENCLATURE EDITOR
SHAUN R. PENNYCOOK
PennycookS@LandcareResearch.co.nz Manaaki Whenua Landcare Research Auckland, New Zealand
BooK REVIEW EDITOR
ELSE C. VELLINGA
bookreviews@mycotaxon.com 861 Keeler Avenue Berkeley CA 94708 U.S.A.
CONSISTING OF I-XII + 368 PAGES INCLUDING FIGURES
ISSN 0093-4666 (PRINT) http://dx.doi.org/10.5248/124.cvr ISSN 2154-8889 (ONLINE)
© 2013. MycoTAxon, LTD.
Iv ... MYCOTAXON 124
MY COTAXON
VOLUME ONE HUNDRED TWENTY-FOUR — TABLE OF CONTENTS
COVER SECTION
CEP PELEE ps tech ono Rt Hl vega eRe eect ASicck OGRE eho RI es Re FeA Ten At EL ees cicck “PA aen ad vii a chy Fea gee The ha a Micah ES I Sea eee Mal SET ed, WAN to Slay rey PU Sry RAS ib SUDMESSION PIO CCHUTES 3 25 vs Pearich Youbet vss Red i 2. cae aps Mayme mechanic ok a gs Cah x OP EPEIOCE LOT Roots B Fhe Ayo. tons hE Se eS Se a oy SY 3 eon ay Re xi
RESEARCH ARTICLES
A new Myrmecridium species from Guizhou, China CHUN- YU JIE, QING-XIN ZHOU, WEN-SHENG ZHAO, YU-LAN JIANG, KEVIN D. Hype, Eric H.C. MCKENZIE & YONG WANG The occurrence of Rutstroemia coracina on Quercus ilex leaves in Spain RICARDO GALAN, FRANCISCO PRIETO-GARCIA, ANGELES GONZALEZ & CARLOS ENRIQUE HERMOSILLA Two species of Cladosporium associated with wood discoloration in Korea YEONGSEON JANG, YOUNG MIN LEE, GyU-HYEOK KIM & JAE-JIN KIM Phialophora avicenniae sp. nov., a new endophytic fungus in Avicennia marina in China YUE-LIAN Liu, PING-GEN XI, XIAO-LAN HE & ZI-DE JIANG Ramichloridium strelitziae associated with sooty blotch and flyspeck on Ravenala madagascariensis in China BAOJUN Hao, WENHUAN LI, CHEN CHEN, Liu GAo, RONG ZHANG, BAOTONG WANG, GUANGYU SUN & MARK L. GLEASON Polycoccum anatolicum sp. nov. on Lepraria incana and a key to Polycoccum species known from Turkey MEHMET GOKHAN HALICI, HATICE EsRA AKGUL, CELALEDDIN OzTURK & Emre KILIG¢ Further additions to the macrolichen mycota of Vietnam UDENI JAYALAL, ANDRE APTROOT, THI THUY NGUYEN, NGUYEN ANH DZUNG, SANTOSH JOSHI, SOON-OK OH & JAE-SEOUN HuR Inonotus niveomarginatus and I. tenuissimus spp. nov. (Hymenochaetales), resupinate species from tropical China Hat-You Yu, CHANG-LIN ZHAO & YU-CHENG Dal Arenariomyces truncatellus sp. nov., an ascomycete on driftwood from the north coast of Zealand, Denmark J. Kocu New ascomycete records from Guatemala RosARIO MEDEL, OSBERTH MORALES, RANULFO CASTILLO DEL MORAL & ROBERTO CACERES
Coleosporium in Europe STEPHAN HELFER
pA
a
39
45
51
61
69
73 87
APRIL-JUNE 2013... V
Septoglomus titan, a new fungus in the Glomeraceae (Glomeromycetes) from Bahia, Brazil BRUNO ToMIO GoTo, ADRIANE FREIRE ARAUJO, ANA CRISTINA FERMINO SOARES, ARAESKA CARENNA DE ALMEIDA FERREIRA, LEONOR COSTA MaIA, CARLA DA SILVA SOUSA & GLADSTONE ALVES DA SILVA 101 Geoglossaceous fungi in Slovakia 5. Geoglossum uliginosum: taxonomy and nomenclature VIKTOR KUCERA, JOHAN NITARE, PAVEL LIZON & JAN GAISLER 111 A new endophytic species of Chaetomium from Jatropha podagrica ROHIT SHARMA, GIRISH KULKARNI, MAHESH S. SONAWANE & YOGESH S. SHOUCHE 117 A contribution to the taxonomy of Lyromma (Lyrommataceae, lichenized Ascomycota) with a species key ADAM FLAKUS & EDIT FARKAS 127 Scolecobeltrania, an interesting new microfungus from Venezuela TERESA ITURRIAGA, ROBERTO FERNANDEZ,RAFAEL F. CASTANEDA-RUIZ, Davip W. MINTER & ALISSON CARDOSO RODRIGUES DA CRUZ 143 A new species of Pisolithus from Spain Maria P. Martin, FATIMA DURAN, CHERDCHAI PHOSRI & Roy WATLING 149 A reassessment of excavated Tuber species from China based on morphology and ITS rDNA sequence data Li FAN, JIN-ZHONG CAo & Yu Li 155 A new species and a new record of the genus Entoloma from China JiaN-RuI WANG & TOLGOR Bau 165 Russula atroaeruginea and R. sichuanensis spp. nov. from southwest China Guo-JiE Li, Qt ZHAO, DONG ZHAO, SHUANG-FEN YUE, SAI-FE1 L1, HUA-AN WEN & XING-ZHONG Liu 173 Five new Terfezia species from the Iberian Peninsula JuaN-JULIAN BoRDALLO, ANTONIO RODRIGUEZ, JUSTO M. MUNoz-MOHEDANO, LaAuRA M. Suz, MARIO HONRUBIA & ASUNCION MorTE 189 Hypochnicium pini, a new corticioid basidiomycete in East Asia YEONGSEON JANG, SUNG WOOK LEE, YOUNG WOON LIM, Jin SUNG LEE, NILS HALLENBERG & JAE-JIN Kim 209 Taxonomic studies on Mucor inaequisporus, isolated for the first time in South America ANDRE Luiz C.M. DE A. SANTIAGO, ANDRE RODRIGUES, ENzO M. CANEDO & EDSON R. FILHO 219 Molecular phylogeny reveals Megacollybia virosa is a Cantharocybe T.K. ARUN KUMAR & P. MANIMOHAN 231 New record of Scedosporium dehoogii from India ROHIT SHARMA, GIRISH KULKARNI, MAHESH S. SONAWANE & YOGESH S. SHOUCHE 239
vI ... MYCOTAXON 124
Inocybe nitidiuscula and its ectomycorrhizae associated with Alnus nitida from Galyat, Pakistan S. ILyas, A. RAZAQ & A.N. KHALID Xerocomus porophyllus sp. nov., morphologically intermediate between Phylloporus and Xerocomus WEN-JUAN YAN, Tai-Hutr Li, MING ZHANG & TING LI Glomus mume and Kuklospora spinosa: two new species of Glomeromycota from China BANG-PING CAI, LIANG-DONG GUO, JUN-YU CHEN & QI-XIANG ZHANG Russula changbaiensis sp. nov. from northeast China Guo-J1E L1, DONG ZHAO, SAI-FEI LI, Hual-JUN YANG, Hua-AN WEN & XING-ZHONG LIU
Studies on three rare coprophilous plectomycetes from Italy FRANCESCO DOVERI, SABRINA SARROCCO & GIOVANNI VANNACCI Morphological and genetic characterisation of Beauveria sinensis sp. nov. from China MING-JUN CHEN, Bo Huana, ZENG-ZHI LI & JOSEPH W. SPATAFORA The lichen genus Fissurina (Graphidaceae) in Vietnam SANTOSH JOSHI, THI , THUY NGUYEN, NGUYEN ANH DzuUNG, UDENI JAYALAL, SOON-OK OH & JAE-SEOUN Hur Studies of North American macrofungi, 1. Validation of Lactarius rubidus comb. nov. and Leccinellum quercophilum sp. nov. MICHAEL Kuo, ANDREW S. METHVEN, ANDREW M. MINNIS, & Roy E. HALLING Cladonia dunensis sp. nov. from southern Brazil, with notes on the genus in beach dune environments EMERSON LuIz GUMBOSKI, FLAVIO BEILKE & SIONARA ELIASARO Coniolepiota spongodes (Agaricaceae, Basidiomycota) in Bangladesh and China Mp. IqBaL HosEN & ZHU L. YANG Pertusaria albiglobosa, a new lichen from China QIANG REN Lichenological notes 6: nomenclatural acts KERRY KNUDSEN & JANA KOCOURKOVA Phylloporia tiliae sp. nov. from China Li-We!I ZHou
NOMENCLATURE Nomenclatural novelties proposed in volume 124
247
255
263
269
2%9
301
309
320
333
341 349
353 361
367
APRIL-JUNE 2013... VII
ERRATA FROM PREVIOUS VOLUMES
VOLUME 122
p-30, table, after Lambertella sp.3 For: Leaf of Pinus sp. | READ: Leaf of Pyrola sp. p-30, table, after Lambertella sp.8 For: Leaf of Pinus sp. READ: Leaf of Pyrola sp.
VOLUME 123
p. ii, lines 12-13 FOR: SEPPO HUHTINEN (2006-2012) Turku, Finland READ: KAREN HANSEN (2013-2017) Stockholm, Sweden p. iv, line 12: FOR: Lichenochora tertia READ: Lichenochora tertia (Phyllochorales): p. iv, line 22: FOR: Jatropha podarica READ: Jatropha podagrica p. vi, line 8: FOR: TEODOR T. DENCHEV, HABIB AHMAD & ABDUL NASIR KHALID
READ: TEODOR T. DENCHEV, ABDUL NASIR KHALID & HABIB AHMAD
vill ... MYCOTAXON 124
PUBLICATION DATE FOR VOLUME ONE HUNDRED TWENTY-THREE MYCOTAXON for JANUARY-MARCH, VOLUME 123 (I-v1 + 1-495) was issued on August 1, 2013
APRIL-JUNE 2013 ... IX
REVIEWERS — VOLUME ONE HUNDRED TWENTY-FOUR
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 volume.
M. Catherine Aime A. Aptroot
Bita Asgari
Timothy J. Baroni Reinhard Berndt Janusz Blaszkowski Wolfgang von Brackel Matias J. Cafaro
Paul EF Cannon Cecilia Cristina Carmaran R.E Castafieda-Ruiz Pedro Crous Yu-Cheng Dai
Kanad Das
FR.M. Dugan
Walter Gams Genevieve M. Gates Matteo Gelardi Sergio P. Gorjon Josep Guarro
Cécile Gueidan Shouyu Guo
Ian R. Hall
Samuel Hammer David L. Hawksworth
G. Sybren de Hoog Vit Hubka Seppo Huhtinen K.D. Hyde Mikael Jeppson Evan Benjamin Gareth Jones Kerry Knudsen Patrick R. Leacock Teresa Lebel James C. Lendemer De-Wei Li Tai-Hui Li Zongqi Liang Runjin Liu Laszl6 Lékés Guo-zhong Lit Robert Licking Marcelo P. Marcelli P. Brandon Matheny Patrick McCarthy Eric H.C. McKenzie Gabriel Moreno Salvatore Moricca Sanjeeva Nayaka Abdul Rehman Niazi
Lorelei L. Norvell Fritz Oehl
Esteri Ohenoja Takamichi Orihara Beatriz Ortiz-Santana Todd W. Osmundson Ka-Lai Pang
Shaun R. Pennycook Evangelina Pérez-Silva Leif Ryvarden
Jaya Seelan Sathiya Seelan Ewald Sieverding
José Ivanildo de Souza Viacheslav Spirin
Iben Margrete Thomsen Giuseppe Venturella Long Wang
A.J.S. Whalley
Pat Wolseley Jian-Ping Xu
Yi-Jian Yao
Xiu-Guo Zhang Zhongyi Zhang Li-Wei Zhou Wen-Ying Zhuang
X ... MYCOTAXON 124
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APRIL-JUNE 2013... XI
FROM THE EDITOR-IN-CHIEF
WELCOME KAREN HANSEN! — We wish to announce - somewhat belatedly - that Sweden's Karen Hansen (Senior Curator, Kryptogambotanik, Naturhistoriska Riksmuseet, Stockholm) has been elected to MycoTaxon’s Editorial Advisory Board. She joins Chair Wen-Ying Zhuang (Beijing), Past Chair Henning Knudsen (Copenhagen), Scott Redhead (Ottawa), Sabine Huhndorf (Chicago), and Peter Buchanan (Auckland) on the 2013 Board. Each member serves a six-year term, standing as Chair in the fifth year and as Past Chair in his or her final year. We apologize profusely for the editorial (!) lapse leading to the omission of Karen’s name from the masthead (on p. ii) of Mycotaxon 123, the first volume for 2013.
MycoTaxon 124 — After some deliberation and in an effort to bring the journal back on schedule this year, we have closed the delayed April-June 2013 volume at 380 pages. The current volume contains 38 papers by 151 authors (representing 26 countries) and revised by 74 expert reviewers.
Within its pages are one new genus and species (Scolecobeltrania from Venezuela) and 33 other taxa new to science representing Arenariomyces from Denmark; Beauveria, Entoloma, Glomus, Inonotus, Kuklospora, Myrmecridium, Pertusaria, Phialophora, Phylloporia, Russula, Tuber, and Xerocomus from China; Chaetomium from India; Cladonia and Septoglomus from Brazil; Coleosporium from Europe; Hypochnicium from East Asia; Leccinellum from the U.S.A.; Lyromma from Bolivia and Brazil; Pisolithus and Terfezia from Portugal and/or Spain; and Polycoccum from Turkey.
In addition to range extensions and/or new hosts for previously named taxa, we also offer new combinations in Aspicilia, Cantharocybe, Lactarius, and Rutstroemia, newly established synonymies and typifications, and conclusions from recent mycorrhizal (Inocybe) and monographic (Fissurina in Vietnam, plectomycetes in Italy, ascomycetes in Guatemala) studies.
The next two volumes will also close ‘early’ to ensure that the four 2013 volumes will not spill over into next year. We shall begin 2014 with a much shorter turn-around time between final submission and publication, and hope eventually to return to publishing each volume at the beginning of a quarter rather than well after!
Warm regards,
Lorelei L. Norvell (Editor-in-Chief) 29 September 2013
ISSN (print) 0093-4666 © 2013. Mycotaxon, Ltd. ISSN (online) 2154-8889
MY COTAXON
http://dx.doi.org/10.5248/124.1 Volume 124, pp. 1-8 April-June 2013
A new Myrmecridium species from Guizhou, China
CHUN- YU JIE“, QING-XIN ZHOU™, WEN-SHENG ZHAO’, YU-LAN JIANG’, KEVIN D. Hype**, Ertc H.C. MCKENZIE° & YONG WANG'*
"Department of Plant Pathology, College of Agriculture, Guizhou University, Guiyang 550025, China
? Institute of Agro-Food Science & Technology, Shandong Academy of Agricultural Sciences, Jinan, Shandong 250100, China
* Department of Plant Pathology, China Agricultural University, Beijing 100193, Peoples Republic of China
* Institute of Excellence in Fungal Research, and School of Science, Mae Fah Luang University, Chiang Rai, Thailand
° School of Science, Mae Fah Luang University, Chiang Rai 57100, Thailand
° Landcare Research, Private Bag 92170, Auckland, New Zealand
* CORRESPONDENCE TO: yongwangbis@aliyun.com OR mppzhaws@cau.edu.cn
ABSTRACT — A strain of Myrmecridium was isolated from farmland soil in Guizhou Province, China. The strain produced distinctive obovoid or fusoid conidia that taper to a subtruncate darkened hilum at the base that distinguished it from other Myrmecridium species. Phylogenetic analysis of combined ITS1-5.8S-ITS2 and large subunit rDNA sequences supported this morphological separation. We describe Myrmecridium obovoideum as new and provide a key to Myrmecridium species.
KEY worps — asexual fungi, hyphomycetes, molecular phylogeny
Introduction
Arzanlou et al. (2007) established Myrmecridium Arzanlou et al. based on its hyaline mycelium with pale to unpigmented, pimple-like denticles that produce obovoid or fusiform conidia, tapering towards a narrowly truncate base with a slightly prominent unpigmented hilum and schizolytic conidial secession. The genus belongs in Sordariomycetes and comprises three species and two varieties (Arzanlou et al. 2007; Crous et al. 2011; Crous et al. 2012).
During a survey of anamorphic ascomycetes, we isolated a strain with characters of Myrmecridium (Arzanlou et al. 2007) from farmland soil in China.
“C.-Y. Jie and Q.-X. Zhou contributed equally to the manuscript and should be considered as joint first authors
2 ... Jie & al.
Combined ITS and LSU rDNA sequences were generated from the strain and analyzed to evaluate the morphological findings. Based on morphology and DNA sequence comparison, the strain is proposed as Myrmecridium obovoideum. In the present paper, we provide detailed illustrations and a description of this new species.
Materials & methods
Morphological and cultural studies
The Myrmecridium specimen — from a farmland soil sample collected from Huaxi District, Guizhou, China, in 2005 —is deposited in the herbarium of the Department of Plant Pathology, Shandong Agricultural University (HSAUP), and ex-type strains are conserved in HSAUP and the Herbarium of Plant Pathology, Guizhou University (HGUP). The specimen was examined in the laboratory and the taxon is described from cultures grown at 25°C on potato dextrose agar (PDA). Conidia and conidiophores were placed in a drop of 85% lactic acid and examined and photographed using a Nikon 80i microscope (Nikon Corporation, Japan) at 400x and 1000x magnification. The taxonomic determinations were made by comparing with descriptions in the literature.
DNA extraction, amplification and DNA sequencing
Genomic DNA was extracted from colonies grown on potato-dextrose agar (PDA), using the Fungal gDNA Kit GD2416 (Biomiga, CA, USA) following the manufacturer's instructions. The universal primers ITS1/ITS4 (White et al. 1990) were used for the ITS region (ITS1-5.8S-ITS2) amplification, and LROR/LR5 were used for a segment of the large subunit rDNA (Vilgalys and Hester 1990). Amplification reactions were performed in a Bio-RAD PTC-200 thermocycler in a 25 ul reaction mixture using the following final concentrations or total amounts: 5 ng DNA, 1 x PCR buffer (20 mM Tris/HCl pH 8.4, 50 mM KCl), 1 uM of each primer, 2.5 mM MgCl, 0.25 mM of each dNTP, 0.5 U of Taq polymerase.
The PCR-amplified DNA fragments were fractionated in 1% agarose gels in 0.5x TBE buffer and visualized by ethidium bromide staining and UV illumination. Sequencing was performed with an ABI PRISM 3730 DNA autosequencer using either dRhodamine terminator or Big Dye Terminator chemistry (Life Technologies™, USA). Sequence data of the isolates used in the study were deposited in GenBank (TaBLE 1). Alignments are available in TreeBASE (www.treebase.org/treebase-web/home.html) under the study ID T3733.
Phylogenetic analyses
Preliminary nucleotide sequence alignments were constructed using Clustal X 1.81 (Thompson et al. 1997). A partition homogeneity test (Farris et al. 1994) was applied to evaluate the feasibility of combining the datasets. Phylogenetic analyses of LSU sequences and combined ITS and LSU rDNA sequence were computed using MP analysis in PAUP* (Swofford 2002). In the MP analyses, trees were inferred using heuristic search option with tree bisection reconnection (TBR) branch swapping and 1000 random sequence additions, maxtrees were 5000, branches of zero length were collapsed and all parsimonious trees were saved. Measures calculated for parsimony
Myrmecridium obovoideum sp. nov. (China) ... 3
included tree length (TL), consistency index (CI), retention index (RI), and rescaled consistence index (RC). Bootstrap analyses (Hillis & Bull 1993) were conducted with 1000 replications.
Results
Phylogenetic analysis
Partition homogeneity tests for combining the two (ITS and LSU) gene regions yielded a P-value of 0.104, and the two gene/regions were combined based on the tree topologies and P-value (Cunningham 1997, Dettman et
Myrmecridium phragmitis JQ044425
Myrmecridium schulzeri EU041770
Myrmecridium banksiae JX069871
Myrmecridium flexuosum EU041768
Myrmecridium obvoideum sp. nov KC136140
Rhodoveronaea varioseptata EU041813
Gondwanamyces wingfieldi JQ844903
Pseudoidriella syzygii JQ044421
Seiridium banksiae JQ044422
Diaporthe musigena JF951138
100
Phomopsis subordinaria GQ922519
Pleurothecium obovoideum EU041784
Carpoligna pleurothecii JQ429149
Ophiostoma stenoceras AF 484476
Veronaeopsis simplex EU041820
— 10
Fic. 1. One of the two equally most parsimonious trees of the analyzed ITS and LSU rDNA region/ genes (284 of 1485 characters were parsimony informative). Bootstrap support values of less than 50% are not shown. The tree is rooted with Pleurothecium obovoideum.
A ... Jie & al. ; Myrmecridium flexuosum EU041825 Myrmecridium obvoideum sp. nov KC136139 100 |_| Myrmecridium schulzeri EU041827
50 Myrmecridium phragmitis JQ044444
74 Myrmecridium banksiae YX069855
Rhodoveronaea varioseptata EU041870 50
Ophiostoma stenoceras DQ836904 Diaporthe musigena JF951158 99 Phomopsis asparagi AF 439634
75 Seiridium banksiae JQ044442
100 Polyscytalum algarvense GQ303318
100 Pseudophloeospora eucalypti HQ599593
Gondwanamyces wingfieldii JQ844902
tat Pleurothecium obovoideum EU041841
Carpoligna pleurothecii JQ429237
Veronaeopsis simplex EU041877 — 10
Fic. 2. The most parsimonious tree of the analyzed LSU rDNA gene (206 of 844 characters were parsimony informative). Bootstrap support values of less than 50% are not shown. The tree is rooted with Veronaeopsis simplex.
al. 2003). The aligned sequence data matrix contained five taxa (including the outgroup Pleurothecium obovoideum) and 1485 (546 ITS and 839 LSU) characters, of which 284 were parsimony informative. Two most parsimonious trees were obtained, with one chosen to represent the topology of the strict
Myrmecridium obovoideum sp. nov. (China) ... 5
consensus tree (Fic. 1; Tree Length (TL) = 380, CI = 0.956, RI = 0.467, HI = 0.044, and RC = 0.446). In this tree, five Myrmecridium species clustered together supported by a 100% bootstrap value. Myrmecridium obovoideum formed a single branch as the sister clade to the other four species with only 56% bootstrap support.
In order to evaluate the relationship of Myrmecridium with other ascomycetes, we also generated a parsimonious tree (Fic. 2) based on LSU rDNA gene region. The alignment with 16 taxa (Veronaeopsis simplex as outgroup) comprised 844 characters, of which 206 were parsimony-informative [Tree Length (TL) = 628, CI = 0.67, RI = 0.68, HI = 0.33, and RC = 0.45]. Only one most parsimonious tree was obtained to represent the topology of the strict consensus tree selected for presentation (Fic. 2). In this tree, the Myrmecridium group had a close relationship with Rhodoveronaea and Ophiostoma supported by a credible bootstrap value (74%), which was consistent with Arzanlou et al. (2007). However, the connection between Rhodoveronaea and Ophiostoma received weak support (<50%). Meanwhile, the placement of M. obovoideum was movable, because it showed a closer relationship with M. flexuosum (de Hoog) Arzanlou et al. supported by 82% bootstrap value, which differed from the analysis combined ITS and LSU rDNA gene regions.
Taxonomy
Myrmecridium obovoideum Jie, Y.L. Jiang, McKenzie & Yong Wang bis, sp. nov. Fig. 3 MycoBank MB 804082
Differs from other Myrmecridium species by its conidia with darkened hilum.
Type: China, Guizhou Province, Guiyang, Huaxi District, isolated from farmland soil, 5 November 2005, Y.L. Jiang (Holotype, HSAUP051001; isotype, HGUP0314; GenBank, KC136140, KC136139).
EryMo_oey: in reference to the conidial shape of this new taxon.
Colonies on PDA at 25°C for 2 weeks reaching 30 mm diam. ‘The surface effuse, grayish. Mycelium mostly superficial, partly immersed is contradictory with lacking aerial mycelium. Conidiophores straight to geniculate- sinuous, unbranched, subhyaline, 1-8-septate, <280 um tall, 3-3.6 um wide. Conidiogenous cells terminal, integrated, cylindrical, pale or reddish brown near the base, gradually becoming paler towards the apex, 50-210 um, often with 1-6 thin septa, forming a rachis with scattered pimple-shaped denticles less than 1 um long, sometimes fertile part slightly inflated. Conidia solitary, aseptate, subhyaline, pale brown, thin-walled, smooth to finely verrucose, surrounded by a wing-like gelatinous sheath, <1.5 um thick, obovoid or fusoid, (7-)8-9(-10) x (3-)3.5(-4.2) um, tapering to a subtruncate, slightly thickened hilum; darkened at rounded end.
6 ... Jie & al.
Fic. 3. Myrmecridium obovoideum (HSAUP051001, holotype) on PDA. a-b. Conidiophores. c. Rachis with pimple-shaped denticles and slightly inflated fertile part. d. Conidia surrounded by wing-like gelatinous sheaths. e-f. The thin septa of conidiogenous cells. Scale bars: a-c, e-f = 35 um; d= 10 um.
Myrmecridium obovoideum sp. nov. (China) ... 7
Discussion Our LSU rDNA sequence analysis clustered Myrmecridium with Rhodoveronaea (sexual state = Annulatascaceae) and Ophiostoma
(Ophiostomataceae) in the Sordariomycetes. Arzanlou et al. (2007) classified Myrmecridium as “Incertae sedis (Sordariomycetes)”, and our results confirm this placement, without any indication of a family affinity.
Only M. schulzeri var. tritici (M.B. Ellis) Arzanlou et al. and M. phragmitis Crous produce septate conidia. Myrmecridium banksiae Crous differs from M. obovoideum by its larger conidia (9-14 x 2.5-3.5 um); and M. flexuosum differs by its smaller conidia (5-9 x 3-4 um). The conidia of M. schulzeri (Sacc.) Arzanlou et al. var. schulzeri (6-12 x 3-4 um) are also larger than those of our species. In Myrmecridium only M. obovoideum has conidia with a darkened hilum, which enlarges the taxonomic criteria of Myrmecridium (Arzanlou et al. 2007). Our ITS and LSU rDNA sequence analyses also confirmed with high bootstrap value that our species differs from other Myrmecridium species. The combined morphological and phylogenetic analysis supports M. obovoideum as a new species.
Key to Myrmecridium species
Las Pile ofconidia ne td atkeneds5 Act, 0 Aeh, chit lead ubai dn Heald tn tnald a aluonttah sal 2 Ibe Hiltom-ofcomidia darketeden 5 fc. 6.5 242.5 nce pale yeh laine Melee Sale ims M. obovoideum 2a; Comidiawithouy septate, .2 Lin. + iden eg dh sees Pieter a Saber a Atnaes garners Abeta Ss Nees 3 2b. Septate conidia frequently-Observed ... guiicieclaicenglaneetlenes M. schulzeri var. tritici ba. Conidia mostly lessthani 10. pint lone: sti. cea cshee Gta sda negate Gea co Bet wed ale 4 Sb, Conidiarip to P4afimislon es. isptea ras atls tay nll len cette tes tres steele M. banksiae dace onidia rounded at the bases) fut. fit.ne PO! Wo hal Wet Weer 1 M. flexuosum 4b. Conidia slightly acuminate at the base......................00 0000. M. phragmitis 4c. Conidia tapering to a subtruncate base................ M. schulzeri var. schulzeri
Acknowledgments
The authors are grateful for pre-submission comments and suggestions provided by Drs. R.E Castafieda-Ruiz and De-Wei Li. This project was supported by the National Science Foundation of China (No. 31060005) and the Doctors’ Funding of Guizhou University (no. 2010014). We are indebted to Prof. Tian-Yu Zhang (Shandong Agricultural University, China) for supplying the culture of this new species.
