YCOTAXON THE INTERNATIONAL JOURNAL OF FUNGAL TAXONOMY & NOMENCLATURE VOLUME 134 (4) OCTOBER-DECEMBER 2019 Rhomboidia wuliangshanensis gen. & sp. nov. (Xu & al.— Fie. 2, p. 657) ISSN (PRINT) 0093-4666 https://doi.org/10.5248/134-4 ISSN (ONLINE) 2154-8889 MYXNAE 134(4): 591-740 (2019) EDITORIAL ADVISORY BOARD KAREN HANSEN (2014-2021), Chair Stockholm, Sweden BRANDON MATHENY (2013-2020), Past Chair Knoxville, Tennessee, U.S.A. ELSE VELLINGA (2019-2022) Oakland, California, U.S.A. XINLI WEI (2019-2023) Beijing, China ToDD OSMUNDSON (2019-2024) La Crosse, Wisconsin, U.S.A. ELAINE MALosso (2019-2025) Recife, Brazil ISSN 0093-4666 (PRINT) ISSN 2154-8889 (ONLINE) MYCOTAXON THE INTERNATIONAL JOURNAL OF FUNGAL TAXONOMY & NOMENCLATURE OCTOBER-DECEMBER 2019 VOLUME 134 (4) http://dx.doi.org/10.5248/134-4 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 MyYcoTAxONn, LTD. © 2019 www.mycotaxon.com & www.ingentaconnect.com/content/mtax/mt P.O. BOX 264, ITHACA, NY 14581-0264, USA Iv ... MYCOTAXON 134(4) MYCOTAXON VOLUME ONE HUNDRED THIRTY-FOUR (4) — TABLE OF CONTENTS 134-4: TABLE OF CONTENTS, NOMENCLATURAL UPDATES, PEERS, EDITORIALS POVIC WONG Perk ols bcs ng ghs_ a tung Sea efor v athe citer sy 3 read RI Us eee Soret get ae vi Nomernclatural novelties ty pifications soo 4. cok ey tp eras keke eens vii COPTICONAG RA es Shox. aSecey gen on eau tats eae Este bod Gallente 8a alee g viii PP ROMISTTCTSAILOT 3, nyc. cess Ses aegy $e stg ents Sea he trv Aes gw cate ce ahh ob ogo eee ix ZOLO Ss WUMISSIOR DPYOCEAURE? 3: Pe tons Fry Sone Py Ra ver oAd oy Dey ly Yo Sha xi RESEARCH ARTICLES Urocystis cumminsii sp. nov., a smut fungus on Themidaceae from Arizona KYRYLL G. SAVCHENKO, SYLENA R. HARPER, Lori M. Carrts, Lisa A. CASTLEBURY 591 Leucoagaricus brunneus sp. nov. from Khyber Pakhtunkhwa, Pakistan Zi1A ULLAH, SANA JABEEN, MUHAMMAD FAISAL, Hasis AHMAD, ABDUL Nasir KHALID 601 Dictyostelids from Jilin Province, China 3: new Cavenderia and Dictyostelium records Pu Liu, SHUNHANG ZHANG, ZHUANG LI*, YuE Zou, XUEPING KANG, Yu Li 613 Records of Aureobasidium harposporum, Sarcophoma miribelii, and Stigmina dothideoides from Turkey MAKBULE ERDOGDU, MERVE ULUKAPI, ALI IHSAN KARAYEL, ZEKIYE SULUDERE 619 Bactrodesmium pulcherrimum sp. nov. from Ecuador FERNANDO ESPINOZA, DAYNET SOSA, LIZETTE SERRANO, ADELA QUEVEDO, FREDDY MAGDAMA, MARCOS VERA, SIMON PEREZ-MARTINEZ, ELAINE MALOSSO, RAFAEL F. CASTANEDA-RUIZ 627 Dendrographium multiseptatum sp. nov. from China L1-Guo Ma, YuE-L1 ZHANG, Bo ZHANG, Kal Qt, CHANG-SONG LI, JUN-SHAN QI 633 Haematomma pluriseptatum sp. nov. from China CONGCONG MIAO, RONG TANG, LINLIN DONG, ZHAOJIE REN, ZUNTIAN ZHAO 637 New records of Didymium inconspicuum, D. karstensii, and D. rugulosporum from China CHAOFENG YUAN, SHU LI, WAN WANG, SHUWEI WEI, QI WANG, Yu LI 643 OCTOBER-DECEMBER 2019 ... V Rhomboidia wuliangshanensis gen. & sp. nov. from southwestern China TaI-MIN Xu, XIANG-Fu Liu, Yu-Hur CHEN, CHANG-LIN ZHAO 649 Filsoniana lhasanensis sp. nov. from Tibet, China XuE-MEI! WEN, HurNISA SHAHIDIN, ABDULLA ABBAS 663 Sarcopodium flocculentum, the correct name for S. macalpinei SHAUN R. PENNYCOOK & PauL M. Kirk) 677 Pteridicolous ascomycetes from a cloud forest in eastern Mexico ROSARIO MEDEL-ORTIZ, YAJAIRA BAEZA, FRANCISCO G. LOREA-HERNANDEZ 681 First sexual morph record of Sarcopodium vanillae NAPALAI CHAIWAN, SAJEEWA S.N. MAHARACHCHIKUMBURA, DHANUSHKA N. WANASINGHE, MINGKWAN DOILoM, RUVISHIKA JAYAWARDENA, KEVIN D. HyDE 707 Notes on rust fungi in China 7. Aecidium caulophylli life cycle inferred from phylogenetic evidence and renamed as Puccinia caulophylli comb. nov. JiInG-XIn Jt, ZHUANG LI, Yu Li, MAKOTO KAKISHIMA 719 Exserticlava aquatica sp. nov., a microfungus from the Brazilian Amazon LUANA TEIXEIRA DO CARMO, D10GO CARELI DOS SANTOS, CAROLINA RIBEIRO SILVA, SHEILA MIRANDA LEAO FERREIRA, THAMARA ARAO FELETTI, Luis FERNANDO PASCHOLATI GuSMAO 731 MycoBioTa (FUNGA) NEW TO THE MYCOTAXON WEBSITE Ascomycota (lichenized and non-lichenized) on Syagrus coronata in the Caatinga biome: new and interesting records for Brazil and South America (suMMARyY) Mata4ra A.L. Dos SANTOS, NILO G. S. FORTEs, TAssio E. FE. Sitva, NaApja S. VITORIA 737 Checklist of Bolivian Agaricales. 1: Species with dark and pink spore prints (suMMaRy) E. MELGAREJO-ESTRADA, M.E. SUAREZ, D. RocaBADO, O. MAILLARD, B.E. LECHNER 739 vi ... MYCOTAXON 134(4) REVIEWERS — VOLUME ONE HUNDRED THIRTY-FOUR (4) The Editors express their appreciation to the following individuals who have, prior to acceptance for publication, reviewed one or more of the papers prepared for this issue. M. Catherine Aime D. Jayarama Bhat Lu-Sen Bian Marcela Eugenia da Silva Caceres Rafael F. Castahteda-Ruiz Vagner G. Cortez Cvetomir M. Denchev Shouyu Guo Tom Hsiang Shah Hussain Sevda Kirbag Klaus Kalb Paul M. Kirk John C. Landolt De-Wei Li José G. Marmolejo Tom May Eric H.C. McKenzie Josiane Santana Monteiro Karen K. Nakasone Nicolas Niveiro Lorelei L. Norvell Cumali Ozaslan Shaun R. Pennycook Marcin Piatek Meike Piepenbring Luis Quijada Scott Redhead Andrea Irene Romero Michelline Lins Silvério Adna Cristina Barbosa de Sousa Steven L. Stephenson Jan Vondrak Xinli Wei Ze-Fen Yu Changlin Zhao OcCTOBER-DECEMBER 2019... NOMENCLATURAL NOVELTIES AND TYPIFICATIONS PROPOSED IN MYCOTAXON 134(4) Bactrodesmium pulcherrimum R.F. Castaneda, F. Espinoza & D. Sosa [MB 830569], p. 629 Dendrographium multiseptatum L.G. Ma & J.S. Qi [MB 832847], p. 634 Exserticlava aquatica L.T. Carmo, C.R. Silva, Careli, S.M. Leao, Feletti & Gusmao [MB 831391], p. 732 Filsoniana lhasanensis X.M. Wen, Shahidin & A. Abbas [FN 570592], p. 669 Haematomma pluriseptatum R. Tang [MB 830618], p. 638 Leucoagaricus brunneus Z. Ullah, Jabeen & Khalid [MB 827985], p. 603 Puccinia caulophylli (Kom.) Jing X. Ji & Kakish. [MB 830631; epitypified: MBT 386779], p. 726 Rhomboidia C.L. Zhao [MB 833318], p.654 Rhomboidia wuliangshanensis C.L. Zhao [MB 833320], p. 656 Urocystis cumminsii Savchenko, Carris & Castl. [MB 830145], p. 595 VII vill ... MyCOTAXON 134(4) CORRIGENDA VOLUME 134-1 p.174, ACKNOWLEDGMENTS FoR: The author would like to thank Prof. Dr. Ertugrul Sesli, Prof. Dr. Ibrahim Tirkekul, and Dr. Shaun Pennycook for their helpful comments and careful review. READ: The author would like to thank Yiiziincii Yil University, Coordination of Scientific Research Projects for financial support (2010-FED-B031 and FYL- 2016-5213) and Prof. Dr. Ertugrul Sesli, Prof. Dr. Ibrahim Tiirkekul, and Dr. Shaun Pennycook for their helpful comments and careful review. MYCOTAXON 134-3 p. vii, line 21 FoR: Marthamyces culmigenus (Ellis & Everh.) P.R. Johnst. READ: Marthamyces culmigenus (Ellis & Langl.) P.R. Johnst. p.496, lines 3-5 FOR: Marthamyces culmigenus (Ellis & Everh.) P.R. Johnst., comb. nov. IF 556322 = Naemacyclus culmigenus Ellis & Everh.., Proc. Acad. Nat. Sci. Philadelphia 45: 151, 1893. READ: Marthamyces culmigenus (Ellis & Langl.) P.R. Johnst., comb. nov. IF 556322 = Naemacyclus culmigenus Ellis & Langl., in Ellis & Everhart, Proc. Acad. Nat. Sci. Philadelphia 45: 151, 1893. [Semi-bold fonts used above to flag corrected terms. ] CORRIGENDA IN CURRENT ISSUE (134-4) Cited below are mistakes or oversights present in approved input files not detected by authors until after PDF conversion. p. 613: Pu Liu and fellow coauthors wish to acknowledge the contributions made to “Dictyostelids from Jilin Province, China 3: new Cavenderia and Dictyostelium records” (MycoTaxon 134: 613-618) by Prof. Zhuang Li (Shandong Provincial Key Laboratory for Biology of Vegetable Diseases and Insect Pests, College of Plant Protection, Shandong Agricultural University, Tai’an 271018, China). The author sequence originally intended should read “Pu Liu, Shunhang Zhang, Zhuang Li, Yue Zou, Xueping Kang, Yu Li.” p-669, line 26 FoR: MK43983 READ: MK439830 OCTOBER-DECEMBER 2019 ... IX FROM THE EDITOR-IN-CHIEF DEADLINES, MYCOTAXON & NOMENCLATURE— The most onerous task your esteemed Editor-in-Chief faces is bringing the year-end issue in ‘on time’ A glance at our publication history suggests that during her tenure, she has failed miserably. Since 2004, she has met only three of her sixteen December deadlines: New Year's Eve 2004, Boxing Day 2007, 30 December 2009. Given this deplorable 37% success rate, it is no surprise that, once again, an October-December MycotTaxon will appear in January. (It could be worse: after adopting electronic submission and new software, we released the last 2005 volume on 11 May 2006!) Reasons/excuses for delays are myriad: time-consuming nomenclatural revision and editorial repair of sloppily prepared submissions (on the misplaced notion that it is quicker for us to do authors’ work for them), outside research commitments, severe health problems, uncontrollable press delays. Nonetheless, this year we were convinced that MycoTaxon 134(4) would be ready for delivery well before 2020. What we did NoT anticipate was the small number of research papers submitted. With several authors not sending us their final papers after their 2019 nomenclatural review, we waited until December 27 before deciding to release the issue with only 140 pages (the smallest issue EVER since 1974) rather than waiting for final submissions that have yet to appear. Frustrating, because removal of required page charges produced a flood of 138 accessions during 2019. What makes the year-end release date so important to MycoTaxon? With nomenclatural priority date-based, the date of publication is dictated by the actual release date and not what is displayed on the cover. That means that taxonomic names published in periodicals must cite both the actual date of publication and the ‘nominal’ date printed in the issue, e.g., following the order, Haematomma pluriseptatum R. Tang, Mycotaxon 134: 638 (2020) but indexed as “Miao, C, Tang R, Dong L, Ren Z, Zhao Z. 2020 (‘2019’). Haematomma pluriseptatum sp. nov. from China. Mycotaxon 134: 637-641. https://doi.org/10.5248/134.637 ” Much less confusing and infinitely more restful to have only ONE date to cite! MyYcOTAXON 134(4) presents 15 papers by 81 authors (representing 15 countries) as revised by 34 expert reviewers and the editors. The 2019 October-December MycoTaxon proposes one new genus (Rhomboidia from China) and eight new species representing Bactrodesmium from ECUADOR; Dendrographium, Filsoniana, Haematomma, and Rhomboidia from CHINA; Exserticlava from BraziL; Leucoagaricus from PAKISTAN; and Urocystis from the U.S.A. We also offer a new combination in Puccinia and epitypification for Puccinia caulophylli. New species range extensions are reported for [ascomycetes] Aureobasidium, Sarcophoma, Stigmina in TURKEY and new records and hosts of fern-associated ascos in eastern MExico and [myxomycetes] Cavenderia, Dictyostelium, and Didymium in CHINA. x ... MYCOTAXON 134(4) Two papers on Sarcopodium [1] explain why S. flocculentum is the correct name for S. macalpinei and [2] discuss the first sexual morph recorded for S. vanillae in Thailand. Another paper treats the full life cycle for the rust Puccinia (= Aecidium) caulophylli. Our small year-end issue closes with the announcements of two mycobiota [recently posted on www.mycotaxon.com] covering [1] new records of ascomycetes on Syagrus coronata in Brazil's Caatinga biome and [2] dark- and pink-spored agarics in Bolivia. Wishing us all health, happiness, illumination, and PEACE in 2020, Lorelei L. Norvell (Editor-in-Chief) 8 January 2020 PUBLICATION DATE FOR VOLUME ONE HUNDRED THIRTY-FOUR (3) MYCOTAXON for JULY-SEPTEMBER 2019 (1—-xIv + 423-590) was issued on October 2, 2019 OCTOBER-DECEMBER 2019 ... XI 2020 MYCOTAXON SUBMISSION PROCEDURE Prospective MycoTaxon authors should download the MycotTaxon 2020 guide, review & submission forms, and MycoTaxon sample manuscript by clicking the ‘file download page’ link on our INSTRUCTIONS TO AUTHORS page before preparing their manuscript. This page briefly summarizes our “4-step’ submission process. 1—PEER REVIEW: Authors first contact peer reviewers (two for journal papers; three for mycobiota/fungae) before sending them formatted text & illustration files and the appropriate 2020 MycoTaxon journal or mycota reviewer comment form. Experts return revisions & comments to BoTH the Editor-in-Chief <editor@mycotaxon.com> and authors. 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After PDF processing, the Editor-in-Chief corrects ONLY PDF editorial/conversion and index entry errors; corrections of all other errors are listed in the Errata of a subsequent issue for no charge. Authors will pay fees for mycobiota uploads, optional open access, and correction of major author errors to the Business Manager <subscriptions@mycotaxon.com> at this time. MyYcoTAaxoN LTD— www.mycotaxon.com The Mycotaxon Webmaster <mycotaxon@gmail.com> posts announcements, subscription & publication information, and author forms & templates on the official MycoTAXxoONn site. Our server also hosts the mycobiota web-page for free download of Fungae (regional annotated species lists). MyYCOTAXON ONLINE— www.ingentaconnect.com/content/mtax/mt The Mycotaxon journal publishes four quarterly issues per year. Both open access and subscription articles are offered. MYCOTAXON ISSN (print) 0093-4666 (online) 2154-8889 Mycotaxon, Ltd. ©2019 October-December 2019—Volume 134, pp. 591-599 https://doi.org/10.5248/134.591 Urocystis cumminsii sp. nov., a smut fungus on Themidaceae from Arizona KYRYLL G. SAVCHENKO"™*, SYLENA R. HARPER’, Lori M. Carris’, Lisa A. CASTLEBURY? Department of Biological Sciences, Butler University, Indianapolis, IN 46208 ? Department of Plant Pathology, Washington State University, Pullman, WA 99164, USA > USDA-ARS, Mycology and Nematology Genetic Diversity and Biology Laboratory, 10300 Baltimore Ave, Beltsville, MD 20705, USA “ CORRESPONDENCE TO: ksavchen@butler.edu ABSTRACT—The morphology and phylogenetic relationships of a species of Urocystis on Dichelostemma capitatum (Themidaceae, Asparagales) collected in the Tucson Mountains in Arizona, United States, were studied using microscopy and ITS rDNA sequence analyses. This is a first record for smut fungi on hosts from Themidaceae. Molecular phylogenetic analyses based on ITS sequence data revealed its basal position in relation to species on Poaceae. As a result, the smut in leaves of Dichelostemma capitatum is described and illustrated here as a new species, Urocystis cumminsii. Key worps—plant pathogens, taxonomy Urocystidales Introduction Urocystis Rabenh. ex Fuckel contains more than 170 species of plant pathogenic smut fungi, found all over the world but more common in temperate areas of both hemispheres (Vanky 2011b). More than 60% of Urocystis species are found on monocotyledons, with Poaceae serving as a major monocotyledonous host family, followed by Juncaceae, Hypoxidaceae, Convallariaceae, Amaryllidaceae, and Hyacinthaceae (Vanky 2011b). The intra-level phylogenetic relationships of Urocystis species have never been analysed and thus far only the graminicolous species from triticoid hosts 592 ... Savchenko & al. have been included in comprehensive phylogenetic analyses (Savchenko & al. 2017). Most of the Urocystis species described recently—e.g., U. achnatheri L. Guo, U. anemones-narcissiflorae Vanky, U. arxanensis L. Guo, U. beckwithiae Vanky, U. circaeasteri Vanky, U. dunhuangensis S.H. He & L. Guo, U. glabella Vanky & R. G. Shivas, U. helanensis L. Guo, U. koeleriae L. Guo, U. phalaridis Vanky, U. puccinelliae L. Guo & H.C. Zhang, U. rostrariae Piatek, U. sinensis L. Guo, U. skirgielloae Piatek, U. wangii L. Guo, and U. xilinhotensis L. Guo & H.C. Zhang—were supported only by morphological and host-specialization data (Guo 2002, 2005, 2006; Guo & Zhang 2004, 2005; He & Guo 2007; Piatek 2006a, 2006b; Vanky 2004, 2005, 2011la; Vanky & Abbasi 2011). The integration of molecular phylogenetic analyses, host plant taxonomy, and morphology provides a natural classification for various smut genera (Bauer & al. 2008; Castlebury & al. 2005; Kruse & al. 2018; Lutz & al. 2008; McTaggart & al. 2012; Savchenko & al. 2013, 2015; Vanky & Lutz 2007), justifying the need of a phylogenetic study of the genus. During a survey of Urocystis species diversity in the United States, we examined a specimen identified as Urocystis sp. on Dichelostemma capitatum (Benth.) Alph. Wood (Themidaceae) from Arizona in the WSP herbarium. Previously, no Urocystis species had been recorded on hosts from this family. The present study aimed to resolve the specific status of the smut on D. capitatum through morphological analysis and determine its phylogenetic affinities within Urocystis. TABLE 1. GenBank sequences used in this study. SPECIES GENBANK NO. REFERENCE Antherospora scillae EF653983 Bauer & al. 2008 A. vaillantii EF653988 Bauer & al. 2008 Urocystis bolivarii KX057771 Savchenko & al. 2017 U. colchici DQ839596 Matheny & al. 2006 U. cumminsii MK575496 This study U. eranthidis JN367299 Kellner & al. 2011 U. fischeri KF668284 Smith & Lutz 2013 U. occulta KX057774, Savchenko & al. 2017 U. trillii HQ239361 Henricot 2010 U. tritici KX057782 Savchenko & al. 2017 Ustilago hordei AY345003 Stoll & al. 2003 Vankya heufleri EF667965 Bauer & al. 2008 V. ornithogali EF635910 Bauer & al. 2008 Urocystis cumminsii sp. nov. (United States) ... 593 Materials & methods The herbarium specimen is deposited in Washington State University Mycological Herbarium, Pullman, WA, United States (WSP). Sorus and spore characteristics were studied using dried herbarium material. Specimens were examined by light microscopy (LM). Pictures of sori were taken with a Canon Power Shot G10 camera. For LM, spores were mounted in 90% lactic acid on a microscope slide, gently heated to boiling point to eliminate air bubbles, and then examined under a Carl Zeiss Axiostar™ light microscope at 1000x magnification and photographed with a Canon Power Shot G10 camera. At least 50 spore balls were measured, and the variation is presented as a range with extreme values given in parentheses. Means and standard deviations (SD) are provided after the spore size ranges. For SEM studies, spore balls were attached to metal stubs by double-sided adhesive tape and coated with gold. Spore surface ornamentation was observed at 15 kV and photographed with a JEOL JSM-6700F scanning electron microscope with a working distance of c. 12-13 mm. Sequences from other species of Urocystis and related genera were obtained from GenBank (TABLE 1). Genomic DNA was isolated from spore balls removed from the herbarium specimen that had been lysed in 1.5 mL tubes for 1 min using FastPrep°24. Tubes were incubated in a water bath for 5 hours at 55 °C, and DNA extracted using DNeasy Plant Mini Kit (QIAGEN) following the manufacturer’s instructions. DNA was amplified in 20 ul aliquots on an Applied Biosystems® GeneAmp 9700 thermal cycler using ITS1 as the forward primer and ITS4 as the reverse primer (White & al. 1990). Standard cycling parameters with an annealing temperature of 57 °C were used for amplification. PCR products were purified with USB ExoSAP-IT according to the manufacturer’s instructions, amplified with the forward and reverse PCR primers with the BigDye® Terminator v3.1 Cycle Sequencing Kit, and sequenced on an ABI PRISM? 3100 Genetic Analyzer. Consensus sequences were assembled, aligned, and edited with Geneious 7.1.8 for MacOS and with MAFFT 6.853 (Katoh & al. 2002, Katoh & Toh 2008) using the L-INS-i option. Maximum Likelihood (ML) was implemented as a search criterion in RAxML (Stamatakis 2014). GTR+I+G was specified as the evolution model in MrModeltest (Nylander & al. 2004). The RAxML analyses were run with a rapid Bootstrap analysis (command -fa) using a random starting tree and 1000 maximum likelihood bootstrap replicates. A Markov Chain Monte Carlo (MCMC) search in a Bayesian analysis (BA) was conducted with MrBayes (Ronquist & Huelsenbeck 2003). Four runs were implemented for 5 million generations. The cold chain was heated to a temperature of 0.25°C. Substitution model parameters were sampled every 500 generations and trees were saved every 1000 generations. Convergence of the Bayesian analysis was confirmed using AWTY (Nylander & al. 2008) and a 594 ... Savchenko & al. burn-in of 18,000 generations was calculated. The ML and Bayesian analyses were run three times to test accuracy. The tree was rooted using Ustilago hordei (Pers.) Lagerh. Results The ITS alignment of 13 sequences (including the outgroup Ustilago hordei) comprised 643 characters including gaps. The different BA and ML analytical runs yielded consistent topologies in respect to well-supported branches (a posteriori probability >90% in most cases). The consensus tree of one run of Bayesian phylogenetic analyses is presented in Fic. 1. The Urocystis sequences fell into two major clades. ‘The first clade comprised species from Poaceae and Themidaceae, and the second clade included species from Cyperaceae, Liliaceae s.l., and Ranunculaceae. The two species from the genus Vankya Ershad clustered together with the second Urocystis clade. Ustilago hordei Antherospora scillae Anth Jo) Antherospora vaillantii a IE Ore Urocystis cumminsii sp. nov. Urocystis bolivarii Urocystis 1 Urocystis occulta Oe 7 is, Urocystis tritici Urocystis eranthidis Vankya heufleri Vankya ornithogali Urocystis 2 Urocystis colchici rae = 2 Urocystis trillii Urocystis fischeri Fic. 1. Bayesian inference of phylogenetic relationships resulting from the analysis of ITS nucleotide sequence data. Numbers on branches are estimates for PPs from Bayesian inference (only probabilities >0.8 are shown). Urocystis cumminsii sp. nov. (United States) ... 595 Fic. 2. Urocystis cumminsii (WSP 68198). A. sori in leaves of Dichelostemma capitatum; B. spore balls seen by SEM; C, D. spore balls seen by LM. Scale bars: A = 2 mm, B = 5 um, C, D = 20 um. Taxonomy Urocystis cumminsii K.G. Savchenko, Carris & Castl., sp. nov. FIG. 2 MB 830145 Differs from Urocystis camassiae by its lighter colored yellowish-brown spores with thicker walls and its smaller thinner walled sterile cells, and by its host specialization on Themidaceae. Type: USA. Arizona, King’s Canyon, 16 km west of Tucson, on Dichelostemma capitatum (as D. pulchellum), 30.03.1981, leg. G.B. Cummins (Holotype, WSP 68198; GenBank MK575496). EryMoLocy: Named after George B. Cummins (1904-2007), an eminent American mycologist, who collected the holotype specimen. 596 ... Savchenko & al. Sor! in leaves as slightly elevated, pustular, elongate areas of various size and shape, sometimes confluent, visible on both sides of the leaf, initially lead- colored and covered by the epidermis which ruptures exposing the powdery, black mass of spore balls. SPORE BALLS globose, subglobose, ovoid to irregular, 20-45 um diam., composed of 1-6 (mostly 3) spores and more or less complete investing layer of sterile cells. Spores subglobose, ovoid, irregularly oblong to elongated, 11-15 x 12-18 um diam. [mean + SD, 13.3 + 2.6 x 15 + 2.9 um], medium yellowish brown, wall 1-1.5 um thick, smooth. STERILE CELLS subglobose, elongated, ovoid, 5-7 x 5-11(-13) um, pale yellow, to almost hyaline, with smooth, 1 um thick wall. Discussion Dichelostemma Kunth is a North American genus of wild hyacinths, closely related to Brodiaea Sm., from the family Themidaceae. Plants from this family are native to Central America and western North America, from British Columbia to Guatemala (Pires & Sytsma 2002, Stevens 2018). No members of Themidaceae were previously known to be parasitized by smut fungi (Vanky 2011b). Our molecular phylogenetic analyses and morphological data have helped resolve the systematic position of Urocystis on D. capitatum. The only possible close relative to U. cumminsii might be another native North American species, U. camassiae Vanky, found on Camassia Lindl. (Agavaceae; Fay & Chase 1996, Pires & Sytsma 2002). However, U. camassiae is distinguished by its darker colored reddish-brown spores with thinner spore walls (0.5-1 um) and its larger (5-17 um) sterile cells with thicker walls (1-2 (-3) um; Vanky 1994). ITS phylogenetic analysis infers that U. cumminsii is sister to the clade of Urocystis species on grasses and separate from species found on hosts from families more closely related to Themidaceae, such as U. colchici (Schltdl.) Rabenh. ex A.A. Fisch. Waldh. and U. trillii H.S. Jacks., indicating multiple inter-family host jumps during the evolution of Urocystis species, similar to those in the closely related genus Thecaphora Fingerh. (Vasighzadeh & al. 2014). Interestingly, Vankya heufleri (Fuckel) Ershad and V. ornithogali (J.C. Schmidt & Kunze) Ershad also clustered within the Urocystis lineage (Fic. 1). Most sequences of Urocystis in GenBank are derived from the LSU region. Unfortunately, our preliminary analysis showed that LSU is not informative for Urocystis phylogenetics. Hence, we based our current phylogeny on ITS data, utilizing the limited number of Urocystis ITS sequences available in GenBank. 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San Diego, Academic Press. https://doi.org/10.1016/B978-0-12-372180-8.50042-1 MY COTAXON ISSN (print) 0093-4666 (online) 2154-8889 Mycotaxon, Ltd. ©2019 October-December 2019—Volume 134, pp. 601-611 https://doi.org/10.5248/134.601 Leucoagaricus brunneus sp. nov. from Khyber Pakhtunkhwa, Pakistan ZIA ULLAH"*?, SANA JABEEN?®3, MUHAMMAD FAISAL’, HaBiB AHMAD‘, ABDUL NASIR KHALID” ' Department of Microbiology and Molecular Genetics & "Department of Botany: University of the Punjab, Quaid-e-Azam Campus-54590, Lahore, Punjab, Pakistan *Department of Botany, Division of Science & Technology, University of Education, Township, Lahore, Pakistan “Islamia College, University Peshawar, Peshawar, Pakistan * CORRESPONDENCE TO: Ziaullah.phd.mmg@pu.edu.pk ABsTRACT—A field survey of macrofungi from the Hindu Kush range of District Swat revealed a fungus morphologically similar to Leucoagaricus but representing a new species, here named L. brunneus. A detailed description and comparison with closely allied taxa are provided. Maximum likelihood analysis based on the internal transcribed spacer rDNA region (ITS) confirm its placement in Leucoagaricus sect. Rubrotincti. Key worps—Agaricaceae, Agaricales, Agaricomycetes, phylogeny, polyphyletic Introduction Leucoagaricus Locq. ex Singer, assigned to the family Agaricaceae (Singer 1986, Vellinga 2004), is represented by over 100 species in the northern and southern hemispheres (Kumar & Manimohan 2009; Ge 2010; Liang & al. 2010; Vellinga 2010; Vellinga & Balsley 2010; Vellinga & al. 2010; Mufioz & al. 2012, 2014; Kumari & Atri 2013; Malysheva & al. 2013; Ye & al. 2014; Ge & al. 2015; Qasim &al. 2015; Dovana & al. 2017; Hussain & al. 2018; Usman & Khalid 2018). From Pakistan only ten Leucoagaricus species have been reported previously: L. asiaticus Qasim & al., L. badius S. Hussain & al., L. lahorensiformis S. Hussain 602 ... Ullah & al. & al., L. lahorensis Qasim & al., L. leucothites (Vittad.) Wasser, L. pabbiensis Usman & Khalid, L. pakistaniensis Jabeen & Khalid, L. serenus (Fr.) Bon & Boiftard. L. sultanii S. Hussain & al., and L. umbonatus S. Hussain & al. (Ahmad & al. 1997, Ge & al. 2015, Qasim & al. 2015, Hussain & al. 2018, Usman & Khalid 2018). The genus is characterized morphologically by small- to medium- sized basidiomata with entire pileus margins, free lamellae, a central, equal to bulbous stipe, a membranous, persistent annulus, a white, cream, or pink spore print white, usually dextrinoid basidiospores, the presence of cheilocystidia, rare occurrence of pleurocystidia, and absence of clamp connections (Singer 1986). Several molecular phylogenies indicate that Leucoagaricus is polyphyletic (Johnson & Vilgalys 1998, Johnson 1999, Vellinga 2004). Vellinga (2003, 2004) showed that Leucoagaricus and Leucocoprinus species cluster together phylogenetically but intermix within a single clade. Because of the large number of species in the clade and relatively limited molecular data, the taxonomic and phylogenetic relationships among many Leucoagaricus and Leucocoprinus species have not yet been resolved. During the present investigation, one Leucoagaricus specimen collected from the Shawar Valley (district Swat, Khyber Pakhtunkhwa, Pakistan) is proposed here as a new species, L. brunneus, supported by our morphological and phylogenetic analyses. Material & methods The mushroom was collected during a forest foray in Shawar valley, Khyber Pakhtunkhwa, Pakistan, during the 2017 monsoon season and photographed at the sampling site. Macrocharacters were recorded from the fresh basidioma, with colors coded according to Munsell’s Soil Color Charts (1975). The specimen was air dried for further analyses and deposited in the Herbarium of University of the Punjab, Quaid- e-Azam Campus, Lahore, Pakistan (LAH). For microscopic studies, fruiting body tissues were mounted in 2% KOH and Congo red (to increase contrast). Basidia, basidiospores, cheilocystidia, and pellicular elements were measured using Scopelmage 9.0. DNA extraction followed Bruns (1995). The internal transcribed spacer region (ITS1-5.8S-ITS2 = ITS) was amplified with the ITS1F/ITS4 primer combination (White & al. 1990, Gardes & Bruns 1993). PCR products were purified and both strands were sequenced by Beijing Genomic Institute (BGI). For phylogenetic analysis, an ITS consensus sequence was generated in BioEdit (Hall 1999) and BLAST searched at NCBI (https://www.ncbi.nlm.nih.gov/); closely related sequences in L. sect. Rubrotincti were retrieved from both database and literature (Hussain & al. 2018). Lepiota subgracilis Kihner was chosen as outgroup (Liang & al. 2010). All sequences were aligned online using PRANK tool Leucoagaricus brunneus sp. nov. (Pakistan) ... 603 (https://www.ebi.ac.uk/goldman-srv/webprank/). Gaps were treated as missing data. The final aligned ITS dataset was phylogenetically analyzed through MEGA 6.0 software. Maximum Likelihood (ML) analysis was performed by selecting Kimura 2-parameter with Gamma Distributed model using Best-Fit Substitution Model approach in MEGA 6.0 (Tamura & al. 2013). Taxonomy Leucoagaricus brunneus Z. Ullah, Jabeen & Khalid, sp. nov. FIGS 1, 2 MB 827985 Differs from Leucoagaricus truncatus by its smaller size, smooth shiny non-squamulose pileus with brown striations, and its longer and narrower elongate to cylindrical basidiospores. Type: Pakistan. Khyber Pakhtunkhwa Province, Swat district, Lower Shawar, under Quercus oblongata D. Don (Fagaceae), 8 July 2017, Zia Ullah LS4 (Holotype, LAH35862; GenBank MH990662). EryMotoey: brunneus (Latin) refers to the brown coloration of the pileus fibrils. Figure. 1. Leucoagaricus brunneus (holotype, LAH35862). Basidioma. Scale bars = 0.5 cm. 604 ... Ullah & al. Ee Figure. 2. Leucoagaricus brunneus (holotype, LAH35862). A. Basidiospores; B. Basidia; C. Cheilocystidia; D. Pileipellis; E. Stipitipellis. Scale bars = 10 um. (Drawing by Sana Jabeen.) PiLteEus 35 mm diam, plane with incurved margins and umbonate center, surface smooth and shiny, dark brown (10YR1/2) from center to lighter brown (10YR2/4) towards margin in the form of radial striations, context cream. Leucoagaricus brunneus sp. nov. (Pakistan) ... 