Persoonia 38, 2017: 38–57 www.ingentaconnect.com/content/nhn/pimj RESEARCH ARTICLE http://dx.doi.org/10.3767/003158517X693174

Taxonomy of Tricholoma in northern Europe based on ITS sequence data and morphological characters

J. Heilmann-Clausen1, M. Christensen2, T.G. Frøslev 3, R. Kjøller 4

Key words Abstract Based on molecular and morphological data we investigated the taxonomy and phylogeny of the ecto­ mycorrhizal genus Tricholoma in northern Europe. Our phylogenetic tree confirmed the presence of at least 72 Agarics well circumscribed species within the region. Of these, three species, viz. T. boreosulphurescens, T. bryogenum biogeography and T. ilkkae are described as new to science, based on morphological, distributional, ecological and molecular cryptic species data. Several other terminal branches represent putative cryptic taxa nested within classical species or species ectomycorrhizal fungi groups. Molecular type studies and/or designation of sequenced neotypes are needed in these groups, before host selection the taxonomy can be settled. In general our phylogenetic analysis supported previous suprageneric classification morphological traits systems, but with some substantial changes. Most notably, T. virgatum and allies were found to belong to sect. phylogeny Tricholoma rather than sect. Atrosquamosa, while T. focale was found to be clearly nested in sect. Genuina rather Tricholomataceae than in sect. Caligata. In total, ten sections are accepted, with five species remaining unassigned. The combination of morphological and molecular data showed pileus colour, pileipellis structure, presence of clamp connections and spore size to be rather conservative characters within accepted sections, while the presence of a distinct ring, and especially host selection were highly variable within these.

Article info Received: 23 January 2015; Accepted: 12 May 2016; Published: 26 August 2016.

Introduction indicated to be probable in Albomagister and Pseudotricho­ loma. Only the latter genus is known to occur in Europe, were The genus Tricholoma is a classic genus of agarics already pro­ Pseudotricholoma metapodium is widespread. posed as a section by Fries (1821) and subsequently erected Tricholoma has a worldwide distribution (Tedersoo et al. 2010), as a genus by Staude (1857). but seems to be most prominent in temperate and subtropical Over the years more than 850 species epithets have been pu­ zones in both the southern and northern hemisphere. All known blished or combined in the genus. Many of these have since species are known or supposed to be ectomycorrhizal (Ryberg been transferred to other genera, including Lepista, Leucopaxil­ & Matheny 2011), mainly with trees in the Pinaceae, Betulaceae lus, Lyophyllum and Melanoleuca based on various deviations, and Fagaceae, but the genus also contains species that are mainly in microscopic characters. Molecular studies have sup- associated with Eucalyptus, Dryas and Helianthemum (Bougher ported the segregation of most of these more modern genera, 1996, Christensen & Heilmann-Clausen 2013). Some species and Tricholoma in its narrow circumscription (e.g. Noordeloos form dual ectomycorrhizal and monotropoid associations linking & Christensen 1999) is supported as a monophyletic genus of trees and monotropoid plants (Leake et al. 2004). The centre ectomycorrhizal fungi within the Tricholomataceae (e.g. Mon- of species richness appears to be in North America. According calvo et al. 2002). According to Ryberg & Matheny (2012), the to Bessette et al. (2013), more than 100 species are reported genus seems to have segregated from its ancestral clade some from this continent while 63 to 88 species are listed from Eu- 60–90 million years ago in the late Cretaceous, possibly with rope (Riva 1988, Bon 1991, Kirby 2012). Several species are Pinaceae as mycorrhizal partners. A recent study (Sánchez- described or reported from Japan, New Zealand and Australia García et al. 2014) surveyed in depth the Tricholomataceae (e.g. Hongo 1988, Bougher 1996, Orlovich & Cairney 2004), but based on several molecular markers and concluded that only the overview of the species diversity in these regions is frag- the genera Albomagister, Corneriella, Dennisiomyces, Leuco­ mentary due to the lack of modern comprehensive treatments. paxillus, Pseudotricholoma, Porpoloma s.str. and Tricholoma Tricholoma species show limited microscopic variation, and are belong to the family, while other previously contained genera characterized by hyaline, subglobose to oblong spores, simple should be assigned to families, including the Lyophyllaceae pileipellis structures and lack of well-differentiated sterile ele- and a poorly resolved residual Tricholomatoid clade. Of the ments, including cystidia. Hence, species identification and genera included in the Tricholomataceae, also Porpoloma partly also the infrageneric classification has mainly been s.str. is proven ectomycorrhizal, while a biotrophic lifestyle is based on macromorphology. Singer (1986) divided the genus in four subgenera, mainly based on pileipellis structure and 1 University of Copenhagen, Natural History Museum of Denmark, Center for Macroecology, Evolution and Climate, Universitetsparken 15, DK-2100 the presence or absence of clamp connections. The four sub- Copenhagen, Denmark; genera were further divided into nine sections, of which three corresponding author e-mail: [email protected]. (Leucorigida, Iorigida and Adusta) do not belong to the genus 2 Hvidtjørnevej 1, DK-4180 Sorø, Denmark. in the current circumscription. In their treatment of the genus, 3 University of Copenhagen, Natural History Museum of Denmark, Center for Geogenetics, Øster Voldgade 5-7, 1350 Copenhagen, Denmark. Noordeloos & Christensen (1999) accepted the four subgenera 4 University of Copenhagen, Institute of Biology, Section of Terrestrial Ecol- suggested by Singer (1986), but with a more narrow definition ogy, Universitetsparken 15, DK-2100 Copenhagen, Denmark. of sections, especially in subg. Tricholoma, in which seven sec-

© 2016-2017 Naturalis Biodiversity Center & Centraalbureau voor Schimmelcultures 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. J. Heilmann-Clausen et al.: Tricholoma in northern Europe 39 tions were accepted. Slightly deviating classification systems connections were explored in more detail, but many other have been proposed by other authors, including Bon (1984a, characters were described for accepted species in Christensen 1991). For a more throughout evaluation see Riva (1988) and & Heilmann-Clausen (2013). Christensen & Heilmann-Clausen (2013). Despite their attractive fruit bodies, and a long mycological tradi- Molecular & phylogenetic methods tion, the overall taxonomy in Tricholoma is still poorly resolved in DNA was extracted from dried specimens by the CTAB-chloro­ Europe. Molecular data have been used to study the taxonomy form method described by Gardes & Bruns (1993). Usually, and phylogeny of some species groups (e.g. Comandini et al. one lamella was taken with a flamed pair of forceps from the 2004, Jargeat et al. 2010, Ota et al. 2012, Moukha et al. 2013), specimens. The internal transcribed spacer (ITS) region was typically resulting in the identification of cryptic diversity within amplified with the primer combination ITS1-F and ITS4 (White previously accepted species. Simultaneously, several species et al. 1990, Gardes & Bruns 1993). The PCR products were se- have been proposed in recent years without a published test quenced by Europhins Genomics (previously MWG-Biotech) or of taxonomic placement based on molecular markers (e.g. Macrogen (www.macrogen.com). Forward and reverse strands Kalamees 2001, Musemeci & Contu 2008, Ferrarese & Zaf- were sequenced using ITS1F or ITS5 (White et al. 1990) and falon 2010, Ludwig 2012), adding to taxonomic confusion in ITS4 as sequencing primers. If sequencing of either the ITS-1 the genus. or ITS-2 region proved difficult, additional sequencing was performed using ITS2 and ITS3 (White et al. 1990) as sequen- Recently, Christensen & Heilmann-Clausen (2013) monograph­ cing primers. Sequence contigs were assembled using Se- ed the genus in northern Europe, backed by sequences of the quencher (v. 3.1). Alignment was done with MAFFT (online v. 7) nuclear ribosomal internal transcribed spacer (ITS). The main using the settings G-INS-i (Katoh et al. 2005), with minor manual objective of the present paper is to present the results of the adjustments in Se-Al (Rambaut 1996) for some sequences underlying scientific studies in a coherent form, with the fol- with incomplete ends or internal gaps. Two alignments were lowing specific aims: produced, one containing all sequences in the dataset, and one 1. to evaluate the monophyly of proposed subgenera and sect- based on a reduced dataset containing only one representative ions as defined by Singer (1986) and Noordeloos & Chris- of each of 72 end-clusters accepted to represent described or tensen (1999); putative species present in northern Europe. For each align- 2. to investigate the congruence between ITS and morphology ment Maximum Likelihood phylogeny estimates were produced in hypothesized sections; and with RAxML v. 8.1.16 (Stamatakis et al. 2008) with 1 000 fast 3. to resolve the taxonomical delimitation of Tricholoma spe- bootstrap replicates and GTR + CAT base substitution model. cies occurring in northern Europe, with a consideration of Both alignments were also subjected to bayesian phylogenetic possibly related taxa occurring in other continents, espe- analyses using MrBayes v. 3.2 (Ronquist et al. 2011) using cially North America. the GTR+I+gamma model (nst = 6, rates = invgamma) with two independent runs of 4 chains for 5 000 000 generations Materials and Methods with sampling every 1 000th generation. Trees from the last 1 000 000 generations from each run (2 000 trees from each Studied material analysis) were summed in a consensus tree with branch fre- Specimens studied for this paper were mainly collected by the quencies corresponding to bayesian posterior probabilities. first two authors during collection trips throughout Europe, since Sequences used in this study are listed in Table 1 including the early 1990s. It was the intention to obtain representative col- source information, geographic origin, and accession numbers. lections of all species present in northern Europe according to Alignments can be obtained from the first author. modern identification books (Gulden 1969, 1992, Noordeloos & Christensen 1999, Christensen & Heilmann-Clausen 2012), but Taxonomic part in addition a number of species recorded from southern Europe were included. Generally, several collections of each species In total we obtained 217 novel ITS sequences for this study, were included in the analysis, and if possible, specimens from while 84 published sequences were downloaded from GenBank different geographical regions were selected. In addition to (67) and Unite (17). The alignment contained 170 unique se- own material we have studied a number of collections, inclu­ quence reads, represented as terminal clusters in the phyloge- ding type-specimens from various public and private herbaria netic tree based on maximum likelihood (Fig. 1). The Bayesian throughout Europe, and for the phylogenetic analyses selected analyses did not contradict the ML phylogeny. Based on tree relevant and trustworthy ITS sequences were downloaded topology these were assigned to 108 putative species hypo­ from GenBank and Unite. Finally, we included a few original theses, of which 27 were represented only by extra-continental ITS sequences kindly provided by Tor Erik Brandrud (Norway) sequences, while seven represented accepted species or spe- and Sven-Gunnar Ryman (Sweden). cies hypotheses only recorded from southern Europe. Of the 81 species hypotheses identified among European sequences, Scoring of morphological characters 72 were selected for scoring of morphological characters and Macromorphological characters were mainly scored on fresh evaluation of previously published infrageneric classification material or more rarely photographs (for details on studied systems (Fig. 2). collections see Christensen & Heilmann-Clausen 2013). Mor­ Below we first evaluate the infrageneric classification system, phological characters were described according to the standard and subsequently the species level taxonomy is reviewed. We terminology published by Knudsen & Vesterholt (2008), while have applied commonly used section names as practical labels colours were recorded according to Kornerup & Wanscher for clades in order to guide readers. We have not intended to (1974). Microscopical characters were recorded from rehydrated resolve the nomenclatural history of each section in depth, specimens in 2 % KOH or 5–10 % NH3. From each collection a as this would require a deeper and quite time-consuming no- minimum of 20 randomly selected spores were measured, avoi- menclatural study on candidate section names. As long as the ding obviously malformed or unripe spores. For this study, data details of the infrageneric classification remain open, due to on pileus and gill colour, pileus surface texture, presence of limited sapling of the genus in North America, and the use of a ring-zone, spore size and the presence/absence of clamp only one molecular marker, we find the time unripe to perform 40 Persoonia – Volume 38, 2017 Notes holotype epitype neotype Outgroup GenBank accession no. LT000199 JF908737 LT000101 LT000102 LT000013 LT000002 AM181413 LT000158 AF377238 LT000001 LT000012 AB036898 AB856037 LT000100 LT000120 LT222031 AY750166 AF377233 LT222024 LT222019 LT000078 AF349701 LT000119 LT000157 LT000127 LT000198 LT000156 LT000155 LT000011 LT000010 LT000118 LT000009 LT000154 LT000008 AB510358 AB699646 LT000135 LT000077 LT000115 LT000153 JN021102 LT000007 LT000006 LT000134 LT000005 HM561970 AB510472 AF377247

UDB000790 UDB000791 UDB000811 UDB001412 UDB011587

UDB001471 UDB001470

UDB001438 UDB002362 UDB001419 UDB000785 UDB001692 UDB000782 UDB000781 UDB000780 UDB001685 UDB001467 UDB011580 UDB001413 UDB001218 UDB018044 UDB001444 UDB002370 UDB001434 UDB000779 UDB002361 UDB001474

Unite accession no. H6002040 LUG F 8450 L0354472 C-F-96201 C-F-96233 C-F-38408 C-F-96240 C-F-59330 C-F-59329 O-64018 O188799 O159872 C-F-59255 C-F-59200 C-F-59014 C-F-96245 L0374886 C-F-96215 C-F-96213 C-F-96212 C-F-35092 C-F-59207 C-F-41884 C-F-35189 C-F-96254 C-F-96234

C-F-96268 C-F-96250 C-F-58885 C-F-59265 C-F-40955 C-F-96223 C-F-41483

Herbarium no.