Literature cited
Arzanlou M, Groenewald JZ, Gams W, Braun U, Shin HD, Crous PW. 2007. Phylogenetic and morphotaxonomic revision of Ramichloridium and allied genera. Studies in Mycology 58: 57-93. http://dx.doi.org/10.3114/sim.2007.58.03
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Crous PW, Summerell BA, Shivas RG, Romberg M, Mel'nik VA, Verkley GJM, Groenewald JZ. 2011. Fungal Planet description sheets: 92-106. Persoonia 27: 130-162. http://dx.doi.org/10.3767/003158511X617561
Crous PW, Summerell BA, Shivas RG, Burgess TI, Decock CA, Dreyer LL, Granke LL, Guest DI, Hardy GEStJ, Hausbeck MK, Hiiberli D, Jung T, Koukol O, Lennox CL, Liew ECY, Lombard L, McTaggart AR, Pryke JS, Roets E Saude C, Shuttleworth LA, Stukely MJC, Vanky K, Webster BJ, Windstam ST, Groenewald JZ. 2012. Fungal Planet description sheets: 107-127. Persoonia 28: 138-182. http://dx.doi.org/10.3767/003158512X652633
Cunningham CW. 1997. Can three incongruency tests predict when data should be combined? Molecular Biology and Evolution 14: 733-740. http://dx.doi.org/10.1093/oxfordjournals. molbev.a025813
Dettman JR, Jacobson DJ, Taylor JW. 2003. A multilocus genealogical approach to phylogenetic species recognition in the model eukaryote Neurospora. Evolution 57: 2703-2720. http://dx.doi.org/10.1554/03-073
Farris JS, Kallersj6 M, Kluge AG, Bult C. 1994. Testing significance of incongruence. Cladistics 10: 315-320. http://dx.doi.org/10.1111/j.1096-0031.1994.tb00181.x
Hillis DM, Bull JJ. 1993. An empirical test of bootstrapping as a method for assessing confidence in phylogenetic analysis. Systematic Biology 42: 182-192. http://dx.doi.org/10.1093/sysbio/42.2.182
Swofford DL. 2002. PAUP*: Phylogenetic analysis using parsimony (*and other methods), version 4.0b10. Sinauer Associates, Sunderland, Massachusetts.
Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG. 1997. The Clustal X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Research 24: 4876-4882. http://dx.doi.org/10.1093/nar/25.24.4876
White TJ, Bruns T, Lee S, Taylor J. 1990. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. 315-322, in: MA Innis et al. (eds). PCR protocols: a guide to methods and applications. Academic Press, New York, U.S.A.
Vilgalys R, Hester M. 1990. Rapid genetic identification and mapping of enzymatically amplified ribosomal DNA from several Cryptococcus species. Journal of Bacteriology 172: 4239-4246.
ISSN (print) 0093-4666 © 2013. Mycotaxon, Ltd. ISSN (online) 2154-8889
MY COTAXON
http://dx.doi.org/10.5248/124.9 Volume 124, pp. 9-20 April-June 2013
The occurrence of Rutstroemia coracina on Quercus ilex leaves in Spain
RICARDO GALAN!*, FRANCISCO PRIETO-GARCIA?’, ANGELES GONZALEZ? & CARLOS ENRIQUE HERMOSILLA‘
” Department of Life Sciences, University of Alcala, Alcala de Henares, 28871 Madrid, Spain ? Marqués de la Valdavia 13 A, 28400, Collado Villalba, Madrid, Spain
° Prado Ibarra 29, 28270, Colmenarejo, Madrid, Spain
* Francisco Cantera 11, 1° izda., 09200 Miranda de Ebro, Burgos, Spain
* CORRESPONDENCE TO: ricardo.galan@uah.es
AxBsTRACT— ‘The authors report new findings of an apparently rare fungus, Rutstroemia coracina (Rutstroemiaceae). The species grew on old fallen leaves of Quercus ilex subsp. ballota at different localities in Guadalajara and Madrid provinces (center of Spain) where it was first collected in 1983 and subsequent years. The apothecia are described and illustrated in detail. Microscopic studies, including microtome sections, show a clear gel layer in the ectal excipulum, which allows assignment in the genus Rutstroemia. Since the type material deposited in PC is so scanty and deteriorated that any future revision will be impossible, an epitype is selected here.
Key worps— Ascomycota, Helotiales, foliicolous fungi, taxonomy
Introduction
Palmer (1994) was the first to report the presence of Rutstroemia coracina from the European continent. This species was initially recorded in Algeria and was documented only by a coloured plate as “Peziza coracina DR. et Lév. in Durieu (1848: pl. 28 fig. 4) reproduced here as Fic. 1, which depicts apothecia (on a Quercus ilex leaf) with egg-shaped spores, paraphyses, and asci. Although no text accompanied the 1848 protologue, the figure caption published in Durieu (1869: 10) identified the substrate as the undersurface of a dried Q. ilex leaf and detailed the individual elements included in the figure. Later Saccardo (1889: 237) provided a brief description as Helotium coracinum, apparently based on the original figure since no microscopic measurements are given and enquiries by J.T. Palmer with herb. PAD (Universita degli Studi di Padova) ascertained that there is no exsiccatum under this epithet in Herb. P.A. Saccardo.
10 ... Galan & al.
The first authentic description seems to be by Dennis (1964: 39-40, fig. 12), who studied the type collection preserved in PC. Reporting ascospores “11-13 x 5-5.5 um’ and an “[e]xcipulum composed of parallel hyphae with glassy walls,” he transferred the species to Rutstroemia.
Spooner (1981: 283, 285, fig. 22) redescribed the species from a few apothecia ona Quercus ilex leaf collected by M.C. Clark in 1977 near Somerset, southwest England, and recombined it in Lanzia, as he failed to find the “glassy walls” found by Dennis. He reported similar spore measurements to those mentioned by Dennis for the type.
In 1992, Palmer visited the Allerford plantation between Bossington and Porlock, Somerset, where Mr. Clark had collected his leaf, and collected large samples of dead Q. ilex leaves from different parts of the wood, which subsequently produced apothecia in natural culture. His measurements for the living spores (in tap water) were 12.4-16.8 (18.4) x 4.8-7.6 (8) um.
Palmer (1994) also restudied the type remnants in PC and found a minute fragment (approximately haft an apothecium) among the stipes of immature apothecia on one of the two spiny leaves of the holotype; although it was not possible to make a detailed study, he managed to draw ovoid spores without any sign of guttulation (measuring 10.6-13.9 x 4.5-6.5 (7.3) um), one ascus (measuring 106 x 12 um), and scarce hyphae suspected to be from the gelatinous layer in the ectal excipulum protruding as obtuse “hairs” (3.3-6.0 um wide) - reproduced here as Fic. 2. In addition, Palmer (1994: 226-228, figs 1-5) made good collections in northern Greece in March 1988, mainly on old, fallen leaves of Quercus coccifera and Q. ilex on Mount Athos and Mount Olympus but also on a leaf of Quercus pubescens (= Q. humilis) on Mt Athos.
This species was collected and identified for the first time in Spain by R.G. in 1983 on old, fallen leaves of Quercus ilex subsp. ballota at Tamajon (Guadalajara province). The identification was later confirmed by Palmer, who had the opportunity to record the species in subsequent years, including collections by Dr. A. Raitviir (Palmer & Galan 1999, Galan 2001). The area is open with scattered trees (intermixed with Juniperus thurifera), and leaf litter is sparse. More recently, during a field trip through Guadalajara as part of the XIII Congress of European Mycologists in Alcala de Henares (September, 1999), several attendees had the opportunity to collect the species in abundance, including Drs. B. Spooner, M.M. Nauta and H.O. Baral, who kindly provided us with herbarium samples. More recent findings of the species by two of us (E.P.G. & A.G.) on old fallen leaves of Quercus ilex subsp. ballota at Colmenarejo (Madrid province) allow us not only to widen the distribution of Rutstroemia coracina in Spain but also to complete its description. In addition, Palmer (pers. comm.) also recorded and identified this species in 1990 on Minorca (Balearic Islands, Spain) growing on Quercus ilex leaves.
Rutstroemia coracina on Quercus (Spain) ... 11
Fic. 1. Illustration of the type of Peziza coracina Durieu & Léveille, Expl. Sci. Algérie, Bot. 1: t. 28, f. 4 (1848).
12 ... Galan & al.
From the beginning, Palmer (1994) found it noteworthy that the greyish green tinges of apothecia in both freshly collected material or that developed in damp chambers were overlooked in Spanish material, which more closely resembled the reddish brown color in the original Algerian plate. That might suggest the existence of two different taxa, but otherwise all specimens appeared to be microscopically identical.
Since the type material preserved in PC allows no further revision, we here select as epitype a very rich Spanish collection (about 30 fallen leaves of Quercus ilex subsp. ballota, bearing ca. 60-70 apothecia) whose habit, colour, and microscopic features (mainly ascospore shape and size) match the holotype very well.
There has been continued controversy since Dennis (1964) reported an “excipulum composed of parallel hyphae with glassy walls.” This was why the taxon was first combined in Rutstroemia and why Spooner (1981) transferred the species to Lanzia, remarking, “I have not been able to find glassy walls.” Spooner later had opportunity to re examine one of the Greek collections (J.T.P. 4418) identified as R. coracina and agreed entirely with Palmer’s identification. He noted the presence of gelatinized tissue, which made him review the British material where he found “a gelatinous matrix to the ectal hyphae, something which I overlooked when referring the species to Lanzia” (B.M. Spooner, in letter). The microtome sections (Fics 6, 7) of English, Greek, and Spanish material show a clear gel layer in the ectal excipulum, although the thickness is not as prominent as in other Rutstroemia spp. and also is somewhat variable in width from collection to collection. Nonetheless, we feel that the species was correctly placed in Rutstroemia by Dennis (1964).
Rutstroemia coracina appears to be known only from Algeria (where the actual type locality is unknown), Britain (Spooner 1981), Greece (Palmer 1994), France (Palmer, pers. comm.), Turkey [K(M) 49709 Spooner, pers. comm.], and Spain.
Materials & methods
Measurements of living cells (marked *) were made in tap water or IKI (1% Lugol’s solution), with dead cells (marked f) in tap water, Melzer’s reagent (MLZ), KOH 2%, and aqueous cresyl blue (CRB) 0.5%. Line drawings were made using a NIKON Labophot-2 either by hand without the aid of a camera lucida (C.E.H.) or with a drawing tube incorporated into the light microscope (R.G.) with magnifications up to 1250x. Photomicrographs were made with using a NIKON Labophot-2 equipped with phase contrast and camera. Macroscopic pictures of fructifications in fresh state showing the general habit were taken in situ (by EP.G.) or at the lab (by C.E.H.). Apothecial colours are cited according to Locquin (1985). Specimens have been deposited in AH and J.T.P. (personal herbarium of the late Mr. J. Terence Palmer, currently deposited in CUP) with additional specimens in H.O.B. (personal herbarium of Hans O. Baral, Tubingen, Germany) and K.
Rutstroemia coracina on Quercus (Spain) ... 13
NATIONAL HERBARIA
J.T.P. Refis WX institute: Museum D'Histo res NaTa@cwe - abbreviation: PC De Paris:-LReorato (ae he Cevetoeamis, PARIS ORTOINAL DETERMINATION: PEZIZA CoRAciN« revid.: = RYTIRRORMA CORACIN'A Leg. Date: Devi Locality: ALG &RIA Sle eon A RE erst ~ < © (6% 20. Zo feziza coraciha DAR er Liv. eet Coops ota ice ker } ae” Deen cee ! 140 , Neletyous in PC - gy ee 3o 4 Ho $0 yo 100 uo Ps : UK 106 ¥l2 prot, : ¥ y .
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Fic. 2. Rutstroemia coracina (holotype, PC): Annotations and drawings by J.T. Palmer showing excipular hyphae and an ascus (up), ascospores, and “hyphoid” hairs (down).
14 ... Galan & al.
Fic. 3. Rutstroemia coracina (AH 7641, epitype) growing in natural habitat. Photo EPG. in situ (bar = 10 mm).
=a Pe. see. ee et” a Ora} Fic. 4. Apothecia of Rutstroemia coracina (C.E.H. 08468) under hi Photo C.E.H. ex situ (bar = 1 mm).
gh
Rutstroemia coracina on Quercus (Spain) ... 15 Taxonomy
Rutstroemia coracina (Dur. & Lév.) Dennis, Persoonia 3: 39 (1964) Fics. 1-7 = Peziza coracina Dur. & Lév., Expl. Sci. Algérie, Bot. 1: t. 28, f. 4 (1848) = Helotium coracinum (Dur. & Lév.) Sacc., Syll. Fung. 8: 237 (1889) = Calycina coracina (Dur. & Lév.) Kuntze, Revis. Gen. Pl. 3(3): 448 (1898) = Ciboria coracina (Dur. & Lév.) Boud., Hist. Class. Discom. Eur.: 106 (1907) = Lanzia coracina (Dur. & Lév.) Spooner, Trans. Br. Mycol. Soc. 76: 283 (1981)
Type: Algeria (Holotype, PC). Spain, Madrid, Colmenarejo, UTM 30TVK1489, 885 m a.s.l., on petioles, lower (more exceptionally upper) faces and veins of fallen leaves of Quercus ilex subsp. ballota, 27.X1.2011, Prieto & Gonzalez, det. R.G. , rev. C.E.H. (Epitype designated here, AH 7641).
APOTHECIA superficial, scattered, cupulate, stipitate, castaneus to reddish- brown, merging from darkened areas of the substrate. pisc slightly concave becoming plane, flesh colour (ie., russus B4f) to yellowish-olivaceous (i.e., venetus S2d), tending to darken with age, 0.5-2.0 mm in diameter. RECEPTACLE cupulate, covered with a sparse and irregular net of brown fibrils, vertically arranged with adpressed ends, sometimes protruding and giving a fringed appearance, colour similar to or concolorous with disc and stipe, sometimes chestnut (with varying reddish brown shades). MARGIN ochre, chestnut to reddish-brown, crenulate. sTIPE equal in length to or longer than disc diameter, reddish-brown often almost black at the base, 0.3-3.0(-6.0) mm long. ECTAL EXCIPULUM composed of three layers: outer zone, 12-38 um thick, formed of some sparse rows of loosely branched, flexuous, brown hyphae (3.7-5.5 um in diam. +) arranged parallel to the surface, with thin walls encrusted with pigment giving a typically banded aspect and protruding as agglutinated “hairs” on flank and stipe or in groups as “clavate hairs” at the margin to form teeth; the medial layer of textura oblita, 15-60 um thick, formed of flexuose, subhyaline to light pale ochre hyphae, arranged parallel or at a low angle to the surface, with very narrow lumina and strongly gelatinized walls; the inner zone of imprecise thickness, formed of prismatic to doliform cells (textura prismatica) and not well delimited from the medullary excipulum, hyphae 5- 10 um (ft) in diameter, with pale chestnut content and thin walls. MEDULLARY EXCIPULUM well developed, of textura intricata and subhyaline cells with thin, smooth walls (3-5 um in diam. ft). SUBHYMENIUM compact, poorly developed and indistinguishable from the medullary area, apart from the brownish colour and highly intricate texture. asci 8-spored, cylindric-clavate, tapering at base to form a slender stalk, emerging from croziers, a round, truncate to fissured apex, with a long pore outlined J+ (deep blue) in both IKI and Melzer’s reagent, 125-152 x11-13 um (*), 110-130(-140) x9-11 um (+). spores biseriate to uniseriate or irregularly biseriate (in dead state), hyaline, ellipsoid to elliptic-
16 ... Galan & al.
fusoid, slightly inequilateral, regularly unicellular, rarely one to three-septate when over-mature, exceptionally producing subglobose microconidia (2-2.5 um * diam.) at one or both apices, with high number of globose lipid bodies (1-2 um) occupying most of the cytoplasm in the living spores (multiguttulate), forming two large distinct apical drops in dead state by coalescence, measuring 14-17(-18) x 6-7.5 um (*), 12-15(-16.8) x (5—)5.5-6.8(—7.2) um (+). PARAPHYSES straight, filiform to subclavate ca. 2-2.5 um diam. (*), enlarged to 4—-5.5 um (*) at the apex, sparsely septate, simple or branched below, equaling or scarcely exceeding the asci in length, bearing brown to reddish-brown vacuolar contents (“refractive vacuolar bodies” sensu Baral 1985) in the living state.
ADDITIONAL SPECIMENS EXAMINED: SPAIN, GUADALAJARA, Tamajon, surroundings
of Ermita del Humilladero, on a fallen leaf of Quercus ilex subsp. ballota in an open
mixed wood (with Juniperus thurifera) with scattered trees and leaf litter sparse on
the ground, 18.XI.1983, Galan, Moreno & Checa, det. R.G., rev. Palmer (AH 6235;
duplicate, J.T.P. as permanent slides); on stromatized petioles and veins of fallen old
Quercus ilex subsp. ballota leaves, 3.X1I.1984, Galan, Moreno & Palmer, det. J.T.P. (J.T.P.
4125); 3.XII.1991, Galan & Raitviir, det. R.G. & J.T.P. (AH 6720; duplicate, J.T.P. 4695).
Cifuentes, between Oter and Carrascosa de Tajo, on fallen leaves of Quercus ilex subsp.
ballota, 25.1X.1999, Nauta & Spooner, det. J.T.P. (J.T.P. 4978; duplicates, AH 7334, K).
Mirabueno, surroundings, 1000 m a.s.l., on side and veins of fallen leaves of Quercus ilex
subsp. ballota lying on ground, 24.1X.1999, Baral & Marson, det. Baral, rev. J.T.P. (J. T-P.
4977; duplicates, AH 7335, H.O.B. 6477E). MAprRID, Colmenarejo, UTM 30TVK1489,
885 ma.s.l., on petioles, lower (more exceptionally upper) faces and veins of fallen leaves
of Quercus ilex subsp. ballota, 18.X1.2011, det. R.G. (AH 7640); 11-XII.2011, det. EP, rev.
C.E.H. (C.E.H. 08468). ENGLAND, SoMERSET, between Allerford and Bossington,
on old, fallen leaves of Quercus ilex sampled, (apothecia developed in damp chamber
from 26.[V—30.V.1992), 20.11.1992, Palmer, det. J.T.P., rev. R.G. (J.T.P. 4713; duplicate,
AH 6813); (J. T-P. 4714; duplicate, AH 6812). GREECE, Maceponlia, Athos Mountain,
surroundings of Monastery Iveron, on stromatized fallen leaves of Quercus ilex subsp.
ilex, 25.11.1988, Palmer, det. J.T.P., rev. R.G. (J.T.P. 4415; duplicate, AH 6604).
CoMMENTS—There are at least three additional Rutstroemia species usually or occasionally reported from decaying Quercus leaves: R. sydowiana (Rehm) W.L. White (widely distributed at the Mediterranean basin) with “allantoid” spores (nearly ovoid, with basally bent beaks); R. petiolorum (Roberge ex Desm.) W.L. White (typically reported on Fagus leaves in Europe but occasionally also appearing on Quercus leaves) with allantoid to reniform spores with small polar gel caps; and R. latispora K.S. Thind & Saini, later transferred to Lambertella (Spooner 1987), found on Quercus incana leaves in the north-west Himalayas, having asymmetrical and much broader ascospores and longer asci than the two previous species (Thind & Saini 1968). In addition Rutstroemia “kalevi” Baral (ined.), which that could be misidentified as R. coracina (despite its Mollisia- like vacuoles in the paraphyses and narrower spores) grows on stromatized veins and petioles of Acer spp. in Central Europe (Baral & Marson 2005).
Rutstroemia coracina on Quercus (Spain) ...
Fic. 5. Rutstroemia coracina (AH 7641, epitype): A. Apothecia in their natural habitat (bar = 10 mm); B. Enlarged view of fresh apothecia (bar = 1 mm); C. Cross section of an apothecium (marginal region) (bar = 50 um); D. Encrusted wall of hyphae of the outer ectal excipulum (bar = 5 um); E. Hymenium showing asci arising from croziers and paraphyses (bar = 50 um); F, apex of a mature ascus with an euamyloid ring (bar = 10 um); G. Paraphyses containing many refractive vacuolar bodies in apical cells (bar = 5 um); H. Free 0-3-septate ascospores, exceptionally forming small conidia on germ tubes (bar = 20 um). Mountants: tap water, except fig. F (IKI). Watercolour by C.E.H.
17
18 ... Galan & al.
Rutstroemia coracina on Quercus (Spain) ... 19
Fic. 6 (left). Rutstroemia coracina: A. Radial section of an apothecium (AH 6812; bar = 200 um); B. Frontal view of the outer ectal excipulum forming marginal teeth (AH 6604; bar = 50 um); C. Radial section of an apothecium at middle flanks showing a three layered ectal excipulum (AH 6812; bar = 50 um); D, E, G, H. Asci with ascospores (AH 6235; bar = 20 um for D, G, H; 25 um for E); F. Mature asci, free spores and tips of paraphyses (AH 7641; bar = 25 um). Mountants: 2% KOH (figs. A, B & C); CRB (figs. D & H); MLZ (figs. E & G); tap water (fig. F). Dead state except fig. F. Photographs by R.G. & C.E.H. (fig. F). Abbreviations: outer (= oee), medial(= mee) and inner (= iee) layers of the ectal excipulum; medullary excipulum (= me).
Fic. 7 (above). Rutstroemia coracina (AH 6812): A. Radial section of an apothecium (bar = 200 tm); B. Enlarged view of an apothecium (bar = 100 um), at flank and margin, showing: the outer (= oee), medial(= mee) and inner (= iee) layers of the ectal excipulum; medullary excipulum (= me); hymenium (= h). Dead state (KOH 2%). Drawings by R.G. & C.E.H.
20 ... Galan & al.
Acknowledgements
We would like to thank Dr. Richard P. Korf for suggestions and improvement in English grammar. Also to A. Pueblas of the “Gabinete de Dibujo y Fotografia Cientifica” at the Alcala University for their invaluable help in the digital preparation of the photographs. We are grateful to Dr. Scott LaGreca (CUP), Dr. J. Rejos (AH), Dr. B.M. Spooner & Dr. B. Aguirre-Hudson (K), Mme J. Perreau (PC) for providing institutional specimens data, and Dr. M.N. Nauta and H.O. Baral for information on Rutstroemia coracina in their personal herbaria. One author (R.G.) extends his gratitude to the late Mr. J.T. Palmer for invaluable help with sclerotiniaceous fungi and a lifetime friendship. Finally, we wish to express our gratitude to Dr. S. Huhtinen and Dr. W.-Y. Zhuang for reviewing the manuscript and for their useful comments.
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Spooner BM. 1987. Helotiales of Australasia: Geoglossaceae, Orbiliaceae, Sclerotiniaceae, Hyaloscyphaceae. Bibliotheca Mycologica 116: 1-711.
Thind KS, Saini SS. 1968. The Helotiales of India VII. Proceedings of the Indian Academy of Sciences — Plant Sciences 67(4): 141-147.
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MYCOTAXON
http://dx.doi.org/10.5248/124.21 Volume 124, pp. 21-29 April-June 2013
Two species of Cladosporium associated with wood discoloration in Korea
YEONGSEON JANG, YOUNG MIN LEE, GYU-HYEOK KIM & JAE-JIN K1m*
Division of Environmental Science & Ecological Engineering, College of Life Sciences & Biotechnology, Korea University 5-1 Anam-dong, Seongbuk-gu, Seoul, 136-701, Korea
*CORRESPONDENCE TO: jae-jinkim@korea.ac.kr
AxBstTRACT — During our previous study on the diversity of molds inhabiting wood, many Cladosporium isolates were collected and phylogenetically identified. Among them were isolates of two species previously unrecorded from Korea. Here, we confirm the isolates as Cladosporium perangustum and C. ramotenellum and provide descriptions and illustrations of cultural and microscopic characters on standard media. We draw attention to a broadening of the range of several characters for C. ramotenellum and to possible alternative phylogenetic relationships of C. perangustum when comparing our multi-gene analysis with previous analyses.
KEY worDs — ascomycetes, phylogeny, taxonomy
Introduction
Cladosporium Link is one of the most widely known (ifunder-studied) molds. It is a cosmopolitan genus with more than 772 names (Dugan et al. 2004) and recently Bensch et al. (2012) recognized a total of 169 Cladosporium species. It has been isolated from air, soil, and many other substrates (Schubert et al. 2007, Bensch et al. 2010, 2012). Members are variously known to be saprobic, endophytic, or plant pathogenic (Schubert et al. 2007, Bensch et al. 2010, 2012). In Korea, about 20 Cladosporium species have been reported (Min 1985, Shin & Braun 1995, Shin & Im 1999, Kwon et al. 2000, 2001, Paul & Yu 2008, Korean Society of Plant Pathology 2009, Bensch et al. 2010). Recently Lee et al. (2012) phylogenetically examined Cladosporium representatives inhabiting wood products in Korea and recognized nine Cladosporium species. Among them were C. perangustum and C. ramotenellum, not previously recorded in Korea. Here, we confirm their identities by morphology and phylogenetic re-analysis and provide detailed descriptions and figures for these two Cladosporium species.
22 ... Jang & al.
Materials & methods
Morphological examination
Four Cladosporium isolates, KUC1462, KUC1767, KUC3027, and KUC5085 studied by Lee et al. (2012) were used in this study. They were inoculated onto SNA (Synthetic nutrient-poor agar: KH,PO, 1 g, KNO, 1 g; MgSO,°7H,O 0.5 g; KCI 0.5 g; glucose 0.2 g; sucrose 0.2 g; agar 20 g; distilled water 1000 ml) and cultivated at 25°C for 7 days. Preparations were mounted in Shear’s solution (CH,COOK 3 g; distilled water 150 ml; glycerin 60 ml; 95% ethanol 90 ml) according to Crous et al. (2009). Microscopic observations were made using an Olympus BX51 light microscope (Olympus, Tokyo, Japan) and photos were taken using the same microscope and an Axio Imager Al microscope (Zeiss, Jena, Germany). For determination of cultural characteristics, colonies were cultivated on PDA (Potato dextrose agar, Difco), MEA (Malt extract agar: malt extract 30 g; agar 15 g; distilled water 1000 ml), and OA (Oatmeal Agar, Difco) at 25°C for 14 days in the dark. The isolates were deposited at the National Institute of Biological Resources, Incheon, South Korea (KB) with the acronym KUC, which refers to the Korea University Culture Collections, Korea University, Seoul, South Korea.
Phylogenetic analysis
DNA sequences of four Cladosporium isolates (KUC1462, KUC1767, KUC3027, KUC5085) from the internal transcribed spacer (ITS), actin (ACT), and translation elongation factor-la (TEF) regions were obtained from Lee et al. (2012). ITS, ACT, TEF sequences were aligned separately with the reference sequences from Bensch et al. (2010) using MAFFT 6.885 (Katoh & Toh 2008). The L-INS-I alignment option was used for all datasets. Each dataset was manually edited with MacClade 4.08 (Maddison & Maddison 2005). Gaps were treated as missing. The best-fit model was applied for each dataset using AIC in MrModeltest 2.3 (Nylander 2004) and the three datasets were combined. Bayesian analysis was performed with the combined dataset using MrBayes 3.2.1 (Ronquist et al. 2012). Two independent runs (four chains each) of 705,000 generations were performed and every 100th tree was sampled. The first 25% of sampled trees were discarded and the remaining 75% were used for construction of a 50% major- rule consensus tree. The graphic representation of the likelihood scores of the sampled trees was checked. The standard deviation of split frequencies is below 0.01 and the potential scale reduction factor (PSRF) was close to 1.0. The tree was viewed via FigTree 1.3.1 (http://tree.bio.ed.ac.uk/software/figtree/).
Taxonomy
Cladosporium perangustum Bensch, Crous & U. Braun, Stud. Mycol. 67: 65. 2010. Fics 1-2
Mycelium internal and superficial; 1.5-4 um wide, septate, sometimes slightly constricted at septa, sometimes intercalary swellings and constrictions present, subhyaline or pale olivaceous-brown, smooth or verruculose. Conidiophores macronematous or micronematous arising terminally and laterally from hyphae, straight or slightly curved, filiform to narrowly cylindrical-oblong, unbranched, occasionally branched, conidiophores 14-142 x 2-3.5 um, pale olivaceous-
Cladosporium species discoloring wood (Korea) ... 23
B Fic. 1. Cladosporium perangustum (KUC1767) grown in 9 cm diam Petri dishes for 14 d at 25°C. A: PDA. B: MEA. C: OA.