605 LAMELLAE free, close to crowded, margins entire, cream. LAMELLULAE absent. STIPE central, 9 x 4-8 mm, narrower towards the apex and wider (<8 mm) towards the base, then again narrowing (<6 mm) at the base, cream with grayish brown (10YR4/4) patches over the central part; annulus superior (at the center of the upper half), white. BasIDIOSPORES [30/1/1] (8.0—)8.2-10.5(-10.6) x (4-)4.1-4.9(-5.1) um, Q = (1.8-)1.9-2.1(-2.2), avQ = 2, elongate to cylindrical in face view, amygdaliform in side view, smooth, germ pore lacking, dextrinoid in Melzer’s reagent, hyaline in KOH. Basip1a 17.3-19 x 7.4—-8.1 um, clavate, smooth, hyaline, 4-spored. CHEILOCYSTIDIA 29.7-32.8 x 8.5-12.7 um, clavate to subclavate, hyaline, without crystals. PLEUROCysTIDIA absent. PILEIPELLIS a cutis with slightly clavate to cylindrical elements, 4.1-6.7 um diam, hyaline in KOH. Stiprripexuis cylindric hyphae, 3.0-5.2 um diam, hyaline in KOH. CLAMP CONNECTIONS absent in all tissues. EcoLtocy & DISTRIBUTION—Saprobic and solitary on humus-rich soil under Quercus oblongata [= Q. incana Roxb., nom. illeg.], at 1200 m elevation, in moist temperate Quercus vegetation. Phylogenetic analysis The 72 ITS sequence dataset (TABLE 1, Fic. 3) comprises 757 positions. Maximum likelihood analysis clusters the local collection in a sister clade with L. truncatus Z.W. Ge & Zhu L. Yang and L. purpureolilacinus Huijsman with a bootstrap support of 72% (Fia. 3). Discussion Leucoagaricus brunneus can be distinguished from L. truncatus based on basidioma size. Leucoagaricus truncatus produces medium to large (40-80 mm) basidiomata (Ge & al. 2015) and is further separated by the orange-white to gray-orange furfuraceous squamules (Ge & al. 2015) on its pileus surface in contrast to the smooth shiny pileus with brown striations that distinguish L. brunneus. Leucoagaricus truncatus is further separated microscopically by its more broadly ovoid and broadly amygdaliform basidiospores (Ge & al. 2015). The ITS sequence analysis also provides strong bootstrap support for L. brunneus as an independent taxon. Leucoagaricus brunneus differs from L. purpureolilacinus, which is characterized by a pinkish brown pileus and presence of crystalliferous cheilocystidia (Vellinga 2001). Pileus morphology also separates Leucoagaricus brunneus with its dark brown umbo and brownish striations running towards the margin from 606 ... Ullah & al. the center from L. serenus and L. crystallifer Vellinga, both characterized by whitish basidiomata and pilei with white-to cream obtuse umbos with obvious striations near the margins. Leucoagaricus serenus and L. crystallifer are further distinguished by their ovoid and broadly amygdaliform TABLE 1. Leucoagaricus species and specimens, and Lepiota subgracilis outgroup, used for ITS phylogenetic analyses. SPECIES VOUCHER COUNTRY GENBANK NO. L. americanus Vellinga 2454 (UCB) USA AY 176407 JRH091509-1 (TENN) USA MF773593 L. asiaticus LAH5872011 Pakistan KP 164972 LAH10012012 Pakistan KP 164971 L. badius LAH SH210 Pakistan KU647734 LAH SH148 Pakistan KU647736 L. bresadolae Bas7981 USA AF295929 MCVE:756 Italy GQ329047 CCBAS802 Czech Rep. LN714565 L. brunneus LAH35862 [T] Pakistan MH990662 L. crystallifer Huijser (L) Germany AF482863 SFC 1010003-02 Spain KY350216 L. dyscritus Vellinga 3532B (UC) USA GU136181 Vellinga 3956 (UC) USA GU136180 L. gaillardii MCVE:16517 Italy GQ329064 MCVE:736 Italy GQ329042 L. griseodiscus MCVE:13719 Italy GQ329059 L. japonicus J. Li221 China KY039572 L. jubilaei Guinberteau 99101101 France AY243635 10115A USA KX258658 L. lacrymans Zhang 599 China KY039574 L. lahorensiformiS FH-SHL2 Pakistan KU647730 LAH SHL2 Pakistan KU647729 L. littoralis MCVE:856 Italy GQ329051 MCVE:702 Italy GQ329041 MCVE:13721 Italy GQ329060 L. cf. majusculus MFLU 09-0164 Thailand HM488764 L. medioflavoides MCVE:2324 Italy GQ329055 L. meleagris IMG 1671 USA KY680786 Vellinga 2095 (L) Netherlands AF482867 Vellinga 1990 (L) Netherlands AY176419 CAW-9 India GQ249888 L. menieri L. nivalis L. pakistaniensis L. purpureolilacinus L. rubroconfusus L. rubrotinctus L. sardous Leucoagaricus sp. L. subcrystallifer L. sublittoralis L. subpurpureolilacinus L. subvolvatus L. sultanii L. truncatus L. umbonatus L. vassiljevae L. viscidulus L. volvatus Lepiota subgracilis herb. Huijser Yang 5792 LAH SJF13 LAH SJF23 MCVE:754 Vellinga 2291 (L) ZT13003 (ZT) gr0557 gr157 KUN:HKAS 54317 KUN:HKAS 54240 AC5195 Thiele 2646 JZB2115002 BAB 4737 Ge 97 Yang 3972 Vellinga 2561 TENN:070790 FLAS-F-60259 M.M. Rogers (UCB) Vellinga 2484 (UC) Ge 878 Ecv2235 Yang 3959 Ge 406 Brand s.n. (L) LAH SH115b LAH SH115 Ge 793 LAH SHL1 LAH SHL8 LE 10350 LE 289338 LE 289432 AC4187 AC1785 Vellinga 1783 (L) HKAS 5802 Netherlands China Pakistan Pakistan Italy Netherlands USA China China China China China Spain Australia China India China China USA USA USA USA USA China Netherlands China China USA Pakistan Pakistan China Pakistan Pakistan Russia Russia Russia Spain Spain Netherlands China Leucoagaricus brunneus sp. nov. (Pakistan) ... 607 KP300879 KY039573 KU647727 KU647728 GQ329045 AF482869 KP300875 KP300877 KP300876 JN944082 JN944081 JX827166 KT992149 AY176432 JN907015 KR154966 KP096237 KP096238 AY176430 MF686514 MF153051 AY176434 GU136182 KP096236 AY176442 KP096234 KP096233 KP300878 KU647732 KU647733 KP096235 KU647737 KU647738 JX133169 JX133170 JX896447 KT992148 KT992150 AY176490 EU416290 608 ... Ullah & al. 69 KP300877 L. rubrotinctus AY 176442 L. sublittoralis KP300876 L. rubrotinctus 69 | JX133169 L. vassiljevae KY 039572 L. japonicus JX133170 L. vassilievae JX896447 L. vassilievae 96 |75 JN944082 L. rubrotinctus GQ329045 L. purpureolilacinus 99 KU647730 L. lahorensiformis KU647729 L. lahorensiformis 4 JN944081 L. rubrotinctus V1 KP300875 L. rubroconfusus 50 JX827166 L. rubrotinctus 63 AY 176432 Leucoagaricus sp. 99 | KP096234 L. subpurpureolilacinus KP096233 L. subpurpureolilacinus 93 | KU647727 L. pakistaniensis KU647728 L. pakistaniensis 92| P24 JN907015 Leucoagaricus sp. 99 KR154966 Leucoagaricus sp. 98 /KU647734 L. badius KU647736 L. badius 99 — KP164972 L. asiaticus KP164971 L. asiaticus KT992148 L. viscidulus 90 99 KU647732 L. sultanii 66 50 KU647733 L. sultanii I 99 — AF482863 L. crystallifer 3p KY350216 L. crystallifer hg -— KP300879 L. menieri g, KP300878 L. subvolvatus 4 KT992150 L. volvatus 46 KP096236 L. subcrystallifer KT992149 L. sardous 99 | GQ329051 L. littoralis GQ329041 L. littoralis 27 GQ329060 L. littoralis 95 | KP096237 Leucoagaricus e 45 Q329059 L. griseodiscus KP096238 Leucoagaricus sp. KY 039573 L. nivalis 78 99 KU647737 L. umbonatus 53 KU647738 L. umbonatus 52 AY 176430 Leucoagaricus sp. 99 |MF686514 Leucoagaricus sp. MF 153051 Leucoagaricus sp. 51 AF482869 L. purpureolilacinus Se ? KP096235 L. truncatus @ MH990662 L. brunneus AY 176434 Leucoagaricus sp. 99 AY243635 L. jubilaei 18 KX258658 L. jubilaei is [197 ,GU136181 L. yee 53 GU136180 L. dyscritus GU 136182 Leucoagaricus sp. 99 ,GQ329064 L. gaillardii 56 GQ329042 L. gaillardii GQ329055 L. medioflavoides “4 KY039574 L. lacrymans 4 og | KY680786 L. meleagris AF 482867 L. meleagris 50 AY 176419 L. meleagris GQ249888 L. meleagris 84 HM488764 L. cf. majusculus AY 176407 L. americanus 81 | _ AF295929 L. bresadolae 56} GQ329047 L. bresadolae MF773593 L. americanus LN714565 L. bresadolae 99 ,AY176490 Lepiota subgracilis —| £U416290 Lepiota subgracilis -——XH 0.02 Ficure. 3. Molecular phylogenetic analysis of ITS sequences of Leucoagaricus species, with Lepiota subgracilis outgroup. The evolutionary history was inferred by using the Maximum Likelihood method based on the Tamura 3-parameter model. The analysis involved 72 nucleotide sequences. All positions containing gaps and missing data were eliminated. There were a total of 448 positions in the final dataset. Our new species is marked with @. Leucoagaricus brunneus sp. nov. (Pakistan) ... 609 basidiospores, contrasting with the elongated to cylindric and amygdaliform basidiospores of L. brunneus. In addition, in L. crystallifer the cheilocystidia have obvious crystals on the surface (Vellinga 2000, 2001) unlike the smooth cheilocystidia of L. brunneus. In comparison with Pakistani Leucoagaricus species, L. brunneus differs from L. asiaticus characterized by oblong to ellipsoid spores and cheilocystidia with crystals at the apex (Ge & al. 2015). Morphologically, its pileus color distinguishes L. brunneus (brown) from L. badius (red), L. pakistaniensis (whitish), L. lahorensis (dark reddish brown), L. lahorensiformis (light orange), L. umbonatus (yellowish pink), and L. sultanii (dark yellow) (Qasim & al. 2015, Hussain & al. 2018). Basidiospore morphology also separates these species from L. brunneus (Qasim & al. 2015, Hussain & al. 2018). All the earlier named Pakistani species have smaller spores except L. lahorensis, whose spores measure more or less the same as L. brunneus (Qasim & al. 2015), and none except L. brunneus present the same basidiospore shape—elongate to cylindrical in face view and amygdaliform in profile: amygdaliform to oblong in L. badius (6.5-7.5 x 4-5 um), ellipsoid to rarely amygdaliform in L. pakistaniensis (7.5-8 x 4.5-5 um), broadly ellipsoid to ellipsoid in L. lahorensis (8-10.6 x 6.4—7.6 um), fusiform to amygdaliform in L. lahorensiformis (6.5-7.5 x 3.5-4 um), amygdaliform to ellipsoid in L. umbonatus (5.5-6.5 x 3.5-4 um), and amygdaliform to ellipsoid in L, sultanii (5.5-7 x 3.5-4.5 um) (Qasim & al. 2015, Hussain & al. 2018). The crystals on the cheilocystidial apices in L. lahorensiformis and L. sultanii also separate both species from L. brunneus. Therefore a combination of morphological differences and a phylogenetic bootstrap support of 72% (Fic. 3) provide convincing support for our new species, Leucoagaricus brunneus. Acknowledgments The work is funded by HEC Research Project number 20-3383/HEC/R&D/14/184. Sincere thanks to Dr. Chang-Lin Zhao (Forestry College, Southwest Forestry University, Yunnan, P.R. China) and Dr. Shah Hussain (Center for Plant Sciences and Biodiversity, University of Swat, Pakistan) for presubmission review of the manuscript. Their comments and suggestions greatly helped to improve the document. We are thankful to Sheraz Khan, Numan Fazal, and Niaz Ali for their help in sampling. Literature cited Ahmad S., Iqbal SH., Khalid AN. 1997. Fungi of Pakistan. Sultan Ahmad Mycological Society of Pakistan, Department of Botany, University of the Punjab, Quaid-e-Azam campus, Lahore. 610 ... Ullah & al. Bruns TD. 1995. Thoughts on the processes that maintain local species diversity of ectomycorrhizal fungi. 63-73, in: HP Collins & al. (eds). The significance and regulation of soil biodiversity. Springer, Netherlands. https://doi.org/10.1007/978-94-011-0479-1_5 Dovana F, Contu M, Angeli P, Brandi A, Mucciarelli M. 2017. Leucoagaricus ariminensis sp. nov., a lilac species from Italy. Mycotaxon 132: 205-216. https://doi.org/10.5248/132.205 Gardes M., Bruns TD. 1993. ITS primers with enhanced specificity of basidiomycetes: application to the identification of mycorrhizae and rusts. Molecular Ecology 2: 113-118. https://doi.org/10.1111/j.1365-294X.1993.tb00005.x Ge ZW. 2010. Leucoagaricus orientiflavus, a new yellow lepiotoid species from southwestern China. Mycotaxon 111: 121-126. https://doi.org/10.5248/111.121 Ge ZW, Yang ZL, Qasim T, Nawaz R, Khalid AN, Vellinga EC. 2015. Four new species in Leucoagaricus (Agaricaceae, Basidiomycota) from Asia. Mycologia 107(5): 1033-1044. https://doi.org/10.3852/14-351 Hussain S, Jabeen S, Khalid AN, Ahmad H, Afshan NUS, Sher H, Pfister DH. 2018. Underexplored regions of Pakistan yield five new species of Leucoagaricus. Mycologia 1-14. https://doi.org/10.1080/00275514.2018.1439651 Johnson J. 1999. Phylogenetic relationships within Lepiota sensu lato based on morphological and molecular data. Mycologia 91: 443-458. https://doi.org/10.2307/3761345 Johnson J, Vilgalys R. 1998. Phylogenetic systematics of Lepiota sensu lato based on nuclear large subunit rDNA evidence. Mycologia 90: 971-979. https://doi.org/10.1080/00275514.1998.12026994 Kumar TKA, Manimohan P. 2009. The genera Leucoagaricus and Leucocoprinus (Agaricales, Basidiomycota) in Kerala state, India. Mycotaxon 108: 385-428. https://doi.org/10.5248/108.385 Kumari B, Atri NS. 2013. New additions of basidiomycetous fungi in Indian mycoflora. Mycosphere 4: 53-59. https://doi.org/10.5943/mycosphere/4/1/4 Liang JF, Yang ZL, Xu J, Ge ZW. 2010. Two new unusual Leucoagaricus species (Agaricaceae) from tropical China with blue-green staining reactions. Mycologia 102: 1141-1152. https://doi.org/10.3852/09-021 Locquin M. 1943a. Etude du développement des spores du genre Leucocoprinus Pat. (Troisiéme partie) suivie de la description d'une espéce nouvelle et dune espéce critique. Bulletin Mensuel de la Société Linnéenne de Lyon 12(5): 75-80. https://doi.org/10.3406/linly.1943.9740 Locquin M. 1943b. Etude du développement des spores du genre Leucocoprinus Pat. (Troisiéme partie) suivie de la description d'une espéce nouvelle et dune espéce critique. Bulletin Mensuel de la Société Linnéenne de Lyon 12(6): 91-96. https://doi.org/10.3406/linly.1943.9747 Malysheva EF, Svetasheva TY, Bulakh EM. 2013. Fungi in the Russian Far East. I. Leucoagaricus lateritiopurpureus and new species of Leucoagaricus (Agaricaceae) with reddish brown basidiocarps. Mikologiya i Fitopatologiya 47: 169-179. Mufioz G, Caballero A, Contu M, Vizzini A. 2012. A new Leucoagaricus species of section Piloselli (Agaricales, Agaricaceae) from Spain. IMA Fungus 2: 117-123. Mufioz G, Caballero A, Contu M, Ercole E, Vizzini A. 2014. Leucoagaricus croceobasis (Agaricales, Agaricaceae), a new species of section Piloselli from Spain. Mycological Progress 13(3): 649-655. https://doi.org/10.1007/s11557-013-0947-x Munsell Soil Color Charts. 1975. Munsell Color Co, Baltimore, MD. Qasim T, Amir T, Nawaz R, Niazi AR, Khalid AN. 2015. Leucoagaricus lahorensis, a new species of L. sect. Rubrotincti. Mycotaxon 130(2): 533-541. https://doi.org/10.5248/130.533 Singer R. 1948. Diagnoses fungorum novorum agaricalium. Sydowia 2: 35-36. Leucoagaricus brunneus sp. nov. (Pakistan) ... 611 Singer R. 1986. The Agaricales in modern taxonomy. 4th ed. Koenigstein: Koeltz Scientific Books. 450 p. Tamura K, Stecher G, Peterson D, Filipski A, Kumar S. 2013. MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Molecular Biology and Evolution 30: 2725-2729. https://doi.org/10.1093/molbev/mst197 Usman M., Khalid AN. 2018. Leucoagaricus pabbiensis sp. nov. from Punjab, Pakistan. Mycotaxon 133(2): 354-363. https://doi.org/10.5248/133.355 Vellinga EC. 2000. Notes on Lepiota and Leucoagaricus. Type studies on Lepiota magnispora, Lepiota barssii and Leucoagaricus americanus. Mycotaxon 76: 429-438. Vellinga EC. 2001la. Leucoagaricus. 85-108, in: ME Noordeloos & al. (eds). Flora Agaricina Neerlandica, Vol. 5. Vellinga EC. 2001b. Notulae ad floram agaricinam neerlandicam—XXXVIII. Leucoagaricus subgenus Sericeomyces. Persoonial7(3): 473-480. Vellinga EC. 2003. Phylogeny and taxonomy of lepiotaceous fungi [Doctoral dissertation]. Leiden: Univ Leiden Press. 259 p. Vellinga EC. 2004. Genera in the family Agaricaceae: evidence from nrITS and nrLSU sequences. Mycological Research 108: 354-377. https://doi.org/10.1017/S0953756204009700 Vellinga, EC. 2010. Lepiotaceous fungi in California, U.S.A. Leucoagaricus sect. Piloselli. Mycotaxon 112: 393-444. https://doi.org/10.5248/112.393 Vellinga EC, Balsley RB. 2010. Leucoagaricus dacrytus — a new species from New Jersey, USA. Mycotaxon 113: 73-80. https://doi.org/10.5248/113.73 Vellinga EC, Contu M., Vizzini A. 2010. Leucoagaricus decipiens and L. erythrophaeus, a new species pair in sect. Piloselli. Mycologia 102: 447-454. https://doi.org/10.3852/09-164 Vellinga E, Sysouphanthong P, Hyde KD. 2011. The family Agaricaceae: phylogenies and two new white-spored genera. Mycologia 103: 494-509. https://doi.org/10.3852/10-204 White TJ, Bruns T, Lee S, Taylor J. 1990. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. 315-322, in: MA Innis & al. (eds). PCR protocols: a guide to methods and applications. Academic Press, San Diego, pp. 315-322. https://doi.org/10.1016/B978-0-12-372180-8.50042-1 Ye Y, Li YK., Liang JF. 2014. Leucoagaricus tangerinus, a new species with drops from Southern China. Mycological Progress 13: 893-898. https://doi.org/10.1007/s11557-014-0974-2 MY COTAXON ISSN (print) 0093-4666 (online) 2154-8889 Mycotaxon, Ltd. ©2019 October-December 2019—Volume 134, pp. 613-618 https://doi.org/10.5248/134.613 Dictyostelids from Jilin Province, China 3: new Cavenderia and Dictyostelium records Pu Liu', SHUNHANG ZHANG', YUE ZOU’, XUEPING KANG’, Yu LI" ' Engineering Research Center of Chinese Ministry of Education for Edible and Medicinal Fungi, Jilin Agricultural University, Changchun, 130118, P. R. China ?Yanbian Academy of Agricultural Sciences, Yanji 133001, P. R. China * CORRESPONDENCE TO: fungi966@126.com ABSTRACT—Cavenderia parvispora, Dictyostelium vermiforme, and D. dimigraforme, isolated from samples of soil collected in Jilin Province, China, represent new records from China. Descriptions and illustrations based on these isolates are provided. Key worps—Cavenderiaceae, cellular slime molds, Dictyosteliaceae, Mycetozoa, taxonomy Introduction Dictyostelid cellular slime molds (dictyostelids), with both animal-like (protozoan) and fungus-like characteristics, are primarily inhabitants of the soil and leaf litter/humus zone of fields and forests, along with animal dung, where they feed mostly on bacteria. These organisms play an essential role in maintaining the balance that exists between bacteria and other soil organisms (Singh 1947; Cavender & Raper 1965a,b; Raper 1973, 1984). Traditional morphological classifications distributed dictyostelids among four genera based on differences in sorophore structure and branching patterns (Kirk & al. 2008). However, a new classification based on unique 18S rRNA sequence signatures was proposed by Sheikh & al. (2018). Jilin Province is situated in the temperate zone of China and is characterized by a monsoon climate (Liu & Li 2014). Previously, a total of 15 species, including two new dictyostelid species, have been reported from Jilin Province (He & Li 614 ... Liu & al. 2008; Liu & Li 2014, 2017). We report another three species in Cavenderia and Dictyostelium for the first time from China. Materials & methods SAMPLING, ISOLATION, CULTIVATION—Samples of soil were collected from three localities in Jilin Province in 2008 and 2009. Each 10-20 g soil sample was placed in a sterile whirl-pack plastic bag for isolation according to Cavender & Raper (1965a). Each sample was weighed and enough ddH,O added for an initial dilution of 1:10. A 0.5 mL aliquot of this dilution was added to each of five duplicate culture plates prepared with hay infusion agar (Raper 1984). After c. 0.4 mL of a heavy suspension of the bacterium Escherichia coli was added to each culture plate, the plates were incubated at 23 °C with a 12h light : 12 h dark cycle. Each plate was examined at least once daily for two weeks after the appearance of initial aggregations. Each isolate recovered from one of the plates was purified and cultivated for taxonomic studies. Spores from these isolates were frozen in HL 5 media (Cocucci & Sussman 1970) and stored at —80 °C in the herbarium of the Mycological Institute of Jilin Agricultural University, Changchun, China (HMJAU). OBSERVATION—The isolates were identified using morphological descriptions (Raper 1984) and molecular characteristics (Sheikh & al. 2018). After we marked the location of each early aggregating clone and sorocarp in a plate, we observed life cycle stages under a Zeiss Axio Zoom V16 dissecting microscope with a 1.5x objective and a 10x ocular. Sorocarps were mounted in water on slides for observation and measurements of spores, sorophores, and sorocarps using a Zeiss Axio Imager A2 light microscope mounted with 10x ocular and 10, 40, and 100x (oil) objectives. Photographs were taken with Zeiss Axiocam 506 color microscope camera. Taxonomy Cavenderia parvispora (H. Hagiw.) S. Baldauf, S. Sheikh & Thulin, Protist 169(1): 20. 2018. PLATE 1A-D When cultured at 23 °C on non-nutrient agar with E. coli, sorocarps gregarious or solitary, unbranched or sparsely irregularly branched, sometimes prostrate, phototropic, normally 0.2-4.4 mm long. Sorophore colorless, sinuose, slender, tapering from bases to tips, consisting of one tier of cells except for the in base and tip, bases clavate, tips clavate. Sori white, globose, commonly 30-150 um diam. Spores hyaline, elliptical, 3.7-6.0 x 1.9-2.9 um, with a few conspicuous polar granules. Aggregations with radiate streams or mound-like. SPECIMEN EXAMINED: CHINA, JILIN PROVINCE, Zuojjia, isolated from soil ($0425) from a broadleaf forest, 9 Sep. 2008 (HMJAU MR056). COMMENTS: Cavenderia parvispora was first isolated from forest soil sampled in Japan (Hagiwara 1989). Its medium-sized sorocarps, thin delicate sorophores with clavate tips, and small elliptical spores readily distinguish this species from morphologically similar dictyostelids. Cavenderia and Dictyostelium spp. new from Jilin (China) ... 615 4 198 @ OVE t. PEGI @ o® = 0% a” ee cy “Fry “aK A/ QO see ee if : _ pee “| ee! Cy =e : . ~ off * < es PLATE 1. Cavenderia parvispora (HMJAU MR056): A. Sorocarps; B. Sorophore base; C. Sorophore tip; D. Spores. Dictyostelium vermiforme (HMJAU MR058): E. Sorocarps; F, G. Vermiform and curved sorogens; H. Sorophore base; I. Sorophore tip; J. Spores. Scale bars: A, E= 2 mm; B, C, H, 1 =5 um; D,J = 10 um; KF G=1 um. 616 ... Liu & al. Dictyostelium vermiforme Vadell & Cavender, Mycologia 99: 118. 2007. PLATE 1E-J When cultured at 23 °C on non-nutrient agar with Escherichia coli, sorocarps clustered or solitary, normally prostrate, 1.0-2.0 mm long. Sorophore colorless, tapering from bases to tips, consisting of one tier of cells except for the base and tip, bases small round or expanded, tips capitate or expanded. Sori white, globose, commonly 50-130 um diam. Spores hyaline, oblong to elliptical, 4.7-7.0 x 2.9-3.9 um, with sparse or prominent polar granules. Aggregations with flattened streams, forming mound-like early sorogens. Sorogens developing rapidly, becoming vermiform and curved, migrating freely to form sinuous and curled late sorogens with prostrate sorophores. SPECIMEN EXAMINED: CHINA, JILIN PROVINCE, National Nature Reserve, isolated from soil (S0083-1) from a broadleaf forest, 10 Jun. 2009 (HMJAU MR058). ComMENTs: Dictyostelium vermiforme was originally isolated from soil/litter sampled in Iguazu Regional Park, Argentina (Vadell & Cavender 2007). This species is characterized by its vermiform and curved early migrating sorogens. Dictyostelium dimigraforme Cavender, J. Gen. Microbiol. 62:115.1970. | PLATE 2 When cultured at 23 °C on non-nutrient agar with Escherichia coli, sorocarps normally solitary, unbranched, erect or inclined, 3.5-10.5 mm long. Sorophore colorless or slightly yellow, tapering from bases to tips, consisting of one tier of cells except for the base and tip, bases round or slightly enlarged, tips capitate. Sori globose, off-white to yellow, 220-380 um diam. Spores hyaline, oblong to elliptical, 7.7-12.0 x 2.9-4.9 um, without polar granules. Aggregations radiating. SPECIMEN EXAMINED: CHINA, JILIN PROVINCE, Chanchung, Jingyuetan National Natural Park, isolated from soil (S0446) from a broadleaf forest, 10 Sep. 2008 (HMJAU MRO059). ComMENTs: Dictyostelium dimigraforme was isolated the first time from surface soil sampled in a tropical forest in the West Indies (Raper 1984). This species is characterized by its yellow sori and round bases, and can be distinguished from D. discoideum by its lack of basal disk. Discussion Our study has raised to 18 the number of dictyostelid species known from Jilin Province. Jilin Province is located in the temperate zone which has rich forests optimal for dictyostelids. The three dictyostelid species (Cavenderia parvispora, Dictyostelium vermiforme, and D. dimigraforme) reported here do not appear to be distributed worldwide (Hagiwara 1989, Vadell & Cavender 2007, Raper 1984). In particular, Dictyostelium vermiforme (originally isolated Cavenderia and Dictyostelium spp. new from Jilin (China) ... 617 ra € PLATE 2. Dictyostelium dimigraforme (HMJAU MR059): A, B. Sorocarps; C. Sorophore base; D. Sorophore tip; E. Spores. Scale bars: A, B = 2 mm; C, D =5 um; E= 10 um. 618 ... Liu & al. from c. 26°S in South America) and D. dimigraforme (originally isolated from c. 11°N in the West Indies) have now been isolated from c. 44°N in Asia. Clearly, the dictyostelids of Jilin Province need to be investigated in more detail. Acknowledgments We wish to express our appreciation to peer reviewers Prof. Steven L. Stephenson (University of Arkansas, U.S.A.) and Prof. John C. Landolt (Shepherd University, U.S.A.) for their valuable comments relating to this manuscript. This study was supported by the National Natural Science Foundation of China (No. 31870015, 31300016), the Science and Technology Research Programs of the Education Department of Jilin Province in the Thirteenth Five-Year Plan (No. JJKH20180671KJ), Science and Technology Development Program of Jilin Province (No. 20180101273JC), and 111 Project (No. D17014). Literature cited Cavender JC, Raper KB. 1965a. The Acrasieae in nature. I. Isolation. American Journal of Botany 52: 294-296. https://doi.org/10.2307/2439943 Cavender JC, Raper KB. 1965b. The Acrasieae in nature. II. Forest soil as a primary habitat. American Journal of Botany 52: 297-302. https://doi.org/10.2307/2439944 Cocucci SM, Sussman M. 1970. RNA in cytoplasmic and nuclear fractions of cellular slime mold amoebas. Journal of Cell Biology 45: 399-407. https://doi.org/10.1083/jcb.45.2.399 Hagiwara H. 1989. The taxonomic study of Japanese dictyostelid cellular slime molds. National Science Museum, Tokyo. He XL, Li Y. 2008. A new species of Dictyostelium. Mycotaxon 106: 379-383. Kirk PM, Cannon PF, Minter DW, Stalpers JA. 2008. Dictionary of the fungi, 10th edition. CABI, UK. Liu P, Li Y. 2014. Dictyostelids from Jilin Province, China. I. Phytotaxa 183(4): 279-283. https://doi.org/10.11646/phytotaxa.183.4.7 Liu P, Li Y. 2017. Dictyostelids from Jilin Province, China. II. Phytotaxa 323(1): 77-82. https://doi.org/10.11646/phytotaxa.323.1.6 Raper KB. 1973. Acrasiomycetes. 9-36, in: GC Ainsworth &al. (eds). The Fungi, vol. IVB. Academic Press, Inc. New York. Raper KB. 1984. The dictyostelids. Princeton University Press, Princeton. Sheikh S, Thulin M, Cavender JC, Escalante R, Kawakami S, Lado C, Landolt JC, Nanjundiah V, Queller DC, Strassmann JE, Spiegel FW, Stephenson SL, Vadell EW, Baldauf SL. 2018. A new classification of the dictyostelids. Protist 169(1): 1-28. https://doi.org/10.1016/j.protist.2017.11.001. Singh BN. 1947. Studies on soil Acrasieae. 1. Distribution of species of Dictyostelium in soils of Great Britain and the effects of bacteria on their development. Journal of General Microbiology 28: 417-429. https://doi.org/10.1099/00221287-1-1-11 Vadell EM, Cavender JC. 2007. Dictyostelids living in the soils of the Atlantic Forest, Iguazu Region, Misiones, Argentina: description of new species. Mycologia 99(1): 112-124. https://doi.org/10.1080/15572536.2007.11832606 MYCOTAXON ISSN (print) 0093-4666 (online) 2154-8889 Mycotaxon, Ltd. ©2019 October-December 2019—Volume 134, pp. 619-625 https://doi.org/10.5248/134.619 Records of Aureobasidium harposporum, Sarcophoma miribelii, and Stigmina dothideoides from Turkey MAKBULE ERDOGDU’, MERVE ULUKAPT?, ALI IHSAN KARAYEL?, ZEKIYE SULUDERE? ' Department of Landscape Architects, Faculty of Agriculture & ? Graduate School of Natural and Applied Sciences: Ahi Evran University, Bagbas1, Kirsehir, Turkey * Department of Biology, Faculty of Science, Gazi University, Teknikokullar, Ankara, Turkey * CORRESPONDENCE TO: merdogdu@ahievran.edu.tr ABsTRACT—During field studies on the microfungi in Mucur District of Kirsehir Province, Aureobasidium harposporum, and Stigmina dothideoides were revealed as new records for Turkey; and Sarcophoma miribelii, previously recorded from northeastern Turkey, is reported from central Turkey. Distinguishing morphological characters of these species are described, and their photographs are provided. Key worps—Ascomycota, Dothideales, leaf pathogenic fungi, Mycosphaerellales, SEM Introduction Aureobasidium Viala & G. Boyer (Saccotheciaceae, Dothideales) contains 24 species (Index Fungorum 2018). The genus is characterized by synchronous conidium production on hyaline conidiogenous cells. Species of Aureobasidium produce variously shaped one-celled conidia from terminal, lateral, or intercalary hyaline conidiogenous cells. Members of this genus occur as saprophytes on a variety of substrates or as parasites on phanerogams (Hermanides-Nijhof 1977). Sarcophoma Hohn. (Dothioraceae, Dothideales) was proposed by Hohnel in 1906 as a monotypic genus, but four species are now accepted (Index Fungorum 2018). 620 ... Erdogdu & al. Stigmina Sacc. (Mycosphaerellaceae, Mycosphaerellales) was established by Saccardo in 1880; c. 80 species are currently accepted in the genus (Index Fungorum 2018). In Turkey, the Stigmina species are poorly known and have not yet been intensively studied; of the four species reported from Turkey— S. carpophila, S. compacta, S. obtecta, S. platani—(Gdobelez 1964, Hiiseyin & al. 2003, Cimen & Ertugrul 2007, Erdogdu & Huseyin 2008), two have been transferred to other genera: S. compacta [= Thyrostroma compactum] and S. platani [= Pseudocercospora platanigena]. Materials & methods Host specimens were prepared following conventional herbarium techniques. Host plants were identified using the Flora of Turkey and East Aegean Islands (Davis 1965-85). Thin fungal sections prepared from host tissue were examined under a Leica DM E light microscope and measured from mounts in tap water. Infected host surfaces were photographed using a Leica EZ4D stereomicroscope. The fungi were identified using relevant literature (for Aureobasidium: Saccardo 1892 (as Gloeosporium), Hermanides-Nijhof 1977; for Sarcophoma: Saccardo 1884 (under Phoma), Morgan-Jones 1971, Aa 1975; for Stigmina: Ellis 1959, Shoemaker & Egger 1982). All examined specimens were deposited in the Mycology Laboratory of Ahi Evran University, Department of Biology, Kirsehir, Turkey (AEUT). For scanning electron microscopy (SEM), 8-10 mm square pieces of infected leaf were mounted on aluminium stubs with double-sided adhesive tape. They were coated with gold using Polaron SC 502 Sputter Coater and were examined with Jeol JSM 6060 scanning electron microscope operated at 5-10 kV in the Electron Microscopy Unit, Faculty of Science, Gazi University, Ankara, Turkey. Taxonomy Aureobasidium harposporum (Bres. & Sacc.) Herm.-Nijh., Stud. Mycol. 15: 151 (1977) PLATE 1 Spots visible on both sides of leaves and twigs, generally leaves beginning to dry from the tip, brown on the upper surface, fuscous chestnut on the lower surface, spots margin dark brown; infected leaves dropping prematurely from the shrub. Conrp1omarta acervular, subepidermal, yellowish, broadly elliptical, 196-330 um diam. CONIDIOGENOUS CELLS erect, densely packed, clavate to subcylindrical, 10-20 x 3.5-6.5 um, forming conidia simultaneously at the apex. Conip1 one-celled, falcate, sometimes elliptical or slightly curved, oil droplet, smooth-walled, 13.7-19.3(-21.8) x 4.4-5.4 um, hyaline. SPECIMEN EXAMINED—TURKEY, Kirsenrr, Mucur District, Tekken Village, 39°02'13”N, 34°13’08”E, 977 m asl, on living leaves and stems of Viscum album L. subsp. album (Santalaceae), 19.06.2013, M. Ulukap1 (AEUT MU1050). Aureobasidium, Sarcophoma, Stigmina spp. in Turkey ... 