L H LUG C C C C C TEB C O O O C C L C C C C C C C C C C C

C C C C C C C Herbarium GenBank This study This study This study Unite GenBank This study GenBank This study This study GenBank GenBank This study This study This study GenBank GenBank This study This study This study GenBank This study This study This study This study This study This study This study This study This study This study This study This study Unite GenBank GenBank This study This study Unite This study Unite This study GenBank This study This study This study This study This study GenBank GenBank GenBank This study Origin of sequence Finland Italy, Trento, Spera val Campanella Trento, Italy, Peura, Raemäki Tervola, Finland, Outer Ostrobothnia, Italy, Probe Brallo, Pavia Italy, Estonia Estonia Denmark, Anholt USA, Califonia Croatia, Istarska, Kanegra Sweden, Öland, Halltorps Haga Japan Japan Croatia, Primorsko-Goranska, Krk Denmark, Møn, Møns Klint USA, Washington USA, Califonia Norway, Aust-Agder, Evje & Hornnes, Dåsvassdalen, Husefjell SW Aust-Agder, Norway, Agnese Cipressa di Toscana, Italy, Gommerud, Vestfossen Buskerud, Øvre Eiker, Norway, Norway, Møre og Romsdal, Norddal Norway, USA, Califonia Norway, Hordaland, Granvin, Urdanes NR Norway, France, Franche-Comte, St. Sifolene Norway, Hedmark, Hornmoen Norway, The Netherlands, Groningen, Verhildersum near Leens The Netherlands, Groningen, Verhildersum Sweden, Medelpad, Borgsjö Slovakia, Presov, Snina Slovakia, Presov, Sweden, Medelpad, Bräcke, Grötingen Sweden, Värmland, Långban S of Lesjöfors Denmark, Sjælland, København, Botanisk Have Denmark, Sjælland, Geel Skov Norway, Hedmark, Hornmoen Norway, Denmark, Jylland, Skagen Klitplantage Estonia Turkey Morocco Sweden, Medelpad, Borgsjö, Bergåsen SÖ Denmark, Jylland, Skivum Krat Estonia Slovenia, Ljubljana Sweden France, Provence Canada, Ontario Nepal, Mustang, Kunjo Sweden, Medelpad, Harrån Denmark, Jylland, Hårup Sande Denmark, Jylland, Sønder Herreds Plantage Slovenia, Central Vino Malaysia Mexico Norway Denmark, Jylland, Elbæk Skov Locality 1996 1997 1996 1991 2001 1998 2001 1997 2008 1996 2010 2003 2005 1998 1998 1994 1995 2003 1997 1997 1996 1995 1998 2000 1995 1995 2001 1999 2005 1997 1994 2002 2000 1999 Collection year Voucher Voucher

JF908737 MEN96112 IK971187 LUG-F8450 UDB011587 AM181413 JHC91-721 AF377238 MC01-200 TL5303 AB036898 AB856037 MC01-209 MC97-227 TEB55008 AY750166 AF377233 MC96-303 O-F64018 O-F188799 AF349701 O-F159872 MC98-120 MC98-020 MEN9491 MC95-102 MC03-251 JHC97-174 JHC97-092 JHC96-244 JHC95-112 MC98-034 JV00-215 UDB011580 AB510358 AB699646 JHC95-049 MC95-159 UDB018044 MC01-201 UDB001218 MC99-060 JN021102 MC05-201 MC97-072 MC94-008 JV02-540 MC00-204 HM561970 AB510472 AF377247 JV99-638 Sequences included in this study. For sequences retrieved from genbank or unite, only accession numbers and country of origin is given, for new sequences obtained for this study, year of collection, locality and fungarium For sequences retrieved from genbank or unite, only accession numbers and country of origin is given, for new obtained this study, Sequences included in this study. sejunctum’ sejunctum’ aff. virgatum’ aff. Table 1 Table data is given. Species names are given as in Fig. 1. Species

boreosulphurescens T.

bonii T. T. batschii T.

T. ‘bakamatsutake’ ‘bakamatsutake’ T. basirubens T.

‘atroviolaceum’ T. ‘aurantium’ T. aurantium T.

T. ‘atrosquamosum’ ‘atrosquamosum’ T. atrosquamosum T.

arvernense T.

T. argyraceum T.

anatolicum T. apium T.

album T.

T. albobrunneum T. T. ‘ T. ‘

T. aestuans T.

T. ‘Sp. Mex1’ ‘Sp. Mex1’ T. acerbum T. Hypsizygus marmoreus J. Heilmann-Clausen et al.: Tricholoma in northern Europe 41 neotype neotype neotype holotype epitype holotype LT000023 LT000140 LT000168 LT000167 LT000075 AF349689 LT000022 LT000021 LT000166 LT000129 AB036895 AF458452 AF458453 AF458456 DQ822834 AF309534 AF377236 LT000139 LT000138 LT000165 LT000020 LT000019 LT000018 LT000017 AF377239 AB738883 holotype AF309523 JF908732 LT000164 AF349693 LT000137 LT000016 LT000200 LT000015 LT000128 LT000074 JN021103 LT000079 LT222034 AF309520 LT000152 KC152249 KC565866 LT222026 LT222020 AY462034 LT000105 LT000104 LT000103 LT000162 LT000014 LT000136 LT000161 LT222030 LT000160 LT222023 LT222032 LT000159 UDB001509 UDB001504 UDB002364 UDB001500 UDB001501 UDB000803 UDB000804 UDB001506 UDB001469 UDB001508 UDB001468 UDB001433 UDB001417 UDB000544 UDB000543 UDB001420 UDB001442 UDB000793

UDB000550 UDB000549 UDB000792 UDB001428 UDB000807 UDB001475 C-F-59084 C-F-96227 C-F-59188 C-F-59031 C-F-59243 C-F-96260 C-F-41444 C-F-27500 C-F-96243 C-F-96226 C-F-96191 C-F-35924 C-F-58890 C-F-96256 C-F-58886 C-F-58898 C-F-59154 C-F-96238 C-F-59068 C-F-59057 C-F-96246 C-F-96265 C-F-96274 OF6702 OF52108 OF160040 C-F-59167 TRgmb00652 C-F-59342 C-F-59341 C-F-59442 C-F-59305 C-F-90092 C-F-96219 C-F-96211 O187683 OF21089 C-F-59441

C C C C C C C C C C C C C C C C C L C C C C C C TROM O O C TR C C C C TEB C C C O TROM C This study This study This study GenBank This study This study This study This study GenBank GenBank GenBank GenBank GenBank GenBank GenBank This study This study This study This study This study This study This study GenBank GenBank GenBank GenBank This study GenBank This study This study This study This study This study This study GenBank This study This study GenBank This study GenBank GenBank This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study Denmark, Jylland, Mols Slovenia, Gorizia, Idria Sweden, Jämtland, Fillstabäcken USA, New Mexico Sweden, Medelpad, Borgsjö France, Franche-Comte, Winkel Denmark, Jylland, Skagen Denmark, Læsø, Træbakke at Holtemmen Træbakke Denmark, Læsø, Japan USA, Oregon USA, Oregon USA, Oregon USA, California USA, Califonia USA, Califonia Sweden, Jämtland, Orrskäret Slovakia, Presov, Rozok Slovakia, Presov, Slovenia, Gorizia, Idria Slovenia, Ljubljana (exhibition) Sweden, Skåne, Balsberget Denmark, Jylland, Bakkerne near Ørsted Denmark, Jylland, Hoverdal USA, Califonia Sweden USA, Califonia Italy Denmark, Jylland, Hoverdal Plantage Norway Denmark, Jylland, Skivum Krat Sweden, Jämtland, Ysjö Sweden, Jämtland, The Netherlands, Drenthe, Boekweitveentje Slovenia, Ljubljana Denmark, Jylland, Borum Denmark Canada, Quebec Slovakia, Presov, Havesova Slovakia, Presov, France, Provence, Massif des Cedres Costa Rica Mexico Algeria Spain, Valencia, E of Gandía, N Els Spain, Valencia, France, Provence Norway, Troms, Storfjord, Lullesletta Troms, Norway, Norway, Nord-Trøndelag, Levanger, Ytterøya Levanger, Nord-Trøndelag, Norway, Norway, Oppland, Øystre Slidre, Heggnes Norway, Sweden, Jämtland, Brunflo Italy, Sardegna, Sassari, Calangianus, Baldo Italy, Italy, Toscana, Monte Soldano Toscana, Italy, Italy, Toscana, Monte Soldano Toscana, Italy, Sweden, Öland, Haltorp Hage Denmark, Jylland, Moesgaard Skov Norway, Oppland, Nord-Aurdal, Mjølkebekken nordre Norway, Slovenia, Ljubljana Sweden, Medelpad, Borgsjö, Julåsen Sweden, Medelpad, Borgsjö, Julåsen Norway, Finnmark, Alta, Kålfjordsbotten Finnmark, Norway, Norway, Troms, Storfjord, Lullesletta Troms, Norway, Sweden, Medelpad, Stöde, V. Västansjö, Kockerabäcken Sweden, Medelpad, Stöde, V. 1996 2000 1997 1995 1998 1998 1999 1997 2003 2000 2001 1996 1996 1995 1994 1995 1997 1995 2001 1996 1996 2003 1999 2007 1999 1995 1979 2006 1997 2006 1996 1996 1994 1995 2006 2001 1995 1995 2004 2007 1995 MC96-235 MC00-225 MC97-151 AF349689 MC95-130 MC98-086 MC98-600 JV99-603 AB036895 AF458452 AF458453 AF458456 DQ822834 AF309534 AF377236 JV97-239 MC03-242 MC00-218 JHC0-1202 C-F35924 MC96-155 MC95-187 AF377239 AB738883 AF309523 JF908732 MC94-027 AF349693 MC95-181 MC97-047 MEN95210 MC01-205 MC96-170 MC96-134 JN021103 MC03-252 MC99049 AF309520 JV07-451 KC152249 KC565866 PH99-519 TROM-F6702 O-F52108 O-F160040 MC97-101 TRgmb00652 MC96-265 MC96-264 CL94-166 MC95-317 TEB22606 MC01-600 JV95-307 JHC95-067 O-F187683 TROM-F21089 SAE9507 japonicum’ cf. japonicum’

T. frondosae type II T.

T. frondosae type I T. T. ‘frondosae’ ‘frondosae’ T.

T. ‘flavovirens’ ‘flavovirens’ T. ‘focale’ T. focale T.

T. filamentosum T.

T. ‘dryophilum’ ‘dryophilum’ T. dulciolens T. equestre T. T. columbetta T.

colossus T.

T. ‘ cingulatum T. T. ‘cedrotorum’ ‘cedrotorum’ T. T. ‘caligatum’ ‘caligatum’ T. caligatum T. ‘caligatum’ T. caligatum T.

T. bryogenum T.

T. bresadolanum T.

boudieri T.

T. borgsjoeënse T.

T. boreosulphurescens (cont.) T. 42 Persoonia – Volume 38, 2017 neotype neotype holotype neotype Notes LT000182 LT000181 LT000179 LT000180 LT000178 TMU62964 AF377211 AF458445 LT000071 LT000029 EU597086 JN021108 AF458448 AF377224 AF309538 LT000028 LT000131 LT000082 LT000081 LT222022 LT000106 LT000177 LT000176 AB036900 HM590876 LT000027 LT000026 LT000025 LT000175 LT000174 LT000173 LT000024 AF377246 LT222029 LT222029 AF377242 LT222028 LT000172 LT222027 AF377229 AB738881 AB738882 LT000065 LT000122 LT000070 LT000080 LT000171 LT000130 LT000121 AB289668 DQ067895 LT000170 LT000169 GenBank accession no. UDB000525 UDB000527 UDB000526 UDB000799

UDB000005 UDB001696 UDB000798 UDB000797 UDB000539 UDB000538 UDB000537 UDB000783 UDB000796 UDB001424 UDB001688 UDB001421 UDB000699

UDB001701 UDB002365 UDB001700 UDB001695 UDB002363 Unite accession no. C-F-59168 C-F-59036 C-F-35209 C-F-96247 C-F-59446 C-F-96230 C-F-96194 C-F-96267 C-F-96266 O167194 TR gmb 00600 C-F-96262 C-F-59159 C-F-59094 C-F-59272 C-F-96242 C-F-35211 C-F-59020 C-F-35182 C-F-59268 F-513823 F-173364 C-F-96261 O74354 C-F-96217 C-F-96251 C-F-96259 C-F-96195 C-F-96193 C-F-59201 C-F-58980 C-F-59395 Herbarium no.