]
Fic. 2. Cladosporium perangustum. A, B. Macro- and micronematous conidiophores (KUC1462). C. Ramoconidia and conidia (KUC1767). Scale bar = 10 um.
brown, asperulate-verruculose towards the base of conidiophores, at the apex smooth, sometimes slightly attenuated towards the apex. Conidiogenous cells integrated, mainly terminal, sometimes also intercalary, narrowly cylindrical-
24 ... Jang & al.
oblong, in intercalary cells loci situated on small peg-like lateral prolongations or just below the septum, 13-36 um long, with 1-4 apically crowded loci. Ramoconidia cylindrical-oblong, up to 44 um long, 2.5-3.5 um wide, rarely septate, base truncate, secondary ramoconidia narrowly ellipsoid to cylindrical- oblong, 8-21 x 2-3 um (av. + SD: 13.7 + 4.1 x 2.5 + 0.2), with 2-4 distal hila, pale olivaceous-brown. Conidia numerous, catenate, in branched chains, branching in all directions, 1-4 conidia in the terminal unbranched part of the chain, small terminal conidia globose, subglobose or ovoid to obovoid, 2-3.5 x 1.5-2 um (av. + SD: 2.9 + 0.4 x 1.8 + 0.2), intercalary conidia ovoid, limoniform to ellipsoid, somewhat fusiform, 4-8.5 x 2-3 um (av. + SD: 5.9 + 1.4 x 2.2 + 0.2), attenuated towards apex and base, with 1-3 distal hila, pale olivaceous-brown. CULTURE CHARACTERISTICS Colonies on PDA attaining 32-38 mm diam after 14 d at 25°C, greenish grey to dull green, reverse olive or dark green, fluffy, floccose or powdery, margins glabrous, whitish, olive grey or pale regular or somewhat undulate, aerial mycelium loosely floccose or felty, occasionally numerous small to large prominent exudates formed, sporulation abundant. Colonies on MEA reaching 20-29 mm diam after 14 d at 25°C, dark green or brown, reverse olive brown, velvety to floccose, margins white to light yellow, narrow to broad, regular to undulate, glabrous, aerial mycelium abundantly formed, sometimes covering most parts of colony surface, loosely to densely floccose or felty, growth habit plane to sometimes elevated, sporulation abundant. Colonies on OA 35-49 mm diam after 14 d at 25°C, grey to dark green, fluffy to felty-floccose, margins colorless or white, up to 2 mm diam, aerial mycelium abundant, covering large parts of the colony surface, dense, low to high, white, sporulating profusely. SPECIMENS EXAMINED: KOREA, GYEONGSANGBUK-DO, Bonghwa-gun, on the surface of Pinus densiflora log, 05 June 2001 (KB, KUC1462; GenBank JN033481, JN033508, JN033536); GYEONGSANGNAM-DO, Pusan, on the surface of Pinus radiata log, August 2000 (KB, KUC1767; GenBank JN033468, JN033495, JN033523); INCHEON, on the
log surface of Picea abies, 16 June 2007 (KUC5085; GenBank JN033460, JN033487, JN033515).
REMARKS — The colony morphology and microscopic features of the isolates agreed well with the description of C. perangustum (Bensch et al. 2010). The fungus has been reported from Africa, Asia, Australasia, Europe, and North America. According to Bensch et al. (2010), Cladosporium exile Bensch et al. and C. scabrellum Bensch et al. are similar to C. perangustum but C. exile has longer and wider conidiophores, wider ramoconidia and conidia, and shorter intercalary conidia. Cladosporium scabrellum differs in having mainly macronematous, wider conidiophores, and wider secondary ramoconidia. Compared to the other species studied by Lee et al. (2012), C. perangustum showed medium discoloration on Pinus densiflora and Pinus radiata.
Cladosporium species discoloring wood (Korea) ... 25
Fic. 3. Cladosporium ramotenellum (KUC3027) grown in 9 cm diam Petri dishes for 14 d at 25°C. A: PDA. B: MEA. C: OA.
Cladosporium ramotenellum K. Schub., Zalar, Crous & U. Braun, Stud. Mycol. 58: 137. 2007. FIGS 3-4 Mycelium internal and superficial, 1.5-4 um wide, septate, without swellings and constrictions, hyaline or subhyaline, smooth. Conidiophores macronematous and micronematous, arising terminally or laterally from branches of plagiotropous hyphae, straight or slightly flexuous, oblong or cylindrical, unbranched, sometimes branches often only as short lateral prolongations, 15-117 x 2-3.5 um, septate, pale olivaceous-brown or brown, smooth to verruculose. Conidiogenous cells integrated, terminal, sometimes also intercalary, cylindrical, 15-49 um long, proliferation sympodial, with few conidiogenous loci, mostly 1-3, loci sometimes situated on small lateral prolongations. Ramoconidia cylindrical-oblong, up to 46.5 um long, 2-3.5 um wide, usually 0-1-septate, pale olivaceous, smooth or verruculose, with a broadly truncate base, secondary ramoconidia subcylindrical to cylindrical- oblong, 12.5-22 x 2-4 um (av. + SD: 18 + 4 x 3.1 + 0.5), pale olivaceous, apex broadly rounded or slightly attenuated towards apex and base. Conidia numerous, catenate, in branched chains, branching in all directions, small terminal conidia numerous, 1-3 conidia in the terminal unbranched part of
Le
Fic. 4. Cladosporium ramotenellum (KUC3027). A, B. Macro- and micronematous conidiophore. C. Ramoconidia and conidia. Scale bars = 10 um.
26 ... Jang & al.
the chain, globose, subglobose or ovoid, obovoid, 3.5-7 x 2-3 um (av. + SD: 4.3 + 0.7 x 2.5 + 0.3), intercalary conidia ellipsoid to subcylindrical, or limoniform, 4-11 x 2.5-4(-4.5) um (av. + SD: 6.6 + 1.6 x 3 + 0.3).
CULTURE CHARACTERISTICS Colonies on PDA reaching 41-47 mm diam after 14 d at 25°C, olive due to abundant sporulation, margins entire, white, glabrous, aerial mycelium absent or sparse, growth flat with a somewhat folded and wrinkled colony center, no exudates, sporulation profuse. Colonieson MEA reaching 46-49 mm diam after 14 d at 25°C, olive, velvety, greyish green to dark green in reverse, margins entire, colorless, glabrous to feathery, aerial mycelium sparse, diffuse, growth flat with slightly elevated colony center, prominent exudates not formed, abundantly sporulating. Colonies on OA attaining 39-41 mm diam after 14 d at 25°C, olivaceous, margin entire, colorless or white, aerial mycelium sparse, growth flat, without exudates, sporulation profuse.
SPECIMEN EXAMINED: KOREA, on the surface of Chromated Copper Arsenate (CCA)-
treated wood product (Pinus radiata), 2003 (KUC3027; GenBank JN033464, JN033491,
JN033519). REMARKS — The colony morphology and microscopic features of our isolate agreed well with the description of C. ramotenellum (Schubert et al. 2007), although it has smaller intercalary conidia and secondary ramoconidia. The fungus previously has been reported only from Slovenia. According to Schubert et al. (2007), C. ramotenellum resembles C. cladosporioides (Fresen.) G.A. de Vries and C. tenellum K. Schub. et al. but differs from C. cladosporioides by having narrower conidiophores and conidia. Cladosporium tenellum possesses conidiophores with numerous conidiogenous loci, and shorter and wider conidia. Also, C. ramotenellum grows faster in culture than C. tenellum. Cladosporium ramotenellum showed a discoloration rate similar to C. perangustum on Pinus densiflora, but a little bit more prominent than C. perangustum on Pinus radiata (Lee et al. 2012).
Phylogeny
The resulting alignment contained 36 taxa (including the outgroup taxon) and 481, 139 and 201 characters (including alignment gaps) were used in the ITS, ACT and TEF partitions, respectively. The model used for ITS was SYM+G model. For ACT and TEE, HKY+I+G model and GTR+G model were used, respectively. For the Bayesian analysis, 10,705 trees were obtained from which the consensus tree and posterior probabilities were calculated (Fic. 5). The species relationships in the tree were sometimes not as well resolved as in previous studies, or resolved differently (Bensch et al. 2010, Bensch et al. 2012). The phylogenetic analysis revealed that Cladosporium perangustum isolates, KUC1462, KUC1767 and KUC5085 were clustered within the C. perangustum species clade with a high posterior probability value (1.0
Cladosporium species discoloring wood (Korea) ... 27
Cladosporium perangustum KUC1767 (JN033468) Cladosporium perangustum KUC1462 (JN033481) Cladosporium perangustum CBS 125996 (HM148121) Cladosporium perangustum CPC 13870 (HM148142) 1p Cladosporium perangustum CPC 11046 (HM148125) Cladosporium perangustum KUC5085 (JN033460) Cladosporium exasperatum CBS 125986 (HM148090) 1f Cladosporium chalastosporoides CBS 125985 (HM148001) Cladosporium hillianum CBS 125988 (HM148097) Cladosporium scabrellum CBS 126358 (HM148195) Cladosporium phyllophilum CPC 13873 (HM148155) Cladosporium licheniphilum CBS 125990 (HM148111) Cladosporium varians CBS 126362 (HM148224) 4 Cladosporium chubutense CBS 124457 (FJ936158) Cladosporium colombiae CBS 274.80B (FJ936159) Cladosporium pini-ponderosae CBS 124456 (FJ936160) Cladosporium asperulatum CBS 126340 (HM147998) Cladosporium myrtacearum CBS 126349 (HM148116) Cladosporium funiculosum CBS 122128 (HM148093) Cladosporium pseudocladosporioides CPC 14992 (HM148192) Cladosporium australiense CBS 125984 (HM147999) ip Cladosporium cucumerinum CBS 176.54 (HM148078) Cladosporium subuliforme CBS 126500 (HM148196) Cladosporium xylophilum CPC 14364 (HM148234) Cladosporium delicatulum CPC 14363 (HM148088) Cladosporium inversicolor CPC 14368 (HM148109) Cladosporium lycoperdinum CBS 274.80C (HM148114) Cladosporium basiinflatum CBS 822.84 (HM148000)
0.81
0.67,
0.59
0.61., | Cladosporium ramotenellum CPC 12047 (EF679385)
Cladosporium ramotenellum KUC3027 (JN033464)
Cladosporium ramotenellum CPC 12043 (EF679384) Cladosporium spinulosum CBS 102044 (EF679387) Cladosporium iridis CBS 138.40 (EF679370)
Cladosporium ossifragi CBS 843.91 (EF679382)
Cercospora beticola CPC 11557 (AY840527)
Fic. 5. 50% majority-rule consensus tree of 10,575 trees resulting from a bayesian analysis of 36 sequences in a combined ITS, ACT, and TEF alignment. Bayesian posterior probabilities 250%
are shown. The tree was rooted to sequences of Cercospora beticola strain CPC 11557. GenBank Accession numbers of ITS sequences are shown in parentheses.
28 ... Jang & al.
p-p.). In our analysis, C. perangustum was sister to C. exasperatum Bensch et al., a result differing substantially from the analysis of Bensch et al. (2010). In our analysis, Cladosporium ramotenellum KUC3027 was monophyletic with C. ramotenellum CPC 12047, but not with C. ramotenellum CPC 12043 and CBS 170.54. Also the monophyly of C. ramotenellum KUC3027 and CPC 12047 was weakly supported (61% posterior probability value). Nevertheless, the cultural and microscopic observations of our C. ramotenellum KUC3027 were consistent with the descriptions of C. ramotenellum (Schubert et al. 2007; Bensch et al. 2010).
Acknowledgments
This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (NRF-2013R1A1A2A10011390) and was funded by the project on survey and excavation of Korean indigenous species of NIBR under the Ministry of Environment, Republic of Korea. We are grateful to Dr. Pedro Crous and Dr. FM. Dugan for their valuable suggestions on the manuscript.
Literature cited
Bensch K, Groenewald JZ, Dijksterhuis J, Starink-Willemse M, Andersen B, Summerell BA, Shin H-D, Dugan FM, Schroers H-J, Braun U, Crous PW. 2010. Species and ecological diversity within the Cladosporium cladosporioides complex (Davidiellaceae, Capnodiales). Stud. Mycol. 67: 1-94. http://dx.doi.org/10.3114/sim.2010.67.01
Bensch K, Braun U, Groenewald JZ, Crous PW. 2012. The genus Cladosporium. Stud. Mycol. 72: 1-401. http://dx.doi.org/10.3114/sim0003
Crous PW, Verkleij GJM, Groenewald JZ, Samson RA (eds). 2009. Fungal biodiversity. CBS Laboratory Manual Series 1. Centraalbureau voor Schimmelcultures, Utrecht, Netherlands.
Dugan FM, Schubert K, Braun U. 2004. Check-list of Cladosporium names. Schlechtendalia 11: 1-103.
Katoh K, Toh H. 2008. Recent developments in the MAFFT multiple sequence alignment program. Briefings Bioinf. 9: 286-298. http://dx.doi.org/10.1093/bib/bbn013
Korean Society of Plant Pathology. 2009. List of plant diseases in Korea, 5th edn. The Korean Society of Plant Pathology, Suwon, Korea.
Kwon J-H, Kang S-W, Park C-S. 2000. Occurrence of sword bean scab caused by Cladosporium cucumerinum in Korea. Mycobiology 28: 54-56.
Kwon J-H, Kang S-W, Park C-S. 2001. Occurrence of strawberry scab caused by Cladosporium herbarum in Korea. Mycobiology 29: 110-112.
Lee YM, Jang Y, Kim G-H, Kim J-J. 2012. Phylogenetic analysis and discoloration characteristics of major molds inhabiting woods. Part 3. Genus Cladosporium. Holzforschung 66: 537-541. http://dx.doi.org/10.1515/hf.2011.184
Maddison D, Maddison W. 2005. MacClade 4: Analysis of phylogeny and character evolution. Version 4.08. Sinauer Associates, Sunderland, MA, USA.
Min KH. 1985. Some undescribed Cladosporium, Alternaria, Curvularia and Eurotium repens in Korea. Kor. J. Mycol. 14: 1-8.
Nylander JAA. 2004. MrModeltest v2. Evolutionary Biology Center, Uppsala University, Uppsala, Sweden.
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Paul NC, Yu SH. 2008. Two species of endophytic Cladosporium in pine trees in Korea. Mycobiology 36: 211-216. http://dx.doi.org/10.4489/MYCO.2008.36.4.211
Ronquist FE, Teslenko M, Mark P van der, Ayres D, Darling A, Hohna S, Larget B, Liu L, Suchard MA, Huelsenbeck JP. 2012. MrBayes 3.2: Efficient Bayesian phylogenetic inference and model choice across a large model space. Syst. Biol. 61: 539-542. http://dx.doi.org/10.1093/sysbio/sys029
Schubert K, Groenewald JZ, Braun U, Dijksterhuis J, Starink MS, Hill CF, Zalar P, Hoog GS de, Crous PW. 2007. Biodiversity in the Cladosporium herbarum complex (Davidiellaceae, Capnodiales), with standardisation of methods for Cladosporium taxonomy and diagnostics. Stud. Mycol. 58: 105-156. http://dx.doi.org/10.3114/sim.2007.58.05
Shin H-D, Braun U. 1995. Cladosporium alliicola sp. nov. on Allium victorialis var. platyphyllum. Kor. J. Mycol. 23: 139-143.
Shin H-D, Lee H-T, Im D-J. 1999. Occurrence of German iris leaf spot caused by Cladosporium iridis in Korea. Plant Pathol. J. 15: 124-126.
ISSN (print) 0093-4666 © 2013. Mycotaxon, Ltd. ISSN (online) 2154-8889
MYCOTAXON
http://dx.doi.org/10.5248/124.31 Volume 124, pp. 31-37 April-June 2013
Phialophora avicenniae sp. nov., anew endophytic fungus in Avicennia marina in China
YUE-LIAN Liu*?, PING-GEN XI, XIAO-LAN HE! & ZI-DE JIANG"
‘College of Natural Resources and Environment, South China Agricultural University, Guangzhou 510642, China
Agricultural College, Guangdong Ocean University, Zhanjiang 524088, China
*CORRESPONDING AUTHOR: Zdjiang@scau.edu.cn
ABSTRACT — Three isolates of an endophytic fungus were obtained from Avicennia marina. Morphological and molecular evidence indicates that they are identical and represent a new species of Phialophora, described here as P. avicenniae. The new species is characterized by brown colonies, branched conidiophores, and catenulate conidia.
Key worps — Herpotrichiellaceae, ITS sequence, taxonomy
Introduction
The morphologically vaguely delimited hyphomycete genus Phialophora Medlar is characterized by dark and slow-growing colonies, pigmented hyphae, solitary or aggregated conidiophores, and flask-shaped phialides usually with a flaring collarette. The type of Phialophora is P. verrucosa Medlar, diagnosed by its slow-growing dark olivaceous colonies and usually pigmented phialides bearing an even darker collarette (Medlar 1915). Phialophora has been regarded as highly polyphyletic, comprising anamorphs of discomycetes, pyrenomycetes, and loculoascomycetes (Gams 2000). Gams & Holubova- Jechova (1976) distinguished sect. Catenulatae with catenate conidia having a truncate base. Some members of this section and other phialophora-like species with catenulate conidia, such as Brachyalara straminea Réblova & W. Gams, Exochalara longissima (Grove) W. Gams & Hol.-Jech., and Infundichalara microchona (W. Gams) Réblova & W. Gams, have been segregated from Phialophora by molecular phylogeny analyses and morphological characters (Bogale et al. 2010; Réblova et al. 2011). Phialophora s. str. is recognized as a member of the Herpotrichiellaceae (Untereiner et al. 1995).
32 ... Liu & al.
We obtained three isolates of an endophytic fungus from twigs of Avicennia marina (Forssk.) Vierh. (Acanthaceae) in Zhanjiang, Guangdong Province, China, in 2007. The taxonomic status of these isolates was determined using morphological and molecular methods, showing them to represent a new Phialophora species. The isolates are stored in the laboratory of the Agricultural College, Guangdong Ocean University, and subcultures at CBS in Utrecht, the Netherlands.
Materials & methods
Isolates
Healthy twigs (0.4-0.8 cm diam.) of Avicennia marina were thoroughly washed in running tap water and then sterilized by washing in 75% ethanol for 2 min and 0.1% mercuric chloride (vw) for 2 min. The twigs were rinsed 3 times in sterile distilled water and then cut into pieces (0.5 cm in length). The small pieces were evenly spaced in Petri dishes (9 cm diam) containing a medium (Arnold et al. 2000) consisting of (per litre distilled water) dextrose 20 g, peptone 10 g and agar 20 g, with addition of 60 mg streptomycin before pouring the plates. The Petri dishes were incubated at 27°C. Hyphal tips were transferred to potato dextrose agar (PDA) dishes.
Morphological examination
Three cultures of every isolate grown on PDA plates were investigated for colony characteristics. Thirty conidia were measured under an OLtympus-BX51 microscope. Scanning electron microscopy (SEM) was used to observe phialide morphology. Specimens were flash-frozen (—196°C) in liquid nitrogen under vacuum for cryo-SEM, transferred to the preparation chamber, and then to the SEM chamber where ice particles were sublimated (—80°C). Samples were sputter-coated with E-1010 in the preparation chamber for 75 sec under 2.0 KV at -170°C. Specimens were viewed under 5 KV at -188°C with a Puitips SL30 scanning electron microscope.
DNA extraction, PCR amplification, DNA sequencing, and phylogenetic analyses
After the isolates were grown and incubated on PDA plates at 27°C for 10 days, the mycelia were scraped off. Genomic DNA was extracted from the mycelia using SDS procedure (Lee & Taylor 1990) with a few modifications. The quality and quantity of DNA were visually assessed by staining with ethidium bromide in 1% agarose gel electrophoresis. The partial ITS sequences were amplified according to Geiser et al. (2005) using primers ITS1 and ITS4 (White et al. 1990) in TP600 (TaKaRa PCR Thermal Cycler Dice). Shanghai Invitrogen Biotechnology Co., Ltd. sequenced the DNA. The ITS sequences of the isolates were aligned using MEGA version 4.1 (Tamura et al. 2007) and manually optimized to ensure positional homology. Gaps were considered as missing data. Seventeen sequences of Phialophora and one sequence of an unidentified ascomycotan species were downloaded from GenBank, and Capronia semiimmersa (Herpotrichiellaceae) was chosen as outgroup. A phylogenetic tree was constructed using the neighbor-joining method of MEGA version 4.1 (bootstrapped 1000 replicates).
Phialophora avicenniae sp. nov. (China) ... 33
Taxonomy
Phialophora avicenniae Yue L. Liu & Z.D. Jiang, sp. nov. Fras. 1-2 MycoBank MB561173 Differs from Phialophora clavispora and P. olivacea by its branching conidiophores and endophytic habit and from P oxyspora by its ovoid to globose conidia and endophytic habit.
Type: China, Guangdong Province, Zhanjiang, endophytic in twig of Avicennia marina, 18 April 2007, Liu Yuelian Q37 (Holotype, CBS 130286 [lyophilized culture]; GenBank, GQ500118).
EryMo.ocy: named after the host genus, Avicennia.
COLONIES initially yeast-like, gradually becoming cottony, brown, reaching 2-3 cm diam. after 5 days at 27°C on PDA plates; reaching 9 cm diam. after 14
Figure 1. Phialophora avicenniae (CBS 1302876). a. Colony; b-f. Conidiophores; g. Phialides; h-l. Conidial chains. Scale bars: b-f, h-] = 10 um; g = 2 um.
34 ... Liu & al.
FiGuRE 2. Phialophora avicenniae. Conidiophore and conidia. Bar = 10 um.
days and changing from brown to black-brown. HyPHAE hyaline to subhyaline, smooth; occasionally brown, verrucose, 1.5-3.0 um wide. CONIDIOPHORES hyaline or brown, branched, rarely simple, septate, smooth, and variable in size. PHIALIDES usually aggregated on conidiophores, flask-shaped or cylindrical, hyaline or brown, smooth and thick-walled, 6.5-11 x 1.8-3.5 um. Collarette somewhat darker than the phialide body, funnel-shaped or flaring, 0.5-1.0 x 0.3-0.5 um. Conip14 catenulate, hyaline or brown, 1-celled, smooth, ovoid to globose, slightly apiculate at the base, 1.6-2.5 x 1.8-3.5 um. CHLAMYDOSPORES absent.
TELEOMORPH: unknown
Hasirat: endophytic in twigs of Avicennia marina.
ADDITIONAL CULTURES EXAMINED: CHINA. GUANGDONG PROVINCE, Zhanjiang,
endophytic in twigs of Avicennia marina, 10 August 2007, Liu Yuelian Q44 (CBS 130287; GenBank, HM055753); Liu Yuelian Q48 (CBS 130288; GenBank, HM055754).
Phialophora avicenniae sp. nov. (China) ... 35
Notes: Phialophora verrucosa, the type of Phialophora, differs from the new species by olivaceous-black colonies and broadly ellipsoidal conidia adhering in slimy heads. The new species resembles P. clavispora W. Gams, P. olivacea W. Gams, and P. oxyspora W. Gams by its catenulate conidia. However, P. clavispora and P. olivacea have simple conidiophores, and P oxyspora has fusiform conidia (Gams & Holubova-Jechova 1976). In addition, the new species is endophytic, while the other three species are so far only known as saprobes.
Phylogenetic analyses ITS rDNA sequences obtained from the isolates of CBS 130286, CBS 130287, and CBS 130288 were submitted to GenBank and received the
P. olivacea AB190379 P. reptans EUS14699
66 P. europaea EF540756 74 P. sessilis EUS14700 54 P. oxyspora AB190870 clade |
@ CBS 130286 GQ500118.1 @ CBS 130287 HM055753.2 98 | @ CBS 130288 HM055754.1 54 Ascomycota sp. FJ037728 P. japonica AB190378 @ P. verrucosa DQ404353 P. clavispora AB190868 P. chrysanthemi AB190387 8 | en P. intermedia AB190399 P. cyclaminis AB190390 so P. geniculata AB190395 1004 P. alba AB190382 ciate I 53 P. atrovirens AB190383 100 P. foetens AB190394 57 P. cinerescens AB190388 91®© P. mustea JN1I23359
Capronia semiimmersa FJ225726 4 0.1
FiGuRE 3. Phylogenetic tree based on neighbor-joining of ITS sequences, showing the relationships among Phialophora avicenniae and related species.
36 ... Liu & al.
accession numbers GQ500118, HM055753, and HM055754. BLast (Basic Local Alignment Search Tool) analysis showed that the ITS sequences are 99% similar to an unidentified strain labeled “Ascomycota sp. GX6-1C” (GenBank FJ037728; isolated as an endophyte from an unknown mangrove host in China) and 79-87% similar to various GenBank isolates of Phialophora spp. A neighbor-joining analysis shows the relationships of the three strains with 17 other species of Phialophora (Fic. 3). The phylogram also shows that the unidentified strain forms a monophyletic clade (bootstrap 98%) with the three P. avicenniae strains, and is probably conspecific.
The Phialophora species shown here form two distinct clades (clade I and clade II) where P. avicenniae is part of clade I (bootstrap 54%), showing a sister relationship with P olivacea, P. reptans, P. sessilis, P europaea, and P. oxyspora.
Acknowledgements
We express our sincere thanks to Prof. W. Gams who revised the manuscript and deposited the strains in CBS. We also thank Prof. Yi-Jian Yao (Key Laboratory of Systematic Mycology and Lichenology, Institute of Microbiology, Chinese Academy of Sciences) for reviewing the manuscript. The authors are grateful to Dr. Martina Réblova (Department of Taxonomy, Institute of Botany, Academy of Sciences, Prihonice, Czech Republic) for providing valuable suggestions. This study was supported by the National Science Foundation of China (No. 30870015).
Literature cited
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Bogale M, Orr MJ, O'Hara MJ, Untereiner WA. 2010. Systematics of Catenulifera (anamorphic Hyaloscyphaceae) with an assessment of the phylogenetic position of Phialophora hyalina. Fungal Biology 114: 396-409. http://dx.doi.org/10.1016/j.funbio.2010.02.006
Gams W. 2000. Phialophora and some similar morphologically little-differentiated anamorphs of divergent ascomycetes. Studies in Mycology 45: 187-199.
Gams W, Holubova-Jechova V. 1976. Chloridium and some other dematiaceous hyphomycetes growing on decaying wood. Studies in Mycology 13: 62-99.
Geiser DM, Lewis-Ivey ML, Hakiza G, Juba JH, Miller SA. 2005. Gibberella xylarioides (anamorph Fusarium xylarioides), a causative agent of coffee wilt disease in Africa, is a previously unrecognized member of the G. fujikuroi species complex. Mycologia 97: 191-201. http://dx.doi.org/10.3852/mycologia.97.1.191
Lee SB, Taylor JW. 1990. Isolation of DNA from fungal mycelia and single spores. 283-287, in: MA Innis et al. (eds), PCR protocols: a guide to methods and applications. Academic Press, New York.
Medlar EM. 1915. A new fungus, Phialophora verrucosa, pathogenic for man. Mycologia 7: 200-203. http://dx.doi.org/10.2307/3753363
Réblova M, Gams W, Stépanek V. 2011. Thenewhyphomycete genera Brachyalara and Infundichalara, the similar Exochalara and species of Phialophora sect. Catenulatae (Leotiomycetes). Fungal Diversity 46: 67-86. http://dx.doi.org/10.1007/s13225-010-0077-6
Phialophora avicenniae sp. nov. (China) ... 37
Tamura K, Dudley J, Nei M, Kumar S. 2007. MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Molecular Biology and Evolution 24: 1596-1599. http://dx.doi.org/10.1093/molbev/msm092
Untereiner WA, Straus NA, Malloch D. 1995. A molecular-morphotaxonomic approach to the systematics of the Herpotrichiellaceae and allied black yeasts. Mycological Research 99: 897-913.
White TJ, Bruns T, Lee S, Taylor JW. 1990. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. 315-322, in: MA Innis et al. (eds). PCR protocols; a guide to methods and applications. Academic Press, New York, USA.
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http://dx.doi.org/10.5248/124.39 Volume 124, pp. 39-45 April-June 2013
Ramichloridium strelitziae associated with sooty blotch and flyspeck on Ravenala madagascariensis in China
BAojJUN HAo*?, WENHUAN LI**?, CHEN CHEN’, Liu GAo’, RONG ZHANG’, BAOTONG WANG', GUANGYU SUN" & MARK L. GLEASON?
'State Key Laboratory of Crop Stress Biology in Arid Areas and College of Plant Protection, Northwest A&F University, Yangling, Shaanxi, 712100, P. R. China
*Department of Plant Pathology, Iowa State University, Ames, Iowa 50011, USA
College of Life Science, Northwest A&F University, Yangling, Shaanxi, 712100, P. R. China
“CORRESPONDENCE TO: sgy@nwsuaf.edu.cn
ABSTRACT — ‘The first report of Ramichloridium strelitziae from China is documented. In a survey of host plants for SBFS fungi, we isolated the fungus from colonies on stems of Ravenala madagascariensis collected from Haikou, Hainan, China. It is distinguished from the other species in the genus by morphological characters and phylogenetic analysis based on ITS sequences.
KEY worps — microfungi, Capnodiales, taxonomy, phylogeny, traveller's palm
Introduction
Sooty blotch and flyspeck (SBFS) are epiphytes that colonize the waxy cuticle of a wide range of plants in humid regions worldwide. SBFS fungi can result in a black or sooty appearance leading to cosmetic damage that can cause significant economic losses (Batzer et al. 2005).