621 we 10k \ xS"6Ge— ) Sam GUFEF | | {OkU ~~ x4,388 Sum GuFEF PLATE 1. Aureobasidium harposporum (AEUT MU1050). A. leaf spots; B. acervuli on leaf; C. acervuli on leaf (SEM); D, E. acervulus on leaf (SEM); E acervulus, vertical section; G. conidia; H, I. conidia (SEM). Notes: Viscum album (European mistletoe) is an evergreen, perennial, epiphytic, hemiparasitic shrub that lives on a wide range of woody plant species (Zuber 2004). European mistletoes can affect their host trees in many ways. Known effects from mistletoe infection include lowering the vigor of the host, inducing premature mortality, reducing quality and quantity of wood grown, reducing fruit production, and predisposing trees to secondary infection by other agents, such as insects or decay fungi (Hawksworth 1983). Biological control of parasites by using plant pathogens has gained acceptance as a practical, safe, and environmentally beneficial management method applicable to agro-ecosystems (Charudattan 2001). The use of biological control agents for weed control has attracted increased attention (Ozaslan & al. 2013; Ozaslan 2016), and diagnosing host weeds and their natural enemies is the foundation stone of a successful biological control program (Ozaslan 2016). Control of European mistletoe is an important consideration for the forest service in Turkey (Yiiksel & al. 2005). Over 20 microscopic fungi live on European mistletoe, but only a few of them cause major damage to the plant 622 ... Erdogdu & al. (Karadzic & al. 2004). Of these, Aureobasidium harposporum, which causes leaf spot disease of European mistletoe, appears to have potential as a biological control agent against of this semi-parasite. In this study, Aureobasidium harposporum on living leaves and stems of Viscum album subsp. album is reported as new to the mycobiota of Turkey. The Turkish specimen agrees with other reports of Aureobasidium harposporum in conidiomata and conidia morphology, the only observable difference being the smaller dimensions of acervuli and conidia. Saccardo (1892) describes 200 um diam. acervuli and 18-20 x 4-5 um conidia, while Hermanides-Nijhof (1977) cites 250-500 um acervuli and (16-)17-21(-23) x 3.5-5 um conidia. XZ66 160)tmh X7> 566 Zharn Aureobasidium, Sarcophoma, Stigmina spp. in Turkey ... 623 Sarcophoma miribelii (Fr.) Héhn., Hedwigia 60: 133 (1918) PLATE 2 = Macrophoma miribelii (Fr.) Berl. & Voglino, Atti Soc. Veneto-Trent. Sci. Nat.10(1): 179 (1886) PYcNIDIA numerous, hypophyllous (rarely epiphyllous), at first covered by the epidermis, later becoming erumpent, scattered, 143-289 um diam., yellowish or brown. Conrp1a single-celled, ovoid, rounded at apex, attenuate at the base, smooth-walled, (8.4—)10-13.6(-14.3) x 6.2-8.7 um, hyaline, with granular contents. SPECIMEN EXAMINED—TURKEY, Kirseuir, Mucur District, Seyfegolti, 39°06’58’N, 34°12'11”E, 1135 m asl, on living leaves and stems of Buxus sempervirens L. (Buxaceae), 16.04.2014, M. Ulukapi (AEUT MU1099). Notes: Our Turkish specimen is morphologically similar to Sarcophoma miribelii specimens described in the literature (Saccardo 1884, Morgan-Jones 1971, Aa 1975). Sarcophoma miribelii has previously been reported (without description or illustration) from Rize Province in coastal northeastern Turkey (Gobelez 1964, Hiiseyin & al. 2005; both as Macrophoma mirbelii [sic]), whereas our new record is from Kirsehir Province in central Turkey at c. 1100 m asl. Sarcophoma miribelii is found worldwide, wherever the Buxus host occurs (Aa 1975). Stigmina dothideoides (Ellis & Everh.) M.B. Ellis, Mycol. Pap. 72: 53 (1959) PLATE 3 SPORODOCHIA erumpent through epidermis, pustulate, 1-4 mm diam., circular to elliptical, blackish. CoNIDIOPHORES densely crowded, arising from stromatic hyphae, pale brown to medium brown, cylindrical to lageniform, 1-2- septate, often branched at the base. Conip1a blastic, solitary and terminal, elliptical to obovoid, at first one-celled, becoming 3-septate, non-constricted or slightly constricted at the septum, rounded at apex, truncated to obtusely rounded at the base, smooth-walled, 25-38 x 11.8-13.9(-14.5) um, golden brown. SPECIMEN EXAMINED— TURKEY, Kirsenir, Mucur District, Gumtiskiimbet Village, 39°05'52”N, 34°12’26”E, 1180 m asl, on stems of Artemisia sp. (Asteraceae), 16.04.2014, M. Ulukapi (AEUT MU1087). Notes: The species was first described by Ellis & Everhart (1891, as Clasterosporium dothideoides), who observed symptoms on stems of Artemisia cana and Shepherdia argentea in Montana, USA; and it has PLATE 2 (left). Sarcophoma miribelii (AEUT MU1099). A. pycnidia on leaf; B. pycnidia on leaf (SEM); C. pycnidium on leaf (SEM); D. pycnidium, vertical section; E. conidia; F. conidia (SEM). 624 ... Erdogdu & al. 15 um Cp - ls PLATE 3. Stigmina dothideoides (AEUT MU1087). Top Lert. conidia and conidiophores; Top RIGHT. conidium and conidiophore; BoTTom. conidia. been recorded on Gaillardia aristata further north in Alberta, Canada (Shoemaker & Egger 1982). Stigmina dothideoides on branches of Artemisia sp. is reported as new to the mycobiota of Turkey. Although our Turkish specimen of Stigmina dothideoides is morphologically similar to specimens described in the literature, it differs slightly in having shorter and narrower 3-septate conidia, compared with 25-46 x 13-19 um 3-septate conidia in Ellis (1959) and 30-38(-42) x (12.3-)14-16.8 um 3(—4)-septate conidia in Shoemaker & Egger (1982). Acknowledgments The authors thank Sevda Kirbag (Department of Biology, Firat University, Elazig, Turkey) and Cumali Ozaslan (Department of Plant Protection, Dicle University, Diyarbakir, Turkey) for pre-submission review. This work was supported by the Ahi Evran University Scientific Research Projects Coordination Unit. (Project Number: PYO-Fen.4003.13.005 and FEF.A4.17.006). Aureobasidium, Sarcophoma, Stigmina spp. in Turkey ... 625 Literature cited Aa HA van der. 1975. The perfect state of Sarcophoma miribelii. Persoonia 8(3): 283-289. Charudattan R. 2001. Biological control of weeds by means of plant pathogens: significance for integrated weed management in modern agro-ecology. Biocontrol 46(2): 229-260. https://doi.org/10.1023/A:101147753 Cimen I, Ertugrul BB. 2007. Determination of mycoflora in almond plantations under drought conditions in Southeastern Anatolia project Region, Turkey. Plant Pathology Journal 6(1): 82-86. https://doi.org/10.3923/ppj.2007.82.86 Davis PH (ed.). 1965-85. Flora of Turkey and East Aegean Islands. Vols 1-9. Edinburgh University Press, Edinburgh. Ellis MB. 1959. Clasterosporium and some allied dematiaceae—phragmosporae. II. Mycological Papers 72.75 p. Ellis JB, Everhart BM. 1891. New species of fungi from various localities. Proceedings of the Academy of Natural Sciences of Philadelphia 43: 76-93. Erdogdu M, Huseyin E. 2008. Microfungi of Kurtbogazi1 Dam (Ankara) and its environment. Ot Sistematik Botanik Dergisi 14(1): 131-150. Gébelez M. 1964. La mycoflore de Turguie. (List of fungi of Turkey). II. Mycopathologia et Mycologia Applicata 23(1): 47-67. https://doi.org/10.1007/BF02049185 Hawksworth FG. 1983. Mistletoes as forest parasites. 317-333, in: M Calder, P Bernhardt (eds). The biology of mistletoes. Australia, Academic Press. Hermanides-Nijhof EJ. 1977. Aureobasidium and allied genera. Studies in Mycology. 15: 141-177. Huseyin E, Selcuk FE, Gaffaroglu M. 2003. Some materials on mitosporic fungi from Turkey I. Hyphomycetes. Botanica Lithuanica 9(2): 151-160. Hiiseyin E, Selcuk F, Gaffaroglu M. 2005. Materials on the micromycetes on box tree (Buxus) and Rhododendron from Turkey. 62-68, in: Proceedings of the XVI Symposium of Mycologists and Lichenologists of Baltic States. 21-25 September, 62-68, Cesis, Latvia. Index Fungorum 2018: http://www.indexfungorum.org. [accessed September 2018]. Karadzi¢ D, Lazarev V, Milenkovi¢ M. 2004. The most significant parasitic and saprophytic fungi on common mistletoe (Viscum album L.) and their potential application in biocontrol. Bulletin Faculty of Forestry, University of Bajna Luka, Serbia 89: 115-126. Morgan-Jones G. 1971. Conidium ontogeny in coelomycetes. I. Some amerosporous species which possess annellides. Canadian Journal of Botany 49(11): 1921-1929. https://doi.org/10.1139/b71-267 Ozaslan C, Hiiseyin E, Erdogdu M. 2013. Microfungi species on the weeds of agro-ecosystem (wheat ecosystem) in Adtyaman City. Mantar Dergisi 4(2): 10-18. Ozaslan C. 2016. Downy mildews species on the weeds of lentil fields in Diyarbakir in Turkey. Scientific Papers. Series A. Agronomy 59: 365-367. Saccardo PA. 1884. Sylloge Spheropsidearum et Melanconiarum. Sylloge Fungorum 3. 860 p. Saccardo PA. 1892. Supplementum universale, pars II. Discomyceteae-Hyphomyceteae. Sylloge Fungorum 10. 964 p. Shoemaker RA, Egger KN. 1982. Stigmina dothideoides. Fungi Canadenses 212. 2 p. Yuksel B, Akbulut S, Keten A. 2005. The damage, biology and control of pine mistletoes (Viscum album ssp. austriacum (Wiesb.) Vollman). Turkish Journal of Forestry 2: 111-124. Zuber D. 2004. Biological flora of Central Europe: Viscum album L. Flora 199(3): 181-203. https://doi.org/10.1078/0367-2530-00147 MY COTAXON ISSN (print) 0093-4666 (online) 2154-8889 Mycotaxon, Ltd. ©2019 October-December 2019—Volume 134, pp. 627-632 https://doi.org/10.5248/134.627 Bactrodesmium pulcherrimum sp. nov. from Ecuador FERNANDO ESPINOZA’, DAYNET SOSA’, LIZETTE SERRANO’, ADELA QUEVEDO’, FREDDY MAGDAMA’, MARCOS VERA’, SIMON PEREZ-MARTINEZ’, ELAINE MALOSSO}, RAFAEL F, CASTANEDA-Rut1Iz?* ' Escuela Superior Politécnica del Litoral, ESPOL, (CIBE), Campus Gustavo Galindo Km. 30.5 Via Perimetral, PO. Box 09-01-5863, Guayaquil, Ecuador ? Universidad Estatal de Milagro (UNEMI), Facultad de Ingenieria, Cdla. Universitaria Km. 1.5 via Milagro-Km26. Milagro 091706, Guayas, Ecuador * Centro de Biociéncias, Departamento de Micologica, Universidade Federal de Pernambuco, Avenida da Engenharia, s/n Cidade Universitaria, Recife, PE, 50.740-600, Brazil ‘Instituto de Investigaciones Fundamentales en Agricultura (INIFAT), Tropical ‘Alejandro de Humboldt’, OSDE, Grupo Agricola, Calle 1 Esq. 2, Santiago de Las Vegas, C. Habana, Cuba, C.P. 17200 * CORRESPONDENCE TO: dasosa@espol.edu.ec ABSTRACT—A new species Bactrodesmium pulcherrimum, found on decaying wood in Ecuador, is described and illustrated. The fungus is distinguished by subnapiform to broadly subturbinate (maize kernel-shaped) 1-septate asymmetrical ochreous-brown conidia. Illustrations of Bactrodesmium novageronense and B. simile and a comparison table of bicellular Bactrodesmium species are also provided. KEY worDs—asexual fungi, freshwater, hyphomycetes, taxonomy, tropics Introduction Saprobic dematiaceous hyphomycetes are highly diverse on plant materials in Central and South American tropical forests, where many new genera and species have recently been discovered (Castafieda-Ruiz & 628 ... Espinoza & al. Fic. 1. Cacao plantation, Guayas Province, Ecuador. Bactrodesmium pulcherrimum sp. nov. (Ecuador) ... 629 al. 2016). During a survey of hyphomycetes associated with plant debris of cacao plantations in the Naranjal, Guayas province, southern Ecuador (Fic. 1), we collected a Bactrodesmium specimen that differs remarkably from all previously described species (Hernandez-Restrepo & al. 2013, Arias & al. 2016). It is described here as a new species, Bactrodesmium pulcherrimum. Materials & methods Samples of decaying plant materials were collected and placed in plastic bags for transport to the laboratory, where they were washed, treated according to Castaneda- Ruiz & al. (2016), and placed in humid chambers. Several attempts to obtain this species in pure culture were unsuccessful after using a flamed needle to transfer conidia to corn meal agar mixed 1:1 with carrot extract and incubating at 25 °C. Mounts were prepared in PVL (polyvinyl alcohol, lactic acid) and measurements were made at 1000x magnification. Microphotographs were obtained with an Olympus BX51 microscope equipped with bright field and Nomarski interference optics. The type specimen is deposited in the Herbarium of Universidade Federal de Pernambuco, Recife, Brazil (URM). Taxonomy Bactrodesmium pulcherrimum RF. Castafieda, F. Espinoza & D. Sosa, sp. nov. Pre; MB 830569 Differs from Bactrodesmium simile by its hemispherical conidial basal cells and maize kernel-shaped conidia. Type: Ecuador, Guayas Province, Guayaquil, Naranjal, 02°41’26”S 79°36’46”W, on decaying wood of Theobroma cacao L. (Malvaceae), 7 March 2018, E. Espinoza & M. Vera (Holotype, URM 91287). Erymo oey: Latin, pulcherrimum, meaning most beautiful. CONIDIOMATA on the natural substrate sporodochial, scattered, pulvinate ochreous to brown, <20 um diam. Mycelium superficial and immersed composed of septate, branched, hyaline, smooth hyphae, 2-3 um diam. CONIDIOPHORES macronematous, mononematous, unbranched or with short lateral branches scorpioid after 1-2 sympodial elongations of the conidiogenous cells, erect, flexuous, 2-3-septate, hyaline, smooth, 30-80 x 4-5 um. CONIDIOGENOUS CELLS monoblastic, integrated or discrete, cylindrical or slightly clavate, sometimes inflated near the conidiogenous loci, determinate or indeterminate with sympodial extensions, hyaline, 10-18 x 4-7 um. Conidial secession rhexolytic. Conrp1A solitary, acrogenous, sometimes acropleurogenous, broadly obovoid to subnapiform, 1-septate, ochreous-brown, smooth, 18-24 x 630 ... Espinoza & al. Fic. 2. Bactrodesmium pulcherrimum (holotype, URM 91287): A, B. Conidia with a basal frill after rhexolytic conidial secession and a detached conidium with a lateral extension of the conidiogenous cell; C. Conidiogenous cells with lateral sympodial extensions, attached conidia; D. Detached conidium and conidiogenous loci indicated by arrows; E. Conidiophores, conidiogenous cells, and conidia; F. Sporodochium. Scale bars: A-E = 10 um; F = 20 um. Bactrodesmium pulcherrimum sp. nov. (Ecuador) ... 631 13-19 um, asymmetrical, with a basal cell hemispherical, 5-7 x 10-12 um, and an apical cell semi-elliptical or subglobose, 12-17 x13-17 um, with markedly pale ochreous-brown lumina. Fic. 3. Bactrodesmium simile (holotype, XAL CB1689): A. Conidia with a basal frill after rhexolytic conidial secession. Bactrodesmium novageronense (holotype, INIFAT C84/101-2): B. Conidia with a basal frill after rhexolytic conidial secession. Scale bars = 10 um. Note: Bactrodesmium simile Arias & al. (Fic. 3A, TABLE 1) is similar to B. pulcherrimum in the conidial sizes, but B. simile has broadly pyriform, obovoid, golden brown to pale olivaceous-brown conidia, with a basal TABLE 1. Comparison of the 1-septate Bactrodesmium species. SPECIES B. novageronense B. pulcherrimum B. simile COLOR Pale brown Ochreous-brown Golden brown to pale olivaceous brown CONIDIAL MORPHOLOGY SHAPE & SIZE (um) Obovoid to subglobose 8-13 x 7-10 Broadly obovoid to subnapiform 18-24 x 13-19 Broadly pyriform to obovoid 19-24 x 12-16 BASAL CELL (um) Hemispherical 3-3.5 x 5-7 Hemispherical 5-7 x 10-12 Cuneiform or obconical 7.5-11 x 5-8.5 REFERENCE Castafieda-Ruiz 1985 This paper Arias & al. 2016 632 ... Espinoza & al. cell cuneiform, 7.5-11 x 5-8.5 um (Arias & al. 2016). Bactrodesmium novageronense R.F. Castaneda (Fic. 3B, TABLE 1) is similar to B. pulcherrimum but can be distinguished by its smaller conidia (8-13 x 7-10 um; Castanieda-Ruiz 1985). Acknowledgments We are indebted to Dr. Josiane Santana Monteiro (Museu Paraense Emilio Goeldi, Belém, Brazil) and Dr. De-Wei Li (The Connecticut Agricultural Experiment Station Valley Laboratory, Windsor, U.S.A.) for their critical reviews. The authors thank Mr. Jairo Gonzalez for providing permission to collect samples. The authors are grateful to Escuela Superior del Litoral (ESPOL), CIBE, for financial support and the International Society for Fungal Conservation for facilities. RFCR is grateful to the Cuban Ministry of Agriculture. We acknowledge the websites provided by Dr. P.M. Kirk (Index Fungorum) and Dr. K. Bensch (MycoBank). Dr. Lorelei Norvell’s editorial review and Dr. Shaun Pennycook’s nomenclature review are greatly appreciated. Literature cited Arias RM, Heredia G, Castafieda-Ruiz RF. 2016. Two new species of Bactrodesmium and Dictyoaquaphila from Mexico. Mycotaxon. 131: 291-295. https://doi.org/10.5248/131.291 Castafeda-Ruiz RE 1985. Deuteromycotina de Cuba. Hyphomycetes 2. Instituto de Investigaciones Fundamentales en Agricultura Tropical, Cuba. 23 p. Castafieda-Ruiz RF, Heredia G, Gusmao LFP, Li DW. 2016. Fungal diversity of Central and South America. 197-217, in: DW Li (ed.). Biology of Microfungi. Springer International Publishing. https://doi.org/10.1007/978-3-319-29137-6_9 Hernandez-Restrepo M, Mena-Portales J, Gené J, Cano J, Guarro J. 2013. New Bactrodesmiastrum and Bactrodesmium from decaying wood in Spain. Mycologia 105: 172-180. https://doi.org/10.3852/12-004 MY COTAXON ISSN (print) 0093-4666 (online) 2154-8889 Mycotaxon, Ltd. ©2019 October-December 2019—Volume 134, pp. 633-635 https://doi.org/10.5248/134.633 Dendrographium multiseptatum sp. nov. from China Li-Guo Ma, YUE-LI ZHANG, Bo ZHANG, Kal QI, CHANG-SONG LI, JUN-SHAN QT Shandong Key Laboratory of Plant Virology, Institute of Plant Protection, Shandong Academy of Agricultural Sciences, Jinan, Shandong, 250100, China “CORRESPONDENCE TO: gi999@163.com ABSTRACT—A new species, Dendrographium multiseptatum, is described and illustrated from specimens collected on rotten twigs in China. This fungus is characterized by synnematous conidiophores with polytretic conidiogenous cells producing obclavate, 6-9-distoseptate, pale brown conidia. KEY wWoRDS—asexual morph, microfungi, hyphomycete, taxonomy Introduction Dendrographium Massee was established by Massee (1892) to accommodate the Brazilian type species D. atrum Massee. The genus is characterized by synnematous, macronematous, unbranched conidiophores with polytretic, integrated, terminal (becoming intercalary), sympodial, cylindrical conidiogenous cells that produce solitary, acropleurogenous, simple, distoseptate conidia (Ellis 1971, Seifert & al. 2011). Species assignment in Dendrographium is based primarily on conidial morphology, including shape, septal number, size, pigmentation, and ornamentation. Six additional species have been described (all from India), but Dendrographium kamatii V.G. Rao with monotretic conidiogenous cells has been recombined as Corynespora kamatii (V.G. Rao) M.B. Ellis (Rao 1963, Ellis 1976). Ghosh & al. (1977) provided a comparison table for the six Dendrographium species (including D. kamatii) reported from India. 634 ... Ma &al. During a survey on wood-inhabiting microfungi in China, a new Dendrographium species was found. As this species did not match any of the currently described species in the genus, it is proposed here as new. Specimens are deposited in the Herbarium of Institute of Plant Protection, Shandong Academy of Agricultural Sciences, Jinan, Shandong, China (HSAAS) and the Mycological Herbarium, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China (HMAS). Taxonomy Dendrographium multiseptatum L.G. Ma & J.S. Qi, sp. nov. FIG. 1 MB 832847 Differs from Dendrographium atrum by its narrower conidiophores and its shorter, broader conidia with fewer distosepta and from D. mitteri by its longer synnemata, and solitary, shorter, 6-9-distoseptate conidia. Type: China, Shandong Province, Taian City, Taishan District, on rotten twigs of an unidentified plant, 22 May 2018, L.G. Ma (Holotype, HSAAS 0415; isotype, HMAS 146095). ETyMOLoGy: multiseptatum, referring to the numerous conidial septa. CoLoNnIEs on the natural substratum effuse, black to dark brown, tufted. Mycelium mostly immersed in the substratum. Synnemata large, black, erect, tapered towards the apex, with dark brown stalks, consisting of parallel conidiophores, 520-1380 um long, 25-85 um diam. at the base. CONIDIOPHORES macronematous, synnematous, mostly adpressed along lower part of the length, divergent towards the upper part, unbranched, erect, straight or flexuous, cylindrical, septate, dark brown, smooth, thick-walled, brown towards the apex, <1400 um long, 3.5-6.5 um diam. CONIDIOGENOUS CELLS integrated, terminal becoming intercalary, polytretic, cylindrical, brown, smooth, thick-walled, 8-24 x 5.0-7.5 um. Conipi1A solitary, acropleurogenous, simple, obclavate, 6—9-distoseptate, smooth, thick-walled, pale brown, 32-66 x 8-11.5 um, tapered towards the apex, with a basal scar, truncate at the base, 2.5-3.5 um diam. CoMMENTS—Dendrographium multiseptatum most closely resembles D. atrum and D. mitteri Syd. in conidial shape. However, D. atrum has broader (6-10 um) conidiophores and longer and narrower conidia with more distosepta (40-130 x 7-9 um, 6-15 distosepta; Massee 1892). Dendrographium mitteri differs by its shorter synnemata (170-250 um) and occasionally catenate, longer (35-110 um), 3-10-septate conidia (Sydow & Mitter 1933). Dendrographium multiseptatum sp. nov. (China) ... 635 Fic. 1. Dendrographium multiseptatum (holotype, HSAAS 0415): A-D. Synnematous conidiophores, conidiogenous cells, and conidia; E. Synnematous conidiophores with conidiogenous cells and conidium; F. Conidia. Acknowledgments The authors express gratitude to Dr. De-Wei Li (The Connecticut Agricultural Experiment Station Valley Laboratory, Windsor, U.S.A.) and Dr. Ze-Fen Yu (Laboratory for Conservation and Utilization of Bio-resources, Yunnan University, Kunming, P.R. China) for serving as pre-submission reviewers and to Dr. Shaun Pennycook for nomenclatural review. This project was supported by Young Talents Training Program of Shandong Academy of Agricultural Sciences (CXGC2018E04), National Key R & D Program of China (2016YFD0300700, 2017YFD0201700), and National Natural Science Foundation of China (31400019). Literature cited Ellis MB. 1971. Dematiaceous hyphomycetes. Commonwealth Mycological Institute, Kew, Surrey, England. Ellis MB. 1976. More dematiaceous hyphomycetes. Commonwealth Mycological Institute, Kew, Surrey, England. Ghosh RN, Pathak NC, Singh MS. 1977. Two new records of stilbaceous fungi from India. Norwegian Journal of Botany 24: 79-81. Massee GE. 1892. Notes on exotic fungi in the Royal Herbarium, Kew. Grevillea 21: 1-6. Rao VG. 1963. A new species of Dendrographium from India. Current Science 32: 473-474. Seifert KA, Morgan-Jones G, Gams W, Kendrick B. 2011. The genera of Hyphomycetes. CBS Biodiversity Series 9. 997 p. Sydow H, Mitter JH. 1933. Fungi indici. I. Annales Mycologici 31: 84-97. MY COTAXON ISSN (print) 0093-4666 (online) 2154-8889 Mycotaxon, Ltd. ©2019 October-December 2019—Volume 134, pp. 637-641 https://doi.org/10.5248/134.637 Haematomma pluriseptatum sp. nov. from China CONGCONG MIAO *!, RONG TANG **4, LINLIN DoNG?, ZHAOJIE REN?, ZUNTIAN ZHAO ** "Key Laboratory of Plant Stress Research, College of Life Sciences & ? Institute of Environment and Ecology: Shandong Normal University, Jinan, 250014, PR. China * Shandong Museum, Jinan, 250014, P. R. China ‘The First High School of Liangshan, Shandong Province, 272600, P. R. China “CORRESPONDENCE TO: ccmjy123@163.com ABSTRACT—A new species, Haematomma pluriseptatum, is described from southwestern China. This corticolous species is characterized by large ascospores with (19-)20-26(-27) septa and by the presence of atranorin, russulone, and pseudoplacodiolic acid. A detailed description of its morphology, high resolution photographs, chemistry, comments, and distribution are provided. Related lichen taxa are discussed, and a key to the species of Haematomma from China is also provided. Key worps—East Asia, Haematommataceae, Lecanorales, lichenized fungi, taxonomy Introduction The crustose Haematomma lichens, with their small but brilliant-red lecanorine apothecia, are often conspicuous on smooth-barked trees in warm-temperate to tropical regions of the world (Staiger & Kalb 1995, Brodo & al. 2008). Haematomma was proposed originally to house two species, H. vulgare A. Massal. and H. ventosum (L.) A. Massal. (Massalongo 1852). The genus is characterized by Haematomma-type asci, blood-red lecanorine apothecia, and transversely septate, submuriform to muriform *CONGCONG MIAO & RonG TANG contributed equally to this work. 638 ... Miao, Tang & al. ascospores (Brodo & al. 2001, Elix 2004, Nelsen & al. 2006, Nash & al. 2004, Smith & al. 2009). Apothecial pigmentation and secondary chemistry play important roles for species delimitation in this genus (Messuti & de la Rosa 2009). The number of spore septa is also used as an important character in separating species (Nelsen & al. 2006). Of the forty Haematomma species reported in the world, twelve have been reported in China (Brodo & al. 2001, Elix 2004, Nelsen & al. 2006, Nash & al. 2004, Smith & al. 2009, Tang & al. 2018). For this study, we examined about two hundred specimens of Haematomma morphologically and chemically, describe a new species, H. pluriseptatum, from China, and present a key to the species of Haematomma from China. Materials & methods Four collections are deposited in the Lichen Section of the Botanical Herbarium, Shandong Normal University, Jinan, China (SDNU), and one is deposited in the Lichen Herbarium of the Kunming Institute of Botany, Kunming, China (KUN). The specimens were examined morphologically and anatomically morphological under an Olympus SZ51 stereomicroscope and Olympus CX21 polarizing microscope. Both thallus and medulla were tested with K (a 10% KOH aqueous solution) and C (a saturated aqueous NaOCl solution). The lichen substances were identified using standardized thin layer chromatography techniques (TLC) with solvent system A and C (Orange & al. 2010). The specimens were photographed using a DP72 camera on Olympus SZX16 and BX61 microscopes. Taxonomy Haematomma pluriseptatum R. Tang, sp. nov. Fie. 1 MB 830618 Differs from Haematomma rufidulum by its ascospores with more septa and its production of pseudoplacodiolic acid. Type: China. Yunnan, Diging Co., Shangri-La, Tianbao Mt., 27°36’37”N 99°53’37”E, alt. 3790 m, on bark of living trees, 17 Aug. 2018, Wang Chunxiao 20181135 (Holotype, SDNU). EryMo.ocy: The epithet pluriseptatum refers to the morphology of the ascospores with many septa. Thallus crustose, corticolous, pale greyish to greenish grey, rugose rimose- areolate to areolate, continuous, thickness 0.6 mm. Soredia and isidia absent. Apothecia scattered, 0.3-0.95(-1.0) mm diam., sessile, convex, constricted at the base, smooth when young, red. Disc orange-yellow to brownish yellow, white pruina present when young but absent when mature; Thalline margin well developed; amphithecium present, 70-90 um thick. Haematomma pluriseptatum sp. nov. (China) ... 639 Fic. 1. Haematomma pluriseptatum (holotype, SDNU (Wang Chunxiao 20181135)). A. Thallus and apothecia; B. Apothecium section; C. K+ reaction (red) of epihymenium and outer exciple; D. Ascus; E. Ascospores. Scale bars: A = 200 um; B, C = 100 um; D = 5 um; E = 10 um. Epihymenium orange, K+ red, 9-20 um thick. Hymenium hyaline, 70-95 um high. Hypothecium hyaline or slightly brown. Paraphyses branched and anastomosing. Asci clavate, containing 8 spores. Ascospores twisted in ascus, filiform, straight or slightly curved, (19-)20-26(-27)-septate, 70-95 x 5-6.5(—7) um. Conidia absent. CHEMISTRY—Cortex K+ yellow, C-, KC-. Atranorin, russulone, and pseudoplacodiolic acid detected by TLC. ADDITIONAL SPECIMENS EXAMINED: CHINA. YUNNAN, Diging Co., Shangri-La, Tianbao Mt., 27°36’37”N 99°53’37’E, alt. 3790 m, on bark of living trees, 17 Aug. 2018, Wang Chunxiao 20181175, 20181209, 20181126 (SDNU); Deqin Co., Yubeng Village, Xiaonong Base Camp. 28°23.92’N 98°46.16’E, alt. 3500 m, on bark of living trees, 14 Sep. 2012, Niu Dongling 12-36322 (KUN-L). COMMENTS—Haematomma pluriseptatum is characterized by the large number of septa (20-27) in each ascospore and the presence of atranorin, russulone and pseudoplacodiolic acid in the thallus. 640 ... Miao, Tang & al. The new ascomycete is morphologically similar to H. rufidulum (Fée) A. Massal., which differs by the occurrence of slightly irregular, white, convex to hemispherical soredia and elongate-fusiform, 8-25-celled, 50-72(-85) x (3.5—)4-6(-—7) um ascospores (Brodo & al. 2008). Secondary metabolites also differ: H. pluriseptatum contains pseudoplacodiolic acid, while H. rufidulum contains placodiolic acid. Haematomma pluriseptatum may also be confused with H. dolichosporum (B. de Lesd.) Kalb & Staiger, which also produces twisted spores in the asci, but H. dolichosporum is readily distinguished by its shorter (42-62 um) spores with fewer (7—9(-11)) septa and the production of placodiolic acid (Staiger & Kalb 1995; Brodo & al. 2008). Key to the species of Haematomma in China 1. Epihymenium with russulone, K+ Ted 25 soe sien seston nie Sn olew slew ales 2 1. Epihymenium with haematommone, K+ purple (entirely dissipating) ........... 8 2 siallastcomtaimi ne UgMte BOR | nse ne hms nadie Rate ree Sad een Bade ea ade eek ae H. fauriei 2. Uhalluslackine-usnic acid.) 25. ebay. Bey ee tie ee tk el MEE in ee ae De geee ts mes 3 SeOMTOCKE 5h nds Awe ade eH eH lng W eiagee nN efegeed ehogee me Ong mee H. fenzlianum Pe EU Dial Raters, onye- dag, nyee apa > aeons teen otis #o genie key ie sn gh mesa ean eM S ees ote ee -: 4, Thallus containing placodiolic acid ............. 0... e eee eee eee H. rufidulum #. balls ekino-placOd iol Ge: dGid. cna ee Ms aren neers Basten Paks ea ede oe Sly or Rodgers Bootes 5 >) thallus lacking sphderophotin ts, atac. eitore a ara eee nea ee hee Paes Pee 5 6 5. Thallus containing sphaerophorin, Sporesto= IA seclledanw to, We M rim oe meh tans Rens Rees CAR a eRe ne ee 7 6. Thallus containing pseudoplacodiolic acid, spores’ 20=27—celled oo n.cey vin eee Reed ee oad Bee EE Be oeS H. pluriseptatum 6. Thallus lacking pseudoplacodiolic acid, spores S=l6scelled) .. is. ese eee ae ee eee es H. puniceum subsp. pacificum Fc SPOVesOa7 ACE 6 x 4 bsg kag t rhea y eAGY 4S MENS MEE EAR H en tie Dy nae H. persoonii 7 Spores 01a xcelled. 28 Anh thal fatter Ma nat Ane nak Met May H. collatum SeSPOTEs MOE SUDEIUTIIOLI, 5 oF 58 ue «Aste eg. ike recom ga-rhc om gro mG aaa oni arec phe e cn eget wie 9 SPS Pores Su DHMITMOLN a ac, tila kn eas Cees earn ies dee tten aeeeeen Rete ss hae H. wattii Re ApOecia: Sorediatee.. M.S ath. a sgeh ese h ead ven tak wonton moe ete H. caperaticum 9 Apothecranorsorediate +5. 2565 28 ls oh Besos eed bs ee hee ema ee et Metgs oe eins 10 10. Thallus containing isoplacodiolic, isopseudoplacodiolic acids ...... H. flexuosum 10. Thallus lacking isoplacodiolic, isopseudoplacodiolic acids ................... 11 Ines poreg celled 7 itr eh Fut i oP RNa aI H. accolens I Sporess eas ceed ie is carte Merde Bevery Rebstes Roden Sods ee Babee hon H. africanum Haematomma pluriseptatum sp. nov. (China) ... 641 Acknowledgments We thank Dr. Klaus Kalb (Lichenological Institute Neumarkt, Germany) and Dr. Shou-Yu Guo (State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, Beijing) for presubmission review. The authors would also like to thank Lisong Wang and Xinyu Wang (Kunming Institute of Botany, CAS, China) for assistance during this study. This work was supported by the National Natural Science Foundation of China Youth Science Foundation (31600100), and Emergency management project of National Natural Science Foundation of China (31750001). Literature cited Brodo IM, Sharnoff SD, Sharnoff S. 2001. Lichens of North America. Yale University Press, New Haven and London, 795 p. Brodo IM, Culberson WL, Culberson CE. 2008. Haematomma (Lecanoraceae) in North and Central America. Bryologist 111: 363-423. https://doi.org/10.1639/0007—2745(2008) 111[363:hlinac]2.0.co;2 Elix JA. 2004. Haematommataceae. 4-10, in: Flora of Australia 56A—Lichens 4. Australian Biological Resources Study/CSIRO Publishing. 222 p. Massalongo AB. 1852. Ricerche sull’ autonomia dei licheni crostosi. Verona: Friziero. 207 p. Messuti MI, de la Rosa IN. 2009. Notes on the genus Haematomma (Ascomycota, Lecanoraceae) in Argentina. Darwiniana 47(2): 297-308. Nash TH, Ryan BD, Diederich P, Gries C, Bungartz F. 2004. Lichen flora of the Greater Sonoran Desert Region, vol. 2. Tempe, Lichens Unlimited. Nelsen MP, Liicking R, Chaves JL, Sipman HJM, Umafa L, Navarro E. 2006. A first assessment of the Ticolichen biodiversity inventory in Costa Rica: the genus Haematomma (Lecanorales: Lecanoraceae). Lichenologist 38: 251-262. https://doi.org/10.1017/s0024282906005573 Orange A, James PW, White FJ. 2010. Microchemical methods for the identification of lichens. 2™ edition. London, British Lichen Society. Smith CW, Aptroot A, Coppins BJ, Fletcher A, Gilbert OL, James PW, Wolseley PA. 2009. The lichens of Great Britain and Ireland. London, British Lichen Society. Staiger B, Kalb K. 1995. Haematomma-Studien: I. Die Flechtengattung Haematomma. Bibliotheca Lichenologica 59. 