C S C C C TURA C C C C C O TR C C C C C C C C C UPS UPS C O C C TURA C C C C C C Herbarium This study This study This study GenBank GenBank GenBank This study This study GenBank GenBank GenBank GenBank GenBank This study This study This study This study This study This study This study This study GenBank GenBank This study This study This study This study This study This study This study Unite GenBank This study GenBank This study This study This study GenBank GenBank GenBank This study This study This study This study This study This study This study GenBank GenBank This study This study This study This study Origin of sequence Sweden, Jämtland, Brunflo Sweden, Lappland, Åsele Lappmark, Risbäck, S slope of arksjöberget Sweden, Medelpad, Borgsjö South Korea USA, Califonia USA, Oregon Sweden, Medelpad, Borgsjö, Julåsen Sweden, Jämtland Canada, British Columbia Canada, Ontario USA, Oregon USA, Califonia China, Yunnan Finland, Koillismaa, Oulanka national park, NWW of biological field station Denmark, Sjælland, Lellinge Skovhusvænge Denmark, Sjælland, Rude Skov Slovakia, Poloniny National Park, Rozok France, Provence, Massif des Cedres France, Provence, Massif des Cedres Norway, Akershus, Bæarum, Borøya Norway, Italy, Trento, Baselga di Piné, Cané Trento, Italy, Japan France Sweden, Gotland Sweden, Jämtland, Østersund Denmark, Jylland, Aarhus Denmark, Jylland, Denmark, Jylland, Aarhus Denmark, Jylland, Denmark, Jylland, Århus Sweden, Medelpad, Borgsjö, Erikslund Sweden, Medelpad, Borgsjö Sweden USA, Califonia Tubbobäcken Sweden, Medelpad, USA, Califonia Denmark, Jylland, Bakkerne near Ørsted Sweden, Gotland, Eksta par, Ekstastrand Sweden, Gotland, Eksta par, Sweden, Uppland, Gräsö par., Djupdal 3 km NE of Gräsö church Sweden, Uppland, Gräsö par., USA, Califonia Spain Spain Sweden, Gotland Norway, Hordaland, Ulvik, Finse Norway, Estonia, Saaremaa Norway, Akershus Norway, Finland, Varsinais-Suomi, Vahto, Seppälä, Ryssänvuori Vahto, Finland, Varsinais-Suomi, France, Franche-Comte, Winkel Åkulla, Valaklitt Sweden, Halland, Varberg, Japan Thailand Slovakia, Poloniny National Park, Nova Sedlica Norway, Hedmark, Sørskogbygdn Norway, Sweden, Jämtland, Fillstabäcken Sweden, Jämtland, Kyckås Locality 1997 1993 1995 1995 2003 2005 1999 2000 2003 1999 1999 2004 2005 1998 1997 1996 1995 2003 1995 1995 1995 1994 2000 2007 1998 2005 2008 1995 2000 1998 2004 2003 1998 1997 1997 Collection year MC97-103 KJ1993 MC95-135 TMU62964 AF377211 AF458445 JHC95-070 MC03-600 EU597086 JN021108 AF458448 AF377224 AF309538 JuV23362F MC99-197 MC00-519 JHC03-020 MC99-056 MC99-053 O-F167194 TRgmb00060 AB036900 HM590876 MC98-603 MC97-060 MC96-172 MC95-152 MC03-229 JHC95-072 MC95-115 UDB000699 AF377246 JHC95-042 AF377242 MC94-046 S-F513823 S-F173364 AF377229 AB738881 AB738882 MC98-602 O-F74354 JV08-364 MC95-103 JuV16997 MC98-078 JHC04-251 AB289668 DQ067895 JHC03-019 MC98-023 MC97-149 MC97-158 Voucher Voucher

Table 1 (cont.) Table

‘moserii’ T. ‘mutabile’ T. olivaceotinctum T.

T. ‘leucophyllum’ ‘leucophyllum’ T. ‘luteomaculosum’ T. ‘magnivelare’ T. matsutake T.

T. lascivum T.

T. josserandii T. T. ‘joachimii’ ‘joachimii’ T.

T. ‘japonicum’ ‘japonicum’ T. ‘joachimii’ T. joachimii T.

T. inocybeoides T.

‘inamoenum’ T. inamoenum T. T. ‘imbricatum’ ‘imbricatum’ T. imbricatum T.

T. ‘huronense’ ‘huronense’ T. ilkkae T.

T. hemisulphureum T.

T. guldeniae T.

T. ‘fulvocastanem’ ‘fulvocastanem’ T. fulvum T.

T. fucatum T. Species frondosae type II (cont.) T.

J. Heilmann-Clausen et al.: Tricholoma in northern Europe 43 neotype epitype neotype epitype LT000049 LT000048 LT000047 LT000110 LT000046 LT000045 AB036899 AF377192 EU819447 LT000044 LT000004 LT000187 holotype HQ184113 LT000146 LT000043 LT000042 LT000087 DQ494700 LT000086 LT000041 LT000040 LT000039 LT000186 LT000185 LT000133 LT000123 LT000038 LT000109 LT000073 LT000072 LT000085 LT000037 LT000125 HQ184102 LT000084 LT000036 LT000108 LT000145 LT000083 LT000035 LT000034 LT000144 LT000033 LT000184 LT000143 AF349686 LT000032 LT000142 LT000202 DQ494699 LT000031 LT000107 LT000132 LT000030 LT000141 LT000183 UDB000786 UDB000530 UDB000532 UDB001431 UDB000548 UDB000547 UDB000784 UDB000542 UDB000541 UDB001498 UDB001507 UDB001689 UDB001505 UDB001699 UDB001697 UDB001694 UDB001693 UDB001499 UDB001432 UDB001439 UDB001418 UDB000795 UDB001503 UDB001472 UDB001429 UDB001409 UDB001686 UDB001698 UDB001410 UDB011624 UDB001502 UDB011581 UDB000802 UDB000520 UDB000521 UDB000522 UDB000801 UDB000523 UDB000524 UDB000800 C-F-35151 C-F-96209 C-F-96205 C-F-58979 C-F-58998 C-F-96255 C-F-58902 C-F-96206 C-F-59399 C-F-96225 C-F-35309 C-F-96210 C-F-96276 C-F-59217 C-F-96218 C-F-96216 C-F-35147 C-F-96198 C-F-96196 C-F-96192 C-F-96188 C-F-23337 C-F-59393 H6002034 H6002032 C-F-59258 C-F-96241 C-F-96263 C-F-96273 C-F-59212 C-F-59324 C-F-96248 C-F-59262 C-F-59053 C-F-58959 C-F-96224 C-F-96202 C-F-96197 C-F-96229 C-F-43780 C-F-96190 C-F-59315 C-F-59427 C-F-59365 C-F-96244 C-F-96208 C-F-96189 C C C C C C C C C C C C C C C C C C C C C C C H H C C C C C C C C C C C C C C C C C C C C C UPS C This study This study This study This study GenBank GenBank GenBank This study This study This study GenBank This study This study This study This study GenBank This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study GenBank This study This study This study This study This study This study This study This study This study This study This study Unite GenBank This study Unite This study This study GenBank This study This study This study This study This study This study This study This study Denmark, Sjælland, Lellinge Skovhusvænge Denmark, Jylland, Trelde Østerskov Trelde Denmark, Jylland, Denmark, Jylland, Trelde Østerskov Trelde Denmark, Jylland, Italy, Toscana, Cipressa di Agnese Cipressa di Toscana, Italy, Japan USA, Califonia USA, Wisconsin Denmark, Jylland, Enemærket Skov Denmark, Jylland, Borum Skov Denmark, Jylland, Fløjstrup Skov France Denmark, Jylland, Nystrup Klitplantage Sweden, Jämtland, Sundsnäs Slovenia, Ljubljana Denmark, Fyn, Tankefuld W. of Svendborg W. Tankefuld Denmark, Fyn, USA, Massachusetts Denmark, Jylland, Trelde Østerskov Trelde Denmark, Jylland, France, Doubs, Forêt Valdahon France, Franche-Comte, Foret de Leval Denmark, Jylland, Fløjstrup Skov Denmark, Sjælland, Frederikskilde Skov Denmark, Sjælland, Sorø Sønderskov Sweden, Halland, Laholm, Blåalt Sweden, Halland, Hylte, Ödegärdet Poloniny National Park, Stuzika Norway, Sogn og Fjordane, Leikanger, Horpa Sogn og Fjordane, Leikanger, Norway, Denmark, Lolland, Ryde Skov Italy, Lazio, Monte Rufenum Italy, Finland, Uusimaa, Hanko, Koverhar, Krogars Finland, Uusimaa, Hanko, Koverhar, Finland, Sompio Lapland, Sodankylä, Jeesiö-Tepsa France, Franche-Comte, Doubs, St.-Julien les Russey France Denmark, Jylland, Stråsø Plantage Portugal, Leiria France, Provence Denmark, Jylland, Fløjstrup Skov Italy, Toscana, Monte Soldano Toscana, Italy, Slovenia, Ljubljana France, Franche-Comte, Doubs, St.-Julien les Russey Denmark, Jylland, Bakkerne near Ørsted Denmark, Sjælland, Ordrup Næs Slovenia, Central Vino Denmark, Lolland, Christianssædeskovene Estonia USA, Califonia Sweden, Halland, Hylte, Ödegärdet Estonia Slovenia, Gorizia, Idria Denmark, Jylland, Ålbæk Klitplantage USA, Massachusetts Slovenia, Idria, Pringle England, Surrey, Norbury Park England, Surrey, Denmark, Jylland, Moesgaard Skov Italy, Toscana, Cipressa di Agnese Cipressa di Toscana, Italy, Slovakia, Presov, Rozok Slovakia, Presov, Denmark, Jylland, Trelde Østerskov Trelde Denmark, Jylland, Sweden, Jämtland, Sällsjö Slovenia, Idria, Mehke Dolina 1995 1993 1993 1996 1995 1995 1994 1993 1995 2000 1994 1993 1998 1998 1987 1997 1995 2004 2004 2003 2000 1992 1996 1992 1988 1998 2003 1999 1999 1998 1996 2004 1998 1996 1994 2000 1992 2004 2000 2004 2001 1998 1997 1996 2003 1993 1981 2001 JHC95-169 JHC93-262 JHC93-224 MC96-314 AB036899 AF377192 EU819447 MC95-187 MC95-182 MC94-007 HQ184113 JHC93-243 MC95-165 MC00-207 JHC94-231 DQ494700 JHC93-263 TF98-098 MC98-059 JV87-682 JHC97-237 JHC95-165 JHC04-439 JHC04-429 JHC03-015 JHC00-049 C-F23337 MC96-376 IK922945 IK881120 MC98-106 HQ184102 MC03228 MC99-044 MC99-089 MC98-048 MC96-345 MC04-600 MC98-116 MC96-156 MC94-082 MC00-206 JHC92-277 UDB011624 AF349686 JHC04-431 UDB011581 MC00-236 JV04-482 DQ494699 JHC01-201 MC98-214 MC97-258 MC96-301 MC03-243 JHC93-261 OP1981 JHC01-200

T. squarrulosum T.

T. ‘sejunctum’ ‘sejunctum’ T. sejunctum T.

T. sciodes T. meleagroides’ ‘scalpturatum forma meleagroides’ T. atrocinctum’ ‘scalpturatum var. T.

T. ‘saponaceum’ ‘saponaceum’ T. scalpturatum T.

T. saponaceum T. T. rufenum T.

T. roseoacerbum T.

pseudotriste’ pseudotriste’ ‘ramentaceum var. T. rapipes T. T. ‘quercetorum’ ‘quercetorum’ T.

T. psammopus T.

portentosum T. populinum T. T. pessundatum T. T. ‘palustre’ ‘palustre’ T. pardinum T.

T. olivaceotinctum (cont.) T. T. orirubens T. 44 Persoonia – Volume 38, 2017 Notes neotype epitype neotype epitype GenBank accession no. LT000067 LT000066 LT000194 LT000076 LT000099 LT000062 LT000061 LT000060 LT222021 LT000093 LT000098 LT000092 LT000201 LT000193 LT000116 LT000149 LT000059 LT000058 LT000057 LT000091 EU439339 EU439340 LT222033 AF377245 LT222025 LT000090 AY462035 AY462038 AY462036 LT000148 LT000192 LT000053 AY462037 LT000191 LT000113 AF377244 EU819448 HQ650743 LT000089 LT000112 LT000051 LT000050 LT000190 LT000124 LT000189 LT000188 LT000088 LT000111 LT000147 LT000003

UDB001691 UDB000805 UDB000814 UDB000535 UDB000534 UDB000808 UDB001446 UDB000813 UDB001445 UDB000533 UDB001425 UDB002368 UDB001411 UDB000812 UDB000536

UDB001440 UDB001416 UDB011543 UDB002367 UDB000809 UDB002366 UDB001684 UDB002398 UDB011582 UDB001427 UDB001426 UDB000529 UDB000531 UDB000528 UDB001414 Unite accession no. H6002036 C-F-96214 C-F-96222 C-F-96220 C-F-96204 C-F-96277 O165767 C-F-96272 L0374887 C-F-96271 C-F-59313 C-F-96253 C-F-96249 C-F-96232 C-F-35154 C-F-35098 C-F-96207 C-F-96275 OF30019 OF288529 C-F-59260 C-F-59062 C-F-59292 C-F-58914 C-F-96237 Missing C-F-96199 C-F-59115 C-F-96200 TR gmb 00062 C-F-96269 C-F-59362 C-F-90094 C-F-96221 C-F-96252 C-F-96258 C-F-59042 C-F-59032 C-F-59238 C-F-59343 C-F-96239 C-F-96235 Herbarium no.