For approximately 70 years, SBFS was viewed as two distinct diseases, each caused by a single species of fungus. Recently, this concept was revised by combining morphological characterization with genetic analysis. Presently, it is evident that SBFS includes a spectrum of intergraded mycelial types and comprises more than 80 species (Batzer et al. 2008, Frank et al. 2010, Gleason et al. 2011, Liet al. 2012, Yang et al. 2010).
In this study, we identified one isolate that was described as the first record of R. strelitziae on stems of Ravenala madagascariensis Sonn. (Strelitziaceae; traveller's palm) from China based on morphological comparison and phylogenetic relationships.
# = Joint first authors
AO ... Hao &al.
Materials & methods
Isolates
Ravenala madagascariensis stems with flyspeck signs (Gleason et al. 2011) were found in Jinniuling Park in Haikou city, Hainan Province (20°00'26"N 110°20'31"E), China, in October 2011. Thalli on the stem were transferred directly from colonies to potato dextrose agar (PDA) slants in a sterile environment and cultured at 25°C for 1 month in darkness (Sun et al. 2003). Hyphal tips were then transferred to malt extract agar (MEA) plates. The isolates were allowed to grow onto an adjacent, sterile cover slip that had been partially inserted into the agar surface at a 60° angle in order to measure and observe fungal structures (Li et al. 2011). Microscopic examination was made after 14 d of incubation. Thirty measurements per relevant microscopic structure were gathered where possible. Colony descriptions (surface and reverse) were made after 2 weeks of growth on MEA plates at 25+1°C in the dark. The isolate acronym HKLRJ-4 was a temporary laboratory number. Representative dried culture and plant specimens were deposited in the Fungal Herbarium of Northwest A&F University (HMUABO), Yangling, Shaanxi Province, China.
DNA extraction, PCR, and phylogenetic analysis
Genomic DNA for polymerase chain reaction (PCR) was obtained according to the protocol of Li et al. (2011). The primer pair ITS1-F and ITS4 was used to amplify the internal transcribed spacer (ITS) region of nuclear ribosomal DNA. The PCR reactions were carried out with Taq polymerase, 1x PCR buffer, 2 mM MgCl, 0.2 mM of each dNTP, 0.4 uM of each primer, and 2 ul of template DNA, and was made up to a total volume of 25 ul with sterile water. Reactions were performed on a Bio-Rad PCR System S1000TM Thermal Cycler. The amplification conditions were: initial denaturation at 94°C for 90 min followed by 35 cycles of denaturation at 94°C for 35 s, annealing at 52°C for 60 s, extension at 72°C for 1 min, and a final extension step at 72°C for 10 min. The PCR products were sequenced by Sangon Biotech Co., Shanghai, China.
The ITS nucleotide sequence generated in this study was added to other sequences with high homology as the result of a BLAST search. Cladosporium bruhnei was used as the outgroup taxon. Sequences were imported into BioEdit 5.0.9.1 (Hall 1999). Preliminary alignments were performed using CLUSTAL-X (Thompson et al. 1997), then manually adjusted. Phylogenetic analysis of aligned DNA sequences was carried out with PAUP version 4.0b 10 for 32-bit Microsoft Windows (Swofford 2003). Heuristic searches were performed with 1,000 random sequence additions. Clade stability was assessed by 1,000 bootstrap replications. Other measures calculated for parsimony analysis included tree length, consistency index (CI), retention index (RI), rescaled consistency index (RC) and homoplasy index (HI).
The ITS sequence described in this study is deposited in GenBank as JX502176, and the alignment and representative tree were deposited in TreeBase (http://purl.org/ phylo/treebase/phylows/study/TB2:S13803).
Results Phylogenetic analyses
The ITS alignment contained 29 taxa (including the outgroup) and 520 characters including alignment gaps. Of these characters, 266 were constant,
Ramichloridium strelitziae, new for China... 41
62 were variable and parsimony-uninformative, and 192 were parsimony- informative. One of the 10 equally most parsimonious trees saved from the maximum parsimony analysis is shown in Fic. 1. From the most parsimonious tree, two major clades were resolved. One clade, with 100% bootstrap support, contained fifteen species in Ramichloridium sensu stricto, Uwebraunia, Dissoconium, and Pseudoveronaea (all in Dissoconiaceae). The other major clade
55 | Ramichloridium apiculatum EU041794
99 | R. mali EF627452
541 'R cucurbitae JQ622087 100
Ramichloridium R. luteum CPC 18962
R. punctatum CPC 18974 Uwebraunia musae CBS 122454
U. australiensis EF394854
100 Uwebraunia
U. commune DQ302948 U. dekkeri EU851919 Dissoconium aciculare CPC 10805
68 F- D. eucalypti CBS 120039 100 Dissoconium
100) LD. aciculare AF 173308 D. proteae CBS 122900 100 [~ _Pseudoveronaea ellipsoidea CPC 18970 Pseudoveronaea Pseudoveronaea obclavata CPC 18972 gf Zasmidium angulare JQ622088
Z. nocoxi CBS 125009
541 __ Ramichloridium cerophilum AF050286
Z. xenoparkii CBS 111089 PA Z. lonicericola CPC 11671 go] 2 citri CPC 10522 95} © Z. anthuriicola CBS 118742 Mycosphaerellaceae 100 Z. scaevolicola HM122240 1007 Ramichloridium streliziae CBS 121711 $3 HKLRJ-4 R. musae EU041801 R. biverticillatum EU041796 40 R. australiense EU041795
Cladosporium bruhnei DQ780401
Fic. 1. One of 10 equally parsimonious trees determined from ITS sequences. Bootstrap support values (>50%) based on 1000 replicates are shown at the node. (CI = 0.6611, RI = 0.8657, RC = 0.5723, HI = 0.3389). The scale bar shows 10 changes. The tree is rooted to Cladosporium bruhnei and the new sequence is presented in bold.
42 ... Hao &al.
had a bootstrap value of 100% including twelve species in Ramichloridium sensu lato and Zasmidium (all in Mycosphaerellaceae). Our isolate (HKLRJ-4) grouped with Ramichloridium strelitziae (Mycosphaerellaceae) with 100% bootstrap support, indicating that they might represent the same species.
Morphology and culture characteristics
Ramichloridium strelitziae Arzanlou, W. Gams & Crous, Stud. Mycol. 58: 74. 2007. Fic. 2
MYcELIvM consisting of smooth, thin-walled, septate, verrucose, pale brown hyphae, 2-3 um wide. CONIDIOPHORES erect, solitary, arising from creeping aerial hyphae, subhyaline, later becoming pale brown, thick-walled, 1-3(-4)- septate, smooth or verruculose, 2 um wide, up to 43 um long. CONIDIOGENOUS cells terminally integrated, (8-)11-38 um long, subhyaline, later turning pale brown, fertile part as wide as the basal part, sympodially proliferating, forming a straight rachis; Scars thickened and darkened, approx. 0.5 um diam. ConipiA (3.5-)4-5 x (1-)2(-3) um, solitary, aseptate, subhyaline, smooth or verruculose, oblong, ellipsoidal to clavate, base truncate with unthickened, non-pigmented hilum.
CULTURAL CHARACTERISTICS — CoLony diameter after 14 d on MEA at 24°C reached 6 mm diam with entire margin and compact, raised, dense aerial mycelium, surface olivaceous-grey, reverse olivaceous-black.
Host CHARACTERISTICS — On stem surface, shiny, black, sclerotium-like bodies, round to oval (120-220 um diam) with no visible mycelial mat, densely arranged (3-5/mm_°). The flyspeck symptom on Ravenala madagascariensis did not physically damage the plants, but greatly reduced their ornamental value.
Based on phylogenetic analysis and morphological characters of the anamorph, we identified isolate HKLRJ-4 as Ramichloridium strelitziae. SPECIMEN EXAMINED: CHINA. HAINAN PROVINCE, Haikou City, Jinniuling Park,
20°00'26"N 110°20'31”E, on stems of Ravenala madagascariensis, Oct 2011, Li WH (HMUABO HKLRJ-4; GenBank, JX502176).
Discussion
The modern circumscription of Ramichloridium is still heterogenous, containing species in two _ different families (Dissoconiaceae and Mycosphaerellaceae), and only species clustering in Capnodiales were considered to be true Ramichloridium (Arzanlou et al. 2007). According to Li et al. (2012) and Arzanlou et al. (2007), R. apiculatum (the generic type), R. cucurbitae, R. indicum, R. luteum, R. mali, and R. punctatum (Ramichloridium s. str.) clustered with Dissoconium species (all now placed in Dissoconiaceae), and other Ramichloridium s. lat. species grouped in Mycosphaerellaceae. Our strain clustered among the mycosphaerellacean species with R. strelitziae.
Ramichloridium strelitziae, new for China... 43
Fic. 2. Ramichloridium strelitziae (HKLRJ-4). A. Signs on stem of Ravenala madagascariensis with close-up view. B. Colony on MEA after 14 days. C-E Conidiophores. G-H. Conidia. Bars: C-F = 10um; G-H = 5um.
Arzanlou et al. (2007) reported Ramichloridium strelitziae from leaves of Strelitzia nicolai in South Africa. The species has also been found in bathrooms and washing machines and is able to degrade surfactants, soap, and shampoo (Hamada & Abe 2010). Our results show that R. strelitziae may also be involved in the sooty blotch and flyspeck complex.
Acknowledgments
This work was supported by National Natural Science Foundation of China (31170015, 31171797), the 111 Project from Education Ministry of China (B07049), Top Talent Project of Northwest A&F University and the earmarked fund for Modern Agro- industry Technology Research System (nycytx-08-04-04). The authors wish to thank Dr Eric H.C. McKenzie (Landcare Research, Private Bag 92170, Auckland, New Zealand) and Professor Zhongyi Zhang (College of Plant Protection, Yunnan Agricultural University, Kunming, Yunnan, China) for reviewing the manuscript.
Literature cited
Arzanlou M, Groenewald JZ, Gams W, Braun U, Shin H-D, Crous PW. 2007. Phylogenetic and morphotaxonomic revision of Ramichloridium and allied genera. Studies in Mycology 58: 57-93. http://dx.doi.org/10.3114/sim.2007.58.03
44 ... Hao &al.
Batzer JC, Gleason ML, Harrington TC, Tiffany LH. 2005. Expansion of the sooty blotch and flyspeck complex on apples based on analysis of ribosomal DNA gene sequences and morphology. Mycologia 97: 1268-1286. http://dx.doi.org/10.3852/mycologia.97.6.1268
Batzer JC, Diaz Arias MM, Harrington TC, Gleason ML, Groenewald JZ, Crous PW. 2008. Four species of Zygophiala (Schizothyriaceae, Capnodiales) are associated with the sooty blotch and flyspeck complex on apple. Mycologia 100: 246-258. http://dx.doi.org/10.3852/mycologia.100.2.246
Frank J, Crous PW, Groenewald JZ, Oertel B, Hyde KD, Phengsintham P, Schroers H-J. 2010. Microcyclospora and Microcyclosporella: novel genera accommodating epiphytic fungi causing sooty blotch on apple. Persoonia 24: 93-105. http://dx.doi.org/10.3767/003158510X510560
Gleason ML, Batzer JC, Sun GY, Zhang R, Diaz Arias MM, Sutton TB, Crous PW, Ivanovic M, McManus PS, Cooley DR, Mayr U, Weber RWS, Yoder KS, Del Ponte EM, Biggs AR, Oertel B. 2011. A new view of sooty blotch and flyspeck. Plant Disease 95: 368-383. http://dx.doi.org/10.1094/PDIS-08- 10-0590
Hall TA. 1999. BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/N'TT. Nucleic Acids Symposium Series 41: 95-98.
Hamada N, Abe N. 2010. Growth characteristics of four fungal species in bathrooms. Biocontrol Science 15: 111-115. http://dx.doi.org/10.4265/bio.15.111
Li HY, Sun GY, Batzer JC, Crous PW, Groenewald JZ, Karakaya A, Gleason ML. 2011. Scleroramularia gen. nov. associated with sooty blotch and flyspeck of apple and pawpaw from the Northern Hemisphere. Fungal Diversity 46: 53-66. http://dx.doi.org/10.1007/s13225-010-0074-9
Li HY, Sun GY, Zhai XR, Batzer JC, Mayfield DA, Crous PW, Groenewald JZ, Gleason ML. 2012. Dissoconiaceae associated with sooty blotch and flyspeck on fruits in China and the United States. Persoonia 28: 113-125. http://dx.doi.org/10.3767/003158512X651157
Sun GY, Zhang R, Zhang Z, Zhang M. 2003. Isolation of sooty blotch and flyspeck fungi from apple surface by picking up the thalli. Acta Phytopathologica Sinica 33: 479-480.
Swofford DL. 2003. PAUP*. Phylogenetic analysis using parsimony (* and other methods). Version 4.0. Sinauer Associates, Sunderland, Massachusetts, USA.
Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG. 1997. The Clustal_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Research 25: 4876-4882. http://dx.doi.org/10.1093/nar/25.24.4876
Yang HL, Sun GY, Batzer JC, Crous PW, Groenewald JZ, Gleason ML. 2010. Novel fungal genera and species associated with the sooty blotch and flyspeck complex on apple in China and the USA. Persoonia 24: 29-37. http://dx.doi.org/10.3767/003158510X492101
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http://dx.doi.org/10.5248/124.45 Volume 124, pp. 45-50 April-June 2013
Polycoccum anatolicum sp. nov. on Lepraria incana and a key to Polycoccum species known from Turkey
MEHMET GOKHAN HAtici’, HATICE Esra AKGUL’, CELALEDDIN OzTURK? & EMRE KILIc?
' University of Erciyes, Faculty of Science, Department of Biology 38039 Kayseri, Turkey * University of Selcuk, Faculty of Science, Department of Biology 42075 Konya, Turkey * CORRESPONDENCE TO: mghalici@gmail.com
ABSTRACT — Polycoccum anatolicum is described as new from the thallus of Lepraria incana on the trunk of a Prunus sp. in western Turkey and represents the first Polycoccum known to occur on Lepraria. The new species is typical with its relatively small subglobose ascomata and verruculose relatively large ascospores. It is compared with similar Polycoccum species, and a key to Polycoccum species known from Turkey is provided.
Key worps — Ascomycota, lichens, lichenicolous fungi, biodiversity
Introduction
Polycoccum Saut. ex Korb. comprises approximately 50 species of lichenicolous fungi (Halici et al. 2007a, 2009; Brackel & Berger 2010; Zhurbenko 2010). The type species of the genus is P sauteri Kérb. with lichenicolous habit on Stereocaulon condensatum Hoftm. Species are characterized by dark perithecioid ascomata, pseudoparenchymatous exciple, fissitunicate asci with brown one-septate ascospores, and a hamathecium of persistent, branched and anastomosing interascal filaments. Following the genus synopsis by Hawksworth & Diederich (1988), Atienza et al. (2003) recognized 37 species. Brackel & Berger (2010) described three Polycoccum species occurring on the lichenised fungus Placopsis.
Attention to the lichenicolous fungi of Turkey has increased since Halici (2008) published the key to the lichenicolous Ascomycota. Of the 181 lichenicolous species known from Turkey (Halici et al. 2012), ten Polycoccum species have been identified (Halic1 2008, Halici et al. 2009; Yazici et al. 2011). In 2012, the second author (EA) collected an interesting specimen of Polycoccum on epiphytic Lepraria incana (L.) Ach. s. lat. from Turkey’s Mediterranean
46 ... Halici & al
Region. After comparison with the described species of the genus, we concluded that the specimen represents a new species, which we describe here. We also provide a key to the eleven Polycoccum species now reported from Turkey.
Material & methods
The type of the new species is deposited in Erciyes University Herbarium Kayseri, Turkey (EUH). Sections were prepared by hand and examined in I [Lugol's iodine (MERCK 9261) with (KI) and without (I) pre-treatment with 10% KOH], 10% KOH, and water. Ascospores were measured in water. Ascospore measurements are given as: (min -) X-sd - X - X+sd (-max.), where ‘min’ and ‘max’ are the extreme values, ‘X’ the arithmetic mean, and ‘sd’ the corresponding standard deviation. The length/breadth ratio of ascospore is indicated as 1/b and given in the same way. The microphotographs were taken with a Leica DFC 420 digital microscope camera with a c-mount interface and with a 5 megapixel CCD. The descriptive notes provided in the key are based on Turkish specimens examined by the first author (MGH), except for Polycoccum evae, which is based on the description given by Calatayud & Rico (1995).
Taxonomy
Polycoccum anatolicum Halici & E. Akgiil, sp. nov. FIGURE 1 MycoBank MB804154
Differs from Polycoccum dzieduszyckii by its 8-spored asci and its Lepraria host.
Type: Turkey, Isparta, Siitciiler, Yazili Canyon, 41°48’N 36°31’E, alt. 290 m, on Lepraria
incana on Prunus sp., 18 July 2012, E.Akgiil (Holotype, EUH (MGH 0.3118)).
Erymo.oey: The epithet refers to Anatolia, the Asian part of Turkey, where the type
specimen was collected. Vegetative hyphae hyaline, scant, very narrow, the infection causing no visible necrosis in host, but eventually bleaching the infected parts. Ascomata perithecioid, subglobose, arising singly, in the early stage immersed with only the ostiole area visible, semi-immersed at maturity, 75-90 um. Ascomatal wall dark reddish brown, darkest in the upper half, 10—15 um thick, and thickening in the ostiole region; composed of several layers of compressed cells, 10-15 x 6-8 um, forming a textura angularis. Hamathecium consisting of septate, branched and anastomosing pseudoparaphyses, 1-2 um thick. Hymenial gelatin K/I-, I-. Asci arising from the lower part of the ascomatal cavity, cylindrical to elongate-clavate, shortly stalked, bitunicate, the apex thickened, 8- spored, (71—)83—91(—103) x (11—)14—18(—21) um (n = 10); wall I-, endoplasm dextrinoid. Ascospores monostichously arranged in the asci, sole-shaped, apices rounded, some tending to taper towards the rounded ends, 1-septate, not or slightly constricted at the septum, upper cell slightly larger, brown, verruculose, non-halonate, some ascospores multi-guttulate, (25—)26—28—29.5(-32) x (8.5—)9-10-11(-13) um (n = 30), I/b = (2.2—)2.52—2.76-3(—3.3). Conidiomata not seen.
Polycoccum anatolicum sp. nov. (Turkey) ... 47
ioe 00pm —| L S00 pm
Fic. 1. Polycoccum anatolicum (Holotype): A, Habitus; B, Ascoma (in water); C, Ascospores (in water); D, Asci showing monostichously arranged ascospores (in water); E, Anastomosed pseudoparaphyses (in methylene blue).
A8 ... Halici & al
ECOLOGY & DISTRIBUTION: Polycoccum anatolicum is currently known only from the type locality in the Western Mediterranean part of Turkey. The new species was collected on Lepraria incana on the trunks of Prunus sp. The species seems to be weakly pathogenic as moderate bleaching was observed in the infected parts of the host thallus. As the host lichen has a wide distribution, P. anatolicum may also have a wide distribution.
Notes: Most Polycoccum species are confined to one host species or genus, and P. anatolicum is the only species known to occur on Lepraria. It is closest to P. dzieduszyckii (Boberski) D. Hawksw., a species known only on Verrucaria spp. (and probably other lichens on limestone). Polycoccum dzieduszyckii has small ascomata (50-100 um) and large verruculose ascospores (25—35(—44) x 8—10(—14) um) (Hawksworth & Diederich 1988, Halici et al. 2007b), but clearly differs from P. anatolicum in having consistently 2-spored asci and infecting a different host genus.
Polycoccum opulentum (Th. Fr. & Almgq.) Arnold, a species known on Polyblastia intercedens (Nyl.) Lonnr., Lecidea plana (J. Lahm) Nyl., and a sterile lichen on calcareous rocks, has similar sized ascospores (25-28 x 11-14 um) but thicker pseudoparaphyses (3 um), larger ascomata (100-170 um), and hymenial gelatin turning blue in iodine (Hawksworth & Diederich 1988; Atienza et al. 2003). Polycoccum squamarioides (Mudd) Arnold, another species known on Placopsis spp., has similar sized ascomata (75-100 um) but narrower ((18—)19-—26 x (5.5—)6—7(—8) um) and smooth-walled ascospores (Hawksworth & Diederich 1988, Brackel & Berger 2010). Polycoccum evae Calat. & V.J. Rico is restricted to Dimelaena oreina (Ach.) Norman and has larger ascomata (100-200 um) and shorter verruculose ascospores ((17—)18—23(—25) x (6—-)7—10(-11) um) (Calatayud & Rico 1995). Polycoccum crassum Vézda, a species known on Peltigera spp., has similar sized verruculose ascospores ((25—)30—32(—36) x 8—10(—11) um) but much larger ascomata (300-500 um) and consistently 4-spored asci (Hawksworth & Diederich 1988; Atienza et al. 2003; Halici et al. 2012).
A key to the Polycoccum species known from Turkey
Mek ASCOSPOLES i OT LOMoee 8 Fhnns Bhat! utattul Mhnsyt Tek mR! etna Leas AL 2 lL. GAscospores S20 Atmos di... a dibete a disnteeg ding ed dintery dente dcnopea’ tin apes diggs Pe) 2 weAscospores'smioothiwalleds .. 034.002 44nctddhed adhe eadead ered den bes 3 2. Ascospores verruculose, 13-14 x 6 um, on Acarospora spp. P. microsticticum 3... “SNOLcatising swellinssion-thehostlichens %. 2.31 d tt het eed he el 4 3. Ascomata c. 200 um, causing swellings on the host lichens.
Ascospores 14—18 x 5—6 um, on Peltigera spp. ........... 0. eee ee P. peltigerae
Polycoccum anatolicum sp. nov. (Turkey) ... 49
4, | Ascomata 100—140 um. Interascal filaments very thin, c. 1.0 um. Ascospores (11—)12—14.5(—15.5) x (4.5—)5-6 um,
On-Xanthoria sp: ON: COaStall VOCKS 0) ssiew-t teers oracuee ofS athe oa ate tw Bat P. teresum 4, | Ascomata 150-235 um. Interascal filaments thicker. Ascospores
(11—)13.5—15(—16) x 6.5—7 um, on Aspicilia cinerea ................ P. aksoyi Bim EVSCIBSZSDOEGA Toe Se tee sok eee to eaten ren ee tay Ee eee Neen Neen ee ER 6 5. Asci 2-spored. Ascospores verruculose, 25—28 x 8-11 um,
on endolithic Verrucaria spp. on hard calcareous rocks ....... P. dzieduszyckii Gimm, ASCISARPOTEd ss kg ea task tees i vleale tele ls + pels § Deals pte Fe OE OG Gt OH ys
6. Asci 4-spored. Ascomata c. 400 um. Hymenial gelatin I+ orange-red. Ascospore cells + equal in size, verruculose, 24—30 x 8-10 um, CP PEIISEPAS DIOL ses tod Nes elo ehioer sey hat hy Spotty When Dee Mon bey eede eittinals P. crassum
Tait eh SOON ALA cee LOG) MRT o,f eR ec erat Mel hey Me Me Ay bel aly et al 8
7. Ascomata 75-90 um. Hymenial gelatin I—. Ascospores verruculose, (25—)26—29.5(—32) x (8.5—)9-11(—13) um, on Lepraria incana ... P. anatolicum
Sepa Vee al alle Fea eet Pattee ARR se, Je Th tcl Bled de lg teil Md eel Wi ssagelt Miacele 9 8. | Ascomata c. 220 um. Hymenial gelatin I+ blue. Ascospores verruculose,
24-27 x 10-13 um, on endolithic Verrucaria spp. ............ P. marmoratum 9. Ascospores verruculose and cells + equal in size ........... 0. ee eee eee eee 10 9. | Ascomata 170-200 um. Ascospores smooth walled and lower cell often
attenuate, 22-23 x 10 um, on Sporastatia testudinea............ P. sporastatiae 10. Ascomata 100-200 um. Ascospores (17—)18—23(—25) x (6—)7—10(—13) pm,
OD TDECLAC TID OP CUIA pseu sot srvtsnbes drstsnben dicts spben-d Wacken drhesekes driep.t-gedhtfebou'y hig boue P. evae 10. Ascomata 180-250 um. Ascospores (25—)28.5—31.5(—34) x (7—)8-9.5 um,
Ol, ACATOSPOTO:CETVING Strate, Bhivarees avs coe timed oe hea or uthhos argent P. acarosporicola
Acknowledgements
The manuscript was reviewed by Kerry Knudsen (USA) and Wolfgang von Brackel (Germany). This study was financially supported by the Selcuk University Scientific Research Project Coordinator with the project coded (BAP-12101025).
Literature cited
Atienza V, Calatayud V, Hawksworth DL. 2003. Notes on the genus Polycoccum (Ascomycota, Dacampiaceae) in Spain, with a key to the species. Lichenologist 35: 125-135. http://dx.doi.org/10.1016/S0024-2829(03)00014-8
Brackel W, Berger F. 2010. Gall-inducing species of Polycoccum (Ascomycota) on the lichen genus Placopsis. Herzogia 23: 195-204.
Calatayud V, Rico VJ. 1995. Polycoccum evae (Dothideales), a new lichenicolous fungus on Dimelaena oreina. Mycotaxon 53: 29-32.
Halici MG. 2008. A key to the lichenicolous Ascomycota (including mitosporic fungi) of Turkey. Mycotaxon 104: 253-286.
Halici MG, Atienza V, Hawksworth DL. 2007a. Two new Polycoccum species from Turkey. Mycotaxon 101: 157-163.
50 ... Halici & al
Halici MG, Candan M, Ozdemir Tiirk A. 2007b. New records of lichenicolous and lichenized fungi from Turkey. Mycotaxon 100: 255-260.
Halic1 MG, Knudsen K, Candan M, Tiirk A. 2009. A new species of Polycoccum (Dothideales, Dacampiaceae) from Turkey. Nova Hedwigia 89: 431-436. http://dx.doi.org/10.1127/0029-5035/2009/0089-0431
Halici MG, Candan M, Ozdemir Tiirk A. 2012. A key to the peltigericolous fungi in Turkey. Mycotaxon 119: 277-289. http://dx.doi.org/10.5248/119.277
Hawksworth DL, Diederich P. 1988. A synopsis of the genus Polycoccum (Dothideales), with a key to accepted species. Transactions of the British Mycological Society 90: 293-312. http://dx.doi.org/10.1016/S0007-1536(88)80101-3
Yazic1 K, Etayo J, Aslan A. 2011. A note about lichenicolous fungi from Ardahan (Turkey). Cryptogamie, Mycologie 32: 429-437.
Zhurbenko MP. 2010. New and interesting lichenicolous fungi from Eurasia. I]. Mycosphere 1: 213-222.
ISSN (print) 0093-4666 © 2013. Mycotaxon, Ltd. ISSN (online) 2154-8889
MY COTAXON
http://dx.doi.org/10.5248/124.51 Volume 124, pp. 51-59 April-June 2013
Further additions to the macrolichen mycota of Vietnam
UDENI JAYALAL ', ANDRE APTROOT *, THI THUY NGUYEN ?, NGUYEN ANH DZUNG 3, SANTOSH JOSHI ', SOON-OK OH ' & JAE-SEOUN Hur **
'Korean Lichen Research Institute, Sunchon National University, Suncheon-540 742, Korea ?ABL Herbarium, Gerrit van der Veenstraat 107, NL-3762 XK Soest, The Netherlands Biotechnology Center, Tay Nguyen University, 567 Le Duan, Buon Ma Thuot City, Vietnam “CORRESPONDENCE TO: jshur1@sunchon.ac.kr
ABSTRACT — Seven new records of foliose lichens are reported from Chu Yang Sin National Park of Dak Lak province in Vietnam. Morphological, anatomical and chemical characteristics are described for Heterodermia obscurata, Hypotrachyna flavida, Leptogium ulvaceum, Parmotrema sancti-angelii, Parmeliella brisbanensis, Physma byrsaeum, and Sticta marginifera.
Key worps — tropical, macrolichens, Parmeliaceae, Da Lat Plateau
Introduction
Although larger organisms are well known in the tropics, the fungi and their lichenized counterparts in tropical forests are less known (Coppins & Wolseley 2002). Aptroot & Sipman (1997) observed that the tropical forest lichens represent about one-third to one-half of the world’s lichen diversity. Vietnam, being a moderately large country with the monsoon tropical climate (VARCC 2009), provides a range of conditions to support lichen growth (Nguyen et al. 2010, 2011).
Although the lichen mycota of Vietnam is underworked, recently many species have been reported or described. Aptroot & Sparrius (2006) provided the first checklist of Vietnam lichens, which has been followed by new records of macrolichens by Giao (2009), some findings on foliicolous lichens by Nguyen et al. (2010, 2011), and reports of some Graphidaceae species by Joshi et al. (2012).