198 p. Tang R, Yan SK, Sun MJ, Zhang LL. 2018. New records of Haematomma and Ophioparma from China. Mycotaxon 133(1): 175-181. https://doi-org/10.5248/133.175 MY COTAXON ISSN (print) 0093-4666 (online) 2154-8889 Mycotaxon, Ltd. ©2019 October-December 2019—Volume 134, pp. 643-648 https://doi.org/10.5248/134.643 New records of Didymium inconspicuum, D. karstensii, and D. rugulosporum from China CHAOFENG YUAN, SHU LI, WAN WANG, SHUWEI WEI, Qi WANG, YU LI Engineering Research Center of Chinese Ministry of Education for Edible and Medicinal Fungi, Jilin Agricultural University, Changchun, Jilin 130118 China * CORRESPONDENCE TO: giwang@jiau.edu.cn ABSTRACT—Didymium inconspicuum, D. karstensii, and D. rugulosporum are reported as new records for China. Morphological characteristics were observed and described by light microscopy and scanning electron microscopy and are discussed and compared with similar species. Key worps—Didymiaceae, Myxogastrea, taxonomy Introduction Didymium was introduced by Schrader (Schrader 1797) and characterized by sporophores with a peridium covered with stellate lime crystals. In contrast, the capillitium is almost always limeless. The lime crystal nature on the peridium separates Didymium from Diderma, in which the lime occurs as amorphous granules (Martin & Alexopoulos 1969). Currently Didymium includes approximately 94 species around the world (Kirk & al. 2008, Lado 2018), of which 35 have been recorded in China (Li & al. 1996, Liu & Chen 1998, Chen 1999, Li 2008, Liu & Chang 2011, Gao & al. 2018). The objectives of this work were to examine some putative Didymium samples, characterize them by light and scanning electron microscopy, and compare them with similar species. 644 ... Yuan & al. Materials & methods Specimen collection and moist chamber culture Didymium rugulosporum was collected from Huzhong Nature Reserve, Heilongjiang Province in 2016. Didymium karstensii was collected from Changbai Mountain, Jilin Province in 2018. Didymium inconspicuum was collected from the desert transition zone of Suwu Township, Mingin County, Gansu Province in 2016. The specimens are deposited in the Herbarium of the Mycological Institute of Jilin Agricultural University, Changchun, China (HMJAU). Morphological identification The morphological characteristics of the sporocarps were observed by light microscope and scanning electron microscope. Each sporocarp was placed on a slide and one drop of 4% KOH added. The slide cover was placed on top and gently tapped, and filter paper used to remove excess solution. The capillitium, calcareous crystals, and spores were observed under a Leica DM 2000 light microscope (LM). Up to 20 spores were randomly selected and their diameters measured. The ultrastructure was observed under the Hitachi SU8010 scanning electron microscope (SEM) operating at 5kV (Martin & Alexopoulos 1969, Zhu & al. 2012, Li & al. 2017). Taxonomy Didymium inconspicuum Nann.-Bremek. & D.W. Mitch., Proc. Kon. Ned. Akad. Wetensch., C 92: 508. 1989 Fic. 1 Sporocarps sessile, scattered, occasionally mingled with — short plasmodiocarps, small and inconspicuous. 0.5-1 mm long, about 0.5 mm high, ovoid, often depressed in the center, grayish white. Columella absent. Peridium membranous, thin, coated with grayish white lime crystals. Capillitium profuse, threads slender and curved, pale brown, 1-1.5 um diam., often branching and anastomosing to an incomplete network, sometimes with flattened and darker colored sections on the branches. Spores dark brown in mass, brown by LM, ovoid to globose, 8-11 um diam., evenly slightly warted. SPECIMEN EXAMINED: CHINA, GANSU PROVINCE, Mingin County, Suwu Township, 38°16'42”N 103°06’17’E, 1360 m, on leaf by moist chamber culture, 16 September 2016, Chaofeng Yuan & Shuwei Wei (HMJAU 10650). CoMMENTs: Didymium inconspicuum is similar to Didymium obducens P. Karst. [= D. fulvum Sturgis] and Didymium ochroideum G. Lister. The spore of D. obducens is tuberculate on one side, while the other side is smooth and the color is pale, but the spore of D. inconspicuum has relatively uniform small warts. The spore of D. ochroideum is 6-8 um diam. whereas the spore of D. inconspicuum is 8-11 1m diam. Didymium spp. new to China... 645 Fic 1. Didymium inconspicuum. A- C. Sporocarps; D. Capillitium (LM); E. Spores (LM); F. Capillitium (SEM); G. Lime crystal (SEM); H. Spores (SEM). Scale bars: A- C = 500 um; D, F= 10 pm; E, G, H=5 pm. Nannenga-Bremekamp (1989) described D. inconspicuum as having white or pale yellow-brown tiny lime crystals, with a coarse capillitium and spores that are 12-14(-15) um diam.; Schnittler & Novozhilov (2000) described this species as having 3-6 um long lime crystals smaller than spores, a pale to pale brown, 1-2(-2.5) um diam. capillitium, and (9.5—)10.5-11.5(-12.5) um diam. spores. Our specimen has white lime crystals about 5-8 um diam., a light brown 1-1.5 um diam. capillitium, and spores 8-11 um diam.; other features are consistent with the published descriptions. Didymium karstensii Nann.-Bremek., Acta Bot. Neerl. 13: 247. 1964 FIG. 2 Sporocarps gregarious, sessile or with a short stalk, white, subglobose, 1 mm wide, up to 0.5 mm high, stalk (when present) calcareous, yellowish white, about 0.2 mm long. Peridium double, outer layer white, with stellate lime crystals about 5 um diam. forming the wrinkled shell; the inner layer membranous and colorless. Columella white, calcareous, slightly thickened at 646 ... Yuan & al. Fic 2. Didymium karstensii. A. Sporocarps; B. Capillitium and swelling (LM); C. Spores (LM); D. Lime crystals (SEM); E. Capillitium (SEM); F. Spores (SEM). Scale bars: A = 1 mm; B-E = 10 um; F=5 um. the base. Capillitium pale brown, threads slender, branching and anastomosing to a network, with dark swellings, 2-3 um diam., attached to the peridium and to the base of the sporocarp. Spores dark brown in mass, brown by LM, densely warted and ridged, 7.5-10 um diam. SPECIMEN EXAMINED: CHINA, JILIN PROVINCE, Changbai Mountain, 42°22’18”N 128°00'14”E, 828 m, on the leaf, 5 September 2018, Chaofeng Yuan & Shuwei Wei (HMJAU 10651). Comments: Didymium karstensii is similar to Didymium squamulosum (Alb. & Schwein.) Fr., but D. karstensii is inconspicuously umbilicate at the base, whereas D. squamulosum is deeply umbilicate at the base. Nannenga-Bremekamp (1964) described stalks of D. karstensii as generally not exceeding half the diameter of the sporocarps, and spinulose spores, 10-12 um diam. Demirel & Kasik (2012) described stalks of D. karstensii generally not exceeding 0.2 mm long, and warted spores, 10-12 um diam. Our specimen had stalks about 0.2 mm long and warted spores, 7.5-10 um across; other features are consistent with the published descriptions. Didymium rugulosporum Kowalski, Mycologia 61: 636. 1969 Fic. 3 Sporocarps sessile, scattered to clustered, globose to subglobose, 1-2.5 mm diam., white. Peridium double, the outer layer thick and fragile, white smooth calcareous shell, stellate lime crystals; the inner layer membranous, thin, colorless and transparent. Columella absent. Capillitium dense, rigid, brown, Didymium spp. new to China ... 647 Fic 3. Didymium rugulosporum. A, B. Sporocarps; C. Capillitium and spores (LM); D. Capillitium and peridium (SEM); E. Lime crystal (SEM); FE. Spores (SEM). Scale bars: A, B = 1 mm; C = 20 um; D = 10 um; E, F=5 um. 1-1.5 um diam., weakly attached to the base and apex of the sporocarps, brown, branched with many colorless swellings. Spores dark brown in mass, purple brown by LM, globose, 17-19 um diam., with large dense warts about 1.5 um high and with some ridges forming a reticulum. SPECIMEN EXAMINED: CHINA, HEILONGJIANG PROVINCE, Huzhong Nature Reserve, 52°06'02”N 123°20'38”E, 168 m, on the bark, 24 September 2016, Wan Wang (HMJAU 10652). Comments: Didymium rugulosporum is similar to Didymium trachysporum G. Lister and Didymium quitense (Pat.) Torrend, which differ by their smaller spores (D. trachysporum 9-10 um diam.; D. quitense is 12-15 um diameter). Kowalski (1969) described spores of D. rugulosporum as (16—)18-20(-22) um diam. Our specimen had spores 17-19 um diam.; other characteristics are consistent with the original description. Acknowledgments We would like to thank Tom Hsiang (University of Guelph, Canada) and Paul M. Kirk (RBG Kew, UK) for presubmission reviews of this manuscript. This work was financed by the National Natural Science Foundation of China (NO.31770011, NO.31370065). Literature cited Chen SL. 1999. Fungal flora of tropical Guangxi, China: a survey of myxomycetes from southwestern Guangxi. Mycotaxon 72: 393-401. 648 ... Yuan & al. Demirel G, Kasik G. 2012. Four new records for Physarales from Turkey. Turkish Journal of Botany 36: 95-100. https://doi.org/10.3906/bot-1010-12 Gao Y, Yan SZ, Wang GW, Chen SL. 2018. Two new species and two new records of myxomycetes from subtropical forests in China. Phytotaxa 350(1): 51-63. https://doi.org/10.11646/phytotaxa.350.1.6 Kirk PM, Cannon PF, Minter DW, Stalpers JA. 2008. Ainsworth & Bisby’s dictionary of the fungi. 10th edition. CAB International Publishing, Wallingford. 771 p. Kowalski DT. 1969. A new coprophillous species of Didymium. Mycologia 61(3): 635-639. https://doi.org/10.2307/3757253 Lado C. (2018). An online nomenclatural information system of Eumycetozoa. Real Jardin Botanico, CSIC. Madrid, Spain. http://www.nomen.eumycetozoa.com (Accessed: September 2018). Li Y. 2008. Flora myxomycetes sinicorum. Science Press. Li HZ, Li Y, Wang Q. 1996 [“1995-96”]. Myxomycetes from China XIII: a new species of Didymium. Mycosystema 8-9: 173-175. Li S, Wang W, Wang W, Wang Q, Li Y. 2017. Morphology and life-cycle of Fuligo leviderma, a newly recorded myxomycete species of China. Mycosystema 36(4): 528-531. https://doi.org/10.13346/j.mycosystema.160122 Liu CH, Chang JH. 2011. Myxomycetes of Taiwan XXIII. The genera Diachea and Didymium. Taiwania 56: 287-294. https://doi.org/10.6165/tai.2011.56(4).287 Liu CH, Chen YE. 1998. Myxomycetes of Taiwan X. Three new records of Didymium. Taiwania 43: 177-184. https://doi.org/10.6165/tai.1998.43(3).177 Martin GW, Alexopoulos CJ. 1969. The myxomycetes. University of Iowa Press, Iowa City. 560 p. Nannenga-Bremekamp NE. 1964. Notes on myxomycetes VIII. A new species of Didymium from the Netherlands. Acta Botanica Neerlandica 13: 246-249. https://doi.org/10.1111/j.1438-8677.1964.tb00155.x Nannenga-Bremekamp NE. 1989. Notes on myxomycetes XXIII. Seven new species of myxomycetes. Proceedings, Koninklijke Nederlandse Akademie van Wetenschappen, Series C, 92: 505-515. Schnittler M, Novozhilov YK. 2000. Myxomycetes of the winter-cold desert in western Kazakhstan. Mycotaxon 74(2): 267-285. Schrader HA. 1797. Nova genera plantarum. Pars prima, Lipsiae. Zhu H, Song XX, Li $, Wang Q, Li Y. 2012. Three new recorded species of myxomycetes in China. Mycosystema 31(6): 947-951. https://doi.org/10.13346/j.mycosystema.2012.06.021 MY COTAXON ISSN (print) 0093-4666 (online) 2154-8889 Mycotaxon, Ltd. ©2019 October-December 2019—Volume 134, pp. 649-662 https://doi.org/10.5248/134.649 Rhomboidia wuliangshanensis gen. & sp. nov. from southwestern China TaI-MIN XU?”, XIANG-Fu Liu3, YuU-HuI CHEN’, CHANG-LIN ZHAO? "Yunnan Provincial Innovation Team on Kapok Fiber Industrial Plantation; ? College of Life Sciences; * College of Biodiversity Conservation: Southwest Forestry University, Kunming 650224, PR. China " CORRESPONDENCE TO: fungichanglinz@163.com ABSTRACT—A new, white-rot, poroid, wood-inhabiting fungal genus, Rhomboidia, typified by R. wuliangshanensis, is proposed based on morphological and molecular evidence. Collected from subtropical Yunnan Province in southwest China, Rhomboidia is characterized by annual, stipitate basidiomes with rhomboid pileus, a monomitic hyphal system with thick-walled generative hyphae bearing clamp connections, and broadly ellipsoid basidiospores with thin, hyaline, smooth walls. Phylogenetic analyses of ITS and LSU nuclear RNA gene regions showed that Rhomboidia is in Steccherinaceae and formed as distinct, monophyletic lineage within a subclade that includes Nigroporus, Trullella, and Flabellophora. Key worps—Polyporales, residual polyporoid clade, taxonomy, wood-rotting fungi Introduction Polyporales Gaum. is one of the most intensively studied groups of fungi with many species of interest to fungal ecologists and applied scientists (Justo & al. 2017). Species of wood-inhabiting fungi in Polyporales are important as saprobes and pathogens in forest ecosystems and in their application in biomedical engineering and biodegradation systems (Dai & al. 2009, Levin & al. 2016). With roughly 1800 described species, Polyporales comprise about 1.5% of all known species of Fungi (Kirk & al. 2008). Currently, there are 46 genomes of polyporalean taxa available from the Joint Genome Institute MycoCosm portal (Grigoriev & al. 2013). 650 ... Xu & al. TABLE 1. Species and sequences used in the phylogenetic analyses SPECIES Abortiporus biennis Antrodiella americana A. semisupina Antrodiella sp. Climacocystis borealis Diplomitoporus flavescens Elaphroporia ailaoshanensis Flabellophora sp. Flaviporus brownii E liebmannii Frantisekia mentschulensis Hypochnicium bombycinum H. lyndoniae Irpex lacteus Ischnoderma benzoinum I. resinosum Junghuhnia crustacea J. micropora Loweomyces fractipes Mycorrhaphium adustum SAMPLE TFRI 274 Gothenburg 3161 FCUG 960 X 418 KH 13318 X 84 CLZhao 595 CLZhao 596 X340 X 1216 X 251 X 249 X 666 BRNM 710170 1377 MA 15305 NL 041031 CBS 431.48 DO 421/951208 KHL 12099 FD-328 X 1127 X 262 Spirin 2652 X 1149 X 1253 X 1250 8024 Dai 10173 GENBANK ACCESSION NO. ITS EU232187 JN710509 EU232182 JN710523 JQ031126 FN907908 MG231568 MG231572 JN710534 JN710537 JN710541 JN710539 JN710540 FJ496728 JN710544 FN552537 JX124704 MH856423 JX109852 JX109841 KP135303 JN710554 JN710553 JN710559 JN710570 JN710569 JN710568 JN710573 KC485537 LSU EU232235 JN710509 EU232266 JN710523 JQ031126 MG231568 MG231572 JN710534 JN710537 JN710541 JN710539 JN710540 JN710544 JX124704 MH867969 JX109852 JX109841 KP135225 JN710554 JN710553 JN710559 JN710570 JN710569 JN710568 JN710573 KC485554 REFERENCES Larsson 2007 Miettinen & al. 2012 Binder & al. 2005 Miettinen & al. 2012 Binder & al. 2013 Miettinen & al. 2012 Wu & al. 2018 Wu & al. 2018 Miettinen & al. 2012 Miettinen & al. 2012 Miettinen & al. 2012 Miettinen & al. 2012 Miettinen & al. 2012 TomSsovsky & al. 2010 Miettinen & al. 2012 Binder & al. 2013 Binder & al. 2005 Vu & al. 2019 Binder & al. 2013 Binder & al. 2013 Floudas & Hibbett 2015 Miettinen & al. 2012 Miettinen & al. 2012 Miettinen & al. 2012 Miettinen et al. 2012 Miettinen & al. 2012 Miettinen & al. 2012 Miettinen & al. 2012 Nigroporus vinosus Panus conchatus P strigellus Physisporinus sanguinolentus P. vitreus Podoscypha venustula Pseudolagarobasidium acaciicola P. belizense Rhomboidia wuliangshanensis Skeletocutis novae-zelandiae Spongipellis spumeus Steccherinum fimbriatum S. ochraceum Trullella dentipora T. duracina T. polyporoides Xanthoporus syringae Rhomboidia wuliangshanensis gen. & sp. nov. (China) ... 651 X 839 8182 BHS2008- 100 X 1234 INPA 243940 BRNM 699576 3163 KHL11959 CBS 486.72 CBS 65684 CBS 115543 CBS 115544 CFMR DLC 04-31 CLZhao 4406 [T] CLZhao 4411 Ryvarden 38641 PRM 891931 BRNM 712630 BRNM 734877 KHL 11905 Ryberg s.n. KHL 11902 X 200 X 290 X 510 X 339 Cui 2177 Gothenburg 1488 N710576 JN710728 JX109857 JN710579 JQ955725 FJ496671 JN710580 JQ031129 MH860538 JN649367 DQ517883 DQ517882 JQ070173 MK860715 MK860716 JN710582 HQ728287 HQ728288 HQ728283 EU118668 EU118669 JQ031130 JN710512 JN710513 JN710602 JN710606 DQ789395 JN710607 N710576 JN710728 JX109857 JN710579 JQ955732 FJ496725 JN710580 JQ031129 MH872244 JN649367 MK860710 MK860711 JN710582 HQ729021 HQ728288 HQ728283 EU118668 EU118670 JQ031130 JN710512 JN710513 JN710602 JN710606 JN710607 Miettinen & al. (012 Miettinen & al. 2012 Binder & al. 2013 Miettinen & al. 2012 Binder & al. 2013 TomSsovsky & al. 2010 Miettinen & al. 2012 Sjokvist & al. 2012 Vu & al. 2019 Binder & al. 2013 Miettinen & Rajchenberg 2012 Miettinen & Rajchenberg 2012 Miettinen & Rajchenberg 2012 Present study Present study Miettinen & al. 2012 Tomsovsky & al. 2010 Tomsovsky & al. 2010 Tomsovsky & al. 2010 Tomsovsky & al. 2010 Larsson 2007 Sjokvist & al. 2012 Miettinen & al. 2012 Miettinen & al. 2012 Miettinen & al. 2012 Miettinen & al. 2012 Miettinen & al. 2012 Miettinen & al. 2012 652 ... Xu & al. Systematics of the Polyporales has benefitted from numerous molecular phylogenetic studies (e.g., Binder & al. 2005, 2013; Larsson 2007; Miettinen & al. 2012; Dai & al. 2015; Choi & Kim 2017). Steccherinaceae Parmasto, one of 18 families recognized in Polyporales (Justo & al. 2017), has been included in several molecular studies (e.g., Binder & al. 2005, 2013; Miettinen & al. 2012; Miettinen & Ryvarden 2016; Justo & al. 2017; Westphalen & al. 2018). Miettinen & al. (2012) published a multigene, molecular phylogenetic study of Steccherinum and allied taxa that clearly delineated Steccherinaceae. They uncovered surprising morphological diversity and plasticity in this family, requiring revision of generic concepts and 15 new genera to accommodate existing and new species. Subsequently, Miettinen & Ryvarden (2016) introduced five new genera, revised one genus, and described two new species that had been identified earlier by Miettinen & al. (2012). Justo & al. (2017), who revised family-level classification in Polyporales, confirmed Steccherinaceae as a distinct lineage in Polyporales that grouped with Cerrenaceae Miettinen & al. and Panaceae Miettinen & al. Ina morphological and molecular study of Neotropical taxa of Junghuhnia and Steccherinum, Westphalen & al. (2018) uncovered a new genus and several new species and reclassified four taxa. Cosmopolitan in distribution, Steccherinaceae has a rich diversity because it is found in boreal, temperate, subtropical, and tropical ecosystems (Nunez & Ryvarden 2001, Dai 2012, Ryvarden & Melo 2014, Dai & al. 2015, Zhou & al. 2016). Many new species in Polyporales have been described from southern, subtropical China (e.g., Li & Cui 2010, Zhao & Wu 2017, Zhao & Ma 2019). Recently, we collected an undescribed taxon from Yunnan Province that could not be assigned to any described genus. We present morphological and molecular phylogenetic evidence that support the recognition of a new monotypic genus in Steccherinaceae—Rhomboidia, typified by R. wuliangshanensis. Materials & methods The specimens studied are deposited at the herbarium of Southwest Forestry University, Kunming, China (SWFC). Macromorphological descriptions are based on field notes. Special colour terms follow Petersen (1996). Micromorphological data were obtained from the dried specimens and observed under a light microscope following Dai (2010). The following abbreviations are used: KOH = 5% potassium hydroxide; CB = cotton blue; CB- = acyanophilous; IKI = Melzer’s reagent; IKI- = non-amyloid and non-dextrinoid; L = mean spore length (arithmetic average of all spores); W = mean spore width (arithmetic average of all spores); Q = variation in Rhomboidia wuliangshanensis gen. & sp. nov. (China) ... 653 the L/W ratios between the specimens studied; (n = a/b) = number of spores (a) from number of specimens (b). HiPure Fungal DNA Mini Kit II was used to obtain genomic DNA from dried specimens, according to the manufacturer’s instructions with some modifications. A small piece (about 30 mg) of dried fungal material was ground to powder with liquid nitrogen. The powder was transferred to a 1.5 ml centrifuge tube, suspended in 0.4 ml of lysis buffer, and incubated in a 65 °C water bath for 60 min. After that, 0.4 ml phenol-chloroform (24:1) was added to each tube and the suspension was shaken vigorously. After centrifugation at 13,000 rpm for 5 min, 0.3 ml supernatant was transferred to a new tube and mixed with 0.45 ml binding buffer. The mixture was then transferred to an adsorbing column (AC) for centrifugation at 13,000 rpm for 0.5 min. Then, 0.5 ml] inhibitor removal fluid was added in AC for a centrifugation at 12,000 rpm for 0.5 min. After washing twice with 0.5 ml washing buffer, the AC was transferred to a clean centrifuge tube, and 100 ml elution buffer was added to the middle of adsorbed film to elute the genomic DNA. The internal transcribed spacer region (ITS) was amplified with primer pairs ITS5 and ITS4 (White & al. 1990). The nuclear large subunit region (LSU) was amplified with primer pairs LROR and LR7 (https://sites.duke.edu/vilgalyslab/rdna_primers_for_fungi/). The PCR procedure for ITS was: initial denaturation at 95 °C for 3 min, followed by 35 cycles of 94 °C for 40 s, 58 °C for 45 s, and 72 °C for 1 min; and a final extension of 72 °C for 10 min. The PCR procedure for LSU was: initial denaturation at 94 °C for 1 min, followed by 35 cycles of 94 °C for 30 s, 48 °C 1 min, and 72 °C for 1.5 min; and a final extension of 72 °C for 10 min. The PCR products were purified and directly sequenced at Kunming Tsingke Biological Technology Limited Company. All newly generated sequences were deposited at GenBank (TABLE 1). Sequencher 4.6 was used to edit the DNA sequence. Sequences were aligned in MAFFT 6 (Katoh & Toh 2008, http://mafft.cbrc.jp/alignment/server/) using the “G-INS-I” strategy and manually adjusted in BioEdit (Hall 1999). The sequence alignment was deposited in TreeBase (submission ID 24216). Xanthoporus syringae (Parmasto) Audet obtained from GenBank was used as an outgroup to root trees following Miettinen & al. (2012) in the ITS+LSU analyses (Fic. 1). The ITS+LSU sequences were analyzed phylogenetically using maximum parsimony, maximum likelihood, and Bayesian inference methods. Maximum parsimony (MP) analyses followed Zhao & Wu (2017), and tree construction was performed in PAUP* version 4.0b10 (Swofford 2002). All characters were equally weighted and gaps were treated as missing data. Trees were inferred using the heuristic search option with TBR branch swapping and 1000 random sequence additions. Max-trees was set to 5000, branches of zero length were collapsed and all parsimonious trees were saved. Clade robustness was assessed using bootstrap (BP) analysis with 1000 replicates (Felsenstein 1985). Tree statistics tree length (TL), consistency index (CI), retention index (RI), rescaled consistency index (RC), and homoplasy index (HI) were calculated for each Maximum Parsimonious Tree generated. Sequences were analyzed using Maximum Likelihood (ML) with 654 ... Xu & al. RAxML-HPC2 through the Cipres Science Gateway (www.phylo.org; Miller & al. 2009). Branch support (BS) for ML analysis was determined by 1000 bootstrap replicates. MrModeltest 2.3 (Posada & Crandall 1998; Nylander 2004) was used to determine the best-fit evolution model for data set for Bayesian inference (BI). Bayesian inference was calculated with MrBayes_3.1.2 using a general time reversible (GTR+G) model of DNA substitution and a gamma distribution rate variation across sites (Ronquist & Huelsenbeck 2003). Four Markov chains were run for 2 runs from random starting trees for 4 million generations (ITS+LSU) in Fic. 1 and trees were sampled every 100 generations. The first 25% of the generations were discarded as burn-in. A majority rule consensus tree of all remaining trees was calculated. Branches that received bootstrap support for maximum likelihood (BS) 275%, maximum parsimony (BP) 275%, and Bayesian posterior probabilities (BPP) 20.95 were considered significantly supported. Molecular phylogenetic results The ITS+LSU (Fic. 1) dataset comprised sequences from 55 fungal specimens representing 34 taxa, including the outgroup taxon. The dataset had an aligned length of 2296 of which 1395 were constant, 201 parsimony- uninformative, and 700 parsimony-informative. MP analysis yielded two equally parsimonious trees (TL = 3897, CI = 0.376, HI = 0.624, RI = 0.627, RC = 0.236). The best-fit model for ITS+LSU alignment estimated and applied in the BI was GTR+I+G, lIset nst = 6, rates = invgamma; prset statefreqpr = dirichlet (1,1,1,1). BI resulted in a similar topology with an average standard deviation of split frequencies equal to 0.006862. Rhomboidia wuliangshanensis forms a monophyletic lineage with strong support (BS = 100%, BP = 100%, BPP = 1) and is sister to the Nigroporus-— Trullella clade (Fic. 1). Taxonomy Rhomboidia C.L. Zhao, gen. nov. MB 833318 Differs from Nigroporus and Trullella by its stipitate to substipitate basidiomata, its orange-brown to reddish brown surface, and its monomitic hyphal system in both context and trama. Fic. 1. Maximum parsimony strict consensus tree illustrating the phylogeny of Rhomboidia wuliangshanensis and related species in the residual polyporoid clade based on ITS+nLSU sequences. Branches are labeled with maximum likelihood bootstrap >70%, parsimony bootstrap proportions >50% and Bayesian posterior probabilities >0.95. Rhomboidia wuliangshanensis gen. & sp. nov. (China) ... 655 G Cerrenaceae + Hyphodermataceae © Inpicaceae © Ischnodermataceae @ Meripilaceae 95/95/1 A Meruliaceae -/-/l & Panaceae - /81/0.95 84/81/1 100/100/1 100/100/1 Physisporinus vitreus CBS 486.72 @ Irpex lacteus CBS 431.48 @ Irpex lacteus DO 421/951208 @ Ischnoderma benzoinum KHL 12099 Ischnoderma resinosum FD-328 © Diplomitoporus flavescens X 84 * Climacocystis borealis KH 13318 @® Panus strigellus TNPA 243940 & Panus conchatus X 1234 & 100/100/1 | Steccherinaceae @ > Fomitopsidaceae WME P, Leo} 0/100/1|Pseudolagarobasidium acaciicola CBS 115543 @ ee ae ae Pseudolagarobasidium belizense CFMR DLC 04-31 @ x Polyporaceae Oe Pseudolagarobasidium acaciicola CBS 115544 @© Hypochnicium lyndoniae NL 041031 + Hypochnicium bombycinum MA 15305 + 95/63/ -| Spongipellis spumeus BRNM 712630 © S782 100/100/1 100/100/1 100/100/1 100/91/1 97/61/1 100/100/1 92/94/1 100/88 1! 1 o9/10071 100/58/0.98 100/100/1 Spongipellis spumeus BRNM 734877 & Spongipellis spumeus PRM 891931 @& 100/99/1)Physisporinus vitreus KHL 11959 (GB) @ Physisporinus vitreus 3163 @ Physisporinus sanguinolentus BRNM 699576 @ Podoscypha venustula CBS 65684 pea Abortiporus biennis TFRI 274 Ht 100/63/ - |Loweomyces fractipes X 1253 na 10010011 roweomves fractipes X 1250 [jj Loweomyces fractipes X 1149 [J 100/100/1) Junghuhnia crustacea X 262 Junghuhnia crustacea X 1127 Jj Flaviporus brownii X 1216 Flaviporus liebmannii X 666 Ij Flaviporus liebmannii X 251 fj 100/100/I' Flaviporus liebmannii X 249 fj 100/100/1 p Bigpirapers ailaoshanensis CLZhao 595 [ij Elaphroporia ailaoshanensis CLZhao 596 | 95/100/1- Trullella duracina X290 Trullella polyporoides X510 Trullella dentipora X 200 B Nigroporus vinosus BHS2008-100 [ij Nigroporus vinosus 8182 Q Nigroporus vinosus X 839 fj 100/100/1; Rhomboidia wuliangshanensis CLZhao 4406 BH Rhomboidia wuliangshanensis CLZhao 441 Ij Flabellophora sp.X340 Mycorrhaphium adustum Dai 10173 [i Mycorrhaphium adustum 8024 [fj 100/99/1 - Junghuhnia micropora Spirin 2652 fj 100/100/1 | l Antrodiella sp. X 418 Pil 100/100/1 Skeletocutis novae-zelandiae Ryvarden 38641 ij Frantisekia mentschulensis BRNM 710170 i | 00/100/1 Fy antisekia mentschulensis 1377 | Steccherinum fimbriatum KHL 11905 ia Antrodiella semisupina FCUG 960 [ij Antrodiella americana Gothenburg 3161 [fj Steccherinum ochraceum KHL 11902 100/100/l' steecherinum ochraceum Rybergs.n. [i Xanthoporus syringae X 339 Oo an Xanthoporus syringae Cui 2177 Xanthoporus syringae Gothenburg 1488 [J 100/100/1 ——i 50 656 ... Xu & al. TYPE SPECIES: Rhomboidia wuliangshanensis C.L. Zhao EryMoLoGy: Rhomboidia (Lat.): referring to the rhomboid pileus of the basidiocarp with the poroid hymenophore. BASIDIOMATA annual, stipitate. Pileus rhomboid, arising from a multiple branched stipe. Pores angular, small, dissepiments thin, entire. Hyphal system monomitic; generative hyphae thick-walled bearing clamp connections, IKI-, CB-; tissues unchanged in KOH. Cystidia absent, fusoid cystidioles numerous; hyphal ends numerous. Basidia barrel-shaped to clavate. Basidiospores broadly ellipsoid, hyaline, thin-walled, smooth, IKI-, CB-. TYPE OF ROT: white rot. Rhomboidia wuliangshanensis C.L. Zhao, sp. nov. Fics 2, 3 MB 833320 Differs from Nigroporus vinosus by its stipitate to substipitate basidiomata and monomitic hyphal structure. Type: China. Yunnan Province: Puer, Jingdong County, Wuliangshan National Nature Reserve, on angiosperm trunk, 6 Oct 2017, CLZhao 4406 (Holotype, SWFC 0004406; GenBank MK860715, MK860710). EryMo.Locy: The specific epithet wuliangshanensis (Lat.) refers to the type locality, Wuliangshan. BASIDIOMATA annual, stipitate to substipitate. Pileus rhomboid, arising from a multiple branched stipe, edges curling slightly inward, 3.5 cm from the base to margin, 4 cm wide, up to 3 mm thick; pileus surface radially striate, slightly brown to orange brown when fresh, drying brown to reddish; the margin acute, entire. Pore surface white when fresh, cream to buff upon drying. Pores angular, 7-9 per mm, dissepiments thin, entire. Context corky, white, thin, up to 0.5 mm thick. Tubes concolorous with pore surface, corky, up to 2.5 mm long. - TYPE OF ROT: white rot. ADDITIONAL SPECIMEN EXAMINED: CHINA. YUNNAN PROVINCE. Puer: Jingdong County, Wuliangshan National Nature Reserve, on angiosperm trunk, 6 Oct 2017, CLZhao 4411 (SWFC 004411; GenBank MK860716, MK860711). Discussion Rhomboidia is supported as a new genus by phylogenetic analyses and morphological characters. It is embedded in Steccherinaceae with strong support. Phylogenetically, Rhomboidia is closely related to Flabellophora G. Cunn., Nigroporus Murrill, and Trullella Zmitr. based on ITS+LSU nuclear RNA gene analyses (Fic. 1), which is similar to the previous multigene sequence- based study (Miettinen & al. 2012). The genera closely related to Rhomboidia Rhomboidia wuliangshanensis gen. & sp. nov. (China) ... 657 Fic. 2. Rhomboidia wuliangshanensis (holotype, SWFC 0004406). Scale bars = 5 mm. 658 ... Xu & al. Leciiiian G (a) t a i Fic. 3. Rhomboidia wuliangshanensis (drawn from the holotype, SWFC 0004406). A. Basidiospores; B. Basidia and basidioles; subiculum. Scale bars: a C. Cystidioles; D. Hyphal ends; E. Hyphae from trama; F. Hyphae from = 5 um; b-f = 10 um. Rhomboidia wuliangshanensis gen. & sp. nov. (China) ... 659 are easily separated morphologically: In Flabellophora basidiomata arise from a submerged pseudosclerotium and develop unilateral pilei with a crust and a coriaceous context (Nufez & Ryvarden 2001). Nigroporus differs from Rhomboidia by developing resupinate to pileate basidiocarps with vinaceous brown to pink or violet pore surface and a dimitic hyphal system (Gilbertson & Ryvarden 1987). In Trullella basidiomata are spathulate and light-coloured, with a monomitic hyphal system in the context but dimitic in the trama (Miettinen & al. 2012, Zmitrovich 2018). Rhomboidia resembles other stipitate genera in Polyporales such as Abortiporus Murrill, Jahnoporus Nuss, and Polyporus P. Micheli ex Adans. Abortiporus, however, has a duplex structure and thick-walled basidiospores (Nunez & Ryvarden 2001). Jahnoporus is characterized by large spindle- shaped basidiospores (Gilbertson & Ryvarden 1987), and Polyporus has a dimitic hyphal system (Bernicchia & Gorjon 2010). Polypores are extensively studied and well-known in North America (Gilbertson & Ryvarden 1987, Zhou & al. 2016) and Eurasia (Nufez & Ryvarden 2001, Bernicchia & Gorjén 2010, Dai 2012, Ryvarden & Melo 2014, Dai & al. 2015), but Chinese polypore diversity is still being explored, especially in subtropical and tropical areas. Rhomboidia wuliangshanensis was collected from Yunnan Province, where many new taxa in Polyporales and Hymenochaetales have been described (e.g., Li & Cui 2010, He & Li 2011, Yu & al. 2013, Yang & He 2014, Zhao & Wu 2017, Zhao & Ma 2019). 