H TURA L C C C C C L O C C C C C C C C C C TROM O C C C C C C C C TR C C C C C C C C C C C C Herbarium This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study GenBank GenBank This study GenBank This study This study This study This study This study This study This study This study This study This study This study Unite GenBank GenBank GenBank This study This study This study This study This study Unite Unite This study This study This study This study This study This study This study This study This study Origin of sequence Finland, Satakunta, Jämijärvi, Hämeenkangas Estonia, Pärnu rajooni, c. 40 km S of Pärnu, Kabli Germany, Baden-Württemberg, Seedorfer Wald, Schwarzwald Baden-Württemberg, Seedorfer Wald, Germany, of Änge Sweden, Jämtland, Lockna, W. France, Provence, Petit Luberon, Massif des Cedres Denmark, Jylland, Staksrode Skov Denmark, Jylland, Trelde Østerskov Trelde Denmark, Jylland, Denmark, Jylland, Klim Bjerg Germany, Bayern, Sperberslohe near Roth Germany, Grua, Olsknappen Oppland, Lunner, Norway, France, Provence, Foret des Caderach France, Provence, Foret des Caderach Holland, Schouwen-Duiveland Sweden, Medelpad, Borgsjö Nepal, Mustang, Lete Slovenia, Ljubljana Denmark, Sjælland, København, Assistens Kirkegård Denmark, Sjælland, København, Denmark, Sjælland, Kongelunden China, Yunnan China, Yunnan Denmark, Jylland, Trelde Østerskov Trelde Denmark, Jylland, Norway France, Doubs, Forêt de Levier Norway, Troms, Storfjord, Lullesletta Troms, Norway, Norway, Oppland, Vang, Uri Oppland, Vang, Norway, France, Franche-Comte, Doubs, St.-Julien les Russey Denmark, Jylland, Løvenholm Skov Denmark, Jylland, Enemærket Skov Sweden, Skåne, Drakamöllan Denmark, Jylland, Kås Hoved Slovenia, Ljubljana Denmark, Lolland, Favrsted Skov Denmark, Jylland, Øjesø Estonia USA, Califonia USA, Wisconsin Canada, British Columbia Sweden, Halland, Halmstad, Nissaström Italy, Trento, Baselga del Bondone Trento, Italy, France, Provence Italy, Toscana, Cipressa di Agnese Cipressa di Toscana, Italy, Denmark, Jylland, Råbjerg Plantage Scotland Estonia Denmark, Læsø, Læsø Klitplantage, Vester Højsandshoved Denmark, Læsø, Læsø Klitplantage, Vester Sweden, Medelpad, Borgsjö Norway, Hedmark, Hornmoen Norway, Sweden, Medelpad, Borgsjö Sweden, Medelpad, Borgsjö France, Franche-Comte, Winkel Italy, Toscana, Monte Soldano Toscana, Italy, Slovenia, Ljubljana Croatia, Primorsko-Goranska, Krk Locality 1993 1990 1996 1997 1999 1995 1993 1993 1995 2005 1999 1999 1998 1995 2004 2001 1995 1995 1993 2006 1996 2008 1998 1996 1995 2007 1994 2001 2007 1996 2008 2005 1999 1996 1998 1996 1995 1998 1995 1995 1998 1996 2001 2001 Collection year Voucher Voucher

IK931613 JuV5271F E3754 JHC97-169 JV99-700 JV95-519 JHC93-222 TL11317 MEN95192 O-F165767 MC99-074 MC99-071 MC98-209 MC95-119 MC05-200 MC01-020 JHC95-172 JHC95-118 EU439339 EU439340 JHC93-260 AF377245 TF06045 TROM-F30019 O-F288529 MC98-109 MC96-162 MC95-188 MC07-001 MC94-023 MC01-204 JHC07-236 MC96-245 UDB011543 AF377244 EU819448 HQ650743 JHC08-049 TRgmb00062 MC99-063 MC96-296 MC98-601 UDB002398 UDB011582 JV96-306 MC95-117 MC98-018 MC95-145 MC95-131 MC98-081 MC96-269 MC01-700 MC01-202 Table 1 (cont.) Table Species

T. ‘ulvinenii’ ‘ulvinenii’ T.

T. triste T. T. ‘tridentinum’ ‘tridentinum’ T.

T. terreum (albinistic) T.

T. ‘terreum’ ‘terreum’ T. terreum T. T. sulphureum clade 3 T.

T. sulphureum clade 2 T.

‘sulphureum’ T. sulphureum clade 1 T. T. sulphurescens clade 2 T.

T. sulphurescens clade 1 T.

sudum T. T. stiparophyllum T.

T. stans T.

T. squarrulosum (cont.) T. J. Heilmann-Clausen et al.: Tricholoma in northern Europe 45

such a study. In the section on species level taxonomy nomen- clatural details are given on all species epithets. neotype Infrageneric classification and congruence with morphology The phylogenetic analysis did not support a clear division of the genus in four subgenera as proposed by Singer (1986) LT000117 FJ197008 LT000097 LT000096 LT000095 LT000197 LT000151 LT000196 LT000195 AF377230 LT000150 LT000094 LT000126 LT000064 AF377240 LT000114 AF377234 LT000063 LT000069 LT000068 and adopted by most subsequent authors. However, ten sec- tions could be reasonably separated based on molecular data and scoring of morphological traits (Fig. 2), with some species remaining unclassified. Pileus colour, pileipellis structure, pre­sence of clamp connections and spores size and shape appeared as rather constant characters supporting the validity

UDB001437 UDB011588 UDB011595 UDB001473 UDB001436 UDB000545 UDB001415 UDB000546 UDB001423 UDB001511 UDB000815 UDB011564 UDB000816 UDB000551

UDB002369 UDB011557 UDB011558 UDB011559

of sections, while the presence of a distinct ring, and especially host selection was variable within sections. It is well known that the ITS region alone is poorly suited for resolving higher level phylogenies (e.g. Frøslev et al. 2005), and our infrageneric classification should be viewed as phylogenetic supported, but C-F-96257 C-F-59249 C-F-59237 C-F-59219 C-F-59398 C-F-96236 C-F-35203 C-F-59017 C-F-96228 C-F-96270 C-F-96264 C-F-96203 TRgmb00651 C-F-96231 preliminary. However, we do trust the sections defined below as relevant hypothetical monophyletic entities that should be

tested in future studies combining a global taxon sampling with C C C C C C C C C C C C TR C TURA TURA multiple molecular markers.

Species with a greyish, radially fibrillose, squamulose to felty cap Species with a dry, grey and a radially fibrillose, squamulose to felty pileipellis quite clearly represent a paraphyletic group, GenBank Unite Unite This study This study This study This study This study This study GenBank This study This study Unite This study This study GenBank This study GenBank This study This study Unite Unite Unite This study This study This study that we here split across four sections; Terrea, Atrosquamosa, Tricholoma and Pardinicutis (Fig. 2). Section Terrea contains species characterized by a dry, felty or squamulose pileipellis, predominantly greyish colours, and spores with a relatively high Q-value. Our concept of the section is narrow, and corresponds­ to stirps Terrea in Singer (1986). Most previous authors, includ- ing Noordeloos & Christensen (1999) operated with a much broader concept of the section, which included also the stirps Virgata and Atrosquamosa ss. Singer (1986). Here, we accept the latter as a separate section, although our ITS phylogeny provide only limited support for monophyly, especially in the maximum likelihood analysis (Fig. 2). The similarity in morpho- logical traits of the assigned species is, however, striking. Spe- cies in the section are morphologically very similar to species in sect. Terrea, but tend to have spores with a higher Q-value, and are characterized by peculiar smells reminding of honey, ground pepper or cedar wood (compared to absent to farina- ceous in sect. Terrea). The species belonging to stirps Virgata in the sense of Singer (1986) (i.e. T. aestuans, T. bresadolanum, T. sciodes and T. virgatum) were in our analysis deeply nested in sect. Tricholoma. As discussed later, this makes good sense Mexico Estonia Estonia New Zealand, Arthurs Pass National Park New Zealand, France, Franche-Comte, Bois Lachat France, Franche-Comte, Winkel France, Franche-Comte, Bois de la Brosse Sweden, Jämtland, Halåsen Slovenia, Ljubljana USA, Califonia Sweden, Medelpad, Björnö, Björkviken Sweden, Medelpad, Borgsjö Estonia Slovenia, Gorizia, Idria France, Provence, Foret des Caderach USA, Califonia Portugal, Leiria The Netherlands Denmark, Sjælland, Suserup Skov Italy, Veneto, Belluno, Meleré Veneto, Italy, Estonia Estonia Estonia Denmark, Lolland, Roden Skov Alastaro, Virttaankangas Finland, Satakunta, Finland, Varsinais-Suomi, Dragsfjärd, Ölmos Finland, Varsinais-Suomi, morphologically. Finally, our analysis supported sect. Pardini­ cutis as a separate section. The members of this section are characterized by a grey, scaly pileipellis, large spores and the 1996 1998 1998 1998 1997 2001 1995 1995 2000 1999 1999 1992 2006 2000 2008 1997 presence of clamp connections, a combination that has lead most modern authors to accept Pardinicutis at the subgenus level.

Species with a reddish brown cap For the reddish brown species our ITS phylogeny showed a FJ197008 UDB011588 UDB011595 MC96-002 MC98-093 MC98-080 MC98-061 MC97-164 MC01-203 AF377230 JHC95-063 MC95-109 UDB011564 MC00-229 MC99-067 AF377240 MC99-047 AF377234 JHC92-299 TRgmb00651 UDB011557 UDB011558 UDB011559 MC00A01 JuV26740F JuV13229F division in three relatively well-supported sections, sect. Cali­ gata, sect. Genuina and sect. Megatricholoma (Fig. 2). The members of sect. Caligata are characterized by an annulate stipe, a whitish, pale brown to dark reddish brown squamose pileus, rather large spores with low Q-value, and a strong per- fumed smell. All European species are associated with conifers, but according to Murata et al. (2013) basal members of the clade from other parts of the world associate with deciduous hosts. The annulate species T. focale has traditionally been included in the section (e.g. Noordeloos & Christensen 1999), but the ‘Uncultured ectomycorrhiza’ ‘Uncultured ectomycorrhiza’ T. viridilutescens type II T. ‘viridiolivaceum’ T.

T. viridilutescens type I T.

T. ‘venenatum’ ‘venenatum’ T. virgatum T.

vaccinum T.

T. ‘ustaloides’ ‘ustaloides’ T. ustaloides T. T. ustale T. T. umbonatum type II T. umbonatum type I T.

T. ‘ulvinenii’ (cont.) ‘ulvinenii’ T. current study shows it to be deeply nested in sect. Genuina, 46 Persoonia – Volume 38, 2017

JV95519_terreum(albinistic)_DK JHC93-222_terreum(albinistic)_DK JHC95-172_terreum_DK TL11317_terreum_DK MC99-071_terreum_FR JHC93-260_terreum_DK MC99-074_terreum_FR MC98-209_terreum_NL 99 MC01-200_terreum_SI 1 O-F-165767_terreum_NO MEN95192_terreum*_DE MC95-119_terreum_SE JHC95-118_terreum_DK MC05-200_terreum_NEPAL JHC91-721_bonii_DK EU439340_"terreum"_CHINA MEN96112_bonii_IT 93 AM181413_bonii_EE 1 LUG F 8450_bonii*_IT E3754_triste*_DE EU439339_"terreum"_CHINA 99 AF377211_"moserii"_USA 1 JHC97-169_triste_SE JuV5271F_triste_EE MC96-134_cingulatum*_DK TERREA MC96-170_cingulatum_DK * MC03-252_cingulatum_SK MEN95210_cingulatum_NL 96 JHC95-112_argyraceum_DK 1 HQ184113_"scalpturatum f. meleagroides”_FR HQ184102_"ramentaceum var. pseudotriste*”_FR MC03-251_argyraceum_SK MEN9491_argyraceum*_NL JHC97-092_argyraceum_SE 51 JHC93-243_"scalpturatum var. atrocinctum”_DK 0.74 JHC97-174_argyraceum_SE JHC96-244_argyraceum_DK MC95-152_inocybeoides_DK 89 MC96-172_inocybeoides_DK 1 MC97-060_inocybeoides_SE MC03-229_inocybeoides_DK 84 JHC95-072_inocybeoides_SE 1 MC95-165_scalpturatum*_SE JHC94-231_scalpturatum_DK * MC00-207_scalpturatum_SI JHC93-263_scalpturatum_DK