The Korean Lichen Research Institute (KoLRI) has initiated collaboration with Tay Nguyen University in Vietnam, which has resulted in the collection of many lichen specimens, which have been deposited in the herbarium of the KoLRI. This paper reports seven taxa (in five lichen families) from Chu Yang Sin National Park, Vietnam.
52 ... Jayalal & al.
Materials & methods
Chu Yang Sin National Park is located in the central highlands of Vietnam in Krong Bong and Lak Districts of Dak Lak Province. This park is situated within the elevations ranging from less than 600 m to 2442 m at the summit of Mount Chu Yang Sin with the area of 58,947 ha. The Park is the largest protected area on the Da Lat Plateau, in the central highlands (Hughes 2010).
The dominant vegetation type in the Park is broadleaved evergreen forest, covering over 65% of the area. This forest is dominated by the vascular plant families Fagaceae, Lauraceae, Meliaceae, and IIliciaceae with a canopy height often in excess of 35 m. The high altitudinal range and varied topography gives rise to high vascular plant diversity in different forest types. Montane and submontane forests are at >900 m and lowland semi evergreen forests at <900 m (Hughes 2010).
Morphological and anatomical investigations were performed under a binocular dissecting (NIKON SMZ645) and light microscope (Zeiss Scope. Al). Measurements of all the microscopic features were made on hand-cut thallus and apothecial sections. The sections were examined by mounting in water, 10% KOH, and Lugol’s iodine solutions. Ascospore measurements were determined in water. Chemical constituents were identified by spot tests and thin layer chromatography performed in solvent systems A (toluene: dioxin:acetic acid 180:45:5) and C (toluene:acetic acid 85:15) (Orange et al. 2010).
Taxonomic descriptions
Heterodermia obscurata (Nyl.) Trevis., Nuovo Gior. Bot. Ital. 1: 114 (1869) Fic.14
Thallus foliose, suborbicular, adnate, 2-6 cm across; lobes narrow, up to 1-2 mm wide, flat; upper surface whitish gray, shiny, epruinose, sorediate; soredia granular, labriform to capitate on lateral and terminal lobes; lower cortex absent; lower surface (medulla) felted with yellowish-brown pigment, K+ purple, marginally rhizinate; rhizines black, simple to squarrosely branched; apothecia not seen.
CHEMISTRY — Cortex K+ (yellow), C-, KC-, P+ (pale yellow); medulla K-, C-, KC-, P-. TLC: atranorin, chloroatranorin, zeorin.
EcoLoGcy & DISTRIBUTION — Found on sandy rock in broad-leaved evergreen forest in Chu Yang Sin National Park. According to Moberg & Nash (2001), this species is growing on tree trunk and mossy rocks in open humid conditions. Mostly cosmopolitan and has been reported from many countries in Africa, Asia, Europe, North America, South America, and Oceania (Aptroot & Feijen 2002, Calvelo & Liberatore 2002, Egea 1996, Eliasaro & Adler 1997, Elix & McCarthy 1998, Fryday et al. 2001, Gowan & Brodo 1988, Hafellner & Tiirk 2001, Knezevic & Mayrhofer 2009, Kurokawa 1960, Llimona & Hladun 2001, Lépez—Figueiras 1986, Osorio 1992, Singh & Sinha 2010, Sipman 1993, Streimann 1986, Tenorio et al. 2002, Wei 1991).
SPECIMEN EXAMINED: VIETNAM. Dak LAK PROVINCE: Chu Yang Sin National Park, on rock, 12°27'57.0"N 108°20'34.9"E, elev. c. 780 m, 20.04.2012, S.O. Oh, J.S. Hur VN120185 (KoLRI).
Macrolichens newly recorded for Vietnam ... 53
REMARKS — Heterodermia obscurata (Physciaceae) is characterized by its robust appearance, labiate soredia, lack of lower cortex, and the rusty-brown pigmented (K+ purple) lower surface or medulla. According to Moberg & Nash (2001), this species is closely related to H. speciosa (Wulfen) Trevis., which differs in having a lower cortex and in lacking yellowish pigmentation. It is also easily confused with H. japonica (M. Sat6) Swinscow & Krog, which has no lower cortex and which is white to violet.
Hypotrachyna flavida (Zahlbr.) Hale, Smithson. Contr. Bot. 25: 37 (1975) Fic. 1B Thallus adnate to loosely attached, coriaceous, green, 4-6 cm across; lobes sublinear, separate, 2-4 mm wide, margins eciliate; upper surface greenish grey, emaculate, plane to convex, continuous or irregularly cracked on older lobes; medulla white; lower surface moderately rhizinate; rhizines long, sparsely dichotomously branched, often projecting beyond the lobe margins; apothecia adnate, 1-3 mm in diameter, disc brown, flat to concave; Asci clavate, 8 spored, spores colour less, 6-8 um; pycnidia not seen. CHEMISTRY — Cortex K-, C-, KC-, P-; medulla K+ (pale yellowish to brown), C-, KC-, P+ (orange-red). TLC: usnic acid, protocetraric acid. EcoLoGy & DISTRIBUTION — Found on a sandy rock in broad-leaved evergreen forest in Chu Yang Sin National Park. Reported as saxicolous species at somewhat higher elevations by Hale (1975). This species has previously been reported from Central and South America (Calvelo & Liberatore 2002, Eliasaro & Adler 1997, Hale 1975, Riddle 1920, Tenorio et al. 2002). SPECIMEN EXAMINED: VIETNAM. DAK LAK PROVINCE: Chu Yang Sin National Park, on rock, 12°28'12.3"N 108°20'59.9”E, elev. c. 763 m, 20.04.2012, S.O. Oh, J.S. Hur VN120120 (KoLRI). REMARKS — Hypotrachyna flavida (Parmeliaceae) is characterized by a saxicolous habit and a medulla containing usnic acid and protocetraric acid. According to Hale (1975), H. flavida is fairly restricted to northern South America. Externally, this species very closely resembles H. protoboliviana (Hale) Hale, which differs in its corticolous habitat and medulla containing barbatic acid.
Leptogium ulvaceum (Pers.) Vain., Ann. Acad. Sci. Fenn., Ser. A, 15(6): 38 (1921) FIG. 1C Thallus foliose, loosely adnate, 4-8 cm across, bluish gray; lobes rotund, 2-8 mm wide, homiomerous, margins entire, surface smooth to somewhat rough; isidia, phyllidia absent; lower surface smooth, etomentose, pale bluish with sparse tufted rhizines; apothecia laminal, shortly pedicellate, 0.5-2 mm diam., disc flat to concave, red brown, exciple smooth to wrinkled, cells paraplectenchymatous and continuous to below the hypothecium; ascospores
54 ... Jayalal & al.
ellipsoid, muriform, 30-40 x 12-18 um, apices acute to elongate; pycnidia submarginal; conidia 2-3 um long.
CHEMISTRY — No chemical detected.
ECOLOGY & DiIsTRIBUTION — Found on a sandy rock in a shady place in broad-leaved evergreen forest in Chu Yang Sin National Park. This species has previously been reported from Asia, Europe, South America, and Oceania (Awasthi 1988, Elix & McCarthy 1998, Hafellner 1995, Marcelli 1991).
SPECIMEN EXAMINED: VIETNAM. DAK LAK PROVINCE: Chu Yang Sin National Park,
on rock, 12°26'51.9"N 108°20'18.1"E, elev. c. 816 m, 21.04.2012, S.O. Oh, J.S. Hur
VN120170 (KoLRI). REMARKS — Leptogium ulvaceum (Collemataceae) is characterized by smooth upper surface lacking vegetative propagules. This species is morphologically similar to L. cochleatum (Dicks.) P.M. Jorg. & P. James, which differs in its periclinally wrinkled thalline exciple.
Parmeliella brisbanensis (C. Knight) P.M. Jorg. & D.J. Galloway, Flora of Australia, 54: 314 (1992) FIG. 1D
Thallus rosette-forming, orbicular, 5-7 cm across, ona thick, black prothallus projecting 1-2 mm beyond lobe apices, closely appressed to substratum; lobes narrow, 0.6-1 mm wide, discrete at margins, flat; margins entire, somewhat thickened, occasionally ascending; upper surface, slightly maculate, smooth, occasionally striate, grayish when wet, whitish gray when dry, isidiate; isidia delicate, simple to finger-like, to 0.5 mm tall and 0.1 mm thick, laminal and marginal; medulla white to blackish, photobiont Nostoc; apothecia, 0.5-2 mm wide, rounded to contorted; exciples thick, thalline, persistent, crenulate- striate, concolorous with thallus; disc flat to concave, red-brown; ascospores simple, ellipsoidal, 10-15 x 8-10 um; pycnidia not seen
CHEMISTRY — No chemical detected.
ECOLOGY & DISTRIBUTION — Found on a sandy rock in a shady place in broad-leaved evergreen forest in Chu Yang Sin National Park. According to Jorgensen and Galloway (1992), Parmeliella brisbanensis also occurs on tree bark in tropical to subtropical rain forests. This species has been recorded from many countries in the tropics, in Asia, Australia, and Oceania (Elix & McCarthy 1998, Jorgensen & Galloway 1992, Singh & Sinha 2010, Sipman 1993, Wolseley et al. 2002).
SPECIMEN EXAMINED: VIETNAM. DAK LAK PROVINCE: Chu Yang Sin National Park,
on rock, 12°28'04.4"N 108°20'39.0"E, elev. c. 622 m, 20.04.2012, S. O. Oh, J. S. Hur
VN120046 (KoLRI). REMARKS: Parmeliella brisbanensis (Pannariaceae) is characterized by its rosette form, black hypothallus, and finger-like isidiate upper surface. According to Jorgensen & Galloway (1992), this species is the isidiate counterpart of the P. mariana (Fr.) P.M. Jorg. & D.J. Galloway complex.
Macrolichens newly recorded for Vietnam ... 55
FiGurE 1: External morphology of Vietnamese lichen specimens (Scale bars = 1 cm). A. Heterodermia obscurata VN120185 (KoLRI); B. Hypotrachyna flavida VN120120 (KoLRI); C. Leptogium ulvaceum VN120170 (KoLRI); D. Parmeliella brisbanensis VN120046 (KoLRI); E. Parmotrema sancti-angelii VN120288 (KoLRI); F. Physma byrsaeum VN120107 (KoLRI); G. Sticta marginifera VN120053 (KoLRI).
Parmotrema sancti-angelii (Lynge) Hale, Phytologia 28: 339 (1974) FIG. 1E
Thallus loosely attached to the substratum, up to 6 cm across; lobes rotund, 4-10 mm wide; margins crinate, ciliate; cilia numerous, simple, 2-4 mm long; upper surface pale grey, emaculate, smooth, sorediate; soralia marginal, soredia granular; medulla white, yellowish brown pigments below; lower surface, with
56 ... Jayalal & al.
relatively narrow, smooth, erhizinate marginal zone; rhizines sparse, simple; apothecia and pycnidia not seen.
CHEMISTRY — Cortex K+ (yellow), C-, KC-, P-; medulla K- C+ (rose), KC+ (red), P-. TLC: atranorin, chloroatranorin, gyrophoric acid and unknown compound.
EcoLoGy & DISTRIBUTION — Found on a sandy rock in an open place in broad-leaved evergreen forest in Chu Yang Sin National Park. A common and widespread species in the pantropical region (Elix 1994), it has been reported from Asia, Australia, South America, and Oceania (Calvelo & Liberatore 2002, Divakar & Upreti 2005, Elix 1994, Elix & McCarthy 1998, Elix & Schumm 2001, Hale 1974, Kurokawa 1993, Wei 1991, Wolseley et al 2002).
SPECIMEN EXAMINED: VIETNAM. DAK LAK PROVINCE: Chu Yang Sin National Park,
on rock, 12°28'04.4"N 108°20'39.0"E, elev. c. 622 m, 20.04.2012, S.O. Oh, J.S. Hur-
VN120288 (KoLRI). REMARKS — Parmotrema sancti-angelii (Parmeliaceae) is characterized by an emaculate upper surface, sorediate margins, yellowish lower medulla, and brown to black lower surface. This species is very close to P indicum Hale, which differs in having norlobaridone and in lacking pigmentation on the lower medulla.
Physma byrsaeum (Ach.) Tuck., Syn. N. Amer. Lich. 1: 115 (1882) FIG. 1F Thallus foliose, rosulate, adnate, 6-8 cm across, 200-350 um thick; lobes radiating, oblong 1-5 mm wide; margins entire, thickened, re-curved at the lobes end; upper surface ridged, somewhat wrinkled, brownish black, lacking isidia; cortex thin, photobiont Nostoc; lower surface pale brown, rhizinate; rhizines of interwoven hyphae forming a cushion-like indumentum, whitish to black; ascomata apothecial, abundant, laminal, sessile, 1-4 mm wide; disc concave to plane, reddish brown; thalline exciple thick, wrinkled, lobed, concolorous with thallus; ascospores simple, ellipsoidal, 12-15 x 10-12 um; episporium, 2-3 um thick; pycnidia laminal, conidia 2-3 um long.
CHEMISTRY — No chemicals detected.
EcoLoGy & DISTRIBUTION — Found on sandy rock in a shady place in broad-leaved evergreen forest in Chu Yang Sin National Park. Physma byrsaeum has been reported from many countries in the paleotropics, including Asia, Australia, Central America, and Oceania (Aptroot & Seaward 1999, Aptroot & Sipman 1991, Elix & McCarthy 1998, Sipman 1993, Tenorio et al. 2002, Verdon 1992, Wolseley et al. 2002).
SPECIMEN EXAMINED: VIETNAM. Dak LAK PROVINCE: Chu Yang Sin National Park, on rock, 12°28'12.3"N 108°20'59.9"E, elev. c. 763 m, 20.04.2012, S.O. Oh, J.S. Hur VN120107 (KoLRI).
REMARKS — Physma byrsaeum (Collemataceae) is characterized by a well- developed cortical layer and wrinkled thalline apothecial exciple. This species
Macrolichens newly recorded for Vietnam ... 57
is morphologically similar to P pseudoisidiatum Aptroot & Sipman, which is isidiate.
Sticta marginifera Mont., Annls Sci. Nat., Bot., Sér. 2 18: 265 (1842) FIG. 1G
Thallus palmate to irregularly branched in clumps, 1-2 cm across, arising from a rooted holdfast; lobes flabellate to irregularly dichotomously branching, 1-3 mm wide, proliferating round to oblong lobules attached to primary lobes by thin terete stalks; margins entire or irregularly notched, thickened below; upper surface grayish glaucous when dry, dark blue-black when wet, thin, papery, smooth, rather fragile, isidiate, phyllidiate and minutely maculate; isidia marginal, sometimes on the upper surface, styliform to coralloid, to 0.5 mm tall; phyllidia marginal, developing from isidia, to 1 mm tall, irregularly lobed; photobiont Nostoc; lower surface white to pale at margins, brown centrally, minutely pubescent; cyphellae scattered, round, to 0.5 mm diam.; pit membrane white; apothecia not seen.
CHEMISTRY — No chemicals detected.
EcoOLoGy & DisTRIBUTION — Found on sandy rock in a shady place in broad-leaved evergreen forest in Chu Yang Sin National Park. According to Galloway (2001), Sticta marginifera prefers humid, deeply shaded habitats in montane rain forests on different substrates. This species has been recorded from Asia, Australia, South America, and Oceania (Awasthi 2007, Calvelo & Liberatore 2002, Elix & McCarthy 1998, Galloway 2001, Streimann 1986).
SPECIMEN EXAMINED: VIETNAM. Dak LAK PROVINCE: Chu Yang Sin National Park,
on rock, 12°28'04.4"N 108°20'39.0"E, elev. c. 622 m, 20.04.2012, S.O. Oh, J.S. Hur VN120053 (KoLRI).
REMARKS — Sticta marginifera (Lobariaceae) is characterized by the presence of secondary lobules and the coralloid isidia. According to Galloway (2001), the similar S. brevipes (Mull. Arg.) Zahlbr. also has similar flabellate lobes with thickened and revolute apices but does not produce secondary lobules.
Acknowledgements
This work was supported by a grant from the National Research Foundation of Korea (#2011-0031494) and the Korea National Research Resource Center Program. Authors are grateful to Ms. Pat Wolseley and Dr. Sanjeeva Nayaka for their valuable comments on manuscript.
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MY COTAXON
http://dx.doi.org/10.5248/124.61 Volume 124, pp. 61-68 April-June 2013
Inonotus niveomarginatus and I. tenuissimus spp. nov. (Hymenochaetales), resupinate species from tropical China
Hatr-You Yu, CHANG-LIN ZHAO & YU-CHENG Dat
Institute of Microbiology, P.O. Box 61, Beijing Forestry University, Beijing 100083, China * CORRESPONDENCE TO: yuchengd@yahoo.com
ABSTRACT — Two new species are described from tropical Yunnan, southwestern China. Inonotus niveomarginatus is characterized by resupinate basidiocarps with distinct white margins, a monomitic hyphal structure, absence of hymenial and hyphoid setae, and yellowish, thick-walled, ovoid to subglobose basidiospores. Inonotus tenuissimus can be distinguished from other Inonotus species by the combination of resupinate habit, a mono- to dimitic hyphal structure, absence of the hymenial and hyphoid setae, and yellowish thick- walled ellipsoid basidiospores.
Key worps — Hymenochaetaceae, polypore, taxonomy, wood-inhabiting fungi
Introduction
Yunnan Province in southwestern China is one of the biodiversity “hotspots” in the world and rich with many vascular plants and fungi. Many wood- decaying fungi have been described from this province (Chen & Cui 2012; Cui et al. 2009; Dai 2010, 2011; Dai et al. 2002, 2009; He & Dai 2012; Li & Cui 2010, 2013; Yuan & Dai 2008; Zhao et al. 2013). During a systematic survey of Inonotus P. Karst. species in southwestern China, two resupinate species were found that lacked hymenial and hyphoid setae and possessed yellowish thick-walled ovoid to subglobose or ellipsoid basidiospores. These prominent characters distinguish the two species from all other known Inonotus species, and they are described here as new.
Materials & methods
The studied specimens were deposited at the herbarium of Institute of Microbiology, Beijing Forestry University (BJFC). The microscopic procedures were follows as in Cui & Decock (2013). Sections were studied at magnifications up to 1000x using a Nikon Eclipse 80i microscope with phase contrast illumination. Drawings were made with the aid of a drawing tube. Microscopic features, measurements, and illustrations were made
62 ... Yu, Zhao & Dai
from the slide preparations stained with Cotton Blue and Melzer’s reagents. Spores were measured from sections cut from the tubes. To present the spore size variation, the 5% of measurements excluded from each end of the range are given in the parentheses; spine lengths are not included in the basidiospore measurements. Abbreviations include IKI = Melzer’s reagent, IKI- = negative in Melzer’s reagent, KOH = 5% potassium hydroxide, CB = Cotton Blue, CB- = acyanophilous, L = mean spore length (arithmetic average of all spores), W = mean spore width (arithmetic average of all spores), Q = variation in the L/W ratios between the specimens studied, and n = number of spores measured from the given number of specimens. Special color terms follow Petersen (1996).
Taxonomy
Inonotus niveomarginatus H.Y. Yu, C.L. Zhao & Y.C. Dai, sp. nov. Fics 1-2 MycoBank MB805435 Differs from other Inonotus species by its resupinate growth habit, absence of hymenial and hyphoid setae, and yellowish, thick-walled, ovoid to subglobose basidiospores. Type: China, Yunnan Province, Xishuangbanna, Jinghong County, Sanchahe Nature Reserve, on fallen angiosperm trunk, 7.VI.2011, Dai 12318 (holotype, BJFC).
ETYMOLOGY: niveomarginatus (Lat.): refers to the white margin of the basidiocarp.
FruitBopy — Basidiocarps annual, resupinate, cushion-shaped, adnate, without odor or taste when fresh, becoming corky upon drying, <4 cm long, 2.5 cm wide, 4 mm thick at centre. Pore surface deep olive and margin white when fresh, dark brown to yellowish brown upon drying; pores circular, 6-8 per mm; dissepiments thin, entire. Subiculum brown, corky, <1 mm thick. Tubes dull brown to blackish brown, corky, <3 mm long.
HyPHAL STRUCTURE — Hyphal system monomitic; generative hyphae simple septate; tissues darkening in KOH.
SUBICULUM — Generative hyphae yellow, thin- to thick-walled with a wide lumen, frequently septate, very rarely branched, more or less straight, interwoven, 3-4.5 um in diam.
TuBEs — Generative hyphae yellowish, thin- to thick-walled, frequently branched and septate, more or less straight, subparallel along the tubes, 2.5-3.5 um in diam. Basidia barrel-shaped, with four sterigmata and a basal simple septum, 13-16 x 6-8.5 um; basidioles in shape similar to basidia, but slightly smaller than basidia.
SpoRES — Basidiospores ovoid to subglobose, yellowish, thick-walled, smooth, occasionally bearing a guttule, IKI-, CB-, (4.5-)4.9-5.7(-6) x (4.2-) 4.5-5.2(-5.5) um, L = 5.35 um, W = 4.95 um, Q = 1.06 (n = 30/1).
REMARKS — Four other Inonotus species — I. costaricensis Ryvarden, I. rigidus B.K. Cui & Y.C. Dai, I. truncatisporus Corner, and I. venezuelicus Ryvarden — also are characterized by resupinate basidiocarps, thick-walled colored spores,
Inonotus spp. nov. (China) ... 63
FiGure 1. Inonotus niveomarginatus (holotype). Basidiocarp.
and a lack of both hymenial and hyphoid setae (Corner 1991; Cui et al. 2011; Ryvarden 1987, 2005). Inonotus costaricensis differs from I. niveomarginatus by having globose and longer basidiospores (5.5-6.5 um long; Ryvarden 1987). Inonotus rigidus differs from I. niveomarginatus by its ellipsoid and smaller
64 ... Yu, Zhao & Dai
(a
———N
Lay) oo
said | \ d
10 um
es 10 pm
FIGURE 2. Inonotus niveomarginatus (holotype) microscopic structures. a: Basidiospores. b: Basidia and basidioles. c: Hyphae from trama. d: Hyphae from subiculum.
spores (3.9-4.5 x 2.9-3.7 um; Cui et al. 2011). Inonotus truncatisporus is distinguished from I. niveomarginatus by its larger pores (4-6 per mm) and truncate basidiospores (Corner 1991). Inonotus venezuelicus has larger pores (3-4 per mm), ellipsoid basidiospores (5-6 x 4.5-5 um), and has been found only in South America (Ryvarden 1987).
Inonotus tenuissimus H.Y. Yu, C.L. Zhao & Y.C. Dai, sp. nov. Figs 3-4 MycoBANnk MB805434
Differs from other Inonotus species by its resupinate basidiocarps, a mono- to dimitic hyphal structure, absence of hymenial and hyphoid setae, and yellowish, thick-walled, ellipsoid basidiospores.
Type: China, Yunnan Province, Puer County, Laiyanghe Forest Park, on fallen angiosperm trunk, 6.V1.2011, Dai 12365 (holotype, BJFC).
ETyMOLOoGy: tenuissimus (Lat.): refers to the very thin subiculum.
Inonotus spp. nov. (China) ... 65
FiGuRE 3. Inonotus tenuissimus (holotype). Basidiocarp.
FRuITBODy — Basidiocarps annual, resupinate, adnate, becoming corky upon drying, <15 cm long, 7 cm wide, 1.5 mm thick at centre. Pore surface vinaceous grey to greyish brown when fresh, pale brown upon drying; pores angular, 3-4 per mm; dissepiments thin, entire. Subiculum brown, corky, up to 0.2 mm thick. Tubes dull brown, corky, <1.3 mm long.
HyPHAL STRUCTURE — Hyphal system mono- to dimitic; generative hyphae, simple septate; tissues darkening in KOH.
66 ... Yu, Zhao & Dai
BW <<‘ d
| mencieniiaael 10 ym
4 a ane aa oe) uae,
Figure 4. Inonotus tenuissimus (holotype) microscopic structures. a: Basidiospores. b: Basidia and basidioles. c: Hyphae from trama. d: Hyphae from subiculum.
SUBICULUM — Generative hyphae yellow, thick-walled, frequently septate, more or less straight, interwoven, 2.5-4.5 um in diam. Some skeletal-like hyphae similar to generative hyphae, rarely septate.
TuBES — Generative hyphae yellowish, thick-walled, occasionally branched, rarely septate, subparallel along the tubes, 3-4 um in diam. Some skeletal-like hyphae similar to generative hyphae, rarely septate. Basidia clavate to barrel- shaped, with four sterigmata and a basal simple septum, 12-18 x 4.5-6.5 um; basidioles barrel-shaped, smaller than basidia.
Spores — Basidiospores ellipsoid, yellowish, thick-walled, smooth, IKI-, CB-, (4-)4.3-5(-5.2) x (3-)3.2-4(-4.2) um, L = 4.8 um, W = 3.6 um, Q = 1.31-1.38 (n = 90/3).
Inonotus spp. nov. (China) ... 67
ADDITIONAL SPECIMENS EXAMINED: CHINA. YUNNAN PROVINCE, PUER COUNTY, Laiyanghe Forest Park, on fallen angiosperm branch, 6.VI.2011, Dai 12245 & Dai 12255 (BJFC); 9.VI.2011, Dai 12330 (BJFC).
REMARKS — Inonotus tenuissimus resembles I. venezuelicus in its resupinate basidiocarps, similar pores, thick-walled and colored spores, and absence of both hymenial and hyphoid setae. However, I. venezuelicus has greyish pores when dry, a distinctly monomitic hyphal structure, larger and ellipsoid to ovoid basidiospores (5-6 x 4.5-5 um), and has been found only in South America (Ryvarden 2005).
Acknowledgments
We express our gratitude to Dr. Li- Wei Zhou (IFP, China) and Mr. Jaya Seelan Sathiya Seelan (Clark University, USA) who reviewed the manuscript. The research was financed by the National Natural Science Foundation of China (Nos. 31070022, 30910103907).
Literature cited
Corner EJH. 1991. Ad Polyporaceas 7. The xanthochroic polypores. Beih. Nova Hedwigia 101: 1-175.
Chen JJ, Cui BK. 2012. Studies on Wrightoporia from China 2. A new species and three new records from South China. Mycotaxon 121: 333-343. http://dx.doi.org/10.5248/121.333
Cui BK, Decock C. 2013. Phellinus castanopsidis sp. nov. (Hymenochaetaceae) from southern China, with preliminary phylogeny based on rDNA sequences. Mycological Progress 12: 341-351. http://dx.doi.org/10.1007/s11557-012-0839-5
Cui BK, Dai YC, Bao HY. 2009. Wood-inhabiting fungi in southern China 3. A new species of Phellinus (Hymenochaetales) from tropical China. Mycotaxon 110: 125-130. http://dx.doi.org/10.5248/110.125
Cui BK, Du P, Dai YC. 2011. Three new species of Inonotus (Basidiomycota, Hymenochaetaceae) from China. Mycological Progress 10: 107-114. http://dx.doi.org/10.1007/s11557-010-0681-6
Dai YC. 2010. Hymenochaetaceae (Basidiomycota) in China. Fungal Diversity 45: 31-343. http://dx.doi.org/10.1007/s13225-010-0066-9
Dai YC. 2011. A revised checklist of corticioid and hydnoid fungi in China for 2010. Mycoscience 52: 69-79. http://dx.doi.org/10.1007/s10267-010-0068-1
Dai YC, Niemela T, Kinnunen J. 2002. The polypore genera Abundisporus and Perenniporia (Basidiomycota) in China, with notes on Haploporus. Annales Botanici Fennici 39: 169-182.
Dai YC, Cui BK, Yuan HS. 2009. Trichaptum (Basidiomycota, Hymenochaetales) from China with a description of three new species. Mycological Progress 8: 281-287. http://dx.doi.org/10.1007/s11557-009-0598-0
He SH, Dai YC. 2012. Taxonomy and phylogeny of Hymenochaete and allied genera of Hymenochaetaceae (Basidiomycota) in China. Fungal Diversity 56: 77-93. http://dx.doi.org/10.1007/s13225-012-0174-9
Li HJ, Cui BK. 2010. A new Trametes species from Southwest China. Mycotaxon 113: 263-267. http://dx.doi.org/10.5248/113.263
Li HJ, Cui BK. 2013. Two new Daedalea species (Polyporales, Basidiomycota) from South China. Mycoscience 54: 62-68. http://dx.doi.org/10.1016/j.myc.2012.07.005
Petersen JH. 1996. Farvekort. The Danish Mycological Society’s colour-chart. Foreningen til Svampekundskabens Fremme, Greve. 6 p.
68 ... Yu, Zhao & Dai
Ryvarden L.1987. New and noteworthy polypores from tropical America. Mycotaxon 38: 525-541.