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Ceriporiopsis kunmingensis sp. nov. (Polyporales, Basidiomycota) evidenced by morphological characters and phylogenetic analysis. Mycological Progress 16: 93-100. https://doi.org/10.1007/s11557-016-1259-8 Zhou LW, Nakasone KK, Burdsall Jr. HH, Ginns J, Vlasak J, Miettinen O, Spirin V, Niemela T, Yuan HS, He SH, Cui BK, Xing JH, Dai YC. 2016. Polypore diversity in North America with an annotated checklist. Mycological Progress15: 771-790. https://doi.org/10.1007/s11557-016-1207-7 Zmitrovich IV. 2018. Conspectus systematis Polyporacearum v. 1.0. Folia Cryptogamica Petropolitana 6: 3-145. MY COTAXON ISSN (print) 0093-4666 (online) 2154-8889 Mycotaxon, Ltd. ©2019 October-December 2019—Volume 134, pp. 663-675 https://doi.org/10.5248/134.663 Filsoniana lhasanensis sp. nov. from Tibet, China XuE-MEI WEN”?, HURNISA SHAHIDIN'?, ABDULLA ABBAS! ‘College of Life Science and Technology, Xinjiang University, Urumai, 830046, P. R. China *Tibet Plateau Institute of Biology, Lhasa, 850001, P. R. China * CORRESPONDENCE TO: hurnisa_xju@sina.com, zxg_lichen@163.com ABSTRACT—A new species, Filsoniana lhasanensis, is described from Tibet, China, where it grows on rocks at altitudes 4200-4432 m. The lichen is characterised by a poorly developed or absent dark orange areolate thallus, lecanorine to zeorine orange to deep-orange aggregated apothecia, and richly branched and anastomosed paraphyses. DNA was obtained for three gene loci—nuclear internal transcribed spacer (ITS), nuclear large subunit (LSU), and mitochondrial small subunit (SSU). Phylogenetic analyses support the taxon as a new species of Filsoniana. Key worps—Himalaya, phylogeny, taxonomy, Teloschistoideae Introduction Teloschistaceae is a widespread and well-delimited family of lichenized fungi, estimated at over 1000 species (Arup & al. 2013). Most species contain anthraquinone (which confers a yellow to orange color) and grow under sun-exposed conditions (Gaya & al. 2015). Recent molecular phylogenetic research supports subdivision of the Teloschistaceae into three subfamilies: Xanthorioideae, Caloplacoideae, and Teloschistoideae (Arup & al. 2013; Gaya & al. 2015). Kondratyuk & al. (2013, 2015), who evaluated Teloschistoideae based on three gene loci (ITS, LSU, SSU), proposed an additional subfamily Brownlielloideae; however this analysis was based on a hybrid sequence dataset comprising ITS and LSU data derived from 664 ... Wen, Shahidin, Abbas the lichen mycobiont and the SSU derived from a fungal contaminant (Vondrak & al. 2018). Many small monophyletic genera established in Brownlielloideae were based solely on DNA data without any distinguishing phenotypic characters, and these genera remain within the Teloschistoideae as proposed by Arup & al. (2013). During our study on Chinese Teloschistaceae, we collected some interesting samples from the suburbs of Lhasa and Shannan in Tibet. The Tibetan plateau is a unique biogeographical region with varied landscapes, altitudinal belts, and alpine ecosystems and is considered a world center of species formation (Zhang & al. 2002). Teloschistaceae in this region have been inadequately studied, with only 15 species reported (Obermayer 2004; Wei 1991, 2017; Poelt & Petutschnig 1992; Poelt & Hinteregger 1993; Wei & Jiang 1986). More species are known from the neighboring Himalayan regions: India, where >74 species of Teloschistaceae have been recorded (Joshi & Upreti 2006, 2011; Joshi & al. 2008, 2009, 2014; Kondratyuk & al. 2018) and Nepal, where 47 species have been discovered (Olley & Sharma 2013). According to available literature, there are still large underexplored areas in the Tibet region, and additional undescribed lichenized fungi species are expected to be discovered in this poorly studied region. Most of crustose Teloschistaceae species have few taxonomically significant morphological characters. Traditional species concepts based in morphology and chemistry are overly simplified for the crustose species in Teloschistaceae. Modern classification methods are essential for accurate taxonomic placement of rare and undescribed species. We combined sequence analyses of three gene loci—ITS, LSU, SsU—with morphological and chemical methods to describe a new species, Filsoniana lhasanensis. This is the first report of the genus Filsoniana in China. Materials & methods Morphology & chemistry Specimens collected from Lhasa and Shannan, Tibet, China, were preserved in Tibet Plateau Institute of Biology, Lhasa, China (XZ) and Lichen Research Center in Arid Zone of Northwest China, Xinjiang University, Urumqi, China (XJU-L). The specimens were examined morphologically using a Shunyu SZM45 dissecting microscope and Nikon Eclipse E100 compound microscope. Colors follow RAL color system (https://www.ralcolorchart.com/) Sections for anatomical details and measurements were made manually and mounted in water. Chemical constituents were identified by thin-layer chromatography using solvent systems C (Orange & al. 2010). Thallus morphological structures were photographed with a Nikon Filsoniana lhasanensis sp. nov. (China) ... 665 TABLE 1. Primers and cycling parameters used for PCR. Locus PRIMERS PCR SETTINGS ITS ITS1F (Gardes & Bruns 1993) 94°C-5 min; 6 cycles of: 94°C-45 sec, 57-52°C 1 ITS4 (White & al. 1990) : (decreasing 1°C per cycle) -55 sec, 72°C-1 min; 30 :_ cycles of: 94°C-45 sec, 51°C-55sec, 72°C-1 min; and | 72°C-7 min LSU { ALIR (Déring & al. 2000) | 94°C-10 min; 6 cycles of: 94°C-1 min, 57-52°C | LR6 (Vilgalys & Hester 1990) 1 (decreasing 1°C per cycle) -1 min, 72°C-1.2 min; 30 | cycles of: 94°C-1 min, 51°C-1 min, 72°C-1.2 min; ' and 72°C-2 min ssu mrSSU1 (Zoller & al. 1999) 94°C-10 min; 6 cycles of: 94°C-45 sec, 57-52°C mrSSU7 (Zhou & Stanosz 2001) : (decreasing 1°C per cycle) —55 sec, 72°C-1 min; 30 1 cycles of: 94°C-45 sec, 51°C-55sec, 72°C-1 min; and 72°C-7 min DS-Fi2 Digital Camera using a Nikon MZ25 and Nikon Eclipse Ci stereomicroscope. All measurements were made using the Nikon Nis elements (D). Morphology and anatomy were described according to Vondrak & al. (2013). DNA extraction, PCR amplification, DNA sequencing Total genomic DNA was extracted using the Fungi Genomic DNA Extraction Kit, following manufacturer's instructions. Molecular data were generated for three loci: the nuclear internal transcribed spacer (ITS), nuclear large subunit (LSU), and mitochondrial small subunit (SSU). Primers and PCR cycling parameters used for amplification are summarized in TABLE 1. Sequence analysis The newly generated sequences were compared to GenBank database sequences using BLASTN search (http://www.ncbi.nlm.nih.gov/BLAST/). All sequences were aligned with sequences of selected representatives of Teloschistoideae obtained from GenBank (TABLE 2) using MAFFT version 7 (Katoh & Standley 2013) with G-INS-i method. Phylogenies were generated using Bayesian inference and Maximum Likelihood (ML) methods. Bayesian inference used the best fitting model as inferred by j Model Test v2.1.6 (Darriba & al. 2012), implemented in MrBayes v3.2.6 (Ronquist & al. 2012) on the Cipres Scientific gateway (Miller & al. 2010), TrNef+I+G for ITS and LSU and HKY+I+G for SSU. Two parallel MCMCMC runs were executed, each using eight chains and 10,000,000 generations, with trees sampled every 1000th generation. Maximum likelihood (RAxML) analyses were performed using RAxMLHPC v.8 on XSEDE (Stamatakis 2014) under the GTRGAMMA model on CIPRES Science Gateway (Miller & al. 2010). Rapid bootstrap analyses were performed with 1000 bootstrap replicates. The phylogenetic tree was drawn using FigTree ver. 1.4.3 (http://tree.bio.ed.ac.uk/software/figtree/). 666 ... Wen, Shahidin, Abbas TABLE 2. Sequences used in the phylogenetic analyses. (Sequences generated for this study are in bold.) SPECIES VOUCHER ITS LSU SSU Brownliella aequata | SK838 [4] | KF264627 | KF264662 { KF264688 ! SK831 [4] ! KF264626 ! KF264661 ! KF264687 B. kobeana ! 130318 [5] ! KT456214 ! KT456229 ! KT456244 ! 120032 [5] ! KT456212 ! KT456227 ! KT456242 B. montisfracti ! SK231 [4] ! KF264625 ! KF264660 ! KF264686 ! SK230 [4] ! KF264624 ! KF264659 ! _ Browniliella sp. SK2013a [4] KF264628 KF264663 ! — Caloplaca sp. ! Casan 242 [3] ! KT291463 ! KT291558 ! KT291508 ! Gaya 110 [3] ! KT291456 ! KT291552 ! KT291499 Elixjohnia bermaguiana ! Kondratyuk 20487 [1] ! KC179299 ! KC179245 ! KC179584 E. gallowayi ! Karnefelt 997504 [1] ! KC179301 ! KC179247 ! KC179586 E. jackelixii ! U1402 [1] ! KC179303 ! KC179248 ! KC179587 ! SK910 [4] ! KF264655 ! KF264683 ! KF264707 Filsoniana australiensis ! SK851 [4] ! KF264631 ! KF264665 ! KF264691 ! SK850 [4] ! KF264632 ! KF264666 ! KF264692 EF. lhasanensis 224, Wen 20110910-05 MK153161 MK439835 MK439831 ! Z19, Wen 20110901-29 ! MK153160 ! MK439834 ! MK439830 E rexfilsonii ! SK859 [4] ! KF264638 ! KF264671 ! — ! SK861 [4] ! KF264636 ! KF264670 ! = Follmannia orthoclada ! Frodén 1772 [1] ! KC179291 ! KC179191 ! _ Haloplaca sp. ! 6-UA2013 [1] ! KC179295 ! KC179203 ! KC179537 Harusavskia elenkinianoides ! SK997 [6] ! KY614404 ! KY614452 ! KY614485 ! SK269 [6] ! KY614405 ! KY614453 ! KY614486 Josefpoeltia parva ! Frédén 1671 [1] ! KC179296 ! KC179204 ! KC179539 J. sorediosa ! Frodén 1593 [1] ! KC179297 ! KC179205 ! KC179540 ! SK992 [4] ! KF264646 ! KF264674 ! KF264697 ! SK991 [4] ! KF264645 ! KF264673 ! KF264696 Kaernefia albocrenulata ! SK245 [4] ! KF264647 ! KF264675 ! KF264698 ! SK246 [4] ! KF264648 ! KF264676 ! KF264699 K. gilfillaniorum ! SK999 [4] ! KF264650 ! KF264678 ! KF264701 ! SK253 [4] ! KF264649 ! KF264677 ! KF264700 K. kaernefeltii ! SK919 [4] ! KF264651 ! KF264679 ! KF264702 ! SK921 [4] ! KF264652 ! KF264680 ! KF264703 Nevilleiella lateritia | SK 261 [6] 1 KY614427 | KY614464 | KY614502 Filsoniana lhasanensis sp. nov. (China) ... 667 SPECIES VOUCHER ITS LSU SSU | SK 878 [6] | KY614426 | KY614463 | KY614501 N. marchantii ! SK D18 [6] ! KY614425 ! KY614462 ! KY614500 Scutaria andina ! PF209 [1] ! KC179298 ! KC179242 ! KC179581 Sirenophila cliffwetmorei ! SK A93 [6] ! KY614438 ! KY614471 ! KY614513 S. eos ! U1408 [1] ! KC179300 ! KC179246 ! KC179585 S. gintarasii ! SK D17 [6] ! KY614437 ! KY614470 ! KY614512 S. maccarthyi ! Karnefelt 977801 [1] ! KC179304 ! KC179249 ! KC179588 Solitaria chrysophthalma ! Cchr157 [3] ! KT291446 ! KT291537 ! KT291484 Stellarangia elegantissima ! PF 81 [1] ! KC179310 ! KC179254 ! KC179593 ! Cele 75 [3] ! KT291454 ! KT291541 ! KT291488 Teloschistes exilis ! Gaya 12 [2] ! JQ301630 ! JQ301577 ! JQ301519 T. flavicans ! AFTOL-ID 315 [2] ! JQ301578 ! JQ301631 ! JQ301520 ! Tflav103 [3] ! KT291472 ! KT291565 ! KT291523 T. hosseusianus Gaya 50 [2] KC179318 ! JQ301579 ! JQ301521 T. hypoglaucus ! PF68 [1] ! KC179319 ! KC179256 ! KC179595 T. sieberianus ! Tsie423 [3] ! EU639655 ! pez ! KT291525 Teloschistopsis bonae-spei ! PF82 [1] ! KC179322 ! KC179257 ! KC179596 Ts. eudoxa ! PF72 [1] ! KC179324 ! KC179258 ! KC179597 Thelliana pseudokiamae SK925 [4] KF264633 KF264667 — ! SK926 [4] ! KF264634 ! KF264668 ! = ! SK 925 [5] ! KT456225 ! KT456240 ! _— ! SK 926 [5] ! KT456226 ! KT456241 ! _— Villophora isidioclada ! USE563 [1] ! KC179325 ! KC179266 ! KC179606 Villophora sp. 50 ! U1342 [1] ! KC179330 ! KC179268 ! — Villophora sp. 40 ! USE574 [1] ! KC179328 ! KC179267 ! KC179607 Wetmoreana decipioides Cadec188 [3] KT291453 KT291540 KT291487 ! SK689 [4] ! KF264644 ! — ! KF264695 W. texana ! SK536 [4] ! KF264658 ! = ! KF264711 ! USE527 [1] ! KC179337 ! KC179273 ! KC179612 W. sp. 52 ! Frédén 1645 [1] ! KC179334 ! KC179270 ! KC179609 W. sp. 54 ! Frédén 1521 [1] ! KC179336 ! KC179272 ! KC179611 Xanthomendoza fallax ! Gaya 33 [2] ! JQ301687 ! JQ301580 ! JQ301633 Xa. ulophyllodes ! T11, Shahidin 145507 ! MK414784 ! MK439836 ! MK439832 ! T22, Shahidin 145449 ! MK414785 ! MK439837 ! MK439833 Xanthoria sp. ! Gaya 31 [2] ! JQ301692 ! JQ301591 ! JQ301532 Sources: [1] Arup & al. (2013); [2] Gaya & al. (2012); [3] Gaya & al. (2015); [4] Kondratyuk & al. (2013); [5] Kondratyuk & al. (2015); [6] Kondratyuk & al. (2017). 668 ... Wen, Shahidin, Abbas Xanthomendoza ulophyllodes Xanthomendoza ulophyllodes Xanthomendoza fallax 1/100) Villophora isidioclada 1/100 illophora sp. 50 Villophora sp. 40 Josefpoeltia parva q- Josefpoeltia sorediosa Josefpoeltia sorediosa 1/80 0.99/99L Josefpoeltia sorediosa 1/100 Teloschistes exilis 1/100 , Teloschistes flavicans 1/94 Teloschistes flavicans Teloschistes sieberianus oor- Teloschistes hosseusianus 1/100 5 Teloschistes hypoglaucus Teloschistes chrysophthalmus 1/100 1/100 Caloplaca sp. + ~ 219 Filsoniana lhasanenses © - 224 Filsoniana Ilhasanenses SK861 Filsoniana rexfilsonii -/88} SK925 Thelliana pseudokiamae SK925 Thelliana pseudokiamae SK926 Thelliana pseudokiamae SK926 Thelliana pseudokiamae SK859 Filsoniana rexfilsonii 0.97/86 Filsoniana australiensis 1/100 SK850 Filsoniana australiensis iy Nevilleiella marchantii 1/9 Nevilleiella lateritia 1/95) 1/99 Nevilleiella lateritia 1/100 Brownliella sp. 1/100 Brownliella aequata Brownliella aequata 1/100, 8rownliella kobeana Brownliella kobeana 1/100 Brownliella montisfracti 1/73 Brownliella montisfracti 1/100; Keernefia gilfillaniorum Kaernefia gilfillaniorum 1/100 ° 6 1/93 thi 1/94 001 1/100 », Kaernefia kaernefeltii SL Kaernefia kaernefeltii O— Kaernefia albocrenulata 99 1/99 Kaernefia albocrenulata / Haloplaca sp. Elixjohnia bermaguiana 1 1/100 as ; R 1/94 1/100 Elixjohnia gallowayi 1/100 Elixjohnia jackelixii 1/90 AGk Bie a 1/1004 Elixjohnia jackelixii 1/99 Sirenophila eos 1/95 Sirenophila maccarthyi Sirenophila gintarasii -/95 1/100 . labbod enophila cliffwetmor 1/100 Teloschistopsis bonae-spei Teloschistopsis eudoxa 1/100 Stellarangia elegantissima 1/70 Stellarangia elegantissima 1/100 Wetmoreana decipioides Wetmoreana decipioides 1/100 1/100 Wetmoreana texana 1/100} Wetmoreana texana 1/95 Wetmoreana sp. 54 -94/90] 0.949 Wetmoreana sp. 52 1/85) 0.95/9 Caloplaca sp. Solitaria chrysophthaima 1/100 Xanthoria sp. Scutaria andina 1/100 , Harusavskia elenkinianoides 1/99 Harusavskia elenkinianoides Follmannia orthoclada 0.95/ ae O09 Fic. 1. The ML tree based on a concatenated 3-loci data matrix. The numbers above each node represent posterior probability (PP) and bootstrap support (BS) values. Only PP values >0.90 and BS values >70% are shown. The species described in this paper is shown in blue font. Results & discussion New ITS, LSU, and SSU sequences were generated in this study. A total of 70 ITS sequences, 67 LSU sequences, and 58 SSU sequences were included in the analyses (Fic. 1). Trees of similar topologies were also generated using the maximum likelihood method and Bayesian approach, Filsoniana lhasanensis sp. nov. (China) ... 669 with Xanthomendoza ulophyllodes and X. fallax as outgroup. Our combined analyses grouped our new species with Filsoniana rexfilsonii and Thelliana pseudokiamae in a monophyletic clade with high support value (BS = 93, PP = 1). The new species is closely related to Thelliana pseudokiamae (S.Y. Kondr. & Karnefelt) S.-Y. Kondr.& al. and E rexfilsonii (S.Y. Kondr. & Karnefelt) S.Y. Kondr. & al.. Almost all other members of Filsoniana have well-developed thalli and occur in Australia. The sequences listed by Kondratyuk & al. (2013) under the name “Squamulea kiamae” were misdetermined; they were derived from specimen Karnefelt 994101 [LD 1101337], which was resequenced by Kondratyuk &al. (2015) and designated as the holotype of a new genus and species, Thelliana pseudokiamae S.Y. Kondr. & al. Therefore all four of the “Squamulea kiamae” sequences represent Th. pseudokiamae, and authentic sequences of S. kiamae are not included in our analyses. Filsoniana kiamae and Th. pseudokiamae differ in the sorelia and lobe size (Kondratyuk & al. 2007, 2015). Our results conflict with Kondratyuk & al. (2015), whose ITS-LSU-SSU combined analyses placed Filsoniana in Brownlielloideae; our analyses place most members of Brownlielloideae in the Teloschistoideae. Taxonomy Filsoniana lhasanensis X.M. Wen, Shahidin & A. Abbas sp. nov. Fic. 2 FN 570592 Differs from Filsoniana kiamae and E rexfilsonii by its reduced thallus, its aggregated adnate apothecia, and its Asian distribution. Type: China. Tibet, Lhasa City, Maizhuokunggar county, Riduo Village, 29°42’35”N 92°05’10’E, alt. 4200 m, 1 September 2011, X. M. Wen 20110901-29 (Holotype, XZ; isotype, XJU-L; GenBank MK153160, MK439834, MK43983). EryMo.ocy: The epithet /hasanensis refers to the region in which the type specimen was collected. THALLUS crustose, areolate, poorly developed or absent, usually confined to the apothecia, 0.1-0.2 mm diam., orange to deep orange [RAL 2011]. CorTEX paraplectenchymatous, 12.9-67.1 um high, composed of thin- walled cells, (2.38-)4.8-7.4(-10.5) um diam. ALGAL LAYER + continuous, 49.5-97.9 um thick. MEDULLA loosely interwoven by thin-walled hyphae under the apothecia, 59.8-86.1 um high. APOTHECIA lecanorine to zeorine, numerous, adnate, always aggregated (2-7) or crowded together, disc margins present, smooth, entire, concolorous with thallus, sun yellow [RAL 1037] to deep orange 670 ... Wen, Shahidin, Abbas [RAL 2011], disc round to irregular, yellow-orange [RAL 2000] to signal orange [RAL 2010], flat to concave, 0.3-0.7 mm diam. EXxcIPLE 9.6-14.8 um thick, consisting of 2-4 layers of cells, upper part of cells spherical, 3.2-6.0 um diam, cells of lower part elongated, 3.5-10.6 x 1.4-3.6 um. HypoTuHEcium thin, 22.3-34.7 um high, composed of several lines of paraplectenchymatous cells, 2.9-5.2 x 2.3-4.6 um. HYMENIUM 52.2-89.5 um high, colorless. PARAPHYSES anastomosed, generally dichotomously branched or trigeminally branched at tips, the 3-4 terminal cells gradually enlarging towards tips, the widened cells nonuniform in size, the second cells 3.5-6.7 um diam., and the third ones 3.2-5.3 um diam. Ascr_ teloschistes-type, 8-spored. Ascospores colorless, ellipsoid, (11.2-)11.7-17.5(-4.7) x 5.8-8.4(-9.3) um, septum 3.3-6.5 um diam. SPOT TESTS—K+ red. CHEMISTRY—Parietin, emodin, and two unknown anthraquinones. EcoLtocy—Filsoniana lhasanensis grows on rocks in open areas and associates with species of Aspicilia, Lecanora, Rusavskia, Physcia, Rinodina, Rhizoplaca, and Acarospora. The new lichen is distributed in arid or semiarid areas of Lhasa city and Shannan city at elevations of 4200-4432 m. ADDITIONAL SPECIMEN EXAMINED: CHINA. TIBET, Shannan City, Nagarze county, Yamzho Yumco, 29°09’34”N 90°30'19’E, alt. 4432 m, 10 September 2011, X.M. Wen 20110910-05 (XZ; GenBank MK153161, MK439835, MK439831). REMARKS: Filsoniana lhasanensis is characterized by a reduced, dark orange to brownish orange thallus and aggregated apothecia. It is morphologically similar to Xanthocarpia ferrarii (Bagl.) Frodén & al., which differs by its narrower septum, larger ascospores, well developed true exciple, zeorine to biatorine apothecia, and classification in Xanthorioideae (Arup & al. 2013). Filsoniana australiensis, F. rexfilsonii, and F. lhasanensis have similar paraplectenchymatous cortex and broad paraphyses (Kondratyuk & al. 2007). However, F. australiensis and E rexfilsonii have well-developed thalli (E australiensis has a placodioid thallus, and FE rexfilsonii has a squamulose thallus) with scattered or rare apothecia, whereas F [hasanensis has a poorly developed thallus consisting of only a few scattered areoles. A summary of the characteristics of FE [hasanensis and morphologically similar species is provided in TABLE 3. Filsoniana lhasanensis sp. nov. (China) ... 671 Fic. 2. Filsoniana lhasanensis (XZ (Wen 20110901-29)): A. Thallus and apothecia; B. Cortex of thallus; C. Cross section of the apothecium; D. Hypothecium; E. Paraphyses branched and enlarged at the tips; F. Paraphyses branched and anastomosed; G. Ascus containing eight ascospores. Scale bars: A = 1 mm; B-F = 10 um; G=5 um. 672 ... 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S9NIABID YOOI UT! snore uo ‘suoHenyis svare uado Ul syooy | IO sadejNs YoY | spoy | pasodxa Jo porayays uy | syoor pasodxy } ASBO[OIF I I I I I I I I I I ‘uml s'9-¢"¢ | Pearasqo JON wm 7-1 3 wrs-z i wm 9-F 1 ‘wreIp Teydas I I I I I Jen : : : : ul ¥'8-8°¢ wm ¢*/-S'°s um /-s°9 um 9-9 saiodsoosy XGZI- LIL | W228 ION | x €C-LT x 0¢C-9T x €I-O1 sdnom | ! spreMmo} pasieyua AyTenpess | ' sja]doip | s]jeo [eururiay p-¢ ay. um ¢-7s]]eo [fo yu ‘tur 7-9 syao un ¢-7 Jao Suryoueiq sdy ‘pasowojseuy PeAtasqo JON yeorde <poyoueig 0} aydunts Jeorde ‘pouspmm ATTes1dy yeorde ‘ojdunts ‘opi s9yIeY soshydeieg a8ueio ysIppar 1 a8UkIO [RUSIS 0} aduRIO ‘JARI ISIP ‘oTISSas -MOTAA OSTP “poyedsa1s3e a8uei0 YsIppor asueio yep 0} adueIO a8ueIO Yep IsIp 0} aeyided ur passourwt ‘gyeupe ‘snorowmnu | ursieul Jodoid 9 ostp | ‘QUIIO}eIg 0} BUTIONZ I ‘pastes AQysis ursreur | ‘QUIIOSZ SAUITJaUIOS J ‘QUIIOSZ 0] JUTIOURIIT ‘Q7e1 OUTIOWIG 1 ‘UOUIWIOD) 1 ‘QUIIOIZ 1 ‘guiiouedey ATUTeYY | epayjody asueio | erjaios } : : ; daap 0} a8uei0 ‘eIpayjode =; wrosyyound 10 erprseyq Aerzayed ; a8ueio ystUMOIG | yuid ystumorg AVIp oy punoze payrnsar Ayensn «+ = Moyyad Jaqy8tzq areI YM ‘padojaasp 1 ‘gsoqnurenbs =; yuId yp ‘ayeqoy, APounsip ‘pedoyaaap Aj100d Jo Juasqy ‘moTad YsTUMoIg ‘SUIBsIe|y Ajr00d 30 yuasqy ‘padopaaap [aM ‘prorpooryd ‘padojaaap [aM snyeqL, I I I I I sisuauvsvY] “I aviuviyopnasd ‘T, MADLsaf "X 1MOS]YXaL + SISUAITDAISND + WILOVAVHD vid VIOYJUVY pure “‘vubIYaY], “vU_IUOS]1T JO Sotdods sIeTIUMTs ATTeOISojoydiour YIM sisuauvsvYy] DULIUOS]1.J JO UOSTIedWIO’) ‘¢ ATAVI, Filsoniana lhasanensis sp. nov. (China) ... 673 Acknowledgments This study was supported by the National Natural Science Foundation of China (31093440, 31260008) and Natural Science Foundation of Xizang Autonomous Region, China (2016ZR-QY-05). The authors are grateful to Dr. Jan Vondrak (Institute of Botany, Academy of Sciences of the Czech Republic, Prihonice) and Prof. Xinli Wei (State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, Beijing) for peer review. Literature cited Arup U, Sechting U, Frédén P. 2013. A new taxonomy of the family Teloschistaceae. Nordic Journal of Botany 31: 16-83. https://doi.org/10.1111/j.1756-1051.2013.00062.x Darriba D, Taboada GL, Doallo R, Posada D. 2012. jModelTest 2: more models, new heuristics and parallel computing. Nature Methods 9(8): 772. https://doi.org/10.1038/nmeth.2109 Doring H, Clerc P, Grube M, Wedin M. 2000. Mycobiont specific PCR primers for the amplification of nuclear ITS and LSU rDNA from lichenised ascomycetes. 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(China) ... 675 Wei JC. 1991. An enumeration of lichens in China. Beijing, International Academic Publishers. 278 p. Wei JC. 2017. An enumeration of lichenized and lichenicolous fungi in China. Beijing, China Forestry Publishing House. 596 p. Wei JC, Jiang YM. 1986. Lichens of Xizang. Beijing, Science Press. 130 p. White TJ, Bruns T, Lee S, Taylor J. 1990. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. 315-322, in: MA Innis & al. (eds). PCR protocols: a guide to methods and applications. New York, NY: Academic Press. https://doi.org/10.1016/B978-0-12-372180-8.50042-1 Zhang BP, Chen XD, Li BL, Yao YH. 2002. Biodiversity and conservation in the Tibetan Plateau. Journal of Geographical Sciences 12(2): 135-143. Zhou S, Stanosz GR. 2001. Primers for amplification of mtSSU rDNA, and a phylogenetic study of Botryosphaeria and associated anamorphic fungi. Mycological Research 105(9): 1033-1044. https://doi.org/10.1016/S0953-7562(08)61965-6 Zoller S, Scheidegger C, Sperisen C. 1999. PCR primers for the amplification of mitochondrial small subunit ribosomal DNA of lichen-forming ascomycetes. Lichenologist 31(5): 511-516. https://doi.org/10.1006/lich.1999.0220 MY COTAXON ISSN (print) 0093-4666 (online) 2154-8889 Mycotaxon, Ltd. ©2019 October-December 2019— Volume 134, pp. 677-679 https://doi.org/10.5248/134.677 Sarcopodium flocculentum, the correct name for S. macalpinei SHAUN R. PENNYCOOK" & PAUL M. KIRK? 'Manaaki Whenua - Landcare Research Private Bag 92170, Auckland 1072, New Zealand ? Royal Botanic Gardens Kew, Surrey, TW9 3DS, UK * CORRESPONDENCE TO: PennycookS@LandcareResearch.co.nz ABSTRACT— The nomenclatural background of the species treated as Sarcopodium macalpinei is briefly outlined, and the requirement to supersede that name with a combination based on Nectriella flocculenta [= Lanatonectria flocculenta] is explained. Key worps—Actinostilbe flocculenta, Actinostilbe macalpinei, Kutilakesopsis macalpinei, ICN, code-compliance In a morphological study, Sutton (1981) synonymised Actinostilbe Petch and Kutilakesopsis Agnihothr. & G.C.S. Barua with the earlier genus Sarco- podium Ehrenb. and published a new combination Sarcopodium macalpinei for the type of Kutilakesopsis. Rossman & al. (1999) proposed Lanatonectria Samuels & Rossman, typified by a sexual morph for which they published the new combination L. flocculenta, additionally proposing a new combination Actinostilbe macalpinei for the conspecific asexual morph. In a multi-gene analysis, Lombard & al. (2015) included sequences from the type species of Sarcopodium (S. circinatum), Actinostilbe (A. vanillae = S. vanillae), and Kutilakesopsis (K. macalpinei = S. macalpinei), demonstrating that they were all included within a monophyletic Sarcopodium clade, thereby confirming the synonymy of these three genera. They also included 678 ... Pennycook & Kirk Lanatonectria as a synonym based on the established synonymy of its type (L. flocculenta) with Sarcopodium macalpinei; consequently, they recombined four of the five Lanatonectria species in Sarcopodium but did not recombine L. flocculenta (presumably because of its existing heterotypic Sarcopodium synonym). Rossman & al. (2016) recommended Sarcopodium as the accepted generic name in preference to Lanatonectria and Actinostilbe, a decision based on priority, widespread use, and the greater number of existing names; but they continued to cite L. flocculenta as S. macalpinei. However, Sarcopodium macalpinei is not the correct name for this species, since it fails to use the epithet of the earliest available legitimate name, as required by ICN (Shenzhen) Art. 11.4 (Turland & al. 2018). The correct name has been proposed by Pennycook & Kirk (2019): Sarcopodium flocculentum (Henn. & E. Nyman) Pennycook & P.M. Kirk, Index Fungorum no. 418: 1. 2019 = Nectriella flocculenta Henn. & E. Nyman, Monsunia 1: 160. 1899 [“1900”] = Nectria flocculenta (Henn. & E. Nyman) Hohn., Sitzungsber. Kaiserl. Akad. Wiss., Wien, Math.-Naturwiss. Cl., Abt. 1, 121: 360. 1912 = Lanatonectria flocculenta (Henn. & E. Nyman) Samuels & Rossman, Stud. Mycol. 42: 138. 1999 = Actinostilbe flocculenta (Henn. & E. Nyman) Rossman, Samuels & Seifert, IMA Fungus 4(1): 46. 2013 = Kutilakesopsis macalpinei Agnihothr. & G.C.S. Barua, J. Indian Bot. Soc. 36: 309. 1957 [as ‘macalpineae’ | = Sarcopodium macalpinei (Agnihothr. & G.C.S. Barua) B. Sutton, Trans. Brit. Mycol. Soc. 76: 99. 1981 = Actinostilbe macalpinei (Agnihothr. & G.C.S. Barua) Seifert & Samuels, Stud. Mycol. 42: 138. 1999 Acknowledgments We wish to thank Tom May (Royal Botanic Gardens, Melbourne) and Scott Redhead (Agriculture and Agri-Food Canada, Ottawa) for refereeing the manuscript. Literature cited Lombard L, van der Merwe NA, Groenewald JZ, Crous PW. 2015. Generic concepts in Nectriaceae. Studies in Mycology 80: 189-245. https://doi.org/10.1016/j.simyco.2014.12.002 Pennycook SR, Kirk PM. 2019. Nomenclatural novelties: S$. Pennycook & P.M. Kirk. Index Fungorum no. 418. 1 p. http://www.indexfungorum.org/Publications/Index%20Fungorum%20n0.418.pdf Rossman AY, Samuels GJ, Rogerson CT, Lowen R. 1999. Genera of Bionectriaceae, Hypocreaceae and Nectriaceae (Hypocreales, Ascomycetes). Studies in Mycology 42. 248 p. Sarcopodium flocculentum (= S. macalpinei) explained ... 679 Rossman AY, Allen WC, Braun U, Castlebury LA, Chaverri P, Crous PW, Hawksworth DL & al. 2016. Overlooked competing asexual and sexually typified generic names of Ascomycota with recommendations for their use or protection. IMA Fungus 7: 289-308. https://doi.org/10.5598/imafungus.2016.07.02.09 Sutton BC. 1981. Sarcopodium and its synonyms. Transactions of the British Mycological Society 76: 97-102. https://doi.org/10.1016/s0007-1536(81)80012-5 Turland NJ, Wiersema JH, Barrie FR, Greuter W, Hawksworth DL, Herendeen PS & al. 2018. International Code of Nomenclature for algae, fungi, and plants (Shenzhen Code) adopted by the Nineteenth International Botanical Congress, Shenzhen, China, July 2017. Regnum Vegetabile 159. https://doi.org/10.12705/Code.2018 MYCOTAXON ISSN (print) 0093-4666 (online) 2154-8889 Mycotaxon, Ltd. ©2019 October-December 2019—Volume 134, pp. 681-705 https://doi.org/10.5248/134.681 Pteridicolous ascomycetes from a cloud forest in eastern Mexico ROSARIO MEDEL-ORTIZ?, YAJAIRA BAEZA?, & FRANCISCO G. LOREA-HERNANDEZ? ‘Centro de Investigacion en Micologia Aplicada, Universidad Veracruzana, Médicos No. 5, Col. Unidad del Bosque, 91010, Xalapa, Veracruz, México. *Facultad de Agronomia, Universidad Veracruzana, Zona Universitaria, 91000, Xalapa, Veracruz, México *Instituto de Ecologia, A.C. Red de Sistemdtica y Biodiversidad, Carretera antigua a Coatepec 351, 91070, Xalapa, Veracruz, México. “CORRESPONDENCE TO: romedel@uv.mx ABSTRACT— Twenty-one ascomycetous taxa collected on ferns in a cloud forest of Veracruz, Mexico are reported and described. Fourteen represent new records for the Mexican mycobiota: Bisporella pteridicola, Crocicreas quinqueseptatum, Dasyscyphella dryina, Durella macrospora, Hamatocanthoscypha helicotricha, Hyaloscypha fuckelii, Lachnum brevipilosum, L. nudipes, Mollisia ventosa, Orbilia auricolor, Protocreopsis pertusa, Seimatosporium tostum, Stictis carnea, and S. radiata. Two collections—Cyathicula and cf. Arachnopeziza—that could not be fully identified may represent new species. The remaining five taxa have previous Mexican records but are reported here from new fern hosts. This study increases to 25 the number of ascomycete species known to occur on ferns in the cloud forests of Mexico. Key worps—Cyatheaceae, Helotiales, Hypocreales, Ostropales, Xylariales Introduction The Ascomycota, with around 90,000 species (Cannon & al. 2018), is the most diverse group in kingdom Fungi. Its diversity significantly complicates gaining knowledge of individual species, especially those with minute fruitbodies restricted to specialized substrates. Such is the case of fungi growing on ferns. Most fungi reported on ferns are saprobes, a group that 682 ... Medel-Ortiz, Baeza, Lorea-Hernandez receives less attention than parasites (Mehltreter et al. 2010). Currently 69 ascomycete species have been recorded in cloud forest of Veracruz (Medel 2013), small in comparison to the over 500 fern species known to inhabit cloud forests (Tejero-Diez & al. 2011). Although the first fern- related fungal study was devoted to rust fungi (Stevenson 1945), subsequent studies have been more focused on ascomycetes and their anamorphs (e.g., Mycosphaerellaceae and Periconiella; Braun 2004, Kirschner & Liu 2014). Other studies have centered on endophytic ascomycetes and fern-associated saprobes (Cannon 1997, Castaheda-Ruiz & Heredia 2000, Del Olmo & Arnolds 2014, Dingley 1972, Medel & Lorea-Hernandez 2008, Samuels & Rogerson 1990). For Mexico there are relatively few studies on the diversity of fern- inhabiting fungi (Castafieda-Ruiz & Heredia 2000, Haines 1980, Medel & Lorea-Hernandez 2008, Medel & al. 2010, Samuels & Rogerson 1990). So far, the seven ascomycete species recorded growing on ferns represent only three genera: Crocicreas Fr. (1), Dimeriella Speg. (1), and Lachnum Retz. (5). The few Ascomycota recorded from these forests contrasting with the great diversity of fern species suggests a high diversity yet to be explored. The present study was conducted in order to provide more information about the diversity of fern-inhabiting fungi in Mexican cloud forests. Materials & methods Sampling sites in central Veracruz state (eastern Mexico) were selected based on the high fern diversity within an area. Three sampling sites were chosen: El Riachuelo, Los Encinales, and Rio Xocoyolapan. Field work was conducted during August-November 2011. Fungi were collected by opportunistic sampling of different fern species, with fronds and rachises presenting ascomata or stromata selected and placed in paper bags (Mueller & al. 2004). Fruiting bodies were identified and processed following standard mycological techniques for Ascomycota (Dennis 1978). Specimens were hand _ sectioned, pretreated with 3% aqueous KOH, and stained with phloxine and Melzer’s reagent; ascal tip bluing reactions are designated by J- (negative, no blue present) and J+ (positive, blue present). Tissues were examined microscopically using a Zeiss Primo Star microscope. Averages, ranges, and Q-calculations (Largent & al. 1977) are based on 30 measurements. Colours are coded according to Kornerup & Wanscher (1978). Ascomata were photographed digitally with a Sony Cyber-Shot camera mounted on a Zeiss Stemi DV4 stereomicroscope. The specimens were identified using specialized literature (Breitenbach & Kranzlin 1984, Carpenter 1981, Dennis 1949, 1956, 1960, 1978). Fungal nomenclature follows Index Fungorum (2019), and host plants were identified following Mickel & Smith (2004). All specimens are Pteridicolous ascomycetes (Mexico) ... 