TROM-F21089_boreosulphurescens_NO IK971187_boreosulphurescens_FI SAE9507_boreosulphurescens_SE * JF908737 _boreosulphurescens_FI * O-F187683_boreosulphurescens*_NO LASCIVA P.P. * MC96-296_sulphurescens type I_IT 80 MC99-063_sulphurescens type I_FR 1 * UDB011543_sulphurescens type II_EE TRgmb00062_sulphurescens type II_IT MC96-162_sulphureum type II_DK JHC07-236_sulphureum type II_DK MC95-188_sulphureum type II_DK MC07-001_sulphureum type II_SE MC94-023_sulphureum type II_DK * MC98-109_sulphureum type II_FR TROM-F30019_sulphureum type II_NO * MC01-204_sulphureum type II_SI O-F288529_sulphureum type II_NO 52 EU819448_"sulphureum"_USA 0.69 JHC95-042_inamoenum*_SE * MC95-115_inamoenum_SE AF377246_"inamoenum"_USA SERICELLA JHC08-049_sulphureum type I_SE 76 MC96-245_sulphureum type I_DK * 1 AF377244_"sulphureum"_USA 99 * JV08-364_hemisulphureum_EE O-F74354_hemisulphureum_NO 1 * * AF377245_sulphureum type III_NO TF06045_sulphureum type III_FR 75 O-F52108_bryogenum_NO MC97-101_"bryogenum"_SE 0.99 O-F160040_bryogenum_NO 95 TROM-F6702_bryogenum_NO 1 HQ650743_"sulphureum"_CANADA UDB011580_album_EE * MC95-159_album_DK 98 MC01-201_album_SI 1 JHC03-020_lascivum_SK * MC00-519_lascivum_DK LASCIVA S.STR. 91 MC99-197_lascivum_DK 1 UDB011582_stiparophyllum_EE 32 * MC95-117_stiparophyllum_SE 0.56 UDB002398_stiparophyllum_UK * JV95-307_borgsjoeense_SE * JHC95-067_borgsjoeense_SE AY750166_"atroviolaceum"_USA TEB22606_”borgsjoeensi”s_NO C-F35924_filamentosum_SE MC00-218_filamentosum_SI * MC03-242_filamentosum_SK * 69 JHC01-202_filamentosum_SI PARDINICUTIS 0.89 AF377229_"huronense"_USA * AF377230_"venenatum"_USA JHC01-201_pardinum_SI

0.05 Fig. 1 Phylogeny inferred from ITS regions for the full dataset, with branch lengths based on the Maximum Likelihood analysis. No notable differences in branching patterns were observed between the Bayesian and the Maximum Likelihood analysis. Maximum Likelihood bootstrap values are indicated above branches, while Bayesian posterior probabilities are indicated below branches. * Denotes 100 % support in both analyses. New sequences obtained for this study are indicated in regular letters, while sequences obtained from GenBank or Unite are given in italics. Species names without quotes represent our in- terpretation of relevant taxa as discussed in this paper. Names in quotes are not interpreted by us, but are given as in the original source, or by the collector. Hypothesized sections are indicated by background shadings with names in capital letters. J. Heilmann-Clausen et al.: Tricholoma in northern Europe 47

JHC95-063_virgatum_SE 65 MC97-164_virgatum*_SE * 0.9 MC01-203_virgatum_SI 99 MC05-201_"aff_virgatum"_NEPAL 1 * MC94-007_sciodes_DK 67 MC95-182_sciodes_DK 0.96 MC96-265_bresadolanum_IT MC96-264_bresadolanum_IT * 66 CL94-166_bresadolanum_SE 1 TRgmb00652_bresadolanum_IT MC97-072_aestuans*_SE * JV02-540_aestuans_DK * 45 MC94-008_aestuans_DK DQ494699_"palustre"_USA 0.84 JHC04-431_portentosum_SE MC98-116_portentosum_FR MC00-206_portentosum_SI 99 AF349686_portentosum_USA 1 MC96-156_portentosum*_DK 22 MC94-082_portentosum_DK 0.59 JHC92-277_portentosum_DK MC98-061_viridilutescens type I_FR 38 * MC98-093_viridilutescens type I_FR 0.6 45 MC98-080_viridilutescens type I_FR 0.57 * MC95-187_sejunctum_DK MC96-314_sejunctum_IT 53 94 JN021108_"leucophyllum"_CANADA 0.93 1 EU819447_"sejunctum"_USA MC96-376_rufenum_IT UDB011595_virdilutescens type II_EE UDB011588_viridilutescens type II_EE * AB036899_"sejunctum"_JAPAN 99 JN021102_"aff_sejunctum"_CANADA TRICHOLOMA 1 AF377192_"sejunctum"_USA 68 JuV13229F_"ulvinenii"_FI 0.99 JuV26740F_"ulvinenii"_FI UDB011557_"ulvinenii"_EE 88 UDB011559_"ulvinenii"_EE 1 AF458453_"flavovirens"_USA 64 AF458452_"flavovirens"_USA 0.96 DQ822834_"flavovirens"_USA MC96-155_equestre_DK 92 MC94-027_equestre_DK 35 1 0.85 MC96-235_frondosae type II_DK * MC00-225_frondosae type II_SI MC97-158_frondosae type II_SE 95 UDB011558_"ulvinenii"_EE 1 IK931613_"ulvinenii"_FI 86 MC95-187_”equestre”_DK 89 0.98 MC95-130_frondosae type I_SE 95 MC97-151_frondosae type I_SE 1 1 AF349689_frondosae type I_USA 87 MC98-086_"frondosae"_FR 61 0.99 83 O-F167194_"joachmii"_NO 0.96 97 0.96 AF458456_"flavovirens"_USA 1 EU597086_"leucophyllum"_CANADA TRgmb00060_joachimii_IT 82 * 96 MC98-603_joachimii_SE 1 1 AB036895_"flavovirens"_JAPAN 88 HM590876_"joachimii"_FR 1 * MC95-181_columbetta*_DK AF349693_columbetta_NO 62 * TRgmb00651_umbonatum type II_IT 1 MC00-A01_umbonatum type I_DK * MC95-103_guldeniae_NO JuV16997_guldeniae_FI MC03-600_matsutake_SE 91 TMU62964_matsutake_SOUTH KOREA 1 JuV23362F_matsutake_FI AF309538_matsutake_CHINA 82 * AB699646_anatolicum_MOROCCO 90 1 AB510358_anatolicum_TR 0.97 AB510472_"Sp. Mex1”_MEXICO 71 AF377224_"magnivelare"_USA 0.98 * KC152249_"caligatum"_MEXICO 41 AF309520_"caligatum"_COSTA RICA 0.47 JV07-451_caligatum_ES * KC565866_caligatum_ALGERIA PH99-519_caligatum_FR CALICATA * AB289668_"fulvocastaneum"_JAPAN DQ067895_"fulvocastaneum"_THAILAND S-F173364_ilkkae_SE 73 S-F513823_ilkkae*_SE 1 * MC98-602_ilkkae_SE AB738882_ilkkae_SP 86 AB738881_ilkkae_SP 1 AB738883_dulciolens*_SE 95 * JF908732_dulciolens_IT 1 AF309523_dulciolens_USA * AB856037_"bakamatsutake"_JAPAN AB036898_"bakamatsutake"_JAPAN MC99-056_josserandii_FR * MC99-053_josserandii_FR 59 AF458445_”mutabile”_USA 0.99 * MC97-149_fucatum*_SE MC98-023_fucatum_NO

0.05

Fig. 1 (cont) 48 Persoonia – Volume 38, 2017

JV97-239_focale*_SE 99 MC98-600_focale_DK 1 JV99-603_focale_DK AF309534_"focale"_USA 52 AF377236_”focale”_USA 26 0.9 MC01-200_batschii_HR 0.62 * AF377238_batschii_USA UDB011587_batschii_EE 88 MC96-303_aurantium_IT 44 0.95 MC97-227_aurantium_DK 0.89 60 AF377233_"aurantium"_USA 37 0.82 JV04-482_pessundatum*_DK UDB011581_pessundatum_EE 44 0.85 MC99-049_"cedrotorum"_FR 0.91 JV99-700_"tridentinum"_FR 49 AF377240_"ustaloides"_USA 0.84 MC00-236_populinum_SI UDB011624_populinum_EE 72 UDB001218_albobrunneum_SE 1 * UDB018044_albobrunneum_EE MC99-060_albobrunneum_FR MC95-131_stans_SE 38 99 AF377239_"dryophilum"_USA GENUINA 0.73 1 MC95-145_stans*_SE MC98-018_stans_NO UDB011564_ustaloides_EE 41 99 MC99-067_ustaloides_FR 0.62 1 MC99-047_ustaloides_PT MC99-044_"quercetorum"_PT 85 JHC03-019_fulvum_SK 1 98 JHC04-251_fulvum_SE 1 MC98-078_fulvum_FR * AF377234_ustale_NL JHC92-299_ustale_DK 63 96 MC04-600_psammopus_SI 0.89 MC99-089_psammopus_FR 0.94 41 MC98-048_psammopus_DK 0.6 MC96-345_psammopus_IT * MC95-109_vaccinum_SE 54 MC00-229_vaccinum_SI 0.84 * UDB000699_imbricatum_SE 80 MC94-046_imbricatum*_DK 33 0.93 AF377242_"imbricatum"_USA 0.72 IK922945_roseoacerbum_FI FJ197008_"Uncult._ectom_MEXICO JN021103_"cf. japonicum_CANADA * IK881120_roseoacerbum_FI AB036900_"japonicum"_JAPAN JV99-638_acerbum_DK MEGATRICHOLOMA 25 * MC00-204_acerbum_SI 0.93 69 AF377247_acerbum_NO 1 * MC01-205_colossus_SI MC97-047_colossus_SE JV00-215_apium_DK * JHC95-049_apium_SE MC98-034_apium_NO MC95-102_arvernense_SE 95 MC98-120_arvernense_FR 1 MC98-020_arvernense_NO AF458448_"luteomaculosum"_USA MC96-269_squarrulosum_IT JHC95-169_squarrulosum_DK MC98-081_squarrulosum_FR JHC93-262_squarrulosum_DK 73 MC01-202_squarrulosum_HR 0.97 MC01-700_squarrulosum_SI JHC93-224_squarrulosum_DK 92 AF349701_"atrosquamosum"_USA 1 * MC01-209_basirubens_HR 74 TL5303_basirubens_SE 0.99 MC97-103_olivaceotinctum_SE* OP1981_olivaceotinctum_SE MC95-135_olivaceotinctum_SE ATROSQUAMOSA 90 JHC95-070_olivaceotinctum_SE 0.99 KJ1993_olivaceotinctum_SE JHC01-200_orirubens_SI MC98-214_orirubens_UK MC03-243_orirubens_SK * MC96-301_orirubens_IT MC97-258_orirubens_DK 83 JHC93-261_orirubens_DK 1 O-F159872_atrosquamosum_NO TEB55008_atrosquamosum_NO O-F64018_atrosquamosum_NO O-F188799_atrosquamosum_NO TF98098_saponaceum_FR JHC04-439_saponaceum_SE JHC95-165_saponaceum_DK MC98-059_saponaceum_FR C-F23337_saponaceum_DK 91 JHC03-015_saponaceum_SK 100 JV87-682_saponaceum_DK JHC97-237_saponaceum_DK JHC04-429_saponaceum_SE JHC00-049_saponaceum_NO CONTEXTOCUTIS * MC03-228_rapipes_DK MC98-106_rapipes*_FR 98 * MC98-601_sudum*_DK 100 JV96-306_sudum_DK MC96-002_"viridiolivaceum"_NEW ZEALAND 95 MC01-600_boudieri*_SI MC95-317_boudieri_DK 100 DQ494700_"saponceum"_USA

HM561970_Hypsizygus_marmoreus (outgroup) 0.05

Fig. 1 (cont) J. Heilmann-Clausen et al.: Tricholoma in northern Europe 49 close to the subannulate T. batschii (Fig. 2). The species lack been assigned to the separate sect. Albata (e.g. Noordeloos a perfumed smell and has no broad scales on the pileus so this & Christensen 1999), but this is not supported by our analysis. makes sense morphologically. Section Megatricholoma was Two less well-supported subclades are formed by T. sejunctum originally erected as a monotypic genus to accommodate and allied species, and by sect. Virgata in the sense of Singer T. colossus (Kost 1984). Based on detailed morphological and (1986) with T. portentosum taking up an intermediate position. ontological studies the genus was by its author suggested to Tricholoma guldeniae appears to be the most deviant and basal be only distantly related to Tricholoma s.str., a view that was member of the section. We previously did not consider it as challenged by Christensen & Noordeloos (1999) who recom- a member of this section (Christensen & Heilmann-Clausen bined Megatricholoma as a section in Tricholoma. Our study 2013), but our current phylogenetic analysis gives reasonable supports this disposition, and gives reasonable support for the support for its inclusion. inclusion of T. acerbum and T. roseoacerbum in the section For five species, viz. T. apium, T. avernense, T. borgejoensis, (Fig. 2). In the preliminary phylogeny presented in Christensen T. fucatum and T. josserandii, our analysis do not support a & Heilmann-Clausen (2013) there was no support for this, but clear assignment to traditionally accepted sections (Fig. 1, 2), the broader taxon sampling and more careful alignment in the and appear to represent deviant lineages. These species are current analysis has changed this. All three species share a very discussed further in the next section. robust and short stipe, close lamellae, and an involute pileus margin, but T. colossus stands apart by its annulate stipe, and Species level taxonomy large spores. Section Genuina in our circumscription include species with a reddish brown and glutinous pileipellis, as well Below we give an overview of the accepted sections, and their as several species with paler brown colours and/or a dry squa- circumscribed species accepted by us to occur in northern mulose pileus. Noordeloos & Christensen (1999) divided these Europe. We compare our results with earlier studies and com- in sect. Imbricata (with a dry fibrillose to squamulose pileipel- ment on further taxa revealed by the analysed ITS data. For lis) and sect. Albobrunnea (with glutinous pelipellis). Both are further details on the ecology, morphology and practical differ­ moderately well supported in our ITS phylogeny, but at present entiation of discussed taxa we refer to Christensen & Heilmann- we prefer to treat them as entities below the section level. Clausen (2013).