Ryvarden L. 2005. The genus Inonotus, a synopsis. Synopsis Fungorum 21: 1-149.
Yuan HS, Dai YC. 2008. Polypores from northern and central Yunnan Province, southwestern China. Sydowia 60: 147-159.
Zhao CL, Cui BK, Dai YC. 2013. New species and phylogeny of Perenniporia based on morphological and molecular characters. Fungal Diversity 58: 47-60. http://dx.doi.org/10.1007/s13225-012-0177-6
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MYCOTAXON
http://dx.doi.org/10.5248/124.69 Volume 124, pp. 69-72 April-June 2013
Arenariomyces truncatellus sp. nov., an ascomycete on driftwood from the north coast of Zealand, Denmark
J. KocuH
GI. Dronninggadrds Allé 12, DK-2840 Holte, Denmark
ABSTRACT — Incubated driftwood from the north coast of Zealand, Denmark, yielded a new species, Arenariomyces truncatellus, belonging to the Halosphaeriaceae, Ascomycota. The species is characterized by globose, spiny ascomata formed on wood with asci deliquescing before spore maturity and 1-septate ascospores with three delicate appendages at each spore end. The species is compared with A. majusculus, A. parvulus, A. trifurcatus, and A. triseptatus. A key to the species is provided.
Key worps — higher marine fungi, Microascales, taxonomy
Introduction
Investigations into the incidence and distribution of higher marine fungi colonizing wood along the Danish coasts were initiated by Héhnk (1955) and Kohlmeyer (1968). Since then more data have been accumulated and published in “A check list of higher marine fungi on wood from Danish coasts” (Koch & Petersen 1996) and further in Petersen (1997), Petersen & Koch (1997a,b), and Koch et al. (2007). Also added (Koch unpublished) are Corollospora angusta Nakagiri & Tokura, C. californica Kohlm. & Volkm.-Kohlm., and C. pseudopulchella Nakagiri & Tokura. In this article, a new species of Arenariomyces Hohnk (Halosphaeriaceae) is described from the north coast of Zealand, Denmark.
Material & methods
Driftwood entrapped among stones and wood from the intertidal zone was collected at Tisvildeleje on the north coast of Zealand, Denmark, in January 2008. Salt content was around 10-20%. The wood was subsequently incubated at 16-19°C and examined periodically for fruiting bodies. From the first formed A. truncatellus ascomata, ascospores were used for inoculation of sterilized pieces of driftwood, on which ascomata developed over a 4-8 month incubation period at 16-19°C ascomata developed.
70 ... Koch
Arenariomyces truncatellus Jorg. Koch, sp. nov. FIGs 1-5 MycoBAnk 50644 Differs from Arenariomyces majusculus by its smaller ascospores with two cells often
conspicuously different in size and with short delicate appendages.
Type: Denmark, North Zealand, Tisvildeleje (UTM 691000E 6217000N), from driftwood (Picea sp.), Jan. 2008, Jorg. Koch 998, incubated nine months in humid chamber (Holotype, CP [piece of conifer wood + slides 1-4]; isotype, CP [remainder of collection Jorg. Koch 998]). [The original material also contained Dictyosporium pelagicum (Linder) G.C. Hughes ex E.B.G. Jones and Halosphaeriopsis mediosetigera (Cribb & W. Cribb) T.W. Johnson. ]
EryMo oey: From the Latin truncatellus = somewhat truncate, referring to the slightly
flattened ends of the ascospores, when appendages are not unfolded. ASCOMATA on wood (rarely seated with subicula on grains of sand) measured in situ 78-131-156 um (n = 19) in diam. Globose-subglobose with the basal part immersed in the outermost layer of the substrate and anchored with brown tortuous hyphae, often in tufts. Ascomata gregarious, black, shiny when young, carbonaceous, brittle, covered with scattered brownish black spines, slightly curved, up to 16 um long and 1.5-2 um wide, but sometimes with a swollen base 4-8 um wide and 4 um high. Immature ascomata from 20 um in diam. are covered with spines up to 8 um long. Neither necks nor ostioles were observed. PERIDIUM membranous, single layered, composed of one layer of flat, on the outside domed, thick-walled cells forming a textura angularis with edges 3-4-6 um, brownish black in transparent light. SustcuLUM composed of hyaline, epidermoid cells merging into the thin-walled pseudoparenchyma, globose or ellipsoidal as free cells (10-20-26 um in diam.), with scattered cytoplasm and small oil globules filling the centre of young ascomata. In some cells a few cytoplasmic strands stretching towards the cell wall indicate pit connections to neighbouring cells.
Asci eight-spored, pyriform, 36-56 x 20-32 um, thin-walled, unitunicate, deliquescing before maturity of the ascospores, developing from the base of the ascocarp. No catenophyses or paraphyses. ASCOSPORES 18-21.4—24 x 8-10.2- 12 um (n = 66) measured in seawater (12%), cylindrical, 1-septate, more or less constricted at the septa, often one cell is shorter and narrower than the other, hyaline, with three subterminal-terminal appendages at each end. APPENDAGES attenuate, straight or curved, delicate, up to 12 um long, about 1 um wide at the base, expanded at right angles to the axis, 120° between each appendage. When not unfolded the spore ends seem slightly flattened.
SUBSTRATE: Driftwood (conifer wood) between stones (intertidal zone).
Discussion Jones et al. (2009) recognized the four species A. majusculus Kohlm. & Volkm.-Kohlm., A. parvulus Jorg. Koch, A. trifurcatus Hohnk, and A. triseptatus
Arenariomyces truncatellus sp. nov. (Denmark) ... 71
Tei ry -
| le - J
~~ , Fic. 1. Arenariomyces truncatellus. Group of ascomata formed on sterilized driftwood inoculated with ascospores. Scale bar = 160 um.
Fics 2-5. Arenariomyces truncatellus. 2. Young asci. 3. Different stages of immature ascospores forming an eight-spored cluster indicating an early deliquescing of the ascus wall. 4. Three ascospores, one germinated after 72 hours in sea water (12%). 5. Part of the peridium with spines and underlying epidermoid flattened cells. Scale bars: 2-4 = 10 um; 5 = 14 um.
Kohlm. in the genus Arenariomyces. The latest, A. truncatellus, is different from these with respect to ascospore dimensions, the uneven sized spore cells, and the short and delicate structure of the appendages. A. truncatellus and A. majusculus ascomata are formed in the outermost part of the wood substrate and bearing spines in contrast to A. parvulus, A. trifurcatus, and A. triseptatus. However, the stability of ascomatal characters in the genus is questioned (on wood / on hard surfaces, spines / no spines). For example, A. trifurcatus ascomata are sometimes seen with a few spines and also directly on wood (Koch unpublished). Further observations of the rare Arenariomyces species could perhaps reveal a similar variability. But general similarities such as the thin-walled pseudoparenchyma of the young ascomata, unitunicate asci that deliquesce early before ascospore maturation, together with the three polar appendages point to A. truncatellus
72 ... Koch
as morphologically well accommodated as a new species in Arenariomyces. It is recognized that examination of A. truncatellus in the electron microscope, especially in regard to appendage development, and at the molecular level is needed to establish its relationship to A. trifurcatus, the type species.
Key to Arenariomyces species
ba-Ascomatamain ly On WOO, of Lie. ade. opal webs hd Ss old by wlth Ate ash talade itty 2 1b. Ascomata mainly.on hard stirfaces.¢< ota. sags sae has blame Os a od ee ee 3 2a ASCOSPOLES ASH MLAS «bt AV oikes dy Foweten dy brwcon dying cdog rs poten dri schon dying op A. majusculus 2bunscospores 214 LO 2a tav: ted aloes 2 ota teed seated See ts es A. truncatellus Day -ASCOSPOLES" SED CALS lay. ots! cot! atest: shade diet ohale do aha late aha lah! aha linia A. triseptatus SH AASCOsPOresel Se ptate ew peels eiucten Bbube 4 aude y Bouin y Mouide y Meeede natauele wa Ry wal 4 Aad, BSCOSPOLES Ox 29 D0 D= Goll trea oan fe weehanll oa eee ele Ree RHEL aul A. parvulus Aby AS COSPOLES 2BASZ, 29 = VG UIT aee sg dives eogdhireseg drt scene eeadrh sea gre aes A. trifurcatus
Acknowledgments
Iam grateful to Iben M. Thomsen and Flemming Rune, both at Forest and Landscape Denmark (Faculty of Life Sciences, University of Copenhagen) for photographic work, and to Iben M. Thomsen for preparing the manuscript for printing. Also I am grateful to Dr. E.B. Gareth Jones and Dr. K.L Pang for reviewing the manuscript and giving valuable comments.
Literature cited
Hoéhnk W. 1955. Studien zir Brack- und Seewassermykologie V. Héhere Pilze des submersen Holzes. Veréffentlichungen des Instituts fiir Meeresforschung in Bremerhaven 3: 199-227.
Jones EBG, Sakayaroj J, Suetrong S, Somrithipol S, Pang KL. 2009. Classification of marine Ascomycota, anamorphic taxa and Basidiomycota. Fungal Diversity 35: 1-203.
Koch J, Petersen KLR. 1996. A checklist of higher marine fungi on wood from Danish coasts. Mycotaxon 60: 397-414.
Koch J, Pang KL, Jones EBG. 2007. Rostrupiella danica gen. et sp. nov. a Lulworthia-like marine lignicolous species from Denmark and the USA. Botanica Marina 50: 294-301. http://dx.doi.org/10.1515/BOT.2007.034
Kohlmeyer J. 1968. Danische Meerespilze (Ascomycetes). Berichten der Deutschen Botanische Gesellschaft 81: 53-61.
Petersen KLR. 1997. Ultrastructural studies of the marine ascomycete Groenhiella bivestia. Botanica Marina 40: 71-75. http://dx.doi.org/10.1515/botm.1997.40.1-6.71
Petersen KLR, Koch J. 1997a. Substrate preference and vertical zonation of lignicolous marine fungi on mooring posts of oak (Quercus sp.) and larch (Larix sp.) in Svanemollen harbour, Denmark. Botanica Marina 40: 451-463. http://dx.doi.org/10.1515/botm.1997.40.1-6.451
Petersen KLR, Koch J. 1997b. Buxetroldia bisaccata gen. et sp. nov., a marine lignicolous halosphaeriacean fungus from coastal waters, Denmark. Mycological Research 101: 1524-1528. http://dx.doi.org/10.1017/S0953756297004383
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MY COTAXON
http://dx.doi.org/10.5248/124.73 Volume 124, pp. 73-85 April-June 2013
New ascomycete records from Guatemala
ROSARIO MEDEL”*', OSBERTH MORALES’, RANULFO CASTILLO DEL MORAL! & ROBERTO CACERES?
"Instituto de Investigaciones Forestales, Universidad Veracruzana, Apartado Postal 551, 91070, Xalapa, Veracruz, México
*Departamento de Microbiologia, Facultad de Ciencias Quimicas y Farmacia, Universidad de San Carlos de Guatemala, Guatemala
* CORRESPONDENCE TO: medel.rosario@gmail.com OR romedel@uv.mx
ABSTRACT — Thirty-two species of ascomycetes were previously recorded from Guatemala. This paper describes 12 newly recorded species, representing five orders and twelve genera. Three previously reported species are redescribed because there are few data in the Guatemalan literature.
Key worps — Helotiales, Hypocreales, Orbiliales, Pezizales, Xylariales, Central America
Introduction
Guatemala is a country of >108,000 km*. The name Quauhtemallan (Guatemala) means “land of trees,’ referring to the original wide extent and diversity of forests. Guatemala has been categorized as biologically mega- diverse (Tolisano & Lopez 2010), but little is known about the fungal diversity of this country and adjacent areas. There have been few studies on fungal diversity, and the knowledge of the ascomycetes is poor; only 33 ascomycetes species have been reported in previous studies (TABLE 1). The present study is part of a project that began in 2007 to document the ascomycete diversity of Guatemala.
Materials & methods
The study was based on a bibliographic revision and additional material collected mainly from oak forests and cloud forests of the west of Guatemala. The following literature was used for identifications: Abbot & Currah (1997); Breitenbach & Kranzlin (1984); Dennis (1954; 1963; 1970; 1978), Glawe & Rogers (1984), Haines & Dumont (1984); Hsieh et al. (2005); Ju & Rogers (1996); Medel (2002) Medel & Calonge, (2004); Méndez-Mayboca et al. (2007); Ramamurthi et al. (1957); San Martin et al. (1998), and
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TABLE 1. Ascomycete species recorded from Guatemala. (New records in bold font.)
Helotiales Bisporella citrina This paper Chlorociboria aeruginascens (Nyl.) Kanouse ex C.S. Ramamurthi et al. Flores et al. (2012)
C. aeruginosa This paper Lachnum abnorme This paper L. brasiliense Haines & Dumont (1984); this paper L. cyphelloides This paper L. virgineum This paper
Leotia lubrica (Scop.) Pers. Sommerkamp & Guzman 1990 Trichoglossum farlowii (Cooke) E.J.Durand Flores et al. (2002, 2012)
Hypocreales Cordyceps melolonthae (Tul. & C. Tul.) Sacc. Flores et al. (2012) C. militaris (L.) Fr. Flores et al. (2012) Hypomyces hyalinus (Schwein.) Tul. & C. Tul. Sommerkamp & Guzman (1990) H. lactifluorum (Schwein.) Tul. & C. Tul. Bran et al. (2003)
Ophiocordyceps gracilis This paper Orbiliales
Hyalorbilia inflatula This paper
Orbilia juruensis This paper Pezizales
Aleuria aurantia (Pers.) Fuckel Cookeina sulcipes (Berk.) Kuntze Gyromitra infula
Helvella acetabulum
H. crispa (Scop.) Fr.
H. elastica Bull.
H. lacunosa Afzel.
H. macropus (Pers.) P. Karst. Melastiza chateri (W.G. Sm.) Boud. Morchella costata (Vent.) Pers.
M. elata Fr.
M. esculenta (L.) Pers.
M. guatemalensis Guzman et al. Otidea onotica (Pers.) Fuckel Phillipsia guatemalensis Paden Pithya cupressina (Batsch) Fuckel Scutellinia scutellata (L.) Lambotte Wynnea americana Thaxt.
Xylariales
Annulohypoxylon thouarsianum Daldinia concentrica
D. fissa Lloyd
D. vernicosa Ces. & De Not. Diatrypella pulvinata
Phylacia poculiformis (Mont.) Mont. Poronia pileiformis
Xylaria cubensis (Mont.) Fr.
X. multiplex (Kunze) Fr.
X. polymorpha (Pers.) Grev.
Sommerkamp & Guzman (1990)
Sommerkamp & Guzman (1990)
Bran et al. (2003): this paper
This paper
Bran et al. (2003)
Sommerkamp & Guzman (1990)
Bran et al. (2003); Sommerkamp & Guzman (1990) Bran et al. (2003); Sommerkamp & Guzman (1990) Flores et al. (2002)
Sommerkamp & Guzman, (1990)
Bran et al. (2003)
Sommerkamp & Guzman (1990); Bran et al. (2003) Guzman et al. (1985)
Flores et al. (2002)
Paden 1977
Sommerkamp & Guzman (1990)
Sommerkamp & Guzman (1990)
Flores et al (2002; 2012)
Sharp (1948), Flores et al. (2012); this paper This paper
Morales et al. (2006)
Bran et al. (2003)
This paper
Sommerkamp & Guzman (1990)
This paper
Sharp (1948)
Sommerkamp & Guzman (1990)
Flores et al. (2012)
Ascomycetes from Guatemala ... 75
Spooner (1987). All material studied was deposited in the mycological collection at XAL herbarium (Instituto de Ecologia, A.C., Veracruz, Mexico) and duplicate specimens were deposited in mycological collection MICG (Universidad de San Carlos, Facultad de Ciencias Quimicas, Guatemala, Guatemala). Microscopic features were examined in KOH 5% and Melzer’s iodine reagent, and at least 20 measurements of ascospores were made per collection. Photographs were taken with a Sony Cyber Shot Camera, and some photographs of spores were cleared of spots, stains and bubbles using Photoshop CS5 software.
Helotiales
Bisporella citrina (Batsch) Korf & S.E. Carp., Mycotaxon 1: 58 (1974) Fic. 1 Apothecia discoid to cupulate shaped, attached to the substrate by a short
stalk. Disc 1 mm diameter. Stalk 0.6 mm long, cylindrical. Hymenium bright yellow and smooth outer surface. Asci cylindrical, 57-80 x 5-7 um, small pore blue in Melzer’s reagent. Ascospores elliptical, smooth hyaline, with 2 guttules, with a single septum when mature, 7-10 x 2-3 um. Paraphyses filiform with rounded tips, 1-1.5 um thick, hyaline, thin walled.
HABITAT — Gregarious, lignicolous; in cloud forest at 1500 m.
MATERIAL EXAMINED — GUATEMALA. CHIMALTENANGO DEPARTMENT, San Andres
Itzapa Municipality, Aldea La Hierbabuena, 27 June 2007, Medel 1451 (XAL). The bright yellow apothecia and ascospore size are diagnostic for this species. Bisporella citrina is widely distributed (Dennis 1970, 1978; Breitenbach & Kranzlin 1984; Medel & Calonge 2004; Méndez-Mayboca et al. 2007). This represents a new record for Guatemala.
Chlorociboria aeruginosa (Oeder) Seaver ex C.S. Ramamurthi, Korf & L.R. Batra, Mycologia 49: 859 (1958) [“1957”] Fic. 2 Apothecia globose to discoid shaped, irregularly distorted, 12 mm diameter, attached to the substrate by a short central stipe; hymenium smooth, blue- green. Stalk cylindrical, 0.5 mm long. Asci cylindrical, 8-spored, (57—) 70-80 x 5-6 um, small blue pore in Melzer’s reagent. Ascospores irregularly fusiform, smooth, hyaline, with 2 guttules, 9-15 x 2-3 um. Paraphyses with rounded tips, 1-1.5 um thick. HABITAT — Gregarious, lignicolous; in Cupressus forest at 1500 m.
MATERIAL EXAMINED — GUATEMALA. GUATEMALA DEPARTMENT, Universidad de San Carlos de Guatemala, University Campus, Parque Ecoldgico Las Ardillas, 28 June 2007, Medel 1495 (XAL); CHIMALTENANGO DEPARTMENT, San Andrés Itzapa Municipality, Aldea la Hierbabuena, 27 June 2007, Medel 1446 (XAL, MICG). According to Dixon (1975) this species is close to C. aeruginascens, which also has the same green color but which differs in smaller ascospores (6-10 x 1.5-2 um). Chlorociboria aeruginosa is distributed in North, South, and Central
76 ... Medel & al.
America, China, India, Japan, and the Philippines (Dixon 1975) and Venezuela (Mardones-Hidalgo & Iturriaga 2011). Despite being a widely distributed species, in Guatemala the species was not known.
Lachnum abnorme (Mont.) J.H. Haines & Dumont, Mycotaxon 19: 10 (1984) Fics 3-4 Apothecia discoid, with white hairs, stipe very small and sometimes almost sessile, 1 mm wide x 0.5 mm tall, hymenium light to dark yellow. Hairs sub- cylindrical with blunt apex, with coarse granules, hyaline, with 2-3 septa, thin walled, (40-)60-88 x (3-)3.5-4 um. Asci cylindrical, 8-spored (75-)87-94 x 7-8 um, small blue pore in Melzer’s reagent. Ascospores straight to s-shaped, hyaline, (38-)40-57 x 2 um. Paraphyses lanceolate, (77-)94-102(-110) x 3-3.5 um, hyaline, 3-4-septate. HaBITAT — Gregarious, lignicolous; in Cupressus forest at 1500 m. MATERIAL EXAMINED — GUATEMALA. GUATEMALA DEPARTMENT, Universidad de San Carlos de Guatemala, University City, Parque Ecolégico Las Ardillas, 28 June 2007, Medel 1486 (XAL). The studied specimen fitted well with the descriptions by Dennis (1963), Haines & Dumont (1984), and Spooner (1987). Lachnum abnorme is widely distributed in tropical regions of the world: Australia, New Zealand, Chile, India, Central America, and Costa Rica (Haines & Dumont 1984); Panama (Piepenbring 2006); and Venezuela (Mardones-Hidalgo & Iturriaga 2011). This is the first record to Guatemala.
Lachnum brasiliense (Mont.) J.H. Haines & Dumont, Mycotaxon 19: 23 (1984) Apothecia scattered on bark, light to dark yellow hymenium, 1.2 mm wide
disc, shallow receptacle, shaped cup with a hairy short stalk, white to pale buff color and a blue-black base. Hairs cylindrical with obtusely rounded tips, (52-)60-75(-80) x 3-4 um septate, thin walled, granulated. Asci cylindrical, (66-—)78-82(-84) x 7-9 um, 8-spored, hyaline, small blue pore in Melzer’s reagent. Ascospores narrowly fusiform, cylindrical, usually slightly curved, occasionally becoming 3-septate, (30—)32-43 x 2.5-3 um. Paraphyses cylindrical, (68-)72-91 x 2-3 um, 1-3-septate.
Hasitat — Gregarious, lignicolous; in cloud forest at 1800 m.
MATERIAL EXAMINED — GUATEMALA. CHIMALTENANGO DEPARTMENT, San Andrés
Itzapa Municipality, Aldea La Hierbabuena, 27 July 2007, Medel 1475 (XAL). The dark to medium yellow ascomata clothed with white hairs and the blue- black base of the stalk are distinctive characters. Lachnum brasiliense is a common species in the tropics and has previously reported from Guatemala (Haines & Dumont, 1984) in pine forest. The material examined was found in cloud forest.
Ascomycetes from Guatemala ... 77
Figures 1-8. Bisporella citrina: 1, ascospores. Chlorociboria aeruginosa: 2, apothecia. Lachnum abnorme: 3, ascospores; 4, apothecia. Lachnum cyphelloides: 5, apothecia. Lachnum virgineum: 6, paraphyses in Congo red. Hyalorbilia inflatula: 7, apothecia. Orbilia juruensis: 8, paraphyses. Bars: 1, 8 = 10 um; 2 = 1 cm; 3, 6 = 20 um; 4, 7 = 2 mm, 5= 1 mm.
Lachnum cyphelloides (Pat.) J.H. Haines & Dumont, Mycotaxon 19: 30 (1984) Fic. 5
Apothecia yellow translucent, disc 1 mm wide x 1.2 mm of height, long stipitate, small white hairs to almost smooth. Hairs cylindrical, with obtusely rounded tips, 48-70 x 3-4 um septate, hyaline, thin wall, granulated. Asci cylindrical 66-67 x 6-7 um, small blue pore in Melzer’s reagent. Ascospores fusiform, 31-40 x 1.5-2 um hyaline. Paraphyses lanceolate, (68-) 72-91 x 2-3 um hyaline, 1-3-septate.
Hasitat — Gregarious, lignicolous; in cloud forest at 1800 m.
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MATERIAL EXAMINED — GUATEMALA. GUATEMALA DEPARTMENT, Universidad de
San Carlos de Guatemala, University Campus, Parque Ecoldégico Las Ardillas, 27 July
2005, Medel 1465-A (XAL); 28 July 2005, Medel 1446 (XAL, MICG). According to Haines & Dumont (1984), L. cyphelloides is covered with white hair, has a yellow pale disc, and its stipe lacks a blue-black base. The species is distributed in Central America, northern South America, and the Lesser Antilles (Haines & Dumont, 1984; Piepenbring 2006). ‘This is first record for Guatemala.
Lachnum virgineum (Batsch) P. Karst., Bidr. Kann. Finl. Nat. Folk 19: 169 (1871) Fic. 6 Apothecia concave smooth disc, 1-1.5 mm in diameter, white to creamy white, and margin enrolled when it dries. Central stipe, covered with white hairs, 1-1.2 mm of height. Hairs cylindrical, with obtusely rounded tips, 75-100 x 4-5 um septate, thin walled, granulated hyaline. Asci cylindrical asci, 8-spored, 34-45 x 4-5um, thin walled, small blue pore in Melzer’s reagent. Ascospores fusiform, 6-8 x 1.5-2 um, thin walled, hyaline. Paraphyses lanceolate, 70-100 x 5-6 um, septate, hyaline, longer than the asci. HaBITaT — Gregarious, lignicolous; in tropical forest at 160 m. MATERIAL EXAMINED — GUATEMALA. ALTA VERAPAZ DEPARTMENT, Coban Municipality, Ecorregién Lachua, 17 September 2005, Lopez y Quezada 2473 (XAL). Lachnum virgineum is characterized by apothecia covered by straight, white granulate hairs. Spooner (1987) reported the species from North and South America, Europe, North Africa, Central Asia, India, Japan, Papua New Guinea, and Australia. This is a new record for Guatemala.
Hypocreales
Ophiocordyceps gracilis (Grev.) G.H. Sung, J.M. Sung, Hywel-Jones & Spatafora, Stud. Mycol. 57:43 (2007)
Stromata with long cylindrical apices terminating in 5 x 3 mm heads, red ochraceous, with dark dotted perithecial ostioles. Stipe yellow-olive. Asci with many individual spores, 250-280 x 6 um, pore not blue in Melzer’s reagent. Ascospores cylindrical, smooth, 7-10 x1-2 um, hyaline, lined chainlike upon each other within the ascus. Paraphyses not observed.
Hasitat — Solitary, on larvae of Lepidoptera; in cloud forest, 1800 m.
MATERIAL EXAMINED — GUATEMALA. CHIMALTENANGO DEPARTMENT, San Andrés
Itzapa Municipality, Aldea La Hierbabuena, 21 June 2007, Medel 1466 (XAL). Our specimen was consistent with the descriptions by Mains (1958), Dennis (1978), and Breitenbach & Kranzlin (1984). Ophiocordyceps gracilis is always associated with lepidopteran larvae. This is the first report for Guatemala.
Ascomycetes from Guatemala ... 79
Orbiliales
Hyalorbilia inflatula (P. Karst.) Baral & G. Marson, Micologia 2000 (Trento): 44 (2000) Fic. 7 Apothecia less than 1.5 mm wide, orange to yellow color, with rolled margin. Asci cylindric-clavate, rounded apex, 8-spored, 21-23(-29) x 3-3.5(-4) um, pore not blue in Melzer’s reagent. Ascospores cylindrical, straight or slightly curved, rounded ends, 4-8 x 0.8-1 um. Paraphyses cylindrical hyaline, 18-21 x 2(-2.5) um, septate, with apices encrusted to form a thin epithecial layer. Hasitat — Gregarious, lignicolous; in cloud forest and Cupressus forest at 1800 m.
MATERIAL EXAMINED — GUATEMALA. CHIMALTENANGO DEPARTMENT, San Andrés Itzapa Municipality, Aldea La Hierbabuena, 27 June 2007, Medel 1467, 1484 (XAL); GUATEMALA DEPARTMENT, Universidad de San Carlos de Guatemala, University Campus, Parque Ecoldgico Las Ardillas, 28 June 2007, Medel 1485-B (XAL, MICG). Our specimen agreed with the description by Spooner (1987). Liu et al. (2006) note that Hyalorbilia inflatula is widely distributed in China. The species has also been recorded from Venezuela (Mardones-Hidalgo & Iturriaga 2011). This is the first report for Guatemala.
Orbilia juruensis Henn., Hedwigia 43: 270 (1904) Fic. 8 Apothecia superficial, sessile or subsessile, disc convex or undulate,
with crenulate margin, translucent orange-yellow, smooth, 1 mm diameter. Receptacle smooth, concolorous or slightly paler than the disc, thin at the margin and thicker towards the centre. Asci cylindric-clavate , broadest at the apex and tapered downwards to a narrow base which is sometimes forked, apex truncate or obtusely rounded, 29-37 x 3.5-4 um, pore not blue in Melzer’s reagent. Ascospores straight or slightly inequilateral to fusoid, hyaline, (6-) 7-9(-10) x 1-1.5 um, not septate. Paraphyses capitate, tips up to 3.5-4 um diameter, 21-30 x 2-3 um, 1-2-septa in the lower part.
HaBITAT — Gregarious, lignicolous; in Cupressus forest at 1500 m.
MATERIAL EXAMINED — GUATEMALA. GUATEMALA DEPARTMENT, Universidad de
San Carlos de Guatemala, University Campus, Parque Ecolédgico Las Ardillas, 28 June
2007, Medel 1491 (XAL). This species is known form South America, Australasia, and Solomon Island (Spooner 1987), Argentina (Wright & Wright 2005), and Panama (Piepenbring 2006). This is the first record for Guatemala.