683 deposited in the mycological collection of the Herbarium, Instituto de Ecologia A.C. (XAL). Specimens discovered in XAL and also examined are noted as “additional specimens examined” in species descriptions where pertinent. Taxonomy cf. Arachnopeziza FIGS 1-3 APOTHECIA sessile, cup-shaped, reddish-golden (6C7) to Pompeian yellow (5C6), <1 mm diam., margin hairy (sparsely pilose), hairs white (1A1), readily dropping from the margin to give a smooth appearance. SUBICULUM white to pale yellowish white (1A1, 2A1), not scattered on the substrate, reticulate or net-like, with hyphae 2-3 um diam., septate, hyaline, thick-walled (<1 um). Ascr cylindrical, 70-90(-100) x 7-9(-10) um, thin- walled, hyaline, 1-2-seriate, pore J+. Ascospores cylindrical-ellipsoid, (13-)14-15 x 5 um (Q = 2.8), smooth, hyaline, 0-1-septate, biguttulate, with sheath and hyaline appendages observed in Cresyl blue. PARAPHYSES filiform, 1-2 um diam., hyaline, septate toward the base. EcraL ExcrpuLUM a textura globulosa to textura angularis, cells 12—25(—40) um diam. HaBItTat—Gregarious, on rachises of Alsophila firma. Known only from these collections. SPECIMENS EXAMINED—MEXICO. VERACRUz: San Andrés Tlalnelhuayocan County, Rio Xocoyolapan, 19°30.89’N 97°00.39’W, 1590 m, 13 October 2011, Cordova 78. ADDITIONAL SPECIMENS EXAMINED: MEXICO. Veracruz: San Andrés Tlalnelhuayocan County, Rancho Agitita Fria, 6 April 2007, Medel 1362; 1363-A. Arachnopeziza Fuckel is characterized by small apothecia growing on a densely reticulate, net-like subiculum covered by hairs, and ellipsoid-fusoid to filiform, 1-7-septate ascospores (Korf 1951). The genus has a worldwide temperate distribution but is uncommon in the tropics (Gamundi & Giaiotti 1994). Although the studied specimens suggest an affinity with Arachnopezizaceae, they do not match any known genus or species covered in the references consulted (Dennis 1949, Huhtinen 1987, Korf 1951). The 0-1-septate spores covered by a hyaline sheath with a small appendage resemble those of Arachnopeziza aurelia (Pers.) Fuckel, which differs in its long hairs, slightly larger spores (17-22 x 3.5-5.5 um, O0-3-septate; Korf 1951). The presence on the apothecial margin of very thick, scarce, and septate hairs that drop off, leaving a smooth margin (as in our specimen; Fic. 1) is a feature unknown in Arachnopeziza. The subiculum is restricted to the apothecial base and not distributed throughout the substrate, unusual in Arachnopeziza (Seaver 1938), but not uncommon in Parachnopeziza Kort. 684 ... Medel-Ortiz, Baeza, Lorea-Hernandez In Parachnopeziza, however, apothecia grow directly from a small stipe immersed in the substrate (Korf 1978) growing directly from a subiculum, as occurs in Arachnopeziza, and certainly in our specimen. Bisporella pteridicola F. Ren & W.Y. Zhuang, Mycosystema 36 (4): 408 (2017). FIGS 4, 5 APOTHECIA superficial, cup-shaped to discoid. Disc concave, yellow (4A6), pastel yellow (2A4), or chicken yellow (2A6), translucent, waxy in appearance, 0.2-0.4 mm diam. Stipe short or absent, concolorous with the disc. Asci claviform (32—)35-40(-45) x 5-6 um, 8-spored, biseriate, apical pore J-, arising from croziers. AScosPoREs fusiform (5 —)8-9(-10) x 2-3 um (Q = 4.3), hyaline, with two oil drops and one septum visible in KOH. PARAPHYSES filiform, slightly enlarged at the apex, 1.5-2 um diam. ECTAL EXCIPULUM a textura angularis, gelatinized, 15-40 um thick, cells 3-6 um diam., protruding from outer surface <3 um. MEDULLARY EXCIPULUM a textura intricata, thin, hyaline, not gelatinized, cells 2-3 um diam. HasitatT—On rachises of unidentified fern as well as on fern rachises and leaf blades of Alsophila firma, Cyathea bicrenata, Sphaeropteris horrida, Pecluma alfredii, and Thelypteris sp. SPECIMENS EXAMINED—MEXICO. VeRAcRUz: San Andrés Tlalnelhuayocan County, Rio Xocoyolapan, 19°30.89’N 97°00.39’W, 1590 m, 13 October 2011, Cordova 80c; 4 November 2011, Castillo del Moral 63, 66; 8 December 2011, Medel & Lorea 2203. El Riachuelo; 19°30.99’N 97°00.39’W, 1626 m, 18 August 2011, Baeza 47a, Cordova 67, 71; 11 November 2011, Medel 2192. Los Encinales, 19°31.10’N 97°00.32’W, 1630 m, 10 August 2011, Baeza 36; 18 August 2011, Baeza 49,55, 57; Medel & Lorea 2158, Cérdova 73, 74. Bisporella Sacc. is a genus easily recognized by its small sessile bright yellow apothecia growing on woody substrata (Carpenter & Dumont 1978a, Dumont 1981). Bisporella pteridicola was described growing on ferns in Asia (Zhuang & al. 2017). Ours is the first report of the species from Mexico. Our collections agree with the description in Zhuang & al. (2017) in color, size, and shape of the apothecia, ascospore measurements, and ecology (growth on ferns). Coenogonium botryosum C. Knight, Syn. Queensl. Fl. 1(Suppl.): 74 (1886). Fics 6, 7 APOTHECIA superficial, discoid, sessile, flattened, 0.5-1 mm diam., pale yellow (3A3), waxy in appearance, margin differentiated from the disc, smooth, yellowish white (3A2), paler than the disc. THaLtus filamentous or felt-like and pilose, with short, upright algal threads resembling hairs, Pteridicolous ascomycetes (Mexico) ... 685 greyish yellow to linden green (2B5-2C5). Hymenium J+. Harrs small and thin, linden green (2C5) to pale yellow (1A3-2A3), filiform, septate. Ascli cylindric-clavate (48—)50-55(-60) x 4-5 um, thin walled, uniseriate. Ascosporss ellipsoid (7-)8-9(-12) x 2 um (Q = 4.2), hyaline, smooth, 0-1-septate, with 4 small guttules. ParApHysEs cylindrical, apices 4-5 um diam., bifurcate, hyaline, septate. ECTAL EXCIPULUM a textura globulosa, with hyaline cells. Hasitat—Gregarious, on fronds of Adiantum andicola and Grammiitis sp. SPECIMENS EXAMINED—MEXICO. VERACRUz: San Andrés Tlalnelhuayocan County, El Riachuelo, 19°30.99’N 97°00.39’W, 1626 m, 8 December 2011, Baeza 70, Medel & Lorea 2211, 2215. Coenogonium Ehrenb. is a tropical and subtropical ascolichen genus found on all continents (GBIF 2019, Rivas-Plata & al. 2006, Uyenco 1963). The filamentous green-pale yellowish thallus attached to the substrate by small filaments as well as the threadlike algae, absence of septa, uniseriate asci, and paraphyses with capitate subglobose apices are characteristic of C. botryosum (Rivas-Plata & al. 2006). Two similar species are C. moniliforme Tuck., which has moniliform algae without septa, and C. implexum Nyl., which has bright yellow thalli and a cottony texture. Known from Neotropics and eastern Paleotropics (GBIF 2019, Rivas-Plata & al, 2006, Uyenco 1963), Coenogonium botryosum was reported previously for Veracruz, Mexico, growing on Ficus (Uyenco 1963); this is the first report of the species on ferns. Crocicreas quinqueseptatum S.E. Carp., Mem. New York Bot. Gard. 33: 160 (1981). Fics 11, 12 APOTHECIA cup-shaped, 0.5-0.8 mm diam. Disc white to yellowish white (2A2, 2A1). MARGIN denticulate, inrolled over the hymenium when dry, almost hyaline or translucent with long teeth; teeth fragile, formed by groups of long, septate, hyaline hyphae, 100-115(-400 um) x 40-48 um, lighter in colour than the rest of the apothecium, white to yellowish white (2A1-2A2), 8-12 teeth per apothecium; breaking easily leaving the apothecia non-denticulate. Stipe cylindrical, thin, concolorous with apothecia. Ascli cylindrical, tapering toward the base, (60-—)80-105 x 6-8 um, thin- walled, apical pore J+, biseriate, base without croziers. AscosporEs fusoid to fusiform with subacute to acute ends, smooth, (20-)26-30 x 3-4 um (Q = 5.5), 5-septate (rarely 6-7), hyaline. PARAPHYSES filiform, unbranched, 686 ... Medel-Ortiz, Baeza, Lorea-Hernandez 1-2 um diam., hyaline. EcraL ExcipuLum single layered, gelatinous, composed of long cells in the inner part, extending outward from the disc to form the teeth. MEDULLARY EXCIPULUM not differentiated. Hasitat—On fallen stems and rachises of Alsophila firma and Cyathea bicrenata. SPECIMEN EXAMINED— MEXICO. VERACRUZ: San Andrés Tlalnelhuayocan County, Los Encinales, 19°31.10’N 97°00.32’W, 1630 m, 10 August 2011, Medel & Lorea 2168. Crocicreas quinqueseptatum was described by Carpenter (1981) as having ascospores with a variable number (4-7) of septa and which are slightly longer and narrower [(28—)30-33(-35) x (2-)2.5-3 um] than those found in our specimen. Other features (outer apothecial surface, morphology, and measurements) are close to C. helios var. parahelios S. E. Carp., which differs by 0-3-septate ascospores, an absent or poorly developed medullary excipulum, and broader apothecia (<1.5 mm; Carpenter 1981). Crocicreas quinqueseptatum has been reported growing on angiosperm stems, especially fallen herbaceous stems of Asteraceae in South America. This is the first report of this species on ferns. Cyathicula sp. Fics 8-10 APOTHECIA cup-shaped, sessile, disc flattened or concave, <1 mm diam., pastel yellow (3A4); margin dentate with 6-10 hyaline teeth per ascoma, teeth composed of groups of long, hyaline, aseptate hyphae that extend beyond the margin, each tooth separated by c. 420-480 um. Ascr cylindric-clavate, (96-) 110-140 x (7-9)10-11 um, 8-spored, biseriate, apical pore J-. ASCOSPORES ellipsoidal-fusiform, 13-16 x 5-6 um (Q = 2.6), smooth, hyaline, 1-3-septate, guttulate. PARAPHYSES filiform, apices simple or branched, 2-3 um diam. at apex, septate; embedded with asci in a gelatinous matrix and separated only with difficulty. ECrAL EXCIPULUM a textura prismatica to textura porrecta, hyphae single-layered, long-celled, subparallel, close together, thick-walled, hyaline, smooth, lacking crystals. MEDULLAR EXCIPULUM little differentiated, hyphae 3-5 um diam., not easily observed due to gelatinous consistency. Hasitat—On rachises of Cyathea bicrenata. Known only from our collections. SPECIMENS EXAMINED—MEXICO. VERACRUz: San Andrés Tlalnelhuayocan County; E] Riachuelo, 19°30.99’N 97°00.39’W, 1626 m, 4 November 2011, Baeza 56; Los Encinales, 19°31.10’N 97°00.32’W, 1630 m, 13 October 2011, Baeza 56a; 4 November 2011, Baeza 61, Castillo del Moral 64. We place our material into Cyathicula based on its cupulate, sessile apothecium with a toothed margin, whitish yellowish to pale brown color, Pteridicolous ascomycetes (Mexico) ... 687 Lair , $10 pm [ 1mm Figs 1-15. cf. Arachnopeziza. 1. Apothecia showing scarce subiculum and hairs (top right); 2. Ascospores in Congo red; 3. Excipulum showing hyphae from the subiculum. Bisporella pteridicola. 4. Apothecia; 5. Asci with ascospores. Coenogonium botryosum. 6. Apothecia; 7. Ascospores. Cyathicula sp.; 8. Apothecia; 9. Cross section of excipulum, 10. One-septate ascospores. Crocicreas quinqueseptatum. 11. Apothecia showing dentate margin; 12. Ascospores. Dasyscyphella dryina. 13. Apothecium; 14. Ascospores. Durella macrospora. 15. Apothecia. 688 ... Medel-Ortiz, Baeza, Lorea-Hernandez septate ascospores, J- asci, and the presence of a gelatinous matrix (Carpenter & Dumont 1978b). Nonetheless, we must also note that the specimens fit diagnostic characters of Crocicreas (Carpenter 1981). The controversy over whether Crocicreas and Cyathicula are congeneric has lasted for decades (Iturriaga & al. 1999, Zheng & Zhuang 2016). According to one classification of Helotiales (Baral & al. 2015), Crocicreas is restricted only to the type species C. gramineum (Fr.) Fr., and Cyathicula is unrelated. Although Wijayawardene & al. (2018) support this distinction, we agree with the conclusion of Zheng & Zhuang (2016) that existing data are insufficient for determining the relationship between Crocicreas and Cyathicula. Molecular analysis is still needed for us to assign our material definitively to a genus. Dasyscyphella dryina (P. Karst.) Raitv., Akad. Nauk Estonskoi S.S.R., Inst. Zool. Bot., Tartu: 72 (1970). Fics 13, 14 APOTHECIA superficial, cup-shaped, stipitate, covered with white (1A1) hairs. Disc 0.8-1 mm diam., yellowish white (2A2) to pale yellow (2A3), smooth. STIPE cylindrical, covered with white (1A1) hairs with small hyaline crystals at the tips. Harrs cylindrical, septate, slightly granular, hyaline, easily visible, giving a distinctive villous appearance to the apothecium. AscI cylindrical, 45-50 x 3-4 um, hyaline, 8-spored, irregularly biseriate, thin walled, apical pore J+. Ascospores cylindric-ellipsoid to fusoid, (6-—)7-10 x 2-3 um, (Q = 4.2), smooth, hyaline, with rounded ends. PARAPHYSES filiform, slightly lanceolate, 1 um diam., hyaline, projecting beyond the asci by 10-20 um. Hasitat—On rachises of Alsophila firma. SPECIMENS EXAMINED—MEXICO. VeRAcRuz: San Andrés Tlalnelhuayocan County, Rio Xocoyolapan, 19°30.89’N 97°00.39’W, 1590 m, 19 August 2011, Medel 2180, Medel 2181. The studied specimen matches well with the concept of Lachnum distinguendum sensu Dennis (1949, 1956, 1960), which is a synonym of Dasyscyphella dryina. Dennis (1956) cited spores 2 um longer than those of our collections, and filiform paraphyses which agreed with our specimen. Rick (1906) described L. distinguendum Rick with broader apothecia (<2 mm diam.), larger asci (50-69 x 7-10 um), larger ascospores (10-15 x 3-4 um), and lanceolate paraphyses (3-4 um diam.). Raitviir (1970) cited slightly longer ascospores (9-12 x 2-3.2 um) and cylindrical paraphyses with pointed tips. Nonetheless, our collection fits better Dennis's concept, because our material has paraphyses that exceed the asci, while Raitviir (2002) described paraphyses not exceeding the asci. Pteridicolous ascomycetes (Mexico) ... 689 Dasyscyphella dryina is known from Europe and South America on Betula, Fagus, and Populus (Dennis 1949, 1956. 1960, Raitviir 1970, Rick 1906). Durella macrospora Fuckel, Jahrb. Nassauischen Vereins Naturk. 23-24: 281 (1870). Fics 15-17 APOTHECIA superficial, cup-shaped, stipe short with a wide base. Disc flat, 0.2-0.5 mm diam., wrinkled, brownish gray (10F2) with dark brown (7F8) tones, hard consistency. Asci cylindrical (90-)100-130 x 11-12 um, hyaline, 8-spored, irregularly biseriate, apical pore J-. Ascosporss ellipsoid- fusiform, (18—)19-22(-23) x 4-5 um (Q = 4.2), smooth, hyaline, 3-septate, constrained at the septum, ends rounded, with 4 small guttules. PARAPHYSES cylindrical, slightly wider at the apex (c. 3 um) brownish to reddish brown in KOH. EcTAL EXCIPULUM Of parallel brown hyphae. Hasitat—On fronds of Sticherus palmatus. SPECIMEN EXAMINED—MEXICO. VERACRUz: San Andrés Tlalnelhuayocan County, El Riachuelo, 19°30.99’N 97°00.39’W, 1626 m, 8 December 2011, Medel 2210. Spores of Durella macrospora have been cited as 15-21 x 3-5.5 um (Breitenbach & Kranzlin 1984, Medardi 2004). Although the spores in our collection are 2 um longer, other morphological characters (black apothecia, hard consistency, ectal excipulum of parallel brown hyphae) agree well with the published descriptions (Breitenbach & Kranzlin 1984, Medardi 2004). Durella macrospora has been previously recorded growing on wood of deciduous trees such as Fagus, Malus domestica, and Quercus sp. (Breitenbach & Kranzlin 1981, Medardi 2004), but not on ferns. Previously known only from Europe (Breitenbach & Kranzlin 1984, Dennis 1956, Medardi 2004). Hamatocanthoscypha helicotricha Huhtinen, Karstenia 29(2): 182 (1990 [“1989”]). Fries 18-20 APOTHECIA cup-shaped, disc concave, pale yellow (3.A3) to pastel yellow (3A4) shades, 0.5 mm diam., covered with fine, white (1A1) to yellowish white (2A1-2A2) short hairs. Stipe cylindrical, short, slightly robust, concolorous with apothecia. Hairs cylindrical, apically coiled to uncinate, 12-32 x 2-4 um, the apex tapering to <1 um diam., hyaline, septate toward to the base, smooth. Ascr cylindric-clavate, apex conical, base attenuate, (42—)45-72 x 6-7 um, hyaline, pore J-, 8-spored, biseriate, base with croziers. Ascospores ellipsoid-fusoid, hyaline, smooth, 7-9(-10) x 2-3 um (Q = 2.6), 1-septate, biguttulate. PARAPHysEs filiform, 2 um diam., narrowing toward the base. 690 ... Medel-Ortiz, Baeza, Lorea-Hernandez Hasitat—On rachises of Cyathea bicrenata and fronds of Sticherus palmatus. SPECIMENS EXAMINED—MEXICO. VeERAcRUZz: San Andrés Tlalnelhuayocan County, El Riachuelo, 19°30.99’N 97°00.39’W, 1626 m, 8 December 2011, Medel & Lorea 2213. Los Encinales, 19°31.10’N 97°00.32’W, 1630 m, 10 August 2011, Baeza 41, Huhtinen (1990) and Raitviir (2004) report somewhat larger (9.8-12.8 x 2.8-4 um) spores than found in our specimens, but the presence of strongly coiled hairs at the apex distinguishes this species. Hamatocanthoscypha helicotricha has been recorded on fern rachises from Colombia and Cuba (Huhtinen 1990, Raitviir 2004); this is the first report of the species for Mexico. Hyaloscypha fuckelii Nannf., Nova Acta Regiae Soc. Sci. Upsal., Ser. 4, 8(2): 273 (1932). Fics 21, 22 APOTHECIUM superficial, sessile, cup-shaped. Disc concave, 0.1-0.3 mm diam., white (1A1), translucent when fresh or when rehydrated in KOH. Receptacle with fine, whitish hairs. Harrs filiform, cylindrical, curved in the apex, thin-walled, slightly granular. Ascr clavate, 50-66(-67) x 4-5 um, hyaline, thin-walled, 8-spored, irregularly biseriate, apical pore J+, base with croziers. AscosporsEs ellipsoid, (5—)6-8(-10) x 2 um, (Q = 3.2), 0-1-septate, smooth, hyaline, with small oil drops. Parapuyses filiform, protruding <9 um beyond asci, narrow, smooth, hyaline, apical cell with small guttules. Hasirat—On rachises of Alsophila firma. SPECIMEN EXAMINED—MEXICO. VerRAcRUz: San Andrés Tlalnelhuayocan County, El Riachuelo, 19°30.99’N 97°00.39’W, 1626 m,10 August 2011, Baeza 38. Our collection agreed with Hyaloscypha fuckelii as described by Huhtinen (1990). The similar H. albohyalina (P. Karst.) Boud. differs in its narrow hairs and smaller spores. Hyaloscypha fuckelii is widely distributed in Europe and North America (Huhtinen 1990) and has been reported growing on monocotyledonous genera such as Juncus, Typha, and Molinia. This is a new record for Mexico and the first report on ferns. Lachnum brevipilosum Baral, Beih. Z. Mykol. 6: 74 (1985). Fics 23, 24 APOTHECIUM superficial, cup-shaped. Disc concave, white (1A1) to pale (2A2), 0.8-1 mm diam. Receptacle covered with whitish hairs visible Pteridicolous ascomycetes (Mexico) ... 691 to the naked eye. Stipe cylindrical, 0.6 mm long, covered by hairs generally concolorous with disc (white, 1A1) except for some with a yellowish gray (4B2) base. Hairs cylindrical, 130-150 x 4-5 um (the apex tapering to <2-3 um), 5-7-septate, hyaline, covered by granules. Asc clavate-cylindrical, 42-50 x 4-5 um, hyaline, pore J+, 8-spored, biseriate, croziers not observed. Ascosporss ellipsoid, smooth, with rounded ends, 8-—10(-12) x 2 um (Q 4.2), hyaline, 1-septate, biguttulate. PaRAPHysES filiform, septate, protruding by 5-10 um above the asci, hyaline. Hasitat—On rachises of Alsophila firma and Cyathea bicrenata. SPECIMENS EXAMINED—MEXICO. VERAcRuz: San Andrés Tlalnelhuayocan County, Rio Xocoyolapan, 19°30.89’N 97°00.39’W, 1590 m, 4 November 2011, Baeza 58. Paraje El Riachuelo, 19°30.99’N 97°00.39’W, 1626 m, 4 November 2011, Cordova 86. The Mexican specimens agree with the morphology of Lachnum brevipilosum as provided by Dennis (1949, as Dasyscyphus brevipilus Le Gal) and Ye & al. (2006), except for the length and width of the hair apices. Lachnum brevipilosum has been reported growing on stems of Clematis, Fagus, and Rubus from Europe and temperate China (Dennis 1949, Ye & al. 2006) This is a new record for Mexico and represents the first report on ferns. Lachnum nudipes (Fuckel) Nannf., Svensk Bot. Tidskr. 22: 124 (1928). Fics 25, 26 APOTHECIUM superficial, cup shaped. Disc concave slightly flattened, yellowish white (4A2) to pale yellow (4A3), 0.3-0.4 mm in diameter. Receptacle densely covered with white (1A1) hairs. Stipe cylindrical, with whitish hairs (1A1), more or less equal to the diameter of the disc. Harrs cylindrical, ornamented with granules, widening at apex to 3-4 um, longer than 80 um, hyaline in KOH. Asc clavate, (35-)38-47 x 4-5 um, 8-spored, biseriate, pore J+, base without croziers. AscosporEs fusiform to elliptical with rounded ends, (5—)6-8(-9) x 1-2(-3) um (Q = 3.1), smooth, hyaline. PARAPHYSES lanceolate, septate towards the base, (50-)68-70 x 3-4 um, clearly extending beyond the asci. Hasitat—On fronds of Lophosoria quadripinnata. SPECIMENS EXAMINED—MEXICO. VERACRUz: San Andrés Tlalnelhuayocan County, Rio Xocoyolapan 19°30.89’N 97°00.39’W, 1500 m, 18 August 2011, Cérdova 72, 13 October 2011, Baeza 51; 4 November 2011, Cordova 65, 84, Castillo del Moral 65. Our material agrees with Dennis's (1949) descriptions of Dasyscypha nudipes (Fuckel) Sacc. var. nudipes and D. nudipes var. minor Dennis [both now treated 692 ... Medel-Ortiz, Baeza, Lorea-Hernandez as synonyms of L. nudipes], but our ascomata and ascospore measurements are closer to his D. nudipes var. minor. Lachnum nudipes has been reported on Epilobium, Filipendula, and Spiraea in Europe (Dennis 1949, as Dasyscypha). This is the first record Lachnum nudipes for Mexico and the first report of its occurrence on ferns. Lachnum pteridophyllum (Rodway) Spooner, Biblioth. Mycol. 116: 470 (1987). Fics 27, 28 Macro and micromorphological characters agree with the description of Medel & Lorea- Hernandez (2008). Hasitat—On rachises of Cyathea divergens var. tuerckheimii, Cyathea sp., Dryopteris sp., and Lophosoria quadripinnata. SPECIMENS EXAMINED—MEXICO. VERACRUZ, San Andrés Tlalnelhuayocan County, El Riachuelo, 13 October 2011, Baeza 29, 52, 53; 4 November 2011, Baeza 60, Medel 2196; 8 December 2011, Medel & Lorea 2214. Los Encinales, 19°31.10’N 97°00.32’W, 1630 m, 10 August 2011, Baeza 36. 19 August 2011, Medel 2179, 2182, 2185. Lachnum pteridophyllum has been reported from Colombia, Dominican Republic, Jamaica, Mexico, Panama, Peru, Puerto Rico, New Guinea, New Zealand, Taiwan, Tasmania, and Venezuela (Haines 1980, Medel & Lorea- Hernandez 2008). Haines (1980) cited this species on Cyathea sp. from Oaxaca (Mexico), and Medel & Lorea-Hernandez (2008) from Veracruz on decaying leaves of Dicksonia sellowiana, Cyathea divergens var. tuerckheimii, and Cyathea spp. We add now two more hosts: Dryopteris sp. and Lophosoria quadripinnata. Lachnum singerianum (Dennis) W.Y. Zhuang & Zheng Wang, Mycotaxon 67: 27 (1998). Fics 29, 30 Macro and micromorphological characters agree with the description of Medel & Lorea- Hernandez (2008). Hasitat—On rachises of Alsophila firma and Dicksonia sellowiana. SPECIMEN EXAMINED—MEXICO. VERACRUz: San Andrés Tlalnelhuayocan County, Rio Xocoyolapan, 19°30.89’N 97°00.39’W, 1590 m; 8 December 2011, Medel 2222. Lachnum singerianum is known from the Andean region in South America from Bolivia to Venezuela and from China, Jamaica, and Mexico (Haines 1980, Medel & Lorea-Hernandez 2008). Haines (1980) earlier reported L. singerianum growing on rachises of ferns, while Medel & Lorea- Hernandez (2008) reported the species on decaying leaves of Alsophila firma and Dicksonia sellowiana in Veracruz. Pteridicolous ascomycetes (Mexico) ... 693 | 20 um -<¥ um Fics 16-31. Durella macrospora. 16. Three-septate ascospores; 17. Asci with 1-septate ascospores. Hamatocanthoscypha helicotricha. 18. Apothecia; 19. Asci and ascospores; 20. Uncinate or coiled hairs. Hyaloscypha fuckelii. 21. Apothecia; 22. Ascospores. Lachnum brevipilosum. 23. Apothecium; 24. Septate hairs. Lachnum nudipes. 25. Apothecium (left) and ascospores (right); 26. Lanceolate paraphysis. Lachnum pteridophyllum. 27. Apothecium; 28. Ascospores. Lachnum singerianum. 29. Apothecium; 30. Septate hair in 5% KOH. Lachnum varians. 31. Apothecia. 694 ... Medel-Ortiz, Baeza, Lorea-Hernandez Lachnum varians (Rehm) M.P. Sharma, Nova Hedwigia 43: 411 (1986). Fics 31, 32 Macro and micromorphological characters agree with the description of Medel & Lorea- Hernandez (2008). Hasitat—On rachises of Cyathea bicrenata and C. divergens var. tuerckheimii and fronds of Adiantum andicola. SPECIMENS EXAMINED—MEXICO. VeRAcRuz: San Andrés Tlalnelhuayocan County, El Riachuelo, 19°30.99’N 97°00.39’W, 1626 m 18 August 2011, Baeza 47b, Cordova 68, 69; 8 December 2011, Baeza 70. ADDITIONAL SPECIMENS EXAMINED: MEXICO. VERACRUZ: Xalapa County, Santuario del Bosque de Niebla, Parque Ecoldgico Francisco Javier Clavijero, 1 September 2013, Medel & Lorea 2259. Lachnum varians is known from Australia, Hawaii, Mexico, New Guinea, New Zealand, South America, and the Caribbean (Haines 1980, Medel & Lorea-Hernandez 2008). The species was previously reported only on decaying leaves of Alsophila firma (Medel & Lorea-Hernandez 2008); the present study adds the new hosts Cyathea bicrenata, C. divergens var. tuerckheimii, and Adiantum andicola. Lachnum virgineum (Batsch) P. Karst., Bidrag Kannedom Finlands Natur Folk 19: 169 (1871). Fics 33, 34 APOTHECIA superficial, cupulate, 1-1.3 mm diam. Disc flat to concave, white to yellowish white (1A1-1A2). Receptacle densely covered with short, broad, white (1A1) hairs. Stipe, thin, covered with hairs concolorous with the apothecia. Hairs cylindrical, 60 x 5-6(-8) um, granulose along the entire length, 3-4-septate, the apex broadening <8 um diam. AscI cylindric-clavate, 42-51x 4-5 um, 8-spored, biseriate, pore weakly blue (J+). AScosPorgs fusiform, ends rounded, with one end sometimes wider than the other, (6—)7-8 x 1-2 um (Q = 4.6). PaRAPHYSES lanceolate 4-5 um diam., 2—3-septate, with small refractive guttules in the apex, extending beyond the asci by 20 um. Hasrirat—On fronds of Phlebodium aureum. SPECIMENS EXAMINED—MEXICO. Veracruz: San Andrés Tlalnelhuayocan County, El Riachuelo, 10 August 2011, Baeza 32; 8 December 2011, Medel 2208. The Mexican specimens fit well the description of Lachnum virgineum (Spooner 1987), especially the shape, color, and size of the apothecium (densely covered by white hairs), spore size, and paraphyses exceeding the asci by 15-20 um. Lachnum virgineum is similar to L. controversum (Cooke) Rehm, which differs in obtuse or slightly swollen hair apexes and very slightly Pteridicolous ascomycetes (Mexico) ... 695 narrower (3.5-4.5 um diam.) paraphyses. Although L. virgineum is generally regarded as lignicolous, we found it growing on fronds of Phlebodium aureum. Known from Australia and Europe (Breitenbach & Kranzlin 1984, Spooner 1987). Mollisia ventosa P. Karst., Bidrag Kannedom Finlands Natur Folk 19: 188 (1871). Fics 35-37 APOTHECIA superficial, sessile, discoid, flat, 0.5-1 mm diam., greenish- yellow (1A7) with dark grey (1F1) to olive gray (1D2) margin. Receptacle greenish gray (29F2), smooth. Ascr (72-)85-115 x 7-8(-10) um, apical pore J+. Ascospores ellipsoid, (9-)12-15(-16) x 1.5-2 um (Q = 7.5), 1-3-septate, guttulate. PARAPHYSES filiform, slender, septate, branched or not. ECTAL EXCIPULUM a textura angularis to textura globulosa, cells thick- walled, brownish, 6-10(-12) x 6-10 um. Hasrirat—On rachises of Cyathea bicrenata. SPECIMENS EXAMINED—MEXICO. VeERaAcRuUz: San Andrés Tlalnelhuayocan County, El Riachuelo, 19°30.99’N 97°00.39’W, 1626 m, 4 November 2011, Medel 2196; 8 December 2011, Baeza 71, Baeza 72. Mollisia ventosa has been reported from Europe, and America (Breitenbach & Kranzlin 1984, Dennis 1950, 1978, Medel & Chac6én 1997). Dennis (1950, 1978) described M. ventosa with spores larger (10-20 x 2-3.5 um) than those in our Mexican collections, which instead match the measurements cited by Breitenbach & Kranzlin (1984). They cited M. ventosa as growing on Alnus and decorticated wood, but not on ferns. Mollisia undulatodepressula (Feltgen) Le Gal & EF Mangenot, the only other Mollisia species known to inhabit cloud forests in Mexico, grows on rotten wood (Medel & Chacén 1997). Orbilia auricolor (A. Bloxam) Sacc., Syll. Fung. 8: 625 (1889). Figs 38, 39 APOTHECIA Superficial, scattered, sessile, cup-shaped. Discconcave,0.8-1mm diam., yellow brownish to yellow gray (4B3-4B5) translucent when rehydrated, margin crenulate. RECEPTACLE concolorous with disc, smooth. Asct cylindric-clavate, 30-40(-42) x 2-4 um, hyaline, thin-walled, 8-spored, biseriate, pore J—, base forked. Ascosporgs curved-filiform, (8—)9-10 x 1 um (Q = 8.8), smooth, hyaline. PARAPHysEs cylindrical, apex capitate, with incrustations, 3-4 um diam., septate, branched at base. ECTAL EXCIPULUM textura globulosa with hyaline cells. 696 ... Medel-Ortiz, Baeza, Lorea-Hernandez Hasitat—On rachises of Cyathea bicrenata and Marattia laxa. SPECIMENS EXAMINED—MEXICO. VERACRUZ: San Andrés Tlalnelhuayocan County, Rio Xocoyolapan 19°30.89’N 97°00.39’W, 1590 m, 8 December 2011, Baeza 67, Medel 2202. The morphology of the apothecia, ascospores, asci, and paraphyses in our Mexican specimens agree with Orbilia auricolor as described by Spooner (1987) and Mo & al. (2005). The asexual stage of O. auricolor has been recorded (as Arthrobotrys oligospora Fresen.) from Xochimilco, Mexico (Chavarria & al. 2010). Orbilia auricolor is known from Asia, Australia, Europe (Breitenbach & Kranzlin 1984, Dennis 1981, GBIF 2019, Mo & al. 2005, Spooner 1987). The species has previously been recorded growing on dead wood and woody stems and other plant matter (Spooner 1987, Mo & al. 2005), but not on ferns. Protocreopsis pertusa (Pat.) Samuels & Rossman, Stud. Mycol. 42: 66 (1999). FIGs 40, 41 PERITHECIA aggregated, globose, sessile, 0.8-1 mm diam., covered by whitish (2A1) to yellowish white (3A2) hairs. Hairs cylindrical, curved to wavy, smooth, thick-walled, hyaline, apices simple or forked, septate. SuBICcULUM of thin whitish (2A1) hyphae, attached to the perithecia. Asci clavate-cylindrical, 75-95(-100) x 10-12 um, thin-walled, hyaline, ascospores irregularly biseriate, 8-spored, base with croziers. ASCOSPORES ellipsoid, (13—)14-16 x 5 um (Q = 2.7), smooth, hyaline, 1-septate, with two large guttules in the ends. PARAPHYSES filiform and hyaline. Hasrrat—On rachises of Alsophila firma and Marattia laxa. SPECIMENS EXAMINED—MEXICO. VeERAcRuz: San Andrés Tlalnelhuayocan County, Rio Xocoyolapan, 19°30.89’N 97°00.39’W, 1590 m, 13 October 2011, Cordova 77; 8 December 2011, Baeza 68, Medel & Lorea 2205. ADDITIONAL SPECIMENS EXAMINED: MEXICO. VERAcRUz: San Andrés Tlalnelhuayocan County, Rancho Agiiita Fria, 6 April 2007, Medel 1362. Rossman & al. (1999) and Chaverri & al. (2010) described Protocreopsis pertusa as having thin white mycelium beneath each perithecium, thick-walled hairs densely united around the perithecial opening, and 1-3-septate spores measuring 13-17 x 4-5 um, all characters agreeing with our Mexican specimens. This species has been reported from tropical to subtropical regions on leaves of Musa, Heliconia, palms, bamboos, and fern rachises. It is a new record for Mexico. Pteridicolous ascomycetes (Mexico) ... 697 aS 40° ; 1b, ‘| ea coe wee ? : FS ae os >= -@¢ te £ ——! A ay ‘ ee Figs 32-48. Lachnum varians. 32. Asci with ascospores. Lachnum virgineum. 33. Apothecia; 34. Hair with enlarged apex. Mollisia ventosa. 35. Apothecia; 36. Excipulum at margin and lower flank; 37. Asci with ascospores. Orbilia auricolor. 38. Apothecia 39. Paraphysis. Protocreopsis pertusa. 40. Apothecia 41. Ascospores. Seimatosporium tostum. 42. Stromata 43. Asci with ascospores, 44. Ascospores. Stictis carnea. 45. Apothecia. 46. Ascospores. Stictis radiata. 47. Apothecia; 48. Asci and ascospores. 