Species with a smooth, white to yellowish cap Section Terrea Species with a smooth, white to yellowish, dry pileipellis and Our detailed phylogenetic tree supports the presence of presence of clamp connections were divided across four clades seven well-differentiated terminal clades in this section, viz. in our tree, which we here assign to three sections; Contex- T. argyraceum, T. cingulatum, T. scalpturatum, T. inocybeoides, tocutis, Sericella and Lasciva (Fig. 2). Members of sect. Con­ T. terreum, T. bonii and T. triste (Fig. 1). All of these are known textocutis are characterized by reddening flesh, a soapy odour, from northern Europe. Many authors (e.g. Huijsman 1968, greenish to greyish colours and small spores, and the section Krieglsteiner 1982, Clémençon 1983, Riva 1988) accepted has long been recognized as a separate entity, often at the T. gausapatum and T. myomyces as distinct species close to subgenus level (e.g. Singer 1986, Noordeloos & Christensen T. terreum, differing by small differences in pileipellis struc- 1999). In contrast most previous authors have not separated ture, veil development and colouration. Based on our quite Sericella and Lasciva at the section level (e.g. Bon 1984a, intensive sampling, we have found no congruence between Singer 1986, Riva 1988, Noordeloos & Christensen 1999). variation in these traits and ITS phylogeny, and we doubt that However, Bon (1984a) and Riva (1988) distinguished two T. gausapatum and T. myomyces as typically interpreted auct. subsections, Sulphurea (corresponding to our sect. Sericella) are taxonomically different from T. terreum. Also material fitting and Lasciva. Our analysis do not suggest the two sections to with T. leucoterreum show no ITS difference to typical T. ter­ be closely related, and they are morphologically well differen- reum, and we interpret this taxon to represent an albinistic form tiated. Thus, species in sect. Sericella are characterized by of T. terreum. In fact, albinism seems to be rather common in very large spores, a strong gas-like odour and white to yellow the section, with albinistic forms and varieties described also colours, while members of sect. Lasciva have small spores, in T. cingulatum and T. scalpturatum (Hermosilla & Sánchez initially whitish to yellowish grey pileus colours, and a strong, 1994, Bidaud & Thévenard 2003). Also T. bonii was originally complex odour combining aromatic flowery, gas-like and rancid described as a species with whitish colours (Basso & Candusso components. A single member of sect. Sericella, i.e. T. inamoe­ 1997), but the type shows 100 % ITS sequence similarity num, was by Noordeloos & Christensen (1999) assigned to a with collections with greyish pileus colours. The taxonomy separate sect. Inamoena, but our analysis clearly shows this and phylogeny of the species group around T. argyraceum section to be redundant. The species assigned to sect. Lasciva was studied in great detail by Jargeat et al. (2010) using is in our analysis divided among two terminal clades (Fig. 2), three molecular markers. The study found very consistent one containing species with non-yellowing context close to phylogenies for all markers, supporting the clear delimitation T. lascivum, the other species with yellowing context. Based of T. argyraceum, T. inocybeoides, T. cingulatum and T. scalp- on morphological similarities, we expect that future multigene turatum as biological species. Especially T. argyraceum was phylogenies will show the two clades to be more closely related shown to encompass forms and varieties described to differ than our current analysis proposes, and at present we prefer in colouration from pure white to strongly coloured. Two re- to keep sect. Lasciva as a single taxonomic unit. cently described species from Europe, T. urbicum and T. dis- tantifoliaceum, have been assigned to the section. They are Species with a radially fibrillose, whitish, greyish, greenish or unknown to us, and type-material should be sequenced to yellow cap compare their relatedness to more classical species. Tricholoma The great majority of species characterized by an innately moseri described from North America clearly also belongs to fibrillose to squamulose pileipellis and whitish, greyish, green- this section, and is close to or even conspecific with T. triste, ish or yellow colours, were joined in one terminal clade in our as suggested by our ITS analysis. Both species share small tree. These are here assigned to sect. Tricholoma in accord- fruit bodies and very long elongate spores. Based on ITS data ance with Noordeloos & Christensen (1999). The T. equestre T. triste is also present in China, which is also the case for group including T. columbetta and T. umbonatum form a well- T. bonii (Fig. 1). supported core clade. Tricholoma columbetta has traditionally 50 Persoonia – Volume 38, 2017

1 3

on 2

lour lum

ps m Subg. Tricholoma Subg. Sericeicutis Subg. pardinicutis Subg. Contextocutis leus surface Host selectiCap CoGill ColourRing/vePi Spore sizeCla virgatum sciodes bresadolanum aestuans portentosum viridilutescens type I TRICHOLOMA sejunctum virdilutescens type II ? ”ulvinenii” equestre frondosae type II frondosae type I 56 joachimii 0.83 umbonatum type II columbetta umbonatum type I guldeniae caligatum 90 matsutake 0.99 ilkkae CALIGATA dulciolens fucatum josserandii SERICELLA sulphureum type I ? hemisulphureum sulphureum type III ? * sulphureum type II inamoenum bryogenum ? lascivum 95 album LASCIVA S.STR. 1 stiparophyllum borgsjoëense

* filamentosum PARDINICUTIS pardinum

batschii focale aurantium populinum pessundatum albobrunneum stans GENUINA ustale 62 ustaloides fulvum 0.99 vaccinum MEGATRICHOLOMA psammopus imbricatum acerbum 63 roseoacerbum 0.98 colossus apium arvernense

rapipes ? 98 saponaceum CONTEXTOCUTIS ? 1 boudieri ? sudum atrosquamosum 80 basirubens orirubens squarrulosum squarrulosum ATROSQUAMOSA 0.96 olivaceotinctum 71 basirubens 0.65 orirubens 1 atrosquamosum

olivaceotinctum LASCIVA P.P.

sulphurescens type I 72 boreosulphurescens 0.99 sulphurescens type II

inocybeoides scalpturatum argyraceum cingulatum TERREA 97 bonii ? 1 terreum triste ? Hypsizygus_marmoreus 0.05 10 μm

Legends: 1Host selection: : Deciduous, : Coniferous, : Coniferous and deciduous, : (within other symbols), Specific host genus 2Ring/velum: : Pseudonannular zone, : Cobwebby, : Membranous, : Cottony to skin like 3Pileus surface: : Glutinuous, : Dry, : (within other symbols), Squarrulose : (within other symbols), Squamose, : (within other symbols), Radially fibrillose J. Heilmann-Clausen et al.: Tricholoma in northern Europe 51

Section Atrosquamosa is widely distributed but rare in Europe. Quite likely, Armillaria Our analysis supports the presence of five well-circumscribed caligata forma gracilis represent an earlier synonym, but as the European species in this section, viz. T. atrosquamosum, T. ori­ name has not been combined in Tricholoma or proposed at the rubens, T. basirubens, T. squarrulosum and T. olivaceotinctum species level, it has limited nomenclatural relevance. (Fig. 1). The European species fall in two distinct clades with The global phylogeny of the section has been studied rather T. squarrulosum and T. orirubens as central species, respec- intensively (Chapela & Garbelotto 2004, Ota et al. 2012, Murata tively. As discussed thoroughly by Christensen & Heilmann- et al. 2013, Gulden et al. 2014). These studies show T. mat- Clausen (2009) we use the name T. atrosquamosum differently sutake (possibly as a species complex) to be present in both than done by Noordeloos & Christensen (1999), and apply it Europe, Asia and North America, while T. dulciolens so far is for a taxon close to T. orirubens, but mainly associated with confirmed from Europe and North America. The other recog- conifers and lacking yellow mycelia. Conversely T. basirubens, nized species appear to be restricted to smaller biogeographic that was first described as a variety to T. orirubens (Bon 1975), regions, i.e. T. anatolicum, T. caligatum and T. ilkkae to Europe is closely related to T. squarrulosum based on our data. The (including adjacent North Africa and Asia Minor), T. bakamat­ section appears to be well represented, but poorly resolved in sutake, T. fulvocastaneum to eastern Asia and T. magnivelare North America. Tricholoma michinganense clearly belongs to and at least two undescribed lineages (one denoted as ‘Mexican this section but appears to be poorly represented in modern magnivelare’ in Gulden et al. (2014), and one or two labelled literature. Bessette et al. (2013) presented three photographs as T. caligatum) in Chapela & Garbelotto (2004), to North labelled as T. squarrulosum, which appear to be somewhat America. These taxa are represented in our dataset, as ‘T. sp. deviant from our concept of this species, based on the slender Mex1.’ from Mexico and ‘T. caligatum’ from Mexico and Costa stipe and occurrence under conifers. The included American Rica, respectively. sequence labelled as ‘T. atrosquamosum’ in our tree, is clearly deviant from included European taxa, but it is unknown if it cor- Section Genuina responds to T. michinganense or T. squarrulosum in the sense Our analysis supports the presence of at least 18 species of Bessette et al. (2013). in this section in Europe, which fall in two more or less well- supported subclades (Fig. 1). Section Pardinicutis The largest subclade contains species with reddish brown col- Our analysis supports five species in this section (Fig. 1), of ours and a glutinous pileipellis, i.e. T. focale, T. batschii, T. pes- which only two, i.e. T. filamentosum and T. pardinum, are known sundatum, T. populinum, T. stans, T. aurantium, T. albobrun­ with certainty from Europe. Bon (1991) included a number of neum, T. fulvum, T. ustale and T. ustaloides, all known from additional taxa in his treatment of the section, including T. tumi- northern Europe. Many authors have accepted T. pseudonicti­ dum, T. cookeanum and T. cedrorum, the latter two being tans as a separate species close to T. fulvum, but differing by described from Morocco. All are unknown to us. At least three less pronounced yellow tinges in the gills and flesh of the North American taxa are described in this section, viz. T. hu­ stipe, and by the association with conifers. Our ITS data do ronense, T. venenatum and T. vernaticum. Our tree supports not support this separation (JHC04-251 and MC98-078 were the two former as distinct species, assuming that the two se- collected under Picea and Abies, respectively, while JHC03-109 quenced specimens have been correctly labelled. was collected under Betula), and hence we treat T. pseudo­ nictitans as a synonym to T. fulvum (see also Christensen & Section Caligata Heilmann-Clausen 2013). On the other hand, collections from Our analysis supports at least ten species in this section (Fig. southern Europe identified as T. cedretorum and T. quercetorum 1), of which only two are known to occur in northern Europe, appear to represent distinct species based on ITS, but we have viz. T. matsutake and T. dulciolens. Two further species, viz. studied too little material and literature to have any opinion on T. caligatum and T. anatolicum occur in southern Europe or the correct nomenclatural judgments regarding these. Further adjacent regions. Kytövuori (1988) made a careful taxonomic European taxa characterized by a reddish brown and glutinuous treatment of the section in Europe, and described T. dulciolens pileus include T. tridentinum, T. ustale var. rufoaurantiacum, as new to science based on collections from Fennoscandia, T. ustaloides var. aurantiodes and T. ezcarayense. The latter while T. anatolicum was recently described from Cedrus forests taxon, T. ezcarayense, however possesses clamp connections in Turkey (Intini et al. 2003). This species has since been re- (Hermosilla & Sánchez 1994), which are otherwise absent in corded from Morocco (Ota et al. 2012), and might well occur in the section and is probably unrelated. The North American southern Europe. A fifth species, T. ilkkae, is here described as sequences included in the tree suggest that T. aurantiacum, new to science. It has long been known from the Swedish island T. batschii, T. focale and T. stans are present also on this con­ of Gotland in the Baltic Sea, but has been identified as either tinent, with T. zelleri and T. dryophilum being potential synonyms T. dulciolens or T. caligatum. Tricholoma ilkkae share small to the two latter species. In contrast, the included American spores with the first mentioned species, and general coloura- sequence assigned to T. ustaloides represents a distinct taxon tion with the latter, so the confusion is not surprising. ITS data, not closely related to our concept of this species. Judging from however, show that T. ilkkae is clearly differentiated from both photos and descriptions in Bessette et al. (2013) T. muricatum, species, showing most affinity to T. dulciolens. We have stud- T. pudorinum ined. and T. transmutans are further North Ameri- ied material of the species only from Sweden, but have seen can taxa in this group, with T. fulvum, T. pessundatum, T. po- photographs of the species from Norway, and ITS sequence pulinum and T. ustale being also recorded as North American. data show that it is also present in Spain (Murata et al. 2013) Grubisha et al. (2012) investigated the phylogeography of T. po- and Turkey (unpubl. data from Nicklas Bergius). Most likely it pulinum in Scandinavia and North America, and found no signs