Pezizales
Gyromitra infula (Schaeff.) Quél., Enchir. Fung.: 272 (1886) Ascoma bilobate or saddle-shaped, generally fused with the stipe. The hymenium covers the whole surface, which is cinnamon to dark brown. The
80 ... Medel & al.
interior is hollow and whitish. The stipe is smooth, often somewhat furrowed or pitted, and finely felty towards the base. Asci cylindrical, (140-)150-250 x 15-17 um. Ascospores elliptical, smooth, with 2 guttules, hyaline, 18-22 x 8-10 um. Paraphyses clavate, 80-100 x 7-10 um. Hasitat — Solitary or gregarious, humicolous; in Pinus-Abies forest, 2900 m. MATERIAL EXAMINED — GUATEMALA. ToTONICAPAN DEPARTMENT, Totonicapan Municipality, Mercado municipal de Totonicapan, 18 September 2001, Morales 295 (MICG); 19 August 2006, Morales 561 (MICG); Totonicapan Municipality, Aldea Panquix, 31 August 2003, Morales 485 (MICG). The bilobate or saddle-shaped ascoma is characteristic of this species (Abbott & Currah 1997). Bran et al. (2003) previously reported this species from Guatemala, and it is also known from Mexico (Medel 2005).
Helvella acetabulum (L.) Quél., Enchir. Fung. 275 (1886)
Ascoma cup-shaped with an irregular margin, 25-50 mm broad; fertile surface dull brown. Lower surface smooth to subglabrous, light brown with conspicuous pallid to cream raised ribs extending from the stipe base to middle of the cup; thick whitish deeply ridged. Asci cylindrical, 200-300 x 14-17 um. Ascospores broadly elliptical, hyaline, 15-21(-22) x 11-15 um, smooth, with one guttule. Paraphyses with rounded tips, 5-6 um thick.
Hasitat — Solitary, humicolous; in pine-oak forest at 2300 m.
MATERIAL EXAMINED — GUATEMALA. CHIMALTENANGO DEPARTMENT, Tecpan
Municipality, Mercado municipal de Totonicapan, 7 October 1999, Morales 47 (MICG) The ridged pale brown apothecium is characteristic of this species (Calonge & Arroyo 1990, Abbot & Currah 1997). The species is known from Canada (Abbot & Currah 1997), China (Zhuang 2004), Mexico (Medel & Calonge 2004; Vite-Garin et al. 2006) and Spain (Calonge & Arroyo 1990). This is the first record for Guatemala.
Xylariales
Annulohypoxylon thouarsianum (Lév.) Y.M. Ju, J.D. Rogers & H.M. Hsieh, Mycologia 97: 861 (2005) Fics 9-10 Stromata sessile, globose, 12 mm wide x 7 mm high, with papillate ostioles in the middle ofa deep ring, surface dark brown to black, carbonaceous, context fibrous woody, radially lined to faintly concentrically zoned grey and brown. Asci not observed. Ascospores inequilateral narrowly elliptical to slightly fusoid, light to medium brown, smooth, 23-27 x 6-7 um, with a germ slit spore length on the flattened side. Paraphyses not seen. Hasitat — Gregarious, lignicolous; in oak forest.
MATERIAL EXAMINED — GUATEMALA. CHIMALTENANGO DEPARTMENT, Tecpan Municipality, Astillero Municipal, 21 July 2007, Medel 1436 (XAL).
Ascomycetes from Guatemala ... 81
FiGurEs 9-14. Annulohypoxylon thouarsianum: 9, ascospores in KOH 5%; 10, stromata. Daldinia concentrica: 11, ascospores in 5% KOH. Diatrypella pulvinata: 12, ascospores in 5% KOH. Poronia pileiformis: 13, ascus with blue pore + in Melzer’s solution; 14, stromata (ostiole and perithecia). Bars: 9 = 20 um; 10, 14 = 1 cm; 11 = 15 um; 12 = 10 um; 13 = 40 um.
The species was first recorded from Guatemala by Sharp (1948) as Hypoxylon malleolus Berk. & Ravenel,, a synonym of A. thouarsianum (Ju & Rogers 1996, Hsieh et al. 2005). This is a common xylariaceous species with greenish pigments in KOH and papillate ostioles in the middle of a deep disc (Ju & Rogers 1996; Medel 2002). The species is also known from Mexico (Pérez-Silva 1987; Medel 2002) and Panama (Piepenbring 2006).
Daldinia concentrica (Bolton) Ces. & De Not., Comm. Soc. Crittog. Ital. 1: 197 (1863) Fic. 11
Stromata, globose circular to oval shaped 20-30 mm wide, sessile and, attached to the substrate, hard surface, reddish-brown to blackish young, and finally black. Asci cylindrical, 100-130 x 7-8 um, uniseriate. Ascospores broadly elliptical to bean shape, dark brown, germ line straight, 13-15 x (6-) 6.5-7 um. Paraphyses not seen.
Hasitat — Gregarious, lignicolous; in oak forest.
MATERIAL EXAMINED — GUATEMALA. CHIMALTENANGO DEPARTMENT, Tecpan Municipality, Astillero Municipal, 21 July 2007, Medel 1423 (XAL).
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The important diagnostic characters of this species are the concentric lines in the stromata, and purple stromatal pigment in 5% KOH (Ju et al. 1997; Rogers et al. 1999). In Guatemala two species have been cited: D. vernicosa (Bran et al. 2003) and D. fissa (Morales et al. 2006). Daldinia concentrica, not previously reported for Guatemala, is also known from Mexico (Pérez-Silva 1978) and Europe, India, New Zealand, Taiwan, and the United States of America (Ju & Rogers 1997).
Diatrypella pulvinata Nitschke, Pyrenomyc. Germ. 1: 72 (1867) Fic. 12 Stromata pulvinate to oval, ostioles 3-5 sulcate, with endostromatic yellow
whitish tissue. Asci multispored. Ascospores 7-9 x 1.5-2 um, slightly curved (allantoid), hyaline, with 2 guttules. Paraphyses not seen.
HaBiTaT — Gregarious, lignicolous; in Cupressus forest at 1500 m.
MATERIAL EXAMINED — GUATEMALA. GUATEMALA DEPARTMENT, Universidad de
San Carlos de Guatemala, University Campus, Parque Ecoldégico Las Ardillas, 28 July
2007, Medel 1489 (XAL). The black sessile and pulvinate stromata with papillate ostioles are typical. Diatrypella pulvinata is known from Costa Rica, Europe, and the United States of America (Glawe & Rogers 1984; Chacén & Humafia 2006). This is the first record for Guatemala.
Poronia pileiformis (Berk.) Fr., Nova Acta R. Soc, Scient. Upsal., Ser. 3, 1: 129 (1851) Fics 13-14
Stromata simple to branched, at the top of the stalk it has a small umbelliform 4-6 mm broad head, beige with dark dots (perithecial ostioles), smooth and leathery, later becoming hard and brittle. Ostioles papillate. Stipe dark brown to black, 60-70 mm high x 1-6 mm wide, smooth or ribbed with a thickened base. Asci cylindrical, 100-130 x 4-5 um, pore blue in Melzer’s reagent. Ascospores ellipsoid, dark brown, 7-11 x 4-5 um, with a straight germ line as long as the length of the spore. Paraphyses not seen.
Hasitat — Gregarious, fimicolous, in tropical forest at 160 m.
MATERIAL EXAMINED — GUATEMALA. ALTA VERAPAZ DEPARTMENT, Coban
Municipality, Ecorregién Lachua, 19 September 2005, Lopez y Quezada 2473 (XAL). This species is distinguished by the simple or branched stromata with a beige head with black papillate ostioles, growing on dung (Dennis 1957; San Martin et al. 1998). It is known from Costa Rica, Mexico, Peru, and Philippines. This is the first record for Guatemala
Discussion With the new records, there are now 44 ascomycete species known from Guatemala (TABLE 1). The species encompass 26 genera of which Helvella
Ascomycetes from Guatemala ... 83
(5) and Morchella (4) provided the most records. The Pezizales was the most diverse order with 11 genera and 18 species. About 50% of the 200 specimens collected during the last survey in 2007 might represent new records or even new species.
The continued study of ascomycetes and other fungi from Guatemala will improve the knowledge of these organisms. This is essential because deforestation threatens to decrease the fungal biodiversity in this country. Recent deforestation data from Guatemala indicate a net 2006-2010 deforestation rate of over 38,000 ha / year (Regalado et al. 2012).
Acknowledgments
The authors thank to M.C. Bran (Universidad San Carlos de Guatemala) for supporting the expedition to collected the studied material of this study. Thanks to Dr. Gaston Guzman and Juan Lara (INECOL) for providing literature and facilities to consult the herbarium, and to the reviewers Dra. Evangelina Pérez Silva (Universidad Nacional Auténoma de México) and Dra. Cecilia Carmaran (Universidad de Buenos Aires), who significantly improved this manuscript. Special thanks to Dr. Shaun Penycook for his critical review.
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ISSN (print) 0093-4666 © 2013. Mycotaxon, Ltd. ISSN (online) 2154-8889
MY COTAXON
http://dx.doi.org/10.5248/124.87 Volume 124, pp. 87-99 April-June 2013
Coleosporium in Europe
STEPHAN HELFER
Royal Botanic Garden Edinburgh, 20A Inverleith Row, Edinburgh EH3 5LR, UK CORRESPONDENCE TO S.helfer@rbge.ac.uk
ABSTRACT — The species of Coleosporium (Uredinales) occurring in Europe are not clearly differentiated morphologically. Most taxa in Europe have been combined into formae speciales (fE.spp.) of Coleosporium tussilaginis by the majority of researchers. This study includes the taxa C. doronici, C. inulae, and C. telekiae as further ff.spp. in C. tussilaginis and redefines the existing ffspp. campanulae-rapunculoidis and senecionis-silvatici. It is hoped that this treatment will establish a pragmatic basis on which future taxonomic work can be built.
KEY worps — rust fungi, cryptic species, host specialism
Introduction
Coleosporium Lév. isa genus of Uredinales G. Winter (rust fungi) with the aecial stage parasitic on two-needle pines (Pinus spp.) and the telial stage on members of a wide range of angiosperm species. Its distribution is mostly limited to the northern hemisphere, where it is widespread and common in Europe, Asia, and North America (Farr & Rossman 2013, GBIF 2013). However, specimens have also been reported from South America (Arthur 1918, Hennen 2005) and New Zealand (McKenzie 1998). Their presence on two-needle pines has not been confirmed in these southern areas and specimens from Chile were reported to lack the teleomorph, possibly as a result (Arthur 1918). In New Zealand it is likely that already infected hosts from the northern hemisphere were planted or intercepted, and there is one reference of introduced Coleosporium rust on an indigenous species but no infections on two-needle pines have been reported (McKenzie 1998, Farr & Rossman 2013).
Léveillé established the genus in 1847. In a short note, he devised a system to bring more clarity into the confusing number of identical names of clearly distinct species and species with multiple names on account of their occurrence on a number of host plant species or genera (Léveillé 1847).
The type species is Coleosporium campanulae (lectotype, Laundon 1975).
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TABLE 1. Distribution of Coleosporium in host plant families in Europe
PLANT FAMILY Host GENERA Host SPECIES PREDOMINANT HOST GENUS Asteraceae 35 110 Senecio (34 host taxa) Campanulaceae 10 82 Campanula (63 host taxa) Orobanchaceae 8 38 Euphrasia (16 host taxa) Ranunculaceae 2 9
Pinaceae 1 10
CASUAL HOSTS & MISDETERMINATIONS NOTES
Tropaeolaceae 1 3 non-native plants Orchidaceae 1 1 border intercept plant Solanaceae 1 1 South American plants [Amaranthaceae] [1] [1] ? misidentified host Amarldccd r Pia seperti [Boraginaceae] [2] [2] = Melampsorella symphyti [Caryophyllaceae] [1] [1] = Caeoma coronariae [Rosaceae] [1] [1] = Pucciniastrum agrimoniae
Most authors agree that the European taxa of Coleosporium are indistinguishable in their morphology (e.g. Sydow & Sydow 1915, Hylander et al. 1953, Gdumann 1959, Wilson & Henderson 1966, Boerema & Verhoeven 1972). Therefore, many authors used the host plant identity to distinguish species (Gdumann 1959, Minkevicius & Ignataviciité 1991, Brandenburger 1994). Many current inventories treat all Coleosporium taxa distinguished in Europe as a single species and synonymise most of the original species into Coleosporium tussilaginis (Encyclopedia of Life 2012, Index Fungorum 2012). However, C. doronici, C. inulae, C. ligulariae, and C. telekiae have never been formally included in C. tussilaginis, although Hylander et al. (1953) did synonymise C. ligulariae with C. senecionis. The three remaining taxa are transferred to C. tussilaginis in the present study.
While some authors do not name the taxa included under C. tussilaginis (e.g. Majewski & Ruszkiewicz-Michalska 2008, Termorshuizen & Swertz 2011), others distinguish and name them (e.g., Gdumann 1959, Boerema & Verhoeven 1972, Encyclopedia of Life 2012, Bahcecioglu & Kabaktepe 2012), most often as formae speciales, i.e., biologically specialised forms (Eriksson 1894, Anikster 1984, Kirk et al 2008; see also the next paragraph below). The latter approach reflects the host specificity of Coleosporium analysed by extensive inoculation experiments (e.g., Klebahn 1904, 1924, Gaumann 1959, EURED names 2012). I follow this approach and extend it to all the European taxa.
Coleosporium in Europe... 89
Formae speciales (ff.spp.) are defined as taxa below the species level which are morphologically not or hardly distinguishable but are separated by host plant specificity. Nomenclature at this rank is not covered by the provisions of the International Code of Nomenclature for algae, fungi, and plants (McNeill et al. 2012: Article 4 Note 4); therefore in this paper I present them as newly named taxa, rather than as “comb. nov.’
This study covers the European taxa of Coleosporium on native plants. In the absence of molecular data, a morphological species concept is applied, extended by the use of discrete host plant ranges, in defining the sub-specific taxa of formae speciales, as described above. The aim is to present a pragmatic and effective basis for specimen sampling for molecular analysis, in order to generate data for further studies.
Materials & methods
The herbaria of Berlin (B), Paris (PC), Copenhagen (CP), St Petersburg (LE), Beltsville (BPI) and Edinburgh (E) were consulted in this research. Furthermore information was acquired from Ziirich and Bern (Helfer et al. 2011) and from databases at Stockholm (Krypto-S 2012), BPI (Farr & Rossman 2013) and National Herbarium Netherlands (NHL 2012). Label information was extracted and compared with accepted host plant taxonomy using the Plant List (2012) database. Geographical information was checked using Fuzzy Gazetteer (Kohlschiitter 2012) and Google Map (©2012 Google) and compiled according to the areas in the Atlas Florae Europaeae delimitations of Europe (Atlas Florae Europaeae 2012). Where reports were from non native or cultivated plants, this is indicated by the use of square brackets [ ]. Type material for C. telekiae, C. doronici, and C. ligulariae was obtained from Berlin (B).
Light microscopy: small portions of herbarium specimens were sectioned by hand and mounted in lactophenol-cotton blue for examination under a Carl Zeiss Axiophot© microscope with Carl Zeiss Axiocam© digital camera equipment. Measurements were achieved using the Axiocam software. Urediniospore dimensions as measured by light microscopy are reported in the following format: (min—) average +/-standard deviation (-max) length x breadth.
Scanning electron microscopy (SEM): small portions of herbarium specimens were removed and fastened to aluminium stubs with adhesive carbon tabs. Prior to sputter coating specimens were examined by a Carl Zeiss Stemi©2000 dissecting microscope, sorus sizes were measured and images taken with Carl Zeiss Axiocam© digital camera equipment as above. Specimens were coated with Palladium for 2min resulting in a deposition of approx 12nm and examined in a Carl Zeiss Supra©55VP SEM at 1-5kV and 5mm working distance. Measurements were achieved using the Axiocam software as above. Morphometric data were subjected to statistical analysis using the statistics package R (R Development Core Team 2011).
Details of the specimens examined are displayed with each taxon. Further information is available from the EURED specimens (2013) list.
Herbarium label images for B, PC, CP, LE and E used in this study can be viewed at the EURED website at E (Helfer et al. 2011). This website is continuously added to and improved.
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Results
The herbarium data searches for European taxa of Coleosporium resulted in 249 host plant relationships (TABLE 1), represented in five families. A further seven taxa in five additional host families had been found on intercepted plants or as single occurrences and four taxa in three families were misinterpretations. In the past more than 50 valid names have been used for these rust taxa (EURED names 2012).
The microscopic examinations of the specimens were inconclusive with respect to urediniospore dimensions and sorus characters: while statistically significant differences could be found between most samples involving various host species there were also significant differences between collections of the same host/fungus relationship (EURED stats 2012). Furthermore, data ranges overlapped widely, indicating that urediniospore dimensions and sorus characters are unsuitable for identification purposes and for the characterisation or delimitation of taxa in this genus (EURED spores 2012). Similarly, spore surface characters observed by SEM revealed no distinctive features. It is therefore concluded that, morphologically, the taxa are all part of the same species.
Taxa present in Europe
Coleosporium tussilaginis (Pers.) Lév., in Orbigny, Dict. Univ. Hist. Nat. 12: 786 (1849) f.sp. tussilaginis PLaTE 1 = Uredo tussilaginis Pers., Syn. meth. fung. 1: 218 (1801). = Ustilago tussilaginis Losa, Anales Jard. Bot. Madrid 5: 126 (1945).
ALTERNATE HOSTS: Pinus halepensis, P. mugo, P. nigra, P. pallasiana, P. pinaster,
P. sylvestris (PLATE 1), [PB echinata, P. ponderosa, P. rigida].
Primary Hosts: Tussilago farfara, [Emilia coccinea, Erechtites glomeratus], E. hieraciifolius,
[Euryops acraeus, E. evansii, Kleinia fulgens, K. grandiflora].
EUROPEAN DISTRIBUTION: pan-European.
SPECIMENS EXAMINED:
(as C. sp.) PC0022213.
(as C. tussilaginis) E00458054, E00458055, E00458056, E00458059, E00458060,
E00458061. UREDINIOSPORE SIZE: (21.7—)28.3+3.2(-39.6) x (15.9-)20.8+ 2.4(-26.6) um.
Notes: Artificial infections were obtained on Tropaeolum minus
(Tropaeolaceae) and Schizanthus grahamii (Solanaceae) (Gaumann 1959); weak infections were also obtained on plants of Senecio vulgaris (Klebahn 1924), calling into question the biological separation of this taxon from f.sp. senecionis discussed below. New specimens in many herbaria are deposited as C. tussilaginis, irrespective of host plant identity.
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PLATE 1 Aeciospore of Coleosporium tussilaginis on Pinus sylvestris. SEM; bar = 2 um.
Coleosporium tussilaginis f.sp. campanulae-rapunculoidis Boerema & Verh., Netherlands J. Pl. Pathol. 78(Suppl. 1): 7 (1972). = Coleosporium campanulae (Pers.) Fr., Summa Vegetabilium Scandinaviae: 512 (1849), as “campanulacearum”. = Coleosporium campanulae-rapunculoidis Kleb., Wirtswechselnde Rostpilze: 365 (1904). = Coleosporium campanulae-rotundifoliae Kleb., Wirtswechselnde Rostpilze: 366 (1904). = Coleosporium campanulae-trachelii Kleb., Wirtswechselnde Rostpilze: 366 (1904). = Coleosporium tussilaginis f.sp. campanulae-rotundifoliae Boerema & Verh., Neth. Jl Pl. Path. 78(Suppl. 1): 7 (1972). = Coleosporium tussilaginis f.sp. campanulae-trachelii Boerema & Verh., Neth. Jl Pl. Path. 78(Suppl. 1): 7 (1972). ALTERNATE HOstTs: Pinus nigra, P. sylvestris.
Primary Hosts: Adenophora liliifolia, [A. triphylla], Asyneuma giganteum, A. limonifolium, Campanula alliariifolia, C. alpina, [C. americana], C. barbata, C. baumgartenii, C. bononiensis, [C. californica], C. carnica, C. carpatica, C. caucasica, C. cervicaria, C. cespitosa, C. cochleariifolia, C. drabifolia, C. erinus, C. glomerata, [C. grandis], C. grossekii, C. hagielia, C. hawkinsiana, C. hispanica, C. incurva, C. isophylla, C. lactiflora, C. lanata, C. latifolia, C. lusitanica, C. lyrata, C. macrorhiza subsp. gypsicola,
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C. medium, C. moesiaca, C. patula, C. pelia, C. persicifolia, C. persicifolia subsp. subpyrenaica, C. pulla, |C. punctata], C. pyramidalis, C. rapunculoides, C. rapunculus, C. rhomboidalis, C. romanica, C. rotundifolia, C. rumeliana, C. rupestris, C. sarmatica, C. saxatilis, C. scheuchzeri, C. serrata, C. sibirica, C. sparsa, C. sparsa subsp. sphaerothrix, C. spathulata, C. speciosa subsp. affinis, C. speciosa, C. stenosiphon, C. stevenii, C. tatrae, C. tomentosa, C. trachelium, C. transsilvanica, C. versicolor, C. wanneri, Jasione montana, Legousia hybrida, L. speculum-veneris, [Lobelia cardinalis, L. xalapensis|, Michauxia campanuloides, M. laevigata, Petromarula pinnata, Phyteuma betonicifolium, P. nigrum, P. orbiculare, P. scheuchzeri, P. spicatum, [Triodanis perfoliata, Wahlenbergia denticulata]. EUROPEAN DISTRIBUTION: Au, Be, Bel, Br, Bu, Cg, Cs, Ct, Da, EU, Fe, Ga, Ge, Gr, He, Ho, Hs, Hu, Is, It, La, Lu, Mo, No, Po, Rm, Rus, Se, Sk, Su, Tu, Uk.
SPECIMENS EXAMINED:
(as C. campanulae) E00159053, E00458064, PC0022266, PC0022293, PC0022305, PC0022308, PC0022317, PC0022324, PC0022354.
UREDINIOSPORE SIZE: (12.0—)20.5+4.8(-30.2) x (9.0-)14.9+3.2(-26.4) um.
Notes: Boerema & Verhoeven (1972) treat this in three separate formae speciales: f.sp. campanulae-rapunculoidis, f.sp. campanulae-rotundifoliae, and fisp. campanulae-trachelii. Gaumann (1959) lists several overlapping host ranges and infections on unrelated angiosperms, such as Tropaeolum minus, and I see no justification to maintain these three special forms.
Coleosporium tussilaginis f.sp. doronici S. Helfer, f.sp. nov. MycoBank MB804294 = Coleosporium doronici Namysl., Spraw. Kom. fizyogr. Akad. Umijetn. Krakowie 45: 125 (1911). ALTERNATE HOSTS: Pinus sp. (presumed). PRIMARY Hosts: Doronicum austriacum, D. columnae. EUROPEAN DISTRIBUTION: Au, Cs, Hu, Po, Rm. SPECIMENS EXAMINED: (as C. doronici) B700015048, B700015049, B700015050, B700015051, B700015052, B700015053, PC0022411, PC0022412. UREDINIOSPORE SIZE: (10.8-)27.8+3.8(-34.1) x (7.0-)19.143.6(-24.8) um. Notes: This appears to be a geographically as well as biologically separate special form on Doronicum sp. (Miller 2003), since its hosts are widely distributed over Europe (Atlas Florae Europaeae 2012). The report in Gaumann (1959) that this rust occurs in Spain is based on a misunderstanding of the geographical name Galicia (“Galizien”), in fact referring to a region in the Tatra mountain border region of Poland.
Coleosporium tussilaginis f.sp. inulae S. Helfer, f.sp. nov. MycoBank MB804295 = Uredo erigerontis Req. ex Duby, Bot. Gall. Edn 2: 893 (1830), as “erigeronis”. = Coleosporium inulae Rabenh., Bot. Zeitung 9: 455 (1851). = Coleosporium carpesii Sacc., Rivista Period. Lav. Regia Accad. Sci. Lett. Arti Padova 24: 208 (1874).
Coleosporium in Europe ... 93
= Coleosporium jasoniae Gonz. Frag., Trab. Mus. Nac. Cienc. Nat. Madrid, é Bot. 9: 23 (1916). = Coleosporium asterisci-aquatici (Sacc.) Syd. & P. Syd., Ann. mycol. 19: 249 (1921) ALTERNATE Hosts: Pinus halepensis, P. sylvestris. PRIMARY HOSTS: Asteriscus aquaticus, Carpesium cernuum, Centaurea hierapolitana, Dittrichia graveolens, D. viscosa, Inula aschersoniana, I. aspera, I. britannica, I. candida, I. conyza, I. ensifolia, I. germanica, I. helenioides, I. helenium, I. helvetica, I. heterolepis,
I. hirta, I. methanaea, I. oxylepis, I. parnassica, [I. royleana], I. salicina, I. spiraeifolia, Jasonia tuberosa.
EUROPEAN DISTRIBUTION: Al, Au, Bl, Bu, Cg, Co, Cs, Ct, Da, Es, Fe, Ga, Ge, Gr, He, Hs, Hu, It, La, Lu, Mk, No, Po, Rm, Rus, Su, Tu, Uk. SPECIMENS EXAMINED: (as C. inulae) E00458051, E00458052, E00458053, E00458063. (as Uredo erigeronis) PC0022413, PC0022415. UREDINIOSPORE SIZE: (21.2—)30.3+3.3(-39.5) x (15.7-)23.4+3.1(-30.7) um.
Notes: A special form on most of the genera of tribus Inuleae Cass. of the Compositae. Significant damage caused by this rust on elecampane (Inula helenium) grown for medicinal use has been reported in Serbia (Pavlovic et al. 2003).
Coleosporium tussilaginis f.sp. melampyri Boerema & Verh., Netherlands J. Pl. Pathol. 78(Suppl. 1): 8 (1972). = Coleosporium melampyri (Rebent.) Kleb., Annls Sci. Nat., Bot., sér. 4, 4: 136 (1854). ALTERNATE HOstTs: Pinus mugo, P. sylvestris.
PRIMARY HOosTs: Melampyrum arvense, M. nemorosum, M. polonicum Soo, M. pratense, [Schizanthus sp.].
EUROPEAN DISTRIBUTION: Au, Bel, Br, Cs, Da, Fe, Ga, Ge, He, Ho, Hs, Hu, It, No, Po, Rm, Rus, Sl, Su.
SPECIMENS EXAMINED:
(as C. euphrasiae) PC0022164.
(as C. melampyri) E00458065, E00458066, E00458067, E00458068.
(as C. tussilaginis) E00458050.
(as C. rhinanthacearum) PC0022434. UREDINIOSPORE SIZE: (20.6—)25.9+2.6(-32.4) x (13.3-)18.942.2(-24.2) um.
Notes: Heavy infections of Melampyrum have been reported in Finland
(Pohjakallio & Vaartaja 1948). This rust was found to infect cultivated Schizanthus sp. (Solanaceae) in Sweden (Hylander et al. 1953) and Norway (Gjaerum 1974); please also see notes to f.sp. rhinanthacearum below.
Coleosporium tussilaginis f.sp. petasitis Boerema & Verh., Netherlands J. Pl. Pathol. 78(Suppl. 1): 8 (1972). = Coleosporium petasitidis Lév., Annls Sci. Nat., Bot., sér. 3, 8: 373 (1847). = Coleosporium petasitis de Bary, Microscopic fungi: 213 (1865).
ALTERNATE HOstTs: Pinus mugo, P. nigra, P. sylvestris.
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PRIMARY HOsTs: Petasites albus, P. frigidus, P. frigidus var. palmatus, P. hybridus, [P japonicus, a naturalized alien in central and northern Europe], P kablikianus, P. paradoxus, P. radiatus, P. spurius.
European distribution: Au, Br, Bu, Cs, Da, Fe, Ga, Ge, Gr, Hb, He, Ho, It, La, No, Po, Rm, Rus, Sk, Su.
SPECIMENS EXAMINED: (as C. petasitis) E00458069, E00458070, E00458071.
UREDINIOSPORE SIZES: (18.4—)26+2.3(-31.6) x (14.3-)20.742.2 (-26.6) um. Notes: Together with f.sp. senecionis, f.sp. tussilaginis, and f.sp. doronici, this rust develops on members of the tribus Senecioneae Cass. of the Compositae.