698 ... Medel-Ortiz, Baeza, Lorea-Hernandez Seimatosporium tostum (Berk. & Broome) Rossman & W.C. Allen, IMA Fungus 7(1): 5 (2016). Figs 42-44 StromartaA shield-like, growing within host tissue, black, grey (30F1) to greenish grey (30F2) pigments are observed in KOH. Two perithecia per stroma. Asctr cylindrical, 62-75 x 5-7(-8) um, thin-walled, apical pore J+, spores 1-2-seriate, without croziers. Ascosporegs ellipsoidal-fusiform, 1-septate, constricted at the septum, smooth, poles subacute to acute, (9-)10-11(-12) x 3-4 um (Q = 2.3). PARAPHYSES filiform, <1 um diam., extending beyond the ascus by 3-5 um, with tiny hyaline guttules at the apex. Hasitat—On rachises of Alsophila firma and Cyathea bicrenata. SPECIMENS EXAMINED—MEXICO. VERACRUZ: San Andrés Tlalnelhuayocan County, Rio Xocoyolapan, 19°30.89’N 97°00.39’W, 1590 m, 8 December 2011, Medel & Lorea 2108; 26 February 2001, Medel & Lorea 2201; Los Encinales, 19°31.10’N 97°00.32’W, 1630 m, 30 March 2014, Medel & Lorea 2379. ADDITIONAL SPECIMENS EXAMINED: MEXICO. Veracruz: San Andrés Tlalnelhuayocan, Rancho Agiiita Fria, 7 February 2007, Medel 1342. Seimatosporium tostum is described by Paulus & al. (2006) as having larger (>14 x 5 um) hyaline ascospores with only one septum, while Brockmann (1976) described North American material (as Discostroma tostum) with 1-6-septate ascospores. The species has been reported growing on stems of Epilobium. Known from North America (Brockmann 1976, Paulus & al. 2006), S. tostum establishes a new record for Mexico and the first report on ferns. Stictis carnea Seaver & Waterston, Mycologia 33: 311 (1941). Fics 45, 46 APOTHECIA immersed in host tissue, hymenium greyish yellow to light yellow (1A5-1B6), 05-0.7 mm diam., surrounded by an elliptical white (1A1) ring. Ascr cylindric-clavate, 110-150 x 6-7 um, 8-spored, hyaline, J-. Ascosporss long filiform, (80-)85-130(-150) x 3 um, tapering to one end, multi-septate, twisted within the ascus. PARAPHYSES filiform, <3 um wide, longer, hyaline, septate. Hasitat—On rachises of Cyathea bicrenata. SPECIMEN EXAMINED—MEXICO. VERACRUz: San Andrés Tlalnelhuayocan County; Rio Xocoyolapan; 19°30.89’N 97°00.39’W, 1590 m, 4 November 2011, Medel 2112. There are two species of Stictis growing on ferns in our study site, S. carnea and S. radiata. ‘The differences in spore length and apothecial morphology of the apothecia are sufficient to separate these two species (Sherwood 1977, Pteridicolous ascomycetes (Mexico) ... 699 Johnston 1983). Stictis carnea has been reported from Asia, North America (including Mexico), and New Zealand (Johnston 1983, Sherwood 1977), growing on dead tissues of many herbaceous and woody plants and the fern Cyathea medullaris (Johnston 1983). This species is a new report from Mexico. Stictis radiata (L.) Pers., Observ. Mycol. 2: 73 (1800 [“1799”]). Fics 47, 48 APOTHECIA deeply sunken in host tissue, hymenium vivid yellow to yellow (3A7-3A8), <1 mm diam., surrounded by an irregular white (1A1) ring. Asc1 cylindric-clavate, 240-290 x (7-)10-12 um, 8-spored, hyaline, narrowed toward the apex, pore J-. Ascospores long filiform, (219-)245-280(-283) x 2-3 um, multiseptate, spirally arranged within the ascus. PARAPHYSES filiform, 1 um diam., slightly widening (<2 1m) at the apex. Hasitat—On _ rachises of Alsophila firma, Cyathea _ bicrenata, Sphaeropteris horrida, and fronds of Polypodium sp. SPECIMENS EXAMINED—MEXICO. VeERAcRuz: San Andrés Tlalnelhuayocan County; Rio Xocoyolapan; 19°30.89’N 97°00.39’W, 1590 m, 4 November 2011, Medel 2199; 8 December 2011, Baeza 69, Medel & Lorea 2200. Paraje El Riachuelo, 8 December 2011; Medel & Lorea 2207. Los Encinales, 19°31.10’N 97°00.32’W, 1630 m, 10 August 2011, Baeza 38b, Medel & Lorea 2169. Diagnostic characters of Stictis radiata include ascomata with a deeply sunken orange yellow hymenium and filiform multiseptate spores with interlaced arrangement within the ascus and variably sized (180-325 x 1.9-2.8(-3) um; Breitenbach & Kranzlin 1984, Johnston 1983, Sherwood 1977). Stictis radiata resembles S. carnea but is distinguished by its shorter and wider ascospores. Known from Mexico, New Zealand, and Australia (Johnston 1983, Sherwood 1977). In New Zealand, S. radiata has been reported on the ferns Cyathea dealbata and Dicksonia squarrosa (Johnston 1983). We found S. radiata also growing on Sphaeropteris horrida and Polypodium sp. Discussion Ferns, which comprise many genera and families, serve as habitat for a large number of fungi. Our study indicates that most of the fungi on ferns belong to Lachnaceae [previously included in Hyaloscyphaceae s.l.], represented by eight species of Lachnum. Haines (1980), Samuels & Rogerson (1980), and Medel & Lorea-Hernandez (2008) previously reported Hyaloscyphaceae s.l. on ferns. Medel (2013), who conducted earlier research on Mexican cloud forest ascomycetes, pointed out that Lachnaceae is “recurrent” on ferns in Cyatheaceae, indicating that some of these fungi may live as endophytes in this fern family. 700 ... Medel-Ortiz, Baeza, Lorea-Hernandez The dominant plant associates in this study belong to the “tree ferns” — Alsophila firma, Cyathea bicrenata, C. divergens var. tuerckheimii, and Sphaeropteris horrida. Herbaceous fern species with short stems at or barely above ground level include Dryopteris sp., Lophosoria quadripinnata, Marattia laxa, and ‘Thelypteris spp. Adiantum andicola and Sticherus palmatus are classed as herbs with underground stems, while Grammitis sp., Phlebodium aureum, Pecluma alfredii, and Polypodium sp., represent a diverse epiphytic fern group. Two fern species hosted the highest number of fungal species: Alsophila firma (11 species) and Cyathea bicrenata (12 species). The ascomycetes identified in this study grew more frequently on petioles and rachises of tree ferns and some herbaceous ferns. Our work stresses the importance of studying the highly diverse small ascomycetes given that new records have been established by every study (Braun 2004, Braun & al. 2013, Cannon 1997, Castaneda-Ruiz & Heredia 2000, Del Olmo & Arnolds 2014, Dingley 1972, Haines 1980, Kirschner & Liu 2014, Medel & Lorea-Hernandez 2008, Samuels & Rogerson 1990, Stevenson 1945); unfortunately, few have studied species from the Western Hemisphere. This paper contributes new fern families and genera to the list of ascomycetes hosts: Dicksoniaceae (genus Lophosoria), Marattiaceae (Marattia), and Polypodiaceae (Grammitis and Pecluma). This is important since there are few studies on the diversity of fungi on ferns, and most have dealt only with genera of tree ferns in Cyatheaceae and Dicksoniaceae. According to the literature surveyed, only seven ascomycete species had previously been registered on ferns from Veracruz. This study adds 18 more records associated with ferns in Mexican cloud forests, bringing the TABLE 1. Ascomycete fungi and fern hosts recorded from Mexico SPECIES Host REFERENCE HELOTIALES Arachnopezizaceae cf. Arachnopeziza Alsophila firma This paper Dermataceae Mollisia ventosa* Cyathea bicrenata This paper Helotiaceae Bisporella pteridicola* Alsophila firma, Cyathea bicrenata, This paper Pecluma alfredii, Sphaeropteris horrida, Thelypteris sp. Crocicreas Cyathea bicrenata This paper quinqueseptatum* Crocicreas sessilis Cyathicula sp. Durella macrospora* Hyaloscyphaceae Dasyscyphella dryina* Hamatocanthoscypha helicotricha* Hyaloscypha fuckelii* Lachnaceae Lachnum brevipilosum* Lachnum fimbriiferum Lachnum nudipes * Lachnum oncospermatum Lachnum pteridophyllum Lachnum singerianum Lachnum varians Lachnum virgineum Paradiopsidaceae Dimeriella polypodii HyPocrEALESs (Bionectriaceae) Protocreopsis pertusa* ORBILIALES (Orbiliaceae) Orbilia auricolor* OSTROPALES Coenogoniaceae Coenogonium botryosum Stictidaceae Stictis carnea* Stictis radiata* XYLARIALES (Sporocadaceae) Seimatosporium tostum Pteridicolous ascomycetes (Mexico) ... 701 Cyathea divergens var. tuerckheimii Cyathea bicrenata Sticherus palmatus Alsophila firma Cyathea bicrenata, Sticherus palmatus Alsophila firma Alsophila firma, Cyathea bicrenata Alsophila firma, Cyathea bicrenata, Cyatheaceae, Dicksonia sellowiana Lophosoria quadripinnata Dicksonia sellowiana Cyathea divergens var. tuerckheimii, Cyathea sp., Dicksonia sellowiana, Dryopteris sp., Lophosoria quadripinnata Alsophila firma, Dicksonia sellowiana Alsophila firma, Cyathea bicrenata, C. divergens var. tuerckheimii, Adiantum andicola Phlebodium aureum Polypodium montigenum, P. madrense Alsophila firma, Marattia laxa Cyathea bicrenata, Marattia laxa Adiantum andicola, Grammitis sp. Cyathea bicrenata Alsophila firma, Cyathea bicrenata, Sphaeropteris horrida, Polypodium sp. Alsophila firma, Cyathea bicrenata. Bold face indicates new hosts for the corresponding fungus * indicates new fungal records for Mexico Samuels & Rogerson 1990 This paper This paper This paper This paper This paper This paper Medel & Lorea- Hernandez 2008 This paper Medel & Lorea- Hernandez 2008 Haines 1980; Medel & Lorea-Hernandez 2008 Medel & Lorea- Hernandez 2008 Haines 1980; Medel & Lorea-Hernandez 2008 This paper Samuels & Rogerson 1990 This paper This paper This paper This paper This paper This paper 702 ... Medel-Ortiz, Baeza, Lorea-Hernandez total to 25 ascomycete species distributed among Helotiales (20 species), Hypocreales (1), Ostropales (3), and Xylariales (1). Taxonomic diversity of Ascomycota reported in similar studies from other regions of the world show a wide diversity of ascomycetes involved in biological interaction with ferns. For example, Del Olmo & Arnolds (2014) reported the primary endophytes to inhabit ferns in Costa Rica belonged to Dothideomycetes, Eurotiomycetes, and Sordariomycetes, while mycosphaerellaceous fungi were more abundant in Taiwan (Kirschner & Liu 2014) and other countries in Asia and Africa (Braun & al. 2013). Only Haines (1980) cited Helotiales (Hyaloscyphaceae s.l.) as the predominant fungi on tropical ferns, further supported by this study where Lachnaceae was the most diverse family represented. To find such diversity of ascomycetes associated with ferns, almost quadrupling the number of species previously known for Mexico, suggests there are several taxa yet to discover. We agree with Kirschner & Liu (2014) that research on ascomycetes growing on ferns is still in its infancy. Additional work is needed to establish the ecology of fern-associated fungi, particularly the recurrence of some taxa on fern hosts. Acknowledgments The first author expresses her gratitude to Gast6n Guzman (+), who was a constant guide during her studies. Thanks to DGI-Universidad Veracruzana for the research assistant fellowship granted to Yajaira Baeza and to Juan Lara Carmona, XAL mycological collection assistant, who processed loans during the our research. Special thanks to Sharon Cantrell, Teresa Iturriaga, and Andrea I. Romero for comments that greatly improved an early version of the manuscript. Luis Quijada (Department of Organismic and Evolutionary Biology, Harvard Herbarium, Cambridge MA, U.S.A.) and José Marmolejo (Facultad de Ciencias Forestales, Universidad Autonoma de Nuevo Leon, México) made valuable suggestions during their expert peer reviews. Special thanks to Shaun Pennycook for his exhaustive revision. Literature cited Baral HO, Haelewaters D, Partel K. 2015. A new attempt to classify the families of the Helotiales. 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Taxonomy of the genus Bisporella (Helotiales) in China with seven new species and four new records. Mycosystema 36(4): 401-420. https://doi.org/10.13346/j.mycosystema.160193 MY COTAXON ISSN (print) 0093-4666 (online) 2154-8889 Mycotaxon, Ltd. ©2019 October-December 2019—Volume 134, pp. 707-717 https://doi.org/10.5248/134.707 First sexual morph record of Sarcopodium vanillae NAPALAI CHAIWAN?”, SAJEEWA S.N. MAHARACHCHIKUMBURA}, DHANUSHKA N. WANASINGHE’, MINGKWAN DOILOM?’, RUVISHIKA JAYAWARDENA’, KEVIN D. HyDE*”” "Center of Excellence in Fungal Research, Mae Fah Luang University, Chiang Rai 57100, Thailand ? Key Laboratory for Plant Diversity and Biogeography of East Asia, Kunming Institute of Botany, Chinese Academy of Science, Kunming 650201, Yunnan, Peoples Republic of China ° School of Life Science and Technology, University of Electronic Science and Technology of China, Chengdu 611731, Peoples Republic of China * CORRESPONDENCE TO: kdhyde3@gmail.com ABSTRACT—Sarcopodium vanillae was isolated from a dead leaf of Dracaena in Chiang Rai Province, Thailand. Combined analyses of ACT, ITS, LSU, and TuB2 sequence data obtained from the cultures derived from single spore isolates confirm that our collections represent S. vanillae. This is the first record of the sexual morph, and the first record of S. vanillae from Dracaena. A description and illustrations of both sexual and asexual stages of S. vanillae are provided. KEY worps—multigene, new host record, Nectriaceae, saprobe, taxonomy Introduction During our research on the diversity of microfungi in Thailand, we recovered isolates from Dracaena in Chiang Rai Province that were typical of Nectriaceae based on our preliminary morphological studies. Further morphological and molecular characterization (multigene-based phylogeny of nuclear ribosomal and protein-coding loci ACT, ITS, LSU and Tus2 sequence data) revealed the taxon as Sarcopodium vanillae. Nectriaceae (Hypocreales; Rossman & al. 1999, Maharachchikumbura & al. 2015) is typified by a characteristic perithecial wall structure and specific anamorphic 708 ... Chaiwan & al. states. Its uniloculate perithecioid ascomata are yellow, orange, red, purple or white, and the asexual morph is phialidic (Rogerson 1970, Rehner & Samuels 1995). The genus Sarcopodium Ehrenb. has had numerous nomenclatural changes since it was first described in 1818 that have been synonymised under Sarcopodium. Cyphina Sacc., had already been synonymised under Sarcopodium by Sutton (1977) and under “one fungus: one name” by Rossman & al. (2016) supported the placement of the (asexual) Actinostilbe and the (sexual) Lanatonectria under the much earlier (asexual) Sarcopodium. Sarcopodium Ehrenb. is closely related to Pseudonectria Seaver. (Rossman & al. 1999). Sutton (1981) synonymized Actinostilbe Petch. under Sarcopodium; and all five species of Lanatonectria Samuels & Rossman (a genus described from a sexual morph) have been transferred to Sarcopodium following phylogenetic analyses (Lombard & al. 2015, Pennycook & Kirk 2019a). The new epithet for the type species (“flocculentum”) has priority over the previous heterotypic synonym, Sarcopodium macalpinei (Pennycook & Kirk 2019b). Some Sarcopodium species have both sexual and asexual morphs, but S. vanillae has been known only from its asexual morph (Sutton 1981; Rossman 1983, 1996, 2000; Rossman & al. 1993, 2013, 2016). This paper describes the first record of a sexual morph for S. vanillae and reports Dracaena as a new host record for this fungus. Materials & methods Sample collection, morphological study and isolation Dead leaves of Dracaena were collected from Chiang Rai Province in Thailand during November 2017. The samples were taken to the laboratory in Zip-lock bags. Micro-morphological structures were observed and photographed with a Canon EOS 600D digital camera fitted to a Nikon Eclipse Ni compound microscope. Single spore isolates were obtained using the method described in Chomnunti & al. (2014). Germinated spores were transferred to potato dextrose agar (PDA) plates and incubated at 25-30 °C. Growth rates and culture characteristics were recorded after three weeks. The morphological characters were measured using Tarosoft (R) Image Frame Work v. 0.9.7 software. Figures were processed with an Adobe Photoshop CS6 Extended v. 10.0 software. The cultures are deposited in Mae Fah Luang University Culture Collection, Chang Rai, Thailand (MFLUCC). Specimens are deposited in the herbarium of Mae Fah Luang University, Chang Rai, Thailand (MFLU). DNA extraction, PCR amplification and sequencing Fungal isolates were grown on PDA for 3-4 weeks at 30 °C. The genomic DNA was extracted from mycelium using the E.Z.N.A. Forensic DNA Kit. Amplification Sexual morph of Sarcopodium vanillae (Thailand) ... 709 TABLE 1. Sequences from Pseudonectria and Sarcopodium species (and Stachybotrys chartarum outgroup) used in the phylogenetic analysis GENBANK ACCESSION NO. TAXON CULTURE NO. ACT ITS LSU TUB2 P. buxi CBS 324.53 KM231171 KM231778 KM231644 KM232037 P foliicola CBS 122566 KM231170 KM231777 KM231643 KM232036 S. circinatum CBS 587.92 KM231180 KM231787 KM231651 KM232046 CBS 100998 KM231179 KM231786 KM231650 KM232045 S. circinosetiferum CBS 100251 KM231175 KM231782 KM231646 KM232041 S. flavolanatum CBS 112283 KM231178 KM231785 KM231649 KM232044 CBS 128370 KM231177 KM231784 KM231648 KM232043 S. macalpinei CBS 115296 KM231176 KM231783 KM231647 KM232042 S. vanillae CBS 100582 KM231173 KM231780 HQ232174 KM232039 MEFLU 17-2595 MK692541 MK608516 MK691503 MK962543 MFLU 17-2597 MK692542 MK685870 MK691502 MK692544 Stachybotrys CBS 129.13 KM231268 KM231858 KM231738 KM232127 chartarum New sequences are set in bold font. primers used were: internal transcribed spacer (ITS)—ITS5 and ITS4 (White & al. 1990); 28S large subunit ribosomal RNA (LSU)—LROR and LRS5 (Rehner & Samuels 1994, Vilgalys & Hester 1990); partial actin gene (ACT)—ACT-512F and ACT-783R (Carbone & Kohn 1999); partial beta-tubulin gene (ruB2)—Bt-2a and Bt-2b (Glass & Donaldson 1995). The PCR mixer comprised 1 ul forward primer, 1 pul reverse primer, 9.5 ul distilled deionized (DD) water, and 12.5 ul mixer. The PCR conditions for ITS and LSU were 3 min at 94 °C; followed by 35 cycles of 30 s at 94 °C, 50 s at 55 °C, and 90 s at 72 °C; and a final elongation step at 72 °C for 10 min. Conditions for ACT were an initial elongation step of 2 min at 95 °C; followed by 35 cycles of 45 s at 95 °C, 45 s at 55 °C, and 1 min at 72 °C; and a final elongation step of 10 min at 72 °C. Conditions for TUB2 were an initial 8 min of 95 °C; followed by 35 cycles of 30 s at 95 °C, 30 s at 55 °C, and 1 min at 72 °C; and a final elongation step at 72 °C for 5 min. The PCR products were purified and sequenced at Shanghai Sangon Biological Engineering Technology and Service Co. Isolates including accession numbers of gene sequences are listed in TABLE 1. Phylogenetic analysis Sequence data of Sarcopodium vanillae and related taxa (TABLE 1) were downloaded from GenBank following Zeng & al. (2018) and Yang & al. (2018). The multiple sequence alignments were produced with MAFFT v. 7 (http://maftft.cbre.jp/ 710... Chaiwan & al. alignment/server/index.html) and Biokdit v. 7.0.5.2 (Hall 1999). The phylogenetic analyses were performed using maximum likelihood (ML) trees and generated using the RAxML-HPC2 on XSEDE (8.2.8) (Stamatakis & al. 2008, Stamatakis 2014) in the CIPRES Science Gateway platform (Miller & al. 2010) using GTR+I+G model of evolution. Parsimony analysis was carried out with the heuristic search option in PAUP v. 4.0b10 (Swofford 2002) using the following parameters: characters unordered with equal weight, random taxon addition, branch swapping with tree bisection-reconnection (TBR) algorithm, branches collapsing if the maximum branch length was zero. Alignment gaps were treated as missing characters in the combined data set, where they occurred in relatively conserved regions. Trees were inferred using the heuristic search option with 1000 random sequence additions, with maxtrees set at 1000. Descriptive tree statistics for parsimony; tree length (TL), consistency index (CI), retention index (RI), relative consistency index (RC) and homoplasy index (HI) were calculated for trees generated. The Kishino-Hasegawa tests (Kishino & Hasegawa 1989) were performed to determine whether trees were significantly different. Bayesian analysis was conducted with MrBayes v. 3.1.2 (Huelsenbeck & Ronquist 2001) to evaluate posterior probability (PP) (Rannala & Yang 1996) by Markov chain Monte Carlo sampling. GTR+I+G was used in the command. Six simultaneous Markov chains were run for 2,000,000 generations with trees sampled every 200th generation. The distribution of log-likelihood scores was examined to determine stationary phase for each search and to decide if extra runs were required to achieve convergence, using the program Tracer 1.4 (Rambaut & Drummond 2007). First 10% of generated trees were discarded and remaining 90% of trees were used to calculate posterior probabilities of the majority rule consensus tree. The phylogenetic tree was figured in FigTree v. 1.4 (Rambaut 2014) and edited using Microsoft Office Power Point 2007. Sequences derived in this study were deposited in GenBank (TABLE 1). Phylogenetic results The combined sequence alignments comprised 12 isolates, with Stachybotrys chartarum (CBS 129.13) as the outgroup taxon. The combined dataset comprised 3065 characters including alignment gaps, of which 844 were derived from ITS, 624 from LSU, 812 from ACT, and 785 from TuB2. The MP analysis for the combined dataset had 415 parsimony informative, 2400 constant, 250 parsimony uninformative characters and yielded a single most parsimonious tree (TL = 1109, CI = 0.812, RI = 0.761, HI = 0.188, RC=0.617; Fic 1). The RAxML analysis of the combined dataset yielded a best scoring tree (Fic. 1) with a final ML optimization likelihood value of -~9266.358949. The matrix had 656 distinct alignment patterns, with 17.28% of undetermined characters or gaps. Estimated base frequencies were as follows: A = 0.220970, C = 0.279441, G = 0.267083, T = 0.232506; substitution rates Sexual morph of Sarcopodium vanillae (Thailand) ... 711 Pseudonectria follicola CBS122566 100/400/1 Pseudonectria buxi CBS324.53 Sarcopodium flavolanatum CBS 112283 100/101 Sarcopodium flavolanatum CBS128370 Sarcopodium circinatum CBS 100998 100/401 Sarcopodium circinatum CBS587.92 99/82/0.99 Sarcopodium circinosetiferum CBS100251 100/100/1 Sarcopodium macalpinel CBS115296 Sarcopodium vanilla CBS100582 100/101 64/100/0.95 Fic. 1. Phylogram generated from RAxML based on combined ITS, LSU, ACT, and Tus2 sequence data. Bootstrap support values for maximum likelihood (ML) 260%, maximum parsimony (MP) 260%, and Bayesian posterior probabilities (PP) 20.95 are indicated as ML/MP/PP at the nodes. Ex-type strains are in bold and the strains from this study are indicated in green. 712 ... Chaiwan & al. AC = 0.730175, AG = 1.879094, AT = 1.313827, CG = 0.798328, CT = 4.086170, GT = 1.000000; gamma distribution shape parameter a = 0.147281. Bayesian posterior probabilities from Bayesian inference analysis were assessed with a final average standard deviation of split frequencies = 0.0056. The phylogenetic tree in this study showed that our strains (Sarcopodium vanillae MFLUCC 17-2595 and MFLUCC 17-2597) grouped in the Sarcopodium clade, and formed a well-supported cluster with S. vanillae (CBS 100582) with 64% ML, 100% MP, and 0.95 PP. Taxonomy Sarcopodium vanillae (Petch) B. Sutton, Trans. Brit. Mycol. Soc. 76: 99 (1981) PLATES 1, 2 = Actinostilbe vanillae Petch, Ann. Roy. Bot. Gard. (Peradeniya) 9(3):327 (1925) SAPROBIC on dead leaves of Dracaena (Asparagaceae). SEXUAL MORPH: ASCOMATA 150-200 um high, 160-240 um wide (x = 173 x 189 um, n = 5), perithecial, subglobose, solitary or in groups, soft-textured, pale yellow or rarely orange, superficial on a leaf or erumpent, with a papillate ostiole. PERIDIUM 15-25 um wide (x = 47 um, n = 5), composed of several layers of white to light orange cells of textura angularis. Asci 36-52 x 3-5 um (x = 44 x 4.5 um, n = 20), 4-spored, unitunicate, cylindrical, rounded at apex, clavate to fusiform, short pedicellate. Ascosporss 8-12 x 3-4.5 um (x = 11 x 3.9 um, n = 20), fasciculate, broadly elongate, 1-septate. ASEXUAL MORPH: Myce.Lium, branched, septate, hyaline, smooth. CONIDIOMATA 200-210 x 220-240 um (x= 205 x 230 um, n= 5), sporodochial, solitary or gregarious, setiferous, yellow to bright yellow or rarely orangish brown, soft-textured, superficial, separate, gregarious or confluent, sessile, attached to the substratum by a small stroma concentrated in the epidermis or outer layers of peridermal tissue, pulvinate, setose. SETAE 100-110 x 5-10 um (x = 105 x 7.5 um, n = 5), septate, unbranched, cylindrical, incurved, erect, very thick-walled, medium to pale golden brown, more or less straight (occasionally slightly bent either at the base or nearer the apex), erect, hyaline, pointed or rounded at the apex. CONIDIOPHORES mononematous, verticillately or penicillately branched, straight or flexuous, smooth, hyaline, short, septate, with 1-4 monochasial branching, compactly arranged, cylindrical, intermixed with long setae. CONIDIOGENOUS cells enteroblastic, monophialidic, integrated, cylindrical or more frequently tapered towards the apices, subulate, widest from middle to base, 10-20 x 1.2-2.7 um (x = 15 x 1.95 um, n = 5), with inconspicuous collarette, hyaline, smooth, Sexual morph of Sarcopodium vanillae (Thailand) ... 713 PLATE 1. Sarcopodium vanillae (MFLU 19-0567, herbarium specimen): A, B. Perithecia on host surface; C-E. Perithecial cross sections; F Crush mount of perithecium; G. Conidioma on perithecium; H, I. Asci, J. Ascospores; K, L. PDA cultures. Scale bars: A, B = 200 um; C-G = 50 um; H, I= 20 um; J = 10 um formed as the ultimate branches of conidiophores and completely covering the external face of the conidiomata. Conrip1a 5-9 x 2.1-2.6 um (x = 6 x 2.5 um, n = 20), cylindrical, 0-1-septate, smooth, hyaline, rounded at both ends, held together in a slimy mass, ellipsoid to oval, straight. CULTURE CHARACTERISTICS: Colony on PDA reaching 30-40 mm diam. after 3 weeks at 25-30 °C; from above, white to yellow at margin, white to 714 ... Chaiwan & al. PLATE 2. Sarcopodium vanillae (MFLU 19-0566, herbarium specimen): A-C. Conidiomata on host surface; D. Conidioma; E-I. Setae; J, K. Setal bases; L. Setal apex; M. Conidia; N, O. PDA cultures. Scale bars: A-C = 200 um; D, J, K = 100 um; E-I, L, M = 50 um. orange in the middle, white at centre; from below, yellow, medium dense, irregular, slightly raised to umbonate, surface slightly rough, dull with umbonate edge, concave at centre, fluffy to floccose, with white tufts at centre. SPECIMENS EXAMINED: THAILAND, CHIANG RAI PROVINCE, Mae Lao District, on dead leaf of Dracaena, 17 November 2017, Napalai Chaiwan NCCR003 (MFLU 19- 0566 [asexual morph]; living culture MFLUCC 17-2595); Napalai Chaiwan NCCR004 (MFLU 19-0567 [sexual + asexual morph]; living culture MFLUCC 17-2597). Discussion Our strains share similar morphological characters with S. vanillae strain CBS 100852, which was isolated from Anthurium sp. in Ecuador (Lombard & al. 2015). The phylogenetic analysis also supports the close relationship. The genus Sarcopodium has both sexual and asexual morphs (Wijayawardene & al. 2017a,b, 2018). The conidial morphology of our strain is similar to S. circinatum (the type species of the genus); however our conidiomata and conidiophores more closely resemble Volutella ciliata (CBS 483.61) Sexual morph of Sarcopodium vanillae (Thailand) ...715 (Lombard & al. 2015). Previously, only the asexual morph has been observed for S. vanillae (Sutton 1981), and this study is the first report of the sexual morph. Sarcopodium vanillae has been reported from Abelmoschus manihot in Papua New Guinea, Citrus nobilis in Brunei, Vanilla planifolia in Sri Lanka, and V. tahitensis in Papua New Guinea (Farr & Rossman 2019). This study provides the first report of S. vanillae from Dracaena, and its first report from Thailand. Acknowledgments N. Chaiwan thanks the Thailand Research Fund (PHD60K0147) and Kunming Institute of Botany for financial support and the molecular laboratory for support. K.D. Hyde thanks the grants entitled: 1 the future of specialist fungi in a changing climate: baseline data for generalist and specialist fungi associated with ants, Rhododendron species and Dracaena species (Grant number: DBG6080013) and 2. The climate changes grant: Impact of climate change on fungal diversity and biogeography in the Greater Mekong Subregion (Grant number: RDG613001). M. Doilom thanks the 5th batch of Postdoctoral Orientation Training Personnel in Yunnan Province and the 64th batch of China Postdoctoral Science Foundation. D.N. Wanasinghe thanks the CAS President’s International Fellowship Initiative (PIFI) for funding his postdoctoral research (number 2019PC0008), the National Science Foundation of China and the Chinese Academy of Sciences for financial support under the following grants:41761144055, 41771063 and Y4ZK111B01. S.C. Karunarathna thanks the CAS President's International Fellowship Initiative (PIFI) for funding his postdoctoral research (number 2018PC0006). The National Science Foundation of China (NSFC) for funding this work under the project code 31750110478. Jianchu Xu thanks the Key Research Program of Frontier Sciences “Response of Asian mountain ecosystems to global change’, CAS, Grant No. QYZDY-SSW-SMC014. All authors are grateful to peer experts D. Jayarama Bhat (Emeritus Professor of Botany. Goa University, India) and Eric H.C. McKenzie (Manaaki Whenua-Landcare Research, Auckland, New Zealand) for their assistance and presubmission review. Literature cited Carbone I, Kohn LM. 1999. 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Molecular phylogeny of Nectria species associated with dieback and canker diseases in China, with a new species described. Phytotaxa 356(3): 199-214. https://doi.org/10.11646/phytotaxa.356.3.2 Zeng ZQ, Zhuang WY, Yu ZH. 2018. New species and new Chinese records of Nectriaceae from Tibet, China. Nova Hedwigia 106: 283-294. https://doi.org/10.1127/nova_hedwigia/2017/0435 MY COTAXON ISSN (print) 0093-4666 (online) 2154-8889 Mycotaxon, Ltd. ©2019 October-December 2019—Volume 134, pp. 719-730 https://doi.org/10.5248/134.719 Notes on rust fungi in China 7. Aecidium caulophylli life cycle inferred from phylogenetic evidence and renamed as Puccinia caulophylli comb. nov. JING-XIN Jr'4, ZHUANG LY, Yu L1’, MAKOTO KAKISHIMA”?4® ' Engineering Research Center of Chinese Ministry of Education for Edible and Medicinal Fungi, Jilin Agricultural University, Changchun, Jilin 130118, China College of Plant Pathology, Shandong Agricultural University, Taian 271000, China > College of Forestry, Beijing Forestry University, Beijing 100083, China ‘University of Tsukuba, Tsukuba, Ibaraki 305-8572, Japan * CORRESPONDENCE TO: *1096314395@qq.com * kakishima.makoto.ga@u.tsukuba.ac.jp ABSTRACT—Spermogonial and aecial stages of Aecidium caulophylli on Caulophyllum robustum (Berberidaceae) were shown by phylogenetic analyses of ITS and 28S sequence data to be identical to a uredinial and telial rust on Milium effusum (Poaceae). A new combination, Puccinia caulophylli is proposed for this species, and an epitype is designated. Key worps—Pucciniaceae, Pucciniales, taxonomy Introduction Aecidium caulophylli was described in 1898 based on spermogonial and aecial stages of a specimen on Caulophyllum robustum (Berberidaceae) collected by V. Komarov in Amur, Siberia, Russian Far East, during June 1895 (Saccardo & Sydow 1902). This rust has been reported from China, Russian Far East, and Japan (Miura 1928, Ito 1950, Tai 1979, Harada 1984, Hiratsuka & al. 1992, Azbukina 2005). However, its uredinial and telial stages have not been clearly demonstrated, although Azbukina (1984, 2005) listed this species as spermogonial and aecial stages of Puccinia brachypodii var. poae-nemoralis (G.H. Otth) Cummins & H.C. Greene [= P. poae-nemoralis G.H. Otth], without any evidence. ji & al. TaN. V8CS8ZIW = €8TS8ZNIIN V8CS8ZIW = C87S8Z IW V8CS8ZIW = [8cS8Z IN V8CS8ZIW =: O8 CS8Z IN V8CS8ZIW = S9TSBZYIN V8CS8ZIW =: 99 TSBZMIN V8CS8ZIW = P9TSBLIN V8CS8ZIW = €9TSBLMIIN V8CS8ZNW = OL7S8ZNIN V8CS8ZIW = 69TSBZIN V8CS8ZIW = 89TS8Z IW. 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(China) ... 721 During an investigation of rust fungi in Jilin Province, China, during 2015-2018, A. caulophylli was observed on C. robustum. Near the infected C. robustum, uredinial and telial stages of Puccinia species were also observed on plants of Poaceae and Cyperaceae. We suspected that this rust may have an heteromacrocyclic life cycle, producing uredinia and telia on one of these plants. Inoculation of plants with spores is an appropriate method to clarify rust fungus life cycles (Ji & al. 2017a,b), but often plant collection may be restricted in conservation areas. As it may be difficult to supply appropriate growth conditions after transplantation, molecular analyses have been applied to clarify rust life cycles (Liu & Hambleton 2013, Ji & al. 2016, Padamsee & McKenzie 2017, Scholler & al. 2019). We report here the results of phylogenetic and morphological analyses using spermogonial/aecial specimens on A. caulophylli and potentially related uredinial/telial specimens on Poaceae and Cyperaceae. Materials & methods Molecular analyses Spermogonial (0) and aecial (I) stages of A. caulophylli on C. robustum were collected in Jiaohe, Jilin Province, China, and used for molecular analyses. Rust specimens on Milium effusum (Poaceae) were collected in several areas in Jilin and Heilongjiang Provinces, China, as preliminary phylogenetic analyses suggested they represented the uredinial (II) and telial (III) stages of A. caulophylli. The specimens were collected during surveys of rust fungi in Jilin Province from 2015 to 2018. For comparative analyses, sequences were included from two life cycles that previously clarified by inoculations (Ji & al. 2017a,b): (1) Puccinia klugkistiana (Dietel) Jing X. Ji & Kakish. on Ligustrum obtusifolium Siebold & Zucc. (0, I) and Cleistogenes hackelii (Honda) Honda (II, II), collected in Changchun, Jilin Province; and (2) PB adenocauli (Syd. & P. Syd.) Jing X. Ji & Kakish. on Adenocaulon himalaicum Edgew. (0, I) and Carex onoei Franch. & Sav. (II, IID). Total genomic DNA was directly extracted from about 200 spores obtained from single sori on the leaves of each specimen, using similar methods reported by Ji & al. (2016, 2019). Specimens used in the experiments were deposited in the Herbarium of Mycology, Engineering Research Center of Chinese Ministry of Education for Edible and Medicinal Fungi, Jilin Agricultural University, China (HMJAU) and all data sequenced in this experiment were deposited at GenBank (TABLE 1). Sequences from the specimens were aligned following Ji & al. (2019). ITS and 28S sequence data retrieved from GenBank were added to phylogenetic analyses. Accession numbers for these data are shown in the phylogenetic trees (Fics 1, 2). Phylogenetic trees were constructed with the sequences of Gymnosporangium yamadae Miyabe ex G. Yamada and G. asiaticum Miyabe ex G. Yamada as outgroup, 722... Ji &al. Puccinia caulophylli HMJAU8620 [Caulophyllum robustum] (SA) Puccinia caulophylli HMJAU8619 [Milium effusum)] (UT) Puccinia caulophylli HMJAU8623 [Milium effusum] (UT) Puccinia caulophylli HMJAU8618 [Milium effusum] (UT) Puccinia caulophylli HMJAU8625 [Milium effusum] (UT) Puccinia caulophylli HMJAU8626 [Milium effusum] (UT) 059/50" Puccinia caulophylli HMJAU8621 [Milium effusum)] (UT) Puccinia caulophylli HMJAU8627 [Milium effusum] (01) Puccinia caulophylli HMJAU8534 [Caulophyllum robustum] (SA) Puccinia caulophylli HMJAU8622 [Milium effusum] (U1) Puccinia caulophylli HMJAU8624 [Caulophyllum robustum] (SA) Puccinia caulophylli HMJAU8531 [Caulophyllum robustum] (SA) osoisyg) -Puecinia caulophylli HMJAU8532 [Caulophyllum robustum) (SA) Puccinia sessilis (AY 217134) Puccinia sessilis (AY 217135) Puccinia hordei (KY764128) [Ornithogahim arabicum] Puccinia recondita (KY798399) [Elymus spicatus] wioonooPuecinia magnusiana (GU058000) [Phragmites sp.] Puccinia magnusiana (KY 764139) [Phragmites sp.] Puccinia triticina (DQ664194) [Triticum aestivum] Puccinia triticina (KY764169) [Triticum aestivum] Puccinia recondita (DQ417424) [Aegilops ovata] ‘Puccinia recondita (AF511082) Puccinia recondita f. sp. secalis (DQ417423) [Secale cereale] Puccinia clavata (KX985761) [Clematis foetida] Puecinia hordei (KX985762) [Holcus lanatius] osgousgPuccinia sorghi (KY764162) [Zea mays] Puccinia sorghi (GU057994) [Zea sp.] Puccina polysora (GU058024) [Zea mays] 'Puccina polysora (MF033480) [Zea sp.] ‘Puccinia brachypodii (KX999868) [Poa annua] ‘Puccinia poae-nemoralis (KY 798384) [Calamagrostis sp.] oossiggjPuccinia coronata (AF426207) [Rhamnus cathartica] 'Puccinia coronata (EU851141) [Holcus lanatus] 4 Puccinia coronata (DQ355448) [Bromus sp.] Puccinia coronata (AB693935) 0.95/°/73 Puccinia graminis (KM249852) [Glyceria maxima] oseomeyPuccinia graminis (KY798389) [Poa annua] 'Puccinia graminis (AF522177) Puccinia adenocauli HMJAU 8628 [Adenocaulon himalaicum] (SA) nooo Puccinia adenocauli HMJAU 8630 [Carex onoei] (UT) Puccinia adenocauli HMJAU 8274 [Carex onoei] (UT) Puccinia adenocauli HMJAU 8629 [Adenocaulon himalaicum] (SA) Puccinia klugkistiana HMJAU 8280 [Cleistogenes hackelii] (U1) Puccinia klugkistiana HMJAU 8633 [Ligustrum obtusifolium] (SA) soon oo| Puccinia klugkistiana HMJAU 8632 [Cleistogenes hackelii] (UT) Puccinia klugkistiana HMJAU 8198 [Ligustrum obtusifolium] (SA) Puccinia klugkistiana HMJAU 8282 [Cleistogenes hackelii] (UT) Puccinia klugkistiana HMJAU 8631 [Ligustrum obtusifolium] (SA) /100/100) Gymnosporangium asiaticum HMJAU 8324 [Pyrus sp.] Gymnosporangium yamadae HMJAU 8096 [Malus baccata] 1/100/100) Fic. 1. Phylogenetic tree constructed by MP method based on sequences of 28S regions of rDNA. Bootstrap values of MP and ML are followed by the Bayesian posterior probabilities (BPP) on the nodes in the topology. Asterisk (*) represents bootstrap values <50% or BPP <0.5 in the topology. Sample data are shown with species name, voucher specimen number or GenBank accession number (in parentheses), and host plant. Sequence data determined in this study are shown in bold face. SA: Spermogonial and aecial stages, UT: Uredinial and telial stages. according to Ji & al. (2019). The alignment and trees were deposited in TreeBase under http://purl.org/phylo/treebase/phylows/study/TB2:S24307 (Fic. 1) and TB2:824308 (Fic. 2). Morphological observations Light (LM) and scanning electron (SEM) microscopy were used to examine morphological characters of rust specimens including the size and shape of sori and spores following Ji & al. (2019). Puccinia caulophylli comb. nov. (China) ... 723 Puccinia caulophylli HMJAU8620 [Caulophyllum robustum] (SA) Puccinia caulophylli HMJAU8619 [Milium effusum] (U1) Puccinia caulophylli HMJAU8618 [Milium effusum] (UT) Puccinia caulophylli HMJAU8532 [Caulophyllum robustum] (SA) Puccinia caulophylli HMJAU8534 [Caulophyllum robustum] (SA) Puccinia caulophylli HMJAU8625 [Milium effusum] (U1) Puccinia caulophylli HMJAU8627 [Milium effusum] (UT) Puccinia caulophylli HMJAU8623 [Milium effusum] (UT) Puccinia caulophylli HMJAU8626 [Milium effusum) (U1) vosioo| Puccinia caulophylli HMJAU8531 [Caulophyllum robustum] (SA) Puccinia caulophylli HMJAU8621 [Milium effusum] (UT) 99/55/62] Puccinia caulophylli HMJAU8624 [Caulophyllum robustum] (SA) Puccinia caulophylli HMJAU8622 [Milium effusum] (UT) Puccinia sessilis (AY 217134) 1100/1001 Pyecinia sessilis (AY 217135) joanoor Puccinia triticina (DQ417417) [Triticum turgidum L. var. durum | o.p/ST/EM Puccinia triticina (DQ417417) [Triticum sp.] Puccinia triticina (KT982695) [Hordeum vulgare] 1007108 Puccinia triticina (DQ460717) [Hordeum vulgare] Puccinia recondita f. sp. secalis (DQ417426) [Secale cereale] M0000 Puccinia recondita (AY956562) [Cerinthe minor] 0.87/#/4 Puccinia coronata (DQ355444) [Holcus lanatus| 0.99/88 Puccinia coronata f. sp. graminicola (HM131240) [Arrhenatherum elatius] Puccinia coronata var. coronata (HM057141) [Calamagrostis epigejos] Puccinia coronata f. sp. avenae (EU014044) [Lolium perenne] Puccinia coronati-hordei (HM131229) [Elymus repens] Puccinia coronati-japonica (HM131317) [Calamagrostis arundinacea] Puccinia coronati-agrostidis (HM131319) [Agrostis stolonifera] Puccinia coronati-calamagrostidis (HM131350) [Elymus sp.] venoqPuccinia sorghi (AY 114291) Puccinia sorghi (HQ154038) [Zea mays] vioggg Puccinia brachypodii (KM 391664) i600} | Puccinia brachypodii (KM 391669) Puccinia brachypodii (GQ457303) Puccinia graminis (AY874140) oom Puccinia graminis f. sp. tritici (DQ 417379) Puccinia graminis (HM131357) [Elymus repens] Puccinia klugkistiana HMJAU 8633 [Ligustrum obtusifolium] (SA) Puccinia klugkistiana HMJAU 8282 [Cleistogenes hackelii] (UT) Puccinia klugkistiana HMJAU 8198 [Ligustrum obtusifolium] (SA) Puccinia klugkistiana HMJAU 8631 [Ligustrum obtusifolium] (SA) sno Puccinia klugkistiana HMJAU 8632 [Cleistogenes hackelii| (UT) 0.98/52/8 Puccinia klugkistiana HMJAU 8280 [Cleistogenes hackelii| (UT) oonoor Puccinia polysora (HQ189433) [Zea mays] Puccinia polysora (HM467909) Puccinia adenocauli HMJAU 8274 [Carex onoei] (UT) Puccinia adenocauli HMJAU 8628 [Adenocaulon himalaicum] (SA) 1100/1001 '— Puccinia adenocauli HMJAU 8629 [Adenocaulon himalaicum] (SA) Puccinia adenocauli HMJAU 8630 [Carex onoei] (UT) 1/100/100 Gymnosporangium asiaticum HMJAU 8324 [Pyrus sp.] Gymnosporangium yamadae HMJAU 8096 [Malus baccata] /54/84) 99} 0.54/52/56 0,98/*/67 /97/100 0.97738763) 0.98/*/86) +/#/8; /1.00/100 Fic. 2. Phylogenetic tree constructed by MP method based on ITS regions of rDNA. Bootstrap values of MP and ML are followed by the Bayesian posterior probabilities (BPP) on the nodes in the topology. Asterisk (*) represents bootstrap values <50% or BPP <0.5 in the topology. Sample data are shown with species name, voucher specimen number or GenBank accession number (in parentheses), and host plant. Sequence data determined in this study are shown in bold face. SA: Spermogonial and aecial stages, UT: Uredinial and telial stages Results & discussion Phylogeny and life cycle The 28S dataset comprised 51 sequences of 50 taxa with 525 total characters, including 430 constant characters, 25 parsimony-uninformative variable characters, and 70 parsimony-informative characters. Parsimony analysis yielded one parsimonious tree with TL = 159, CI = 0.679, RI = 0.864 724 ... Ji & al. and RC = 0.587. Bayesian analysis resulted in average standard deviation of split frequencies of 0.005869. The final ITS dataset comprised 51 sequences of 50 taxa with 820 total characters, of which 336 were parsimony- informative. Parsimony analysis yielded one parsimonious tree with TL=995, CI = 0.623, RI = 0.840 and RC = 0.523. Bayesian analysis resulted in average standard deviation of split frequencies of 0.004251. Tree topologies formed by MP, ML, and MCMC methods were identical among trees. The phylogenetic trees generated through Bayesian analysis are shown in Fiac..1 (28S) and! Fie. 2(ITS): Both 28S and ITS phylogenetic trees placed spermogonial and aecial stages on C. robustum (HMJAU 8531, 8532, 8534, 8620, 8624) and uredinial and telial stages on M. effusum (HMJAU 8618, 8619, 8621, 8622, 8623, 8625, 8626, 8627) within a monophyletic clade (Fries 1, 2). All stages of P. klugkistiana and P adenocauli were placed in separate monophyletic clades, confirming that heteroecious life cycles of rust fungi can be revealed by phylogenetic analyses as demonstrated by Liu & Hambleton (2013), Padamsee & McKenzie (2017), and Scholler & al. (2019). The phylogenetic analyses also supported the life cycle connection between the rust on C. robustum and the rust on M. effusum. Although the Caulophyllum/Milium rust was shown to be phylogenetically close to P. sessilis W.G. Schneid. ex J. Schrot., P. triticina Erickss., and P. recondita Roberge ex Desm., these three rusts inhabit different host plants. This rust is also genetically distant from P brachypodii G.H. Otth and P. poae-nemoralis, two other telial rusts reported on Milium spp. (Cummins 1971, Zhuang & al. 1998, Azbukina 2005). Therefore, we conclude that this rust is distinct from other species. Morphology & taxonomy From the phylogenetic analyses, the spermogonia and aecia on A. caulophylli and the uredinia and telia on M. effusum are produced by one and the same heteromacrocyclic rust species. LM and SEM observations showed that overall morphology of the rust on C. robustum is identical with that of A. caulophylli described by Saccardo & Sydow (1902), Ito (1950), Harada (1984), and Hiratsuka & al. (1992) (Fics 3, 54,B). The morphologies of uredinial and telial specimens on M. effusum were similar to each other (Fics 4, 5c-£), and the two-celled teliospores refer the rust to the genus Puccinia (Cummins & Hiratsuka 2003). Specimens on Milium effusum are phylogenetically close to P. sessilis, but urediniospores on M. effusum are bigger than those of P. sessilis (20-32 Puccinia caulophylli comb. nov. (China) ... 725 ae > AS ye ey S ay, 4 io FAM ae Fic. 3. Puccinia caulophylli on Caulophyllum robustum: spermogonial and aecial stages. A. Yellow lesions on the leaves producing spermogonia and aecia; B. Aecia produced around spermogonia on lower surface of the plant; C. Catenulate aeciospores surrounded by peridium in a vertical section of an aecium; D. Vertical section of a spermogonium; E. Aeciospores. Scale bars: C, D = 30 um; E = 15 um. x 19-25 um), and teliospores are smaller than those of P. sessilis (32-58 x 13-20 um). Additionally, the rust on M. effusum has uredinial paraphyses that have not been reported in P. sessilis (Hiratsuka & al. 1992, Zhuang & al. 1998). PB. brachypodii var. poae-nemoralis occurs on M. effusum (Cummins 1971, Zhuang & al. 1998) and its telial structures and teliospores are 726 ... Ji & al. morphologically similar to the present rust fungus. However, urediniospores of the current rust are bigger than those of P. brachypodii var. poae-nemoralis (18-25 x 15-23 um). The aecial stage of P brachypodii var. poae-nemoralis occurs on Berberis spp. (Cummins 1971). Aecidium caulophylli is a legitimate name under the INTERNATIONAL CODE OF NOMENCLATURE FOR ALGAE, FUNGI, AND PLANTS (Shenzhen Code, Art. F.8, 2018). However, the application of an asexual name, such as Aecidium, to a sexual species can cause confusion (Ono 2016). Therefore, we propose a new combination in Puccinia for A. caulophylli. The holotype specimen on C. robustum has only spermogonial and aecial stages of the rust; because the uredinial and telial stages define the rust genus, we designate an epitype specimen on M. effusum. Puccinia caulophylli (Kom.) Jing X. Ji & Kakish., comb. nov. Figs 3-5 MB 830631 = Aecidium caulophylli Kom., in Jaczewski & al., Fungi Rossiae Exsicc. 4: no 176, 1898. Types: Russian Federation, Russian Far East, Siberia, Amur, Mt. Burejenses, stages 0, I on Caulophyllum robustum Maxim. [= C. thalictroides subsp. robustum (Maxim.) Kitam.], June 1895, leg. V. Komarov (holotype, LE; isotype, NY 00610976). China, Jilin Province, Yanbian, Erdaobaihe, stages II, III on Milium effusum L., 2 September 2018, leg. J.X. Ji & M. Kakishima (epitype designated here, HMJAU 8627; MBT 386779). SPERMOGONIA amphigenous, pale yellow to yellowish brown, subepidermal, type 4 of Cummins & Hiratsuka (2003). Azcta hypophyllous, yellow, subepidermal, erumpent, Aecidium-type with firmly connected peridia. AECIOSPORES Catenulate, subglobose, ovate to ellipsoid, 14-24 x 13.5-20 um (av. 18.5 x 15.5 um), walls hyaline, 0.5-2 um thick (av. 1 um), densely verrucose. UreEDINIA mostly hypophyllous, pale yellow to cinnamon-brown, subepidermal, erumpent, with abundant peripheral and intermixed paraphyses. PARAPHYSES cylindric to capitate, 16.5-41 x 3-9.5 um (av. 30 x 5 um), walls hyaline, 0.5-1.5 um thick (av. 1 um). UREDINIOSPORES pedicellate, globose to subglobose, 22-35 x 21-30.5 um (av. 30 x 27 um), walls hyaline or pale yellow, 1-3.5 um thick (av. 1.5 um), echinulate, germ pores obscure. TELIA mostly hypophyllous, dark brown to black, subepidermal, covered by epidermis, without paraphyses. TELIOSPORES 2-celled by transverse septum, borne singly on pedicels, clavate to oblong, with round to obtuse apex and attenuate towards base, 28.5-40.5 x 9.5-17 um (av. 35.5 x 13 um), walls pale brown to dark brown, 0.5-1.5 um thick at sides (av. 1 um), 1-5 um thick at apex (av. 2.5 um), smooth; pedicels short, hyaline. Puccinia caulophylli comb. nov. (China) ... 727 Fic. 4. Puccinia caulophylli on Milium effusum: uredinial and telial stages. A. Uredinia and telia produced on the leaves; B. Pale yellow uredinia (U) on lower leaf surface; C. Dark brown telia on lower leaf surface; D. Echinulate urediniospores; E. Vertical section of uredinium with urediniospores and paraphyses (P); F. Teliospores; G. Vertical section of telia covered by host epidermis. Scale bars: D, E = 30 um; F, G = 20 um. 728 ... Ji & al. Fic. 5. Puccinia caulophylli observed by SEM. A. Aecium with catenulate aeciospores and peridium; B. Aeciospore with densely verrucose surface; C. Uredinium with urediniospores and paraphyses (P); D. Echinulate urediniospore; E. Vertical section of a telium covered by host epidermis. Scale bars: A = 30 um; B, D = 5 um; C, E= 20 um. ADDITIONAL SPECIMENS EXAMINED—Stages 0, I on Caulophyllum robustum: CHINA: JILIN PROVINCE, Jilin, 23 June 2015 (HMJAU 853); 24 June 2015 (HMJAU 8531, HMJAU 8534); 29 June 2017 (HMJAU 8620); 1 July 2018 (HMJAU 8624). Stages II, III on Milium effusum: CHINA: JILIN PROVINCE, Jilin, 12 September 2017 (HMJAU 8622); 1 July 2018 (HMJAU 86232). Yanbian, 28 July 2015 (HMJAU 8618).; Baishan, 2 September 2018 (HMJAU 8625); 3 September 2018 (HMJAU 8626). HEILONGJIANG PROVINCE: Wuchang, 5 July 2016 (HMJAU 8619); 9 September 2017 (HMJAU 8621). Puccinia caulophylli comb. nov. (China) ... 729 HostTs & DISTRIBUTION—Stages 0, I on Caulophyllum robustum: China (Miura 1928, Tai 1979), Japan (Ito 1950, Harada 1984, Hiratsuka & al. 1992). Stages II, UI on Milium effusum: China. Acknowledgments This work was financed by the Fungal Flora in Jilin Province (20130206073NY). We thank Dr E.H.C. McKenzie (Manaaki Whenua Landcare Research, Auckland, New Zealand) and Dr C.M. Denchev (Bulgarian Academy of Sciences, Sofia, Bulgaria) for critical reading of the manuscript and suggestions. We also thank Dr H. Koba (College of Arts and Sciences, J. F Oberlin University, Tokyo, Japan) for identification of M. effusum. Literature cited Azbukina ZM. 1984. The manual of rust fungi in Soviet Far East. Nauka, Moscow. (In Russian) Azbukina ZM. 2005. Rust fungi. Cryptogamic plants, fungi and mosses of the Russian Far East, vol. 5. Dalnauka, Vladivostok. (In Russian) Cummins GB. 1971. The rust fungi of cereals, grasses and bamboos. Springer-Verlag, New York. Cummins GB, Hiratsuka Y. 2003. Illustrated genera of rust fungi, 3 ed. American Phytopathological Society, St. Paul, Minnesota. Harada Y. 1984. Materials for the rust flora of Japan IV. Transactions of Mycological Society of Japan 25: 287-294. Hiratsuka N, Sato S, Katsuya K, Kakishima M, Hiratsuka Y, Kaneko S, Ono Y, Sato T, Harada Y, Hiratsuka T, Nakayama K. 1992. The rust flora of Japan. Tsukuba-shuppankai, Tsukuba. Ito S. 1950. Mycological flora of Japan, vol. 2, no 3. Yokendo, Tokyo. Ji JX, Li Z, Wan Q, Li Y, Kakishima M. 2016. Notes on rust fungi in China 1. Autoecious life cycle of Puccinia tatarinovii on Prenanthes. Mycotaxon 131: 653-661. https://doi.org/10.5248/131.653 Ji JX, Li Z, Wan Q, Li Y, Kakishima M. 2017a. Life cycle of Aecidium klugkistianum on Ligustrum and its new combination, Puccinia klugkistiana. Mycoscience 58: 307-311. https://doi.org/10.1016/j.myc.2017.01.004 Ji JX, Li Z, Wan Q, Li Y, Kakishima M. 2017b. Notes on rust fungi in China 3. Puccinia adenocauli comb. nov. and its life cycle and new host. Mycotaxon 132: 141-148. https://doi.org/10.5248/132.141 Ji JX, Li Z, Li Y, Kakishima M. 2019. Two new species of Pucciniastrum producing dimorphic sori and spores from northeast of China. Mycological Progress 18: 529-540. https://doi.org/10.1007/s11557-018-1460-z Liu M, Hambleton S. 2013. Laying the formation for a taxonomic review of Puccinia coronata s.l. in a phylogenetic context. Mycological Progress 12: 63-89. https://doi.org/10.1007/s11557-012-0814-1 Miura M. 1928. Flora of Manchuria and east Mongolia 3. Cryptogams, Fungi. Minamimanshutetsudo, Dalian. Ono Y. 2016. Phakopsora hornotina, an additional autoecious rust species on Meliosma in the Philippines and the Ryukyu Islands, Japan. Mycoscience 57: 71-78. https://doi.org/10.1016/j.myc.2015.09.003. Padamsee M, McKenzie EHC. 2017. The intriguing and convoluted life of a heteroecious rust fungus in New Zealand. Plant Pathology 66: 1248-1257. https://doi.org/10.1111/ppa.12672 730 ... Ji & al. Saccardo PA, Sydow P. 1902. Supplementum universale, pars 5. Sylloge fungorum 16. 1291 p. Scholler M, Lutz M, Aime MC. 2019. Repeated formation of correlated species in Tranzschelia (Pucciniales). Mycological Progress 18: 295-303. https://doi.org/10.1007/s11557-018-1417-2 Tai FL. 1979. Sylloge fungorum sinicorum. Science Press, Beijing. Zhuang JY, Wei SX, Wang YC. 1998. Flora fungorum sinicorum, vol.10, Uredinales (1). Science Press, Beijing. MY COTAXON ISSN (print) 0093-4666 (online) 2154-8889 Mycotaxon, Ltd. ©2019 October-December 2019—Volume 134, pp. 731-735 https://doi.org/10.5248/134.731 Exserticlava aquatica sp. nov., a microfungus from the Brazilian Amazon LUANA TEIXEIRA DO CARMO’, DIOGO CARELI DOS SANTOS?, CAROLINA RIBEIRO SILVA”, SHEILA MIRANDA LEAO FERREIRA’, THAMARA ARAO FELETTI’, Luis FERNANDO PASCHOLATI GUSMAO”? ' Universidade Estadual de Feira de Santana, Programa de Pés-graduagao em Botanica, Av. Transnordestina s/n, Novo Horizonte, 44036-900, Feira de Santana, Brazil ? Universidade Federal de Pernambuco, Centro de Biociéncias, Depto de Micologia, Av. Prof. Nelson Chaves, s/n, Cidade Universitaria, 50670, Recife, Pernambuco, Brazil * Correspondence to: lgusmao@uefs.br ABSTRACT—A new species, Exserticlava aquatica, collected on submerged decaying twigs in Para State in the Brazilian Amazon, is described and illustrated. The microfungus is characterized by monoblastic conidiogenesis with repeated percurrent conidiophore extensions and conidiogenous cells with a slightly swollen apex that does not protrude beyond the ruptured outer wall. Key worps—Ascomycota, Chaetosphaeriaceae, conidial fungi, freshwater fungi, taxonomy Introduction Exserticlava S. Hughes is characterized by distoseptate conidia and a hyaline apical swelling of the conidiogenous cell formed by the inner wall disrupting a pigmented outer wall (Hughes 1978). Tsui & al. (2001) reviewed the morphology of the genus and provided a key to five species; subsequently two more species were described: E. yunnanensis and E. manglietiae (Cai & Hyde 2007, Ren & al. 2012). Exserticlava is associated with the perithecial genus Chaetosphaeria Tul. & C. Tul. (Reéblova & Seifert 2003, Seifert & al. 2011: 204). During a survey of microfungi associated with twigs in estuary areas, an interesting specimen was collected and is described here as a new Exserticlava species. 732 ... Carmo & al. Material & methods Samples of submerged decaying twigs were washed in river water to remove sand and placed in plastic bags. In the laboratory, the samples were processed according Castafeda-Ruiz & al. (2016) and regularly examined for 40 days under stereomicroscope for observation of the reproductive structures. Slide mounts were prepared in PVL (polyvinyl alcohol, lactic acid, and phenol) and/or in lactic acid. Microphotographs were obtained using a Olympus BX51 microscope with Nomarski interference optics. The type specimen has been deposited in the Herbarium of Universidade Estadual de Feira de Santana, Brazil (HUEFS). Taxonomy Exserticlava aquatica L.T. Carmo, C.R. Silva, Careli, S.M. Ledo, Feletti & Gusmao, sp. nov. PLATE 1 MB 831391 Differs from Exserticlava triseptata by its monoblastic conidiogenesis, its conidiogenous cells with a slightly swollen apex that does not protrude beyond the ruptured outer wall, and its smaller oblong-obtuse conidia. Type: Brazil, Parad State: Belém, Mosqueiro island, 1°03’45”S 48°20'14”W, on submerged decaying twigs of unidentified plant, 2.V1.2018, coll. L.T. Carmo (Holotype, HUEFS249946). EryMo.oey: the specific epithet refers to the aquatic habitat from which the species was collected. CoLonigs effuse, brown. MyceLium superficial and immersed, composed of septate, branched, smooth, pale brown hyphae. CoNnrIDIOPHORES macronematous, mononematous, unbranched, erect, straight or slightly flexuous, smooth, thick-walled, dark brown to paler toward the apex, slightly swollen at the base, surrounded by pseudo-parenchymatous cells forming a brown stroma 22.5-27 um wide, (6-)8-13-septate, 330-455 x 6.5-7.5 um. CONIDIOGENOUS CELLS monoblastic, enteroblastic, percurrent and progressive leading to the production of up to 7 conidiogenous cell extensions, integrated, terminal, clavate, slightly swollen at the apex, without conspicuous expansion of the inner wall and not protruding from the fragmented and pigmented outer wall of conidiogenous cells, smooth, pale brown, 18.5-46.5 x 5-7.5 um. Conidial secession schizolytic. Conip1a acrogenous, holoblastic, solitary, smooth, oblong-obtuse, thick-walled, 3-distoseptate, sometimes with conspicuous PuaTE 1. Exserticlava aquatica (holotype, HUEFS 249946): A. General aspect of conidiophore with attached conidium; B. Conidiogenous cell development; C. Conidium attached to conidiogenous cell; D. Conidiogenous cell; E. Proliferating conidiogenous cells; F. Conidiogenous cell producing a conidium after extension; G-I. Conidia. Scale bars: A = 50 um; B-I = 10um. Exserticlava aquatica sp. nov. (Brazil) ... 733 734 ... Carmo & al. aperture on distosepta, pale brown, 21-26.5 x 10-11.5 um; conidia sometimes produced within intercalary old conidiogenous cells. SEXUAL MORPH: not observed. Note: Exserticlava triseptata, E. keniensis, and E. vasiformis most closely resemble E. aquatica by producing 3-distoseptate conidia; the four other accepted species are 1-distoseptate (Tsui & al. 2001, Rao & de Hoog 1986). However, E. triseptata and E. keniensis have brown elliptic-obovoid conidia and the conidiogenous cells extend forming a hyaline swelling that produces conidia; and E. vasiformis is distinguished by vigorous inner wall extension of funnel-shaped conidiogenous loci (Hughes 1978, Matsushima 1975, Tsui & al. 2001). Exserticlava aquatica is easily separated from the other Exserticlava species by its pale brown conidia, its several percurrent conidiogenous cell extensions, and the distinctive crown-like appearance of the outer wall of the conidiogenous cell; the new species is further characterized by monoblastic conidiogenous cells and an absence of conspicuous inner wall expansion to fragment the pigmented outer conidiogenous cell wall. Exserticlava triseptata and E. vasiformis have been widely recorded in Brazil, including in the Amazon region (Cruz & al. 2008). Acknowledgments We are indebted to Dr. De-Wei Li and Dr. Rafael F. Castafieda Ruiz for critical review of the manuscript. LTC and TAF thank the Programa de Pds-graduacao em Botanica (PPGBot/UEFS), the Nacional Council for Scientific and Technological Development (CNPq), and the “Coordenacgao de Aperfeigoamento de Pessoal de Nivel Superior (CAPES). DCS and CRS thank the Programa de Pés-graduacao em Biologia de Fungos (PPGBF/UFPE). LFPG is grateful to the CNPq (Proc. 312984/2018-9). Literature cited Cai L, Hyde KD. 2007. Anamorphic fungi from freshwater habitats in China: Dictyosporium tetrasporum and Exserticlava yunnanensis spp. nov. and two new records for Pseudofuscophialis lignicola and Pseudobotrytis terrestris. Mycoscience 48(5): 290-296. https://doi.org/10.1007/s10267-007-0369-1 Castafieda-Ruiz RF, Heredia G, Gusmao LFP, Li DW. 2016. Fungal diversity of Central and South America. 197-217, in: DW Li (ed.). Biology of microfungi... New York, Springer International Publishing. https://doi.org/10.1007/978-3-319-29137-6_9 Cruz ACR, Hernandez-Gutiérrez A, Gusmao LFP. 2008. O género Exserticlava (fungo anamorfo—Hyphomycetes) no Brasil. Revista Brasileira de Botanica 31(2): 357-361. https://doi.org/10.1590/S0100-84042008000200018. Hughes SJ. 1978. New Zealand fungi 25. Miscellaneous species. New Zealand Journal of Botany16(3): 311-370. https://doi.org/10.1080/0028825X.1978.10425143 Exserticlava aquatica sp. nov. (Brazil) ... 735 Matsushima T. 1975. Icones microfungorum a Matsushima lectorum. Kobe, Published by the author. 209 p. Rao V, de Hoog GS. 1986. New or critical hyphomycetes from India. Studies in Mycology 28. 84 p. Réblova M, Seifert KA. 2003. Six new species of Chaetosphaeria from tropical rain forests in Thailand and redescription of Chaetosphaeria hiugensis. Sydowia 55: 313-347. Ren SC, MaJ, Zhang XG. 2012 [“2011”]. Two new species of Exserticlava and Spiropes on decaying wood from Guangdong, China. Mycotaxon118: 349-353. https://doi.org/10.5248/118.349 Seifert K, Morgan-Jones G, Gams W, Kendrick B. 2011. The genera of hyphomycetes. CBS Biodiversity Series 9. 997 p. Tsui CK, Goh TK, Hyde KD. 2001. A revision of the genus Exserticlava, with a new species. Fungal Diversity 7: 135-143. MYCOTAXON ISSN (print) 0093-4666 (online) 2154-8889 Mycotaxon, Ltd. © 2019 October-December 2019—Volume 134, pp. 737 https://doi.org/10.5248/134.737 Regional annotated mycobiotas new to the Mycotaxon website ABSTRACT—Mycotaxon is pleased to add to our ‘web-list’ page the following new annotated species distribution list under South America (Brazil): “Ascomycota (lichenized and non- lichenized) on Syagrus coronata in the Caatinga biome: new and interesting records for Brazil and South America” by Maiara A.L. dos Santos, Nilo G.S. Fortes, Tassio E.F. Silva, Nadja S. Vitoria. This brings to 133 the number of free access Fungae now available on our website: http://www.mycotaxon.com/mycobiota/index.html SOUTH AMERICA Brazil Matra A.L. DOs SANTOS, NILO G.S. FoRTES, TAssio E.F. Sitva, NADJA S. VITORIA. Ascomycota (lichenized and non-lichenized) on Syagrus coronata in the Caatinga biome: new and interesting records for Brazil and South America. 10 p. ABSTRACT—The Caatinga biome occupies most of the semiarid region of northeastern Brazil, with varied landscapes and notable endemism. Among the plants having significant importance in the Caatinga environment is the palm tree Syagrus coronata, which is known as the “life-saving plant” due to its high socio- biological and economic value. To better understand the mycota of the Arecaceae, collections were undertaken in the municipalities of Paulo Afonso and Nova Gloria within the Raso da Catarina eco-region in the drylands (“sertao”) of Bahia State, Brazil. Twenty species of Ascomycota were identified during the present work: three are new records for South America (Diplodia galiicola, Seimatosporium corni, and Wojnowiciella viburni); eleven are new records for Brazil (Anthostomella caricis, Caryospora callicarpa, C. putaminum, Chaetomium subaffine, Diatrype bermudensis, Diatrypella persicae, Didymosphaeria massarioides, Eutypella fraxinicola, Munkovalsaria donacina, Oedohysterium sinense, and Pleospora calvescens); while six are new records for Bahia State (Dirinaria confusa, Lecanora achroa, Phaeosphaeria sp., Pleospora herbarum, Polymeridium julelloides, and Saccardoella macrasca). Syagrus coronata represents a new botanical host for all taxa identified here. Key worps—Pezizomycotina, semiarid, taxonomy MYCOTAXON ISSN (print) 0093-4666 (online) 2154-8889 Mycotaxon, Ltd. © 2019 October-December 2019—Volume 134, pp. 739 https://doi.org/10.5248/134.739 Regional annotated mycobiotas new to the Mycotaxon website ABSTRACT—MycotTaxon is pleased to add a new annotated species distribution list to our 134 previously posted free access fungae. The 22-page “Checklist of Bolivian Agaricales. 1: Species with dark and pink spore prints.” by E. Melgarejo-Estrada, M.E. Suarez, D. Rocabado, O. Maillard, and B.E. Lechner may be downloaded from our website via http://www.mycotaxon.com/mycobiota/index.html SOUTH AMERICA Bolivia E. MELGAREJO-EsTRADA, M.E. SUAREZ, D. ROCABADO, O. MAILLARD, B.E. LECHNER. Checklist of Bolivian Agaricales. 1: Species with dark and pink spore prints. 22 p. ABSTRACT—We provide a literature-based checklist of Agaricales reported from Bolivia. In this first contribution, 101 species belonging to 28 genera and 9 families are listed. Pluteaceae, Agaricaceae and Hymenogastraceae are the most species- abundant families. Key worps—Basidiomycota, distribution, diversity, Gasteromycetes, Neotropics, macromycetes, South America Exserticlava aquatica sp. nov. (Carmo & al.— Pate 1, p. 733)