← Fig. 2 Phylogeny inferred from ITS regions for a reduced dataset, based on representative sequences for 72 well-circumscribed species or species hypoth- eses. Branch lengths and branching patterns are based on the Maximum Likelihood analysis. Branching patterns were similar in the Bayesian analysis, except for sect. Atrosquamosa, were the alternative configuration is shown as an insert. Maximum Likelihood bootstrap values are indicated above branches, while Bayesian posterior probabilities are indicated below branches for proposed sections indicated with background shadings and names in capital letters. * Denotes 100 % support in both analyses. Coloured bars show the affiliation to subgenera in the sense of Singer (1986). Host selection and six different morphological traits are scored using different symbols, to illustrate their distribution across the phylogeny, and to the proposed sections. 52 Persoonia – Volume 38, 2017 of recent intercontinental gene flow in this specific partner of unknown which of the three additional lineages in our tree Populus spp. Based on the molecular clock approach they corresponds to T. sulphureum s.str. Hence they are labelled estimated divergence between European and American popula- as type I to III in correspondence with Comandini et al. (2004) tions to have happened between 1 and 1.7 million years ago. and Christensen & Heilmann-Clausen (2013). A large number A smaller, and slightly less well-supported subclade contain of varieties have been described in T. sulphureum (see Bon species with a dry, squamulose pileus. Our analysis include 1991 for an overview). Some of these might correspond to the only three European species in this group, viz. T. psammopus, presently cryptic species in our tree. According to our phylo- T. vaccinum and T. imbricatum, but Moreau (2011) presented genetic tree, T. inamoenum occurs also in North America, at and discussed two additional central-European taxa, viz. T. in- least based on ITS data. Another species from this continent odermeum and T. subfusipes. Both appear to be well delimited that clearly belongs to this section is T. odorum. species, close to T. imbricatum and T. vaccinum, respectively, Section Contextocutis (= section Rigida) but with slightly different macroscopic characters (Moreau 2011) This section contains at least four European taxa in our and deviant ITS data (P.-A Moreau pers. comm.). Ecologically, tree, viz. T. saponaceum, T. sudum, T. rapipes (comb. nov.) T. subfusipes differs from T. imbricatum by being associated with and T. boudieri (Fig. 1, 2). Of these, the two latter are normally Larix rather than Pinus. A third species, T. pseudoimbricatum, not differentiated from T. saponaceum at the species level, described from Denmark is by us regarded as a synonym to but we find that differences in morphology and ITS sequence T. imbricatum (for details see Christensen & Heilmann-Clausen data warrant their distinguishing. A large number of further 2013). The included North American sequence of T. imbricatum varieties have been described in the section, mainly based on is quite deviant from the two European sequences, and might pileus colours and surface texture of the stipe (see Bon 1991). represent a distinct species. Both T. imbricatum and T. vaccinum Our studies indicate that these characters are quite plastic are illustrated with several photographs from various American characters with limited taxonomic relevance (Christensen & states in Bessette et al. (2013). The variation in colouration, Heilmann-Clausen 2013). Based on the collections studied stature and pileipellis structure is quite striking, and suggests by us, T. saponaceum and T. boudieri are associated with the presence of several additional species on the continent. deciduous hosts, while T. rapipes and T. sudum are associ- Section Megatricholoma ated with conifers (Christensen & Heilmann-Clausen 2013). This relatively well-supported section was not accepted in We are far from convinced that these preferences are strict, Christensen & Heilmann-Clausen (2013) but as mentioned and await future studies testing host selection and taxonomy above the present analysis has lead us to treat it in a wider in the group. Our current analysis indicates that our concept of sense than done previously, by including the non-annulate spe- T. boudieri could cover more than one species, and we would cies T. acerbum and T. roseoacerbum beside the types species not be surprised if more dedicated studies would prove the exis­ T. colossus. Thus, our concept­ of the section includes three well tence of additional species in the section in Europe. The two known species in Europe (Fig. 1, 2), with T. robustum represent- included extralimital sequences from New Zealand and North ing a tentative fourth, badly known member (see Christensen & America represent further independent species in the section, Heilmann-Clausen 2013). Tricholoma roseoacerbum appears which judging from the photographs given in Besseette et al. to be remarkably widely distributed, with almost perfect ITS (2013) contains several different species in North America. sequence matches connecting collections and environmental Section Lasciva samples from Finland, Japan, Canada and Mexico. Tricholoma In our tree this section is split across two subclades, containing japonicum probably represents the oldest valid name for this a total of five species (Fig. 1, 2) in northern Europe, viz. T. lasci- species, with T. radotinense representing a further potential vum, T. album, T. stiparophyllum, T. sulphurescens and T. boreo- synonym. Also T. manzanitae described from North America sulphurescens. The latter is described as new to science in this belongs to this group, judging from the presentation in Bessette paper. A sequence labelled a T. sulphurescens from Estonia, et al. (2013). appears to represent a further, undescribed species. The taxo­ nomy of the section was discussed in detail by Christensen & Section Sericella Noordeloos (1999) who neotypified T. lascivum, T. album and This section contains six well-separated European end- T. stiparophyllum. Tricholoma albidum and T. farinaceum in the clusters in our tree. Two additional sequences appear to re­ sense of Bessette et al. (2013) appear to represent North present distinct taxa occurring in Canada and the USA (Fig. American members of this section. 1). Across continents, only three of these can be assigned to well-known species, i.e. T. inamoenum, T. hemisulphureum Section Tricholoma and T. sulphureum, while a forth, T. bryogenum is described as This section contains at least 13 species in northern Europe, new to science in this paper. Already Comandini et al. (2004) viz. T. virgatum, T. sciodes, T. bresadolanum, T. aestuans, T. por- reported the presence of cryptic diversity within the section, tentosum, T. sejunctum, T. viridilutescens, T. equestre, T. fron- but mainly concluded that T. bufonium, described to differ from dosae, T. joachimii, T. columbetta, T. umbonatum and T. gul­ T. sulphureum by more reddish to purplish pileus colours, deniae, but several included subclades have complex ITS could not be readily separated from T. sulphureum. Our studies sequence patterns, and remain poorly resolved in our tree. partly confirm pileus colours to be poorly suited to differenti- This is especially the case in the T. equestre group, but also ate taxa within the section (Christensen & Heilmann-Clausen T. sejunctum/viridilutescens and T. umbonatum represent spe- 2013), but we are not convinced that the cryptic taxa detected cies complexes based on our phylogeny. The complex phylo­ by the phylogenetic analysis are truly indistinguishable from geny of T. equestre s.lat. was noted previously by Horton (2002) T. sulphureum s.str. The colours of the lamellae and basal based on North American specimens, and have been confirmed mycelium appear to be promising characters in this respect, by subsequent studies, dealing with the group across the north- but we also expect differences in ecology and biogeography. ern hemisphere (Moukha et al. 2013). Even before molecular At least this is the case for T. bryogenum that differs from phylogenies were available, a number of taxa were proposed T. sulphureum s.lat. by its habitat in boreal coniferous forests, but often synonymized in this group, with T. equestre, T. aura­ the dull yellow colours and the whitish basal mycelium. No tum and T. flavovirens representing classical names. Kala­mees modern type exists of T. sulphureum, and hence it remains (2001) studied the group based on ecological and morpho- J. Heilmann-Clausen et al.: Tricholoma in northern Europe 53 logical characters. He described two new species, T. fron- are additional European species described in this group and dosae and T. ulvinenii, and at the same time assigned T. aura­ represent further candidate names (or synonyms) for T. viri- tum and T. flavovirens as synonyms to T. equestre. The study dilutescens type I and II. Types of the mentioned taxa have not was not supported by molecular sequences, and unfortunately been studied by us. we have been unsuccessful in our attempts to extract DNA from Also T. viridifucatum and T. luridum are characterized by green- the types of the two new species. In our analysis collections ish to olivaceous pileus colours and both may belong to this labelled as T. frondosae form three groupings in the tree, but group, even if the squamulose stipe in the former and the grey- we are quite convinced that T. frondosae type I match the type, ish lamellae in the latter are deviant. We have been unsuc- as all collections have been characterized by warm colours cessful in obtaining sequence data for these two taxa. Chapon and small spores as emphasized in the diagnosis. Tricholoma (2011) compared T. viridifucatum with a further taxon denoted frondosae type II have larger spores and more greenish colours as T. coryphaeum. This species might well belong to this and seem to represent an undescribed species. A further col- group but could also be part of the T. equestre complex. For lection from France (MC98-086) is only distantly related to the a nomenclatural discussion on this name see Christensen T. equestre core group based on ITS data. & Heilmann-Clausen (2013). Judging from descriptions and Collections labelled as T. ulvinenii fall in two distinct clusters photos in Bessette et al. (2013), also the North American taxa within the poorly resolved core T. equestre clade, and it is un- T. davisiae and T. subluteum belong to this group, probably known if any of these represent the type concept of this species. together with T. muscarium described from Japan (e.g. Hongo Collections identified as T. ulvinenii by Kuulo Kalamees (viz. 1988). Comprehensive studies using a global sampling strategy, UDB011557–UDB011559) are separated on both groups, indi- type studies and multiple molecular markers are needed to cating them to be difficult to separate based on morphological resolve the taxonomy of this difficult species complex. characters. In our simplified overview tree (Fig. 2) T. ulvinenii Species with a grey or yellow, radially fibrillose dry pileus (sect. is represented by the upper terminal clade containing four col- Virgata ss. Singer 1986) form a relatively well-resolved sub- lections labelled with this name, and illustrated in Christensen clade in our tree, with the glutinous T. portentosum having a & Heilmann-Clausen (2013: 103). more distant position. According to our data, T. bresadolaum is Collections labelled as T. joachimii appear on three widely heterogeneous in the ITS region, and in general we would not divided branches in the tree. We have not studied the type of be surprised if a more comprehensive sampling would show this species, and as far we know no type sequence is available the subclade to contain more species in Europe. Several ad- to test the correct position of this species in the phylogeny. ditional species have been described in the group, including The included extra-liminal sequences add to the complexity of T. lilacinocinereum, T. sciodellum and T. vinaceogriseum, and the groups. A global analysis including multiple genetic mark- some authors also distinguish T. hordum as a separate species ers, and renewed attempts to sequence type collections is close to T. sciodes (but see Christensen & Heilmann-Clausen needed to resolve the taxonomy of the group, which also con- 2013). The subclade seems to be richly represented in North tains T. chrysophyllum, described from southern Europe (Riva America. Judging from Bessette et al. (2013) at least T. acris, 1988), and T. intermedium, described from North America. T. argenteum, T. atrodiscus, T. palustre and T. pullum belongs Somewhat surprisingly, the two whitish species with radially here. The included sequence of the latter species is close to fibrillose pileipellis, T. columbetta and T. umbonatum seem to T. aestuans. Finally, the sequence of T. aff. virgatum from Nepal have a basal position to the T. equestre complex. As already clearly represent a separate taxon close to T. virgatum. emphasized by Christensen & Heilmann-Clausen (2013) col- Unassigned species lections labelled as T. umbonatum fall in two distinct subclades. Apart from the species that are assigned to the ten hy- We are quite convinced that type II, as illustrated in Chris- pothesized sections discussed above, five sequenced North- tensen & Heilmann-Clausen (2013), corresponds to the original European species, viz. T. arvernense, T. josserandii, T. fucatum, concept of this species as presented by Clémençon & Bon in T. borgsjoeënse and T. apium, remain unclassified at section Bon (1984b). Judging from Bessette et al. (2013) also T. sub­ level (Fig. 2). Despite the presence of clamp connections, T. ar- resplendens belongs to this species group. vernense has traditionally been assigned to the clampless The T. sejunctum/viridilutescens group is another poorly sect. Tricholoma (e.g. Riva 1988, Noordeloos & Christensen resolved subclade in sect. Tricholoma. Except for the South- 1999), but in our tree it forms an isolated cluster with a se- European taxon T. rufenum that has a grey pileus, the European quence identified as T. luteomaculosum from North America members of this group are characterized by greenish to yel- (Fig. 1). This species is characterized by a greyish, fibrillose lowish pileus colours. In our recent monograph, we accepted to squamulose pileus and yellowing flesh and in contrast to only two species viz. T. sejunctum and T. viridilutescens to T. arvernense, it is described to lack clamp connections (Ovrebo occur in northern Europe, but the current analysis shows that 1986). Smith (1942), who described the species, pointed out our concept of T. viridilutescens as presented in Christensen its similarity with T. scalpturatum, which lead Singer (1986) to & Heilmann-Clausen (2013) circumscribes two well separated regard it as a possible member of sect. Terrea. This placement species based on ITS. These are here denoted as T. viridilute­ is not supported by our analysis. The second unclassified scens type I and II, respectively. Further collections from North species, T. josserandii, has traditionally been considered as a America labelled as T. sejunctum and T. leucophyllum represent close relative to T. virgatum (Bon 1984a, Riva 1988), but this further distinct terminal branches. In our data T. viridilutescens is disapproved by our analysis. Instead, the species clusters type I is represented by two collections from France, quite closely with two sequences from North America identified as close to the type locality in the Austrian Alps, and it might well T. mutabile, and more distantly so with a third unassigned spe- represent T. viridilutescens s.str. Type II is represented by two cies, T. fucatum, that has traditionally been assigned to sect. collections from Estonia, that have high ITS similarity to col- Tricholoma (e.g. Riva 1988, Noordeloos & Christensen 1999). lections from Canada and Japan. Tricholoma subsejunctum All three species are characterized by slender fruit bodies described from eastern North America is a relevant candidate with a cylindrical stipe, a radially fibrillose pileipellis and large name for these collections. We have studied the type collection spores. Based on photographs and descriptions in Bessette et of T. subsejunctum which is in poor condition and unlikely to al. (2013), the North American T. aurantio-olivaceum appears to yield usable ITS data. Tricholoma eosinobasis and T. clavocystis be a close relative of T. fucatum, together with T. olivaceobrun­ 54 Persoonia – Volume 38, 2017 neum. All the above species might form an evolutionary lineage cal, warm brown hyphae, 50–300 × 5–15(–20) μm, pigment worth accepting at the section level. A further deviant lineage is not incrusting. Clamp connections absent. formed by T. borgsjoeënse that clusters closely with a sequence Ecology & Habitat — Ectomycorrhizal with Pinus and pos- of T. atroviolaceum from North America. Both species share sibly Picea, mainly in forests on calcareous ground. a dark grey, felty to squamulose pileus, greyish lamellae and Known distribution — Central Sweden (holotype), Norway, large spores. When describing T. borgsjoeënse, Jacobsson et Spain and Turkey; most likely widespread in Europe. al. (2006) assigned the species to sect. Terrea, but the current Additional material examined. Sweden, Gotland, Eksta Par., Ekstastrand, phylogenetic analysis indicates it to be only distantly related to coniferous forest on old beach walls, 9 Oct. 1998 (MC98-602, C-F-96261); ibid., this section. The large spores and the quite special pileipellis 18 Sept. 2000 (UPS-F013888); ibid., associated with T. aurantium, T. fracti- structure (see Christensen & Heilmann-Clausen 2013: 20) sup- cum, Hydnum albidum, Hygrophorus latitabundus and Sarcodon fuligineovio­ port the isolated position among European Tricholoma species. laceous, 3 Oct. 2009, Irene Anderson & Michael Krikorev (MKR 091003-4, Interestingly, our analysis points to substantial variation in the IMG: 100/1208-11); ibid., 29. Sept. 2011 (TF2011-201); Uppland, Gräsö par., ITS region in T. borgsjoeënse, with the Norwegian collection Djupdal 3 km NE of Gräsö church (Grid: RN1648667 x 6697072), in needle bed under Picea abies in old Picea/Pinus forest, 2 Oct. 2007, Gillis Aronsson deviating considerably from the two Swedish collections that (UPS-F173364); Uppland, Börstil par., the turnaround on NW Tvärnö (Grid: both stem from the type locality. RN1648788 x 6681279), under Picea abies and Pinus sylvestris in older, Finally, T. apium appears to be isolated among the species grass-dominated forest on old slag heap, 13 Sept. 2007 (UPS-F173264); analysed. Noordeloos & Christensen (1999) classified it in ibid., 24 Sept. 2009, Gillis Aronsson (UPS-F173265). sect. Imbricata (here a part of sect. Genuina). The species Notes — According to the phylogenetic analysis the new spe- do show some morphological resemblance to species in this cies is close to T. dulciolens. Both species share small spores, section, and might have a basal position in it, as suggested by but T. ilkkae is easily distinguished from T. dulciolens by the the maximum likelihood analysis. shorter stem and much darker pileus scales and stipe girdles. Another similar species is T. caligatum, which is distinguished New combinations by larger spores and by slightly darker, more contrasting pileus scales and stipe girdles. In addition, the two species differ in Tricholoma rapipes (Krombh.) Heilm.-Claus. & Mort.Chr., habitat and distribution, as T. caligatum seems to be a strictly comb. nov. — Mycobank MB816908 Mediterranean species, in contrast to T. ilkkae, which so far is known from more temperate environments. Finally, T. matsutake Basionym. Agaricus rapipes Krombh. (1836: 22). differs by duller colours, larger fruit bodies and larger spores. Description in Christensen & Heilmann-Clausen (2013). Armillaria caligata var. gracilis, as illustrated by Bresadola (1927), matches well with T. ilkkae, but we don’t know if authen- tical material exists that could prove this. A potential synonymy Diagnoses and descriptions of new species will not have nomenclatural consequences as Armillaria caligata var. gracilis has never been combined as a species epithet. Tricholoma ilkkae Mort.Chr., Heilm.-Claus., Ryman & Niclas Bergius, sp. nov. — MycoBank MB816909; Fig. 3a, b Tricholoma bryogenum Mort.Chr., Heilm.-Claus. & Vauras, Etymology. Latin ‘ilkkae’ in honour of the Finnish mycologist Ilkka Kutö- sp. nov. — MycoBank MB816910; Fig. 3c vuori. Etymology. From Greek ‘βρύον’ (moss) and ‘γεννώ’ (born), referring to Holotype. Sweden, Gotland, Eksta par, Ekstastrand, coniferous forest do­ the habitat in mossy Picea-forests. minated by Pinus sylvestris, with scattered Picea abies on old beach ridges, 21 Sept. 2000, leg. Svengunnar Ryman 9080 (UPS, F-513823). Holotype. Sweden, Jämtland, Brunflo, under Picea abies on rich soil, among mosses, 4 Sept. 1997, Morten Christensen MC97-101 (C-F59167). Diagnosis — A medium-sized to large Tricholoma, with a dis- tinct ring, and fawn to dark brick, confluent fibrillose patches Diagnosis — A small to medium-sized Tricholoma, with dull yel- on the pileus and girdles on the stipe. Mycorrhizal with Pinus low colours on stipe, lamellae and pileus margin and a strong, and possibly Picea. chemical smell. Differing from the closely related T. sulphureum by the duller colours, white basal mycelium and by the occur- Pileus 40–100 mm, at first hemispherical to convex with involute rence in mossy Picea forests. margin, later convex to flattened, often with low, broad umbo, in central part soon breaking up into appressed, ± confluent Pileus 30–100 mm, at first conical, bell-shaped or convex, scales, which are fawn, orange brown to dark brick, on a cream soon low convex to plane, with or without a low umbo, smooth, to straw yellow background; scales gradually or more abruptly glossy, at margin whitish chrome to pale chrome, with age thinning out towards the marginal zone, which is typically white and towards centre darker, pinkish buff to ochraceous orange. to straw yellow or pale mouse grey; margin in young fruit bodies Lamellae adnate to deeply emarginate, rather broad and thick, shaggy due to remnants of veil, but soon ± smooth to somewhat medium spaced to rather distant, lemon yellow to lemon chrome felty-costate. Lamellae emarginate, with even to somewhat or honey, more saturated than the margin of the pileus. Stipe eroded edges, whitish, with age sometimes with orange brown 50–130 × 8–25 mm, cylindrical or slightly club-shaped, smooth spots, rather close to medium spaced. Stipe 50–100 × 15–25 or more often distinctly fibrillose, at base often with white tomen- mm, cylindrical or tapering downwards, often somewhat rooting, tum, straw yellow, pale yellow to light chrome, darkest and most with a distinct, persistent, cuff-like, cottony-woolly ring, whitish yellow towards base, with age often duller, pale cream to cream, and granulose or slightly fibrillose above ring, below ring with with a ± fibrillose brownish covering; basal mycelium whitish to irregular, fawn, orange brown to dark brick confluent girdles and faintly yellowish. Flesh rather firm, coloured more or less like the patches on a whitish background, basal part occasionally with surface; smell strong, tar- or gas-like as in T. sulphureum, after a weak greenish tinge. Flesh whitish; smell sweetish, perfumed cutting more farinaceous; taste unpleasant, mild, farinaceous- fruity, similar to Inocybe corydalina or Hemipholiota heteroclita; rancid to slightly bitter. Spores 8.2–14.4 × 4.7–8.4 μm, average taste unknown. Spores 4.5–6.7 × 3.9–5.5 μm, average 5.1–6.0 9.4–12.3 × 5.6–7.5 μm, ellipsoid to elongate or amygdaliform, × 4.4–4.9 μm, predominantly broadly ellipsoid, Q = 1.0–1.5, Q = 1.3–2.0, average 1.62–1.69. Basidia 35–60 × 7.5–10.0 average 1.15–1.31. Basidia 30–40 × 6–8 μm, 4-spored. μm, mainly 2-spored. Cheilocystidia not observed. Pileipellis Cheilocystidia not observed. Pileipellis cutis made up of cylindri- an interwoven cutis with individual hyphal elements generally J. Heilmann-Clausen et al.: Tricholoma in northern Europe 55