Coleosporium tussilaginis f.sp. pulsatillae Boerema & Verh., Netherlands J. Pl. Pathol. 78(Suppl. 1): 8 (1972). = Uredo tremellosa var. pulsatillae F. Strauss, Ann. Wetterauischen Ges. Gesammte Naturk. 2: 89 (1811) [“1810”]. = Coleosporium pulsatillae (F. Strauss) Fr., Summa Vegetabilium Scandinaviae: 512 (1849), as “pulsatillarum”. ALTERNATE Hosts: Pinus sylvestris. PRIMARY HOSTS: Anemone alpina, A. halleri, A. montana, [A. patens], A. pratensis, A. pulsatilla, A. slavica, Pulsatilla grandis, P. nigricans. EUROPEAN DISTRIBUTION: Au, Cs, Da, Fe, Ga, Ge, He, Ho, Hu, It, La, No, Po, Rm, Rus, Sk, Su, Uk. SPECIMENS EXAMINED: (as C. pulsatillae) E00458057, E00458062. UREDINIOSPORE SIZE: (20.8-)28.7+4.2(-40.3) x (15.5-)19.6+2.2(-24.4) um. Notes: This special form occurs on members of the Ranunculaceae. Despite its morphological likeness, many authors (Braun 1981, 1982, Su et al. 2012) consider this as a separate species. I concur with Boerema & Verhoeven (1972) in the treatment as a special form, as no reliable distinction can be made using morphometric methods.
Coleosporium tussilaginis f.sp. rhinanthacearum Boerema & Verh., Netherlands J. Pl. Pathol. 78(Suppl. 1): 8 (1972). = Coleosporium rhinanthacearum (DC) Lév., Annls Sci. Nat., Bot., sér. 3 8: 373 (1847). = Coleosporium flavum Bonord., Abh. Naturf. Ges. Halle: 186 (1860). = Coleosporium euphrasiae (Schumach.) G. Winter, Rabenh. Krypt.-Fl., Edn 2, 1(1): 246 (1881) [“1884”]. ALTERNATE HOstTs: Pinus mugo, P. sylvestris. Primary HOSTS: Bartsia alpina, |B. lutea, B. trixago], B. viscosa, Euphrasia x reuteri, E. arctica, E. brevipila, E. hirtella, E. liburnica, E. micrantha, E. minima, E. nemorosa, E. parviflora, E. pectinata, E. picta subsp. kerneri, E. rostkoviana, E. rostkoviana vat. fennica, E. salisburgensis, [E. striata], E. tricuspidata, Odontites litoralis, O. purpureus, O. vulgaris, Orthantha lutea, Parentucellia viscosa, Pedicularis palustris, Rhinanthus
Coleosporium in Europe ... 95
aestivalis, R. alectorolophus, [R. apterus], R. glaber, R. glacialis, R. pulcher subsp. alpinus, R. riphaeus, R. serotinus.
EUROPEAN DISTRIBUTION: Au, Be, Bel, Br, Bu, Cg, Cs, Da, Fe, Ga, Ge, Hb, He, Ho, Hs, Hu, It, La, Lu, No, Po, Rm, Rus, Sk, Su, Uk.
SPECIMENS EXAMINED:
(as C. euphrasiae) PC0022172, PC0022176, PC0022178, PC0022200, PC0022420.
(as C. flavum) PC0022202.
(as C. rhinanthacearum) E00458082, PC0022434, PC0022459.
UREDINIOSPORE SIZE: (19.2—)22.9+2.0(-28.4) x (14.1-)17.941.7(-22) um.
Notes - Besides f.sp. melampyri this is the second forma specialis on these more advanced members of the Orobanchaceae (Tank et al. 2006). In many herbaria the specimens of rusts on Rhinanthus, Euphrasia, Melampyrum and related hosts are labelled or arranged under any of the names Coleosporium euphrasiae, C. melampyri, C. rhinanthacearum, C. tussilaginis.
Coleosporium tussilaginis f.sp. senecionis-silvatici Boerema & Verh., Netherlands J. Pl. Pathol. 78(Suppl. 1): 9 (1972) = Coleosporium senecionis f.sp. senecionis-silvatici Wagner ex Gaum., in Beitr. KryptogFlora Schweiz 12: 124. 1959. = Coleosporium senecionis (Pers.) Fr., Summa Vegetabilium Scandinaviae: 512 (1849). = Coleosporium cacaliae G.H. Otth, Mitt. naturf. Ges. Bern: 179 (1865). = Coleosporium ligulariae Thiim., Bull. Soc. Imp. Nat. Moscou 52: 140 (1877).
ALTERNATE Hosts: Pinus halepensis, P. mugo, P. nigra, P. pinaster, P. pinea, P. sylvestris.
PRIMARY HOSTS: Adenostyles alliariae, A. alpina, [Delairea odorata, Farfugium japonicum var. giganteum, Hasteola suaveolens], Jacobaea alpina, J. arnautorum, J. erucifolia subsp. praealta, J. paludosa, [Ligularia fischeri, L. macrophylla], L. sibirica, Pallenis maritima, Parasenecio hastatus, Pericallis cruenta, [P. hybrida, Senecio candicans], S. carpetanus, [S. congestus], S. doria, S. doronicum, S. duriaei, |S. elegans], S. gallicus, |S. glaberrimus], S. hercynicus, [S. inaequidens], S. incrassatus, S. jacobaea, S. leucanthemifolius, S. leucanthemifolius subsp. vernalis, [S. leucostachys], S. lividus, S. macrophyllus, [S. moorei], S. nemorensis, S. ovatus, [S. pulcher], S. pyrenaicus, [S. scandens, S. smithii], S. squalidus, S. subalpinus, S. sylvaticus, [S. tristis], S. umbrosus, [S. viravira, S. viscidulus], S. viscosus, S. vulgaris, Tephroseris palustris.
EUROPEAN DISTRIBUTION: Au, Az, Be, BI, Br, Bu, Cg, Co, Cs, Da, Fe, Ga, Ge, Gr, Hb, He, Ho, Hs, Hu, It, La, Lu, No, Po, Rm, Rus, Sk, Su, Uk.
SPECIMENS EXAMINED:
(as C. cacaliae) PC0022228.
(as C. ligulariae) B700015041, B700015042, B700015044, B700015045, B700015046, B700015047.
(as C. senecionis) E00458072, E00458073, E00458074, E00458075, E00458076, E00458077.
UREDINIOSPORE SIZE: (18.6—)25.7£3.2(-37.1) x (11.8-)18.742.2(-26.4) um.
Notes: This is probably a pan-European form. Its distinctness from f.sp. tussilaginis needs to be confirmed by detailed host range studies or molecular investigations (see notes there).
96 ... Helfer
Coleosporium tussilaginis f.sp. sonchi Boerema & Verh., Netherlands J. Pl. Pathol. 78(Suppl. 1): 9 (1972). = Uredo tremellosa var. sonchi F. Strauss, Ann. Wetterauischen Ges. Gesammte Naturk 2: 90 (1811) [“1810”]; see Hylander et al. (1953). = Coleosporium sonchi Lév., Annls Sci. Nat., Bot., sér. 4 2: 190 (1854). = Coleosporium sonchi-arvensis (Pers.) Lév., in Orbigny, Dict. Univ. Hist. Nat. 12: 786 (1849). ALTERNATE HOstTs: Pinus sylvestris. PRIMARY HOSTS: Aposeris foetida, Arnoglossum atriplicifolium, Crepis palaestina, C. tectorum, Dendroseris litoralis, Lactuca muralis, Lapsana communis, Picris cupuligera,
Sonchus arvensis, S. asperl, S. brachyotus, S. oleraceus, S. palustris, S. tenerrimus, S. uliginosus.
EUROPEAN DISTRIBUTION: Au, Bel, BH, Br, Bu, Cg, Cs, Da, Fe, Ga, Ge, Hb, He, Ho, Hs, It, La, Lu, No, Po, Rm, Rus, Sk, Su, Uk.
SPECIMENS EXAMINED: (as C. sonchi) E00458078, E00458079. (as C. sonchi-arvensis) E00458080.
UREDINIOSPORE SIZE: (15.9—)21.8+2.5(-29.6) x (9.6-)17.2+3.0(-23.9) um. Notes: This forma specialis is restricted to the tribus Cichorieae Lam. & DC. of the Compositae.
Coleosporium tussilaginis f.sp. telekiae S. Helfer, f.sp. nov. MycoBank MB804296 = Coleosporium telekiae Thim., Fungi austriaci exsiccati no. 850 (1873) ALTERNATE HOsTs: Pinus sp. (presumed) Primary Hosts: Telekia speciosa, Xerolekia speciosissima. EUROPEAN DISTRIBUTION: Au, Bu, Cs, [Fe,] Ge, It, [No,] Po, Rm, Sk, Uk. SPECIMENS EXAMINED: (as C. telekiae) B700004933, B700010610, B700010611, B700015019, B700015020, B700015021, B700015022, B700015023, B700015024, B700015026, B700015027, B700015030, B700015033, B700015034, B700015035, B700015037, B700015038, B700015039, B700015040, E00458058. UREDINIOSPORE SIZE: (15.8—)22.6+3.0(-31.8) x (10.6-)17.2+2.4(-24.1) um. Notes: As well as in its natural range in Eastern Central Europe (Kokes 2004, Dietrich 2009), this rust was recently found on cultivated Telekia speciosa in Western Norway (Gjaerum et al. 2008).
Discussion
The present study used the host plant taxonomy of the Plant List (2012) database and revised the host plant spread to at least 239 taxa in 55 genera represented in 4 angiosperm families as well as ten Pinus species as alternate hosts (TABLE 1). The most common host family is the Compositae, represented by 96 genera and 706 taxa worldwide and 35 genera and 110 taxa in Europe (Farr & Rossman 2013). Kenneth & Palti (1984), who published a detailed study of
Coleosporium in Europe ... 97
the fungus-plant interactions of this family, found that globally Coleosporium was most widely distributed on the tribe Heliantheae with 14 genera affected followed by Senecioneae with 11 genera. Members of Heliantheae are not native in Europe, leaving Senecioneae as the most important host tribe, as is confirmed in this study.
To date few molecular investigations into Coleosporium have been published. A total of 32 sequences is currently deposited in GenBank (GeneBank 2013), mostly of the Asian/North American species C. asterum (Dietel) Syd. & P. Syd. Maier et al. (2003) compared four accessions of Coleosporium using the nuclear large subunit ribosomal DNA. In their neighbor joining analysis C. asterum (synonym Stichopsora asterum Dietel) was clearly separated from C. cacaliae, C. campanulae, and C. tussilaginis, which were not resolved.
Recent molecular studies have revealed the presence of cryptic species among morphologically indistinguishable specimens of Melampsora Castagne (Bennett et al. 2011, Milne et al. 2012), a rust genus of comparable host diversity, and it is probable that similar speciation is taking place in Coleosporium.
Conclusions
The genus Coleosporium is well defined within the Uredinales. Its taxa, however, are seriously confused. While host specificity appears to restrict the host range in some taxa, it does not seem to do so in others. Molecular taxonomic investigation of Coleosporium taxa is only in its infancy, and new initiatives are urgently needed to deal with the relationships in this genus.
Acknowlegements
The author acknowledges the support of the herbaria in B, CP, LE, PC, Z+ZT, BERN for access to specimens. The study was financially supported by SYNTHESYS (for visits to B [DE-TAF-924] and PC [FR-TAF-5178]) as well as a travel grant from the British Society for Plant Pathology to visit CP. Thanks are also expressed to Dr Reinhard Berndt and Prof. Salvatore Moricca for helpful comments during the peer review process and to the Nomenclature Editor.
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ISSN (print) 0093-4666 © 2013. Mycotaxon, Ltd. ISSN (online) 2154-8889
MY COTAXON
http://dx.doi.org/10.5248/124.101 Volume 124, pp. 101-109 April-June 2013
Septoglomus titan, a new fungus in the Glomeraceae (Glomeromycetes) from Bahia, Brazil
BRUNO ToMIO GOTO!" »,ADRIANE FREIRE ARAUJO?, ANA CRISTINA FERMINO SOARES’, ARAESKA CARENNA DE ALMEIDA FERREIRA’, LEONOR Costa Malta}, CARLA DA SILVA SOUSA? & GLADSTONE ALVES DA SILVA;
‘Departamento de Botanica, Ecologia e Zoologia, CB, Universidade Federal do Rio Grande do Norte, Campus Universitario, 59072-970, Natal, RN, Brazil
*Nucleo de estudo em Microbiologia Aplicada, CCA, Universidade Federal do Recéncavo da Bahia, Centro, S/N, CEP 44380-000, Cruz das Almas, BA, Brazil
*Departamento de Micologia, CCB, Universidade Federal de Pernambuco, Av. Prof. Nelson Chaves, S/N, CEP 50670-420, Cidade Universitaria, Recife, PE, Brazil
“CORRESPONDENCE TO: brunogoto@hotmail.com & adrianebiologa@yahoo.com. br
ABSTRACT — A new fungus of the Glomeraceae found in the rhizosphere of Agave sisalana is described under the epithet Septoglomus titan. It forms large glomerospores, (243-)265 x 325(-400) um in diameter. They generally are subglobose and have three spore wall layers: a semi-persistent short lived sub-hyaline to yellow-brown outer layer (0.5-1.4 um thick), an adherent smooth light-yellow to orange-brown middle layer (2.5-5.1 um), and a laminate smooth thick orange brown to dark red brown innermost layer (12.8-19.2 um). Spore size and colors of the spore wall layers separate this species from other yellow-brown to dark- brown species.
Key worps — Glomerales, Agavaceae, Glomus group A, crop system, rDNA
Introduction
Approximately 230 species of arbuscular mycorrhizal fungi (AMF; Glomeromycota) have been described, of which 118 exhibit glomoid spore development (sensu lato; Oehl et al. 2011a, Estrada et al. 2011). Historically, identification of the glomoid species was considered extremely difficult because of the limited morphological characters (Morton 1988), and the molecular approach has been used to solve the hypothetical separation into different genera, families, and orders (Walker & Schiifler 2004, 2007, Schiifler & Walker 2010). For this reason recent taxa from different glomeromycotan groups have been poorly described morphologically (Walker & SchiiBler 2004, Schiifler & Walker 2010), including groups with diversified morphological data sets such as Acaulospora, Kuklospora, and Ambispora (Kaonongbua et al. 2010, Kriiger et
102 ... Goto & al.
al. 2011, SchiiBler & Walker 2010). Recently Oehl et al. (2011a,b,c) combined significant morphological characters with concomitant molecular data to reorganize the Glomeromycota, especially the groups with glomoid spore development. This newly available morphological data set is being successfully used to identify distinct groups with glomoid spores (Estrada et al. 2011, Furrazola et al. 2011, Goto et al. 2012).
Glomoid species are very commonly found in Brazilian agrosystems (Silva et al. 2007, Goto et al. 2010, Maia et al. 2010, Nobre et al. 2010) with reports of many undescribed species (Carrenho et al. 2010) but with new descriptions rarely available. Studies on AMF diversity in Agave sisalana (common name: sisal) crop systems in Bahia State, Brazil, have revealed an undescribed species forming large spores with glomoid development, which we describe here as Septoglomus titan.
Material & methods
Study areas, soils sampling and soil parameters
Study areas were in the municipalities of Sao Domingos located at 12°48'S 41°37'W, and Ourolandia located at 10°58'S 41°05'W, both in the semi-arid region of Bahia State, in the Northeast Brazil. This semi-arid region is characterized by tropical dry climate (type Am of K6ppen-Geiger; Kottek et al. 2006), with mean annual temperature of 30°C and precipitation of 400-600 mm. The vegetation was agriculturally managed sisal, a perennial culture of 40-year-old systems, in a Caatinga biome characterized by low tree stratum without a continuous canopy, trees and shrubs with thin stems and small or composite leaves, deciduous in the dry season (Queiroz 2006). The predominant local geology is characterized by a granite-greenstone formed by the Santa Luz complex, Archaean basement, and comprises an assemblage of migmatitic gneisses with subordinate granitoids. The fungus was found in soil samples collected around Agave sisalana roots in farms of both municipalities. Soil characteristics in Ourolandia were: pH (CaCl) = 6.4, organic matter = 13 g dm’, available P = 18 mg dm’ and in Sao Domingos were: pH (CaCL,) = 6.3, organic matter = 32 g dm’, and available P = 58 mg dm’.
AMF bait cultures
The native AMF communities were cultured for two cycles (5 months each) with Sorghum bicolor, Brachiaria decumbens, and Panicum miliaceum in 500 mL pots filled with autoclaved sand substrate (250 g per pot) mixed with the natural field soil as AMF inoculum (250 g per pot), at the greenhouse of the Center for Agricultural, Environmental and Biological Research, Universidade Federal do Recéncavo da Bahia, Cruz das Almas, Brazil. Additionally, multiple glomerospores from the new species were separated and used as infective propagules in single species cultures on S. bicolor. The new fungus could successfully be propagated in bait cultures together with Acaulospora scrobiculata, Ambispora appendicula, Claroideoglomus etunicatum, Entrophospora infrequens, Funneliformis mosseae, Glomus intraradices, G. sinuosum, Paraglomus brasilianum, and P. occultum. However, no single species cultures were obtained from the fungus. Glomerospores isolated exclusively from the trap cultures were used for morphological and molecular analyses.
Septoglomus titan sp. nov. (Brazil) ... 103
Morphological analyses
Spores of the new species were extracted as described in Sieverding (1991). Trap cultures were established according to Goto et al. (2012) modified by using Hoagland’s solution without phosphorus. The spores were thereafter mounted in polyvinyl alcohol- lacto-glycerin (PVLG), in PVLG + Melzer’s reagent, and in water (Brundrett et al. 1994, Spain 1990). The terminology used is based on recent papers (Furrazola et al. 2011, Goto & Maia 2006, Goto et al. 2012, Estrada et al. 2011)
Molecular analyses
DNA EXTRACTION: DNA was extracted from three single spores of the new species. Individual spores on a slide with a drop (5-10 ul) of ultrapure water were crushed with a needle. The resulting material was used directly in the PCR reactions.
AMPLIFICATION AND SEQUENCING: DNA extract was used as template for a semi- nested PCR using the primers ITS3 (White et al. 1990) - 28G2 (Silva et al. 2006) and LR1 (van Tuinen et al. 1998) - 28G2 consecutively. PCR reactions were carried out in a volume of 50 ul, containing 75 mM Tris-HCl pH 8.8, 200 mM (NH,),SO,, 0.01% Tween 20, 2 mM MgCl, 200 uM each dNTPs, 1 uM of each primer, and 2 units of Taq" DNA polymerase (Fermentas); cycling parameters were 5 min at 95°C (1 cycle), 45 s at 94°C, 1 min at 55°C, 1 min at 72°C (40 cycles), and a final elongation of 7 min at 72°C following the last cycle. The amplified products were purified with a PureLink PCR Purification Kit (Invitrogen), following the manufacturer’s instruction and sequenced. Sequencing was provided by the Human Genome Research Center (Sao Paulo, Brazil).
SEQUENCE ALIGNMENT: Querying the National Center for Biotechnology Information databases with the BLASTn program, we verified that the sequences obtained from S. titan were affiliated with the Glomerales (Glomeromycota) before phylogenetic analysis. The AM fungal sequences (partial LSU rRNA) obtained in our laboratory were aligned with other glomeromycotean sequences from the GenBank using the program ClustalX (Larkin et al. 2007) and edited with the BioEdit program (Hall 1999) to obtain a final alignment. The sequences were deposited at GenBank under the accession numbers JQ312667-312669.
PHYLOGENETIC ANALYSES: Maximum parsimony (MP) analysis with 1000 bootstrap replications was performed using the Phylogenetic Analysis Using Parsimony (PAUP) program version 4 (Swofford 2003). Bayesian (two runs over 1 x 10° generations with a burnin value of 2500) and maximum likelihood (1000 bootstrap) analyses were executed, respectively, in MrBayes 3.1.2 (Ronquist & Huelsenbeck 2003) and PhyML (Guindon & Gascuel 2003), launched from Topali 2.5. The model of nucleotide substitution (GTR + G) was estimated using Topali 2.5 (Milne et al. 2004). Sequences from Claroideoglomus claroideum and C. etunicatum were used as outgroup.
Results
Molecular analyses
Phylogenetic analyses of LSU rRNA gene sequences (Fic. 1) revealed that the new fungus is a Septoglomus species (former Glomus group Aa3 sensu Oehl et al. 2011a).
104 ... Goto & al.
Claroideoglomus etunicatum AJ623310
C. claroideum AJ271929
100 100 1.00
0.1
Glomus sinuosum FJ461846 G. intraradices AF396797 G. intraradices AY842577 G. proliferum FM992402 G. proliferum FM992398 G. manihotis AM158947 G. clarum AJ510243 G. clarum AJ510242
Septoglomus deserticola AJ746249
55 53 0.73 72 S. constrictum FJ461827 84 70 1.00 83 S. xanthium AJ849467 D.99) S. titan JQ312667 100 100 1.00) S. titan JQ312669 3 S. titan JQ312668 1.00 Funneliformis coronatus AF 145739 F. mosseae AY639273 100 F. mosseae AF 145735 100 1.00
F. fragilistratus AF 145747
F. caledonius AJ628059
F. caledonius AJ510239
F. coronatus AF 145740
F, geosporus AF 145742
F. geosporus AF 145743
Fic. 1. Phylogenetic tree of the Glomeraceae based on LSU rDNA analysis and rooted by Claroideoglomus claroideum and C. etunicatum. Sequences are labeled with database accession numbers. Support values are from maximum parsimony (MP), maximum likelihood (ML) and bayesian analyses, respectively. Septoglomus titan sequences are in bold. Only topologies with 250% bootstrap values are shown. (Consistency Index = 0.62; Retention Index = 0.79).
Septoglomus titan sp. nov. (Brazil) ... 105
Fics 2-10. Septoglomus titan. 2. Spore formed on subtending hypha (sh) with three spore wall layers (SWL1, SWL2, SWL3) continuous with subtending hyphal wall layers. 3-6. Spore wall with three layers in young and mature spore (SWL1, SWL2 and SWL3). [3, 4. Young spores with inner layer (SWL3) less pigmented than outer layers (SWL1 and SWL2). 5, 6. Mature spores with inner layer (SWL3) more pigmented than outer layers (SWL1 and SWL2)] 7, 8. Subtending hyphae slightly funnel-shaped to slightly constricted and all three spore wall layers continuous with subtending hyphal wall (SW). 9, 10. Detail of septum formed by sublaminae of laminated inner layer (SWL3) typical of Septoglomus.
Taxonomic analyses
Septoglomus titan B.T. Goto & G.A. Silva, sp. nov. Fics 2-10 MycoBank MB 564321
Differs from Septoglomus africanum, S. deserticola, and S. xanthium by its larger spores with 3-layered walls; and from S. constrictum by its thicker 3-layered spore walls.
TyPE: BRAZIL: Bahia, near Ourolandia, in soil cultivated with Agave sisalana, 20.Apr.2011, B.T. Goto, permanent PVLG slides (Holotype, URM83528; GenBank JQ312667, JQ312668, JQ312669. Isotypes, URM83529; URM83530; Z+ZT Myc 15119).
ETyMoLoey: titan = in reference to the large spores formed by the new species.
GLOMEROSPORES are yellow brown to orange brown when young to dark red brown at maturity, formed terminally on hyphae, and globose to sub-globose (243-)265-375(-400) um or (rarely) oblong to irregular, (225-)245-285 x 308-325(-400) um.
106 ... Goto & al.
SPORE WALL is (15.3—)17.9-23.0(-—25.6) um thick in total, comprising three smooth layers (SWL1-3). SWL1 is sub-hyaline to light yellow, thin (0.5-1.4 um) observed in young and mature spores or in the subtending hyphal wall. SWL2 is pigmented, yellow brown to orange brown, unit (2.5-5.1 um); darker than SWL3 in young spores, SWL2 acquires pigmentation during maturation. The SWLS3 is dark yellow to yellow brown in young spores becoming orange brown to dark red brown in mature spores, laminated,12.8-19.2 um thick. Spore wall layers are continuous with the subtending hyphal wall layers, and the SWL2 and SWL3 pigments continue into the subtending hyphal wall. The three layers do not stain when exposed to Melzer’s reagent.
SUBTENDING HYPHA (sh) generally present, single, straight, cylindrical to constricted, light yellow to orange brown, 17.9-33.2 um wide (mean = 25.4 um) at the spore base. The subtending hyphal wall is 7.6-8.9(-10.2) um thick (mean 8.2 um) near the spore base, tapering to approximately 2.5-5.2 um about 100 um distant from the spore base; occlusion formed by bridging septum arising from sublaminae of SWL3, although the pore at the subtending hypha sometimes appears partly open.
GERMINATION by regrowth of the subtending hypha.
GLOMEROSPORE DEVELOPMENT deduced from spores found in different developmental stages. The sub-hyaline hyphal wall layer differentiates into sub-hyaline to yellow, evanescent (SWL1) and unified (SWL2) spore wall layers after which the laminate layer 3 differentiates with increasing number of developing sublaminae and becomes pigmented. Finally, the pore is not closed by introverted thickening of SWL3 but by a bridging septum arising from the laminate wall layer at the spore base.
ARBUSCULAR MYCORRHIZA FORMATION associated with roots of Sorghum bicolor (L.) Moench.
DISTRIBUTION — Currently known only from agricultural soils mainly cultivated with Agave sisalana Perrine (Agavaceae) in Ourolandia and Sao Domingos, Bahia State (Brazil).
ADDITIONAL COLLECTIONS EXAMINED: BRAZIL: Banta, near SAo Domingos, in soil cultivated with Agave sisalana, UFRN1895 UFRN1896 UFRN1897.
Discussion
Sequence analyses of the partial LSU rRNA region confirm with high support that the new species represents a Septoglomus that differs from the four other described Septoglomus species (Fic. 1). The morphology of the subtending hyphal structure (Fics 3-8) also supports the new species in Septoglomus as circumscribed by Oehl et al (201 1a).
Septoglomus africanum (Btaszk. & Kovacs) Sieverd. et al. S. deserticola (Trappe et al.) G.A. Silva et al., and S. xanthium (Blaszk. et al.) G.A. Silva et al.
Septoglomus titan sp. nov. (Brazil) ... 107
differ from S. titan by their smaller spores with 2-layered walls [S. africanum (60-)87(—125) um (Blaszkowski et al. 2010); S. deserticola 47-54(-115) x 38-52(-102) um (Trappe et al. 1984); and S. xanthium 20-55 x 45-100 um (Blaszkowski et al. 2004)]. Although Septoglomus constrictum (Trappe) Sieverd. et al. also has large (150-330 um) spores, it differs from S. titan by its thinner, 2-layered spore walls with a total thickness of 7-15 um (Trappe 1977, Oehl et al. 201:1a).
Septoglomus titan is easily distinguished from all other glomoid non- Septoglomus species by spore wall structure. A few dark species such as Glomus tenebrosum (‘Thaxt.) S.M. Berch and Funneliformis geosporus (T.H. Nicolson & Gerd.) C. Walker & A. Schiiéler resemble S. titan. Glomus tenebrosum has spores with only two spore wall layers (Thaxter 1922, Trappe 1977, Walker 1982). Funneliformis geosporus has spores with three wall layers, but the inner layer is flexible and thin (<1.0 um), whereas in S. titan the SWL3 is laminated and thick.
Goto et al. (2012) noted that the spore wall of G. trufemii B.T. Goto et al. was one of the thickest (7.4-15.5 um) among the glomoid-spored glomeromycotan species, but S. titan has even thicker spore walls.
Acknowledgements
The authors acknowledge Dr. Janusz Blaszkowski (Department of Plant Protection, West Pomeranian University of Technology, Szczecin, Poland) and Dr. Fritz Oehl (Agroscope Reckenholz-Tanikon ART, Zurich, Switzerland) for reviewing the manuscript and making helpful comments and suggestions and appreciate the corrections by Dr. Shaun Pennycook (Nomenclature Editor) and suggestions by Dr. Lorelei Norvell (Editor-in-Chief). This work was supported by: Conselho Nacional de Desenvolvimento Cientifico e Tecnolégico (CNPq) through Protax (Programa de Capacitacao em Taxonomia); PPBio (Programa de Biodiversidade do Semiarido) and INCT-HV (Herbario Virtual da Flora e dos Fungos); FACEPE (Fundacao de Amparo a Ciéncia e Tecnologia do Estado de Pernambuco); UFRN (Programa de Pés-Graduacgao em Sistematica e Evolugao); UFPE (Programa de Pés-Graduacao em Biologia de Fungos) and CAPES-PNPD (Coordenacao de Aperfeigcoamento de Pessoal de Nivel Superior - Programa Nacional de Pés-Doutorado). The Graduate Program in Microbiologia Agricola da Universidade Federal do Recéncavo da Bahia is acknowledged for providing technical support.
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Geoglossaceous fungi in Slovakia 5. Geoglossum uliginosum: taxonomy and nomenclature
VIKTOR KUCGERA’ , JOHAN NITARE’, PAVEL LIZON! & JAN GAISLER?
‘Institute of Botany, Slovak Academy of Sciences, Dubravskad cesta 9, SK-845 23, Bratislava, Slovakia *Skogsstyrelsen, SE-551 83 Jonkdping, Sweden >Crop Research Institute Prague,