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AR TICLE Calocybella, a New Genus For IMA FUNGUS · 6(1): 1–11 (2015) doi:10.5598/imafungus.2015.06.01.01 Calocybella, a new genus for Rugosomyces pudicus (Agaricales, ARTICLE Lyophyllaceae) and emendation of the genus Gerhardtia Alfredo Vizzini1, Giovanni Consiglio2, Ledo Setti3, and Enrico Ercole1 1Department of Life Sciences and Systems Biology, University of Torino, Viale P.A. Mattioli 25, I-10125 Torino, Italy; corresponding author e-mail: [email protected] 2Via Ronzani 61, I-40033 Casalecchio di Reno (Bologna), Italy 3Via C. Pavese 1, I-46029 Suzzara (Mantova), Italy Abstract: Calocybella is a new genus established to accommodate Rugosomyces pudicus. Phylogenetic Key words: analyses based on a LSU-ITS sequence dataset place Calocybella sister to Gerhardtia from which it differs Agaricomycetes morphologically in the presence of clamp-connections and reddening context. The genus Gerhardtia is Calocybe emended to also include taxa with smooth spores. According to our morphological analysis of voucher Lyophyllaceae material, Calocybe juncicola s. auct. is shown to be Calocybella pudica. Lyophyllum FORTHCOMING MEETINGS FORTHCOMING tricholomatoid clade LSU and ITS sequences taxonomy Article info: Submitted: 12 January 2015; Accepted: 10 March 2015; Published: 23 March 2015. INTRODUCTION cutting or bruising, and red-violaceous after applying a drop of NH3 or KOH, and verruculose spores. Since these features The generic name Rugosomyces, typified by Agaricus appeared aberrant within Rugosomyces, they established the onychinus, was established by Raithelhuber (1979) for the new subsect. Rubescentes of sect. Rugosomyces for it. As lyophylloid species (taxa with siderophilous basidia) previously this puzzling taxon combines features of several genera within placed in Calocybe with a collybioid habit, bright colourations Lyophyllaceae, the taxonomic position of this species has been (vacuolar pigment) and a pileipellis consisting of inflated, greatly debated and was far from clear. Contu & Ortega (2001) pseudoparenchymatic elements. Later, the circumscription provided SEM micrographs of the spores showing evident of the genus was emended and broadened by Bon (1991a) Rhodocybe-like verruculose ornamentation, and elevated to include, besides the taxa with a more or less subcellular subsect. Rubescentes to the rank of section. They recognized pileipellis (sect. Rugosomyces), also species with a an affinity of the species with taxa placed in the genus trichodermic pileipellis formed by short, catenulate elements Gerhardtia, which, however, are devoid of clamp-connections. (sect. Carneoviolacei), mixed parietal and intracellular Contu & Consiglio (2004) recombined Rugosomyces pudicus pigments and distinct from Calocybe which was restricted to into Lyophyllum which they employed in a broad sense to the species with paler colorations, a cutis-like pileipellis and a include Calocybe. Arnolds (2006), following the statements tricholomatoid habit. A few years later, the genus Rugosomyces of Hofstetter et al. (2002) and Moncalvo et al. (2002) based was monographed by Bon (1999) and accepted by Kalamees on molecular markers, did not recognize Rugosomyces as (1995, 2004, 2012a) and Horak (2005). Phylogenetic analyses independent from Calocybe and recombined R. pudicus into of Lyophyllaceae (Hofstetter et al. 2002, 2014) based on Calocybe. nuclear and mitochondrial rDNA sequences showed that the Picillo & Contu (2009) reported R. pudicus also from a generic concept based only on morphological characters littoral site in Latium (Sabaudia), and adopted the concept of (Singer 1986, Bon 1999, Consiglio & Contu 2002, Kalamees Arnolds (2006), highlighting affinities with Calocybe and not 2004, 2012a–c) was artificial and should be re-framed; in with Tephrocybe, due to the vacuolar pigment. Finally, Vizzini particular, Rugosomyces species form a single clade together et al. (in Vizzini 2014) combined R. pudicus into Gerhardtia with species of Calocybe. Rugosomyces pudicus, a striking because of the spore ornamentation. species of Rugosomyces sect. Rugosomyces, was not As this rare enigmatic species had not been studied included in the molecular work by Hofstetter et al. (2002, 2014). molecularly so far, the aim of this paper was to determine It was described by Contu & Bon (2000) on the basis of a its phylogenetic position within Lyophyllaceae based on collection from Sardinia (Italy) and characterized by a Collybia- LSU and ITS rDNA analyses as well as to fully describe and or Callistosporium-like habit, the context turning blood-red on illustrate it on the basis of recent collections. © 2015 International Mycological Association You are free to share - to copy, distribute and transmit the work, under the following conditions: Attribution: You must attribute the work in the manner specified by the author or licensor (but not in any way that suggests that they endorse you or your use of the work). Non-commercial: You may not use this work for commercial purposes. No derivative works: You may not alter, transform, or build upon this work. For any reuse or distribution, you must make clear to others the license terms of this work, which can be found at http://creativecommons.org/licenses/by-nc-nd/3.0/legalcode. Any of the above conditions can be waived if you get permission from the copyright holder. Nothing in this license impairs or restricts the author’s moral rights. (44) VOLUME 6 · NO. 1 1 Vizzini et al. ARTICLE 2 bold. bootstrap support > 70 %. For each sequenced taxon the Genbank/UNITE number is given. Newly sequenced collections are in > 0.95 and ML probability posterior Bayesian indicate branches Thickened branches. the on shown are % 50 ≥ values MLB and 0.7 ≥ bold) (in values BPP taxa. Fig. 1. Fig. Sphagnurus palustre AF223199 Tricholomella constricta AF223186 Bayesian phylogenetic analysis based on the LSU sequences of sequences LSU the on based analysis phylogenetic Bayesian Sagaranella gibberosa AF223198 0.96/70 0.98/93 Sagaranella tylicolor AF223195 0.83/50 Sagaranella tylicolor AF223192 Sagaranella gibberosa AF223196 0.79/82 Sagaranella gibberosa AF223197 1/96 Ossicaulis lignatilis AF261397 Ossicaulis lachnopus HE649956 0.75/- Ossicaulis lignatilis AF261396 0.90/64 Hypsizygus tessulatus DQ917664 0.95/- Tephrocybe sp. AY207304 0.99/60 Hypsizygus ulmarius AF042584 1/100 Asterophora parasitica AF223191 Asterophora lycoperdoides AF223190 Calocybe gangraenosa AF223202 1/100 Lyophyllum ochraceum AF223185 1/89 0.99/100 Lyophyllum favrei AF223184 Lyophyllum favrei AF223182 1/88 Calocybe ionides AF223179 0.95/70 1/81 Calocybe obscurissima AF223181 0.81/- Calocybe gambosa AF223177 1/100 Calocybe persicolor AF223176 0.94/61 Calocybe carnea AF223178 Calocybe cyanea AF261400 0.89/- Calocybe naucoria AF223180 Rugosomyces onychinus AM946467 Calocybe s.l. 0.92/61 Myochromella inolens AF223201 1/93 Myochromella boudieri DQ825430 Myochromella boudieri AF223206 Myochromella boudieri AF223204 0.92/75 Myochromella boudieri AF223205 Myochromella 1/72 1/99 Gerhardtia pseudosaponacea KJ461911 1/99 Gerhardtia highlandensis EF535275 0.74/85 Gerhardtia borealis AY232716 1/99 Gerhardtia borealis AM946449 Gerhardtia sp. EF421091 Gerhardtia borealis AMB 15993 KP858009 Gerhardtia 1/100 Calocybella pudica AMB 15997 KP858008 Calocybella pudica AMB 15996 KP858007 Lyophyllaceae Calocybella pudica AMB 15995 KP858006 Calocybella pudica AMB 15994 KP858005 Calocybella Tephrocybe rancida AF223203 0.89/62 Blastosporella zonata EU708337 0.77/- Termitomyces medius AY127796 1/99 0.80/52 Termitomyces subhyalinus AF223174 Termitomyces globulus AY232695 1/100 Termitomyces mammiformis AY232701 Termitomyces mammiformis AY232703 , with Termitomyces clypeatus AF261398 0.86/51 Termitomyces letestui AY127800 0.88/- Termitomyces titanicus AY232715 Entoloma sinuatum Termitomyces singidensis AY232713 0.86/58 Termitomyces aurantiacus AY232690 Termitomyces heimii AF042586 Termitomyces robustus AY232711 0.79/- Termitomyces striatus AY232714 0.96/76 Termitomyces eurrhizus AY232694 Termitomyces schimperi AY232712 0.99/75 Termitomyces microcarpus AY127798 0.73/54 Termitomyces microcarpus AY232708 Termitomyces entolomoides AY232693 0.73/77 1/86 Termitomyces microcarpus AY232707 Podabrella microcarpus AF042587 and 1/100 Arthromyces matolae EU708336 Arthromyces matolae EU708335 0.90/64 Arthromyces claviformis EU708334 E. prunuloides 0.97/93 Lyophyllum anthracophilum AF223213 Lyophyllum atratum KJ461895 1/100 1/100 Lyophyllum fumosum HM572542 1/100 Lyophyllum fumosum AY207229 1/98 0.98/79 Lyophyllum decastes AY207228 1/94 Lyophyllum decastes AF042583 Lyophyllum ambustum AF223214 0.86/71 Lyophyllum ambustum AF223215 0.99/78 AF042581 as outgroup as Lyophyllum semitale Lyophyllum caerulescens AF223209 Lyophyllum moncalvoanum KJ461891 0.92/100 Entoloma sinuatum AY691891 1/100 Entoloma sinuatum KC710154 Entoloma prunuloides AY700180 0.01 expected changes per site IMA FUNGUS Calocybella gen. nov. and Gerhardtia (Lyophyllaceae) Samples sequenced for the present study. Table 1. ARTICLE Species GenBank accession no. Source and country LSU ITS Calocybella pudica KP858005 KP858000 AMB 15994, Lido di Ostia (RM), Latium, Italy Calocybella pudica KP858006 KP858001 AMB 15995, Sabaudia (LT), Latium, Italy Calocybella pudica KP858007 KP858002 AMB 15996, Bassa del Bardello (RA), Emilia-Romagna, Italy Calocybella pudica KP858008 KP858003 AMB 15997, Bassa del Bardello (RA), Emilia-Romagna, Italy Gerhardtia borealis KP858009 KP858004 AMB 15993, Castelìr, Bellamonte (TN), Trentino-Alto Adige, Italy MATERIALS AND METHODS DNA extraction, PCR amplification,
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