a b

c d Fig. 3 Fruitbodies of Trichloma species. a. Tricholoma ilkkae (holotype); b. Tricholoma ilkkae, older specimens (UPS-F173364); c. Tricholoma bryogenum (holotype); d. Tricholoma boreosulphurescens (holotype). — Scale bars = 1 cm.

50–150 × 3–6 μm; subpellis poorly differentiated. Clamp con­ Holotype. Norway, Finnmark, Alta, Kåfjordsbotten, S of Hesteskovattnet, nections not observed, apparently absent. under Betula, 18 Aug. 2004, Per Marstad 197-04 (O-F187683). Ecology & Habitat — Ectomycorrhizal with Picea and pos- Diagnosis — A medium-sized to large Tricholoma, with whitish sibly Pinus, mainly in rich, mixed forests on calcareous soils. colours and strongly yellowing context. Mycorrhizal with Betula Most records are from moist, eutrophic depressions, or spring- and possibly Picea in boreal and subalpine forests on calcare- fed slopes with abundant bryophytes, but there are also some ous soils. Morphologically very similar to T. sulphurescens, but records from drier soils. with substantial differences in the mitochondrial ITS region and Known distribution — Central Sweden, Norway and Finland; a different ecology and distribution range. most likely widespread in Fennoscandia, and possibly in the mountains of central Europe. Pileus 30–100 mm, at first bell-shaped to convex, soon low convex to plane or slightly depressed, often irregularly wavy, Additional material examined. Finland, Koillismaa, Kuusamo, Iivaara, with or without a low umbo, dry and dull, very finely velutinate, E slope, S of Saunakunnas, near Isokorpi, forest with mainly Picea abies and without radial structure, white when young, becoming pale scattered Pinus sylvestris, Alnus incana and Betula, eutrophic depression, 29 Aug. 2007, Jukka Vauras (25068, TURA); Perä-Pohjanmaa, Rovaniemi chrome, ochraceous or yellowish brown with age, especially rural commune, Jaatila, Jaatilanvaara, near Kylmäojao brook, fairly rich, in central part, strongly yellowing when touched, after some gently W-sloping, spring-fed forest with Picea abies, Betula, Alnus incana, time fading to ochre. Lamellae adnate to emarginate, medium Populus tremula and Pinus sylvestris, 11 Aug. 1999, Jukka Vauras (15082F, broad, medium spaced to rather crowded, whitish to cream TURA); ibid., 19 Aug. 1999, Jukka Vauras (15223F, TURA). or pale chrome, with age becoming lemon yellow to honey, Notes — The new species is distinguished from T. sulphure­ especially near the edges or when damaged. Stipe 50–100 × um mainly by its habitat, the dull yellow colours and the whitish 10–20 mm, ± cylindrical, mostly widened at base, more rarely basal mycelium. The difference in coloration is distinct even in tapering, smooth, but mostly finely floccose to squamulose at exsiccata, which are typically pale buff in T. bryogenum, but top, at base often velutinate, at first white to whitish chrome, cinnamon to greyish brown in T. sulphureum. A further difference staining lemon yellow to pale chrome, especially when touched, may be the absence of clamp connections in T. bryogenum, slowly fading to clay buff reddish brown. Flesh rather firm, white to cream, staining lemon yellow to sulphur yellow after but we are not certain if this character difference is truly stable. cutting; smell strong, at first recalling lemons, then complex Tricholoma bryogenum is quite similar to T. odorum described nauseating, combining aromatic flowery, gas-like and rancid from North America, but the latter taxon has more crowded components; taste first mild, but after a while somewhat acrid lamellae. to bitter. Spores 4.5–7.6 × 3.9–6.0 μm, average 5.6–6.4 × 4.2–5.1 μm, predominantly broadly ellipsoid, Q = 1.0–1.5, aver- Tricholoma boreosulphurescens Mort.Chr. & Heilm.-Claus., age 1.20–1.25. Basidia 25–35 × 5.5–8 μm, mainly 4-spored. sp. nov. — MycoBank MB816911; Fig. 3d Cheilocystidia not observed. Pileipellis an irregularly interwoven Etymology. From latin ‘borealis’ (northern) combined with the species cutis with individual hyphal elements generally 50–200 × 4–10 epithet of Tricholoma sulphurescens, a closely related and morphologically μm; subpellis poorly differentiated. Clamp connections present almost similar relative with a southern distribution in Europe. at some septa. 56 Persoonia – Volume 38, 2017

Ecology & Habitat — Ectomycorrhizal with Betula and pos- T. aurantium, T. batschii, T. bonii, T. dulciolens, T. focale, T. fron- sibly Picea on calcareous soils in rich, mixed Picea dominated dosae, T. inamoenum,T. matsutake, T. portentosum, T. roseo­ forests and in subalpine Betula forests near the timber line. acerbum T. stans and T. triste have almost exact ITS similarity Known distribution — Seemingly with an eastern distribution across two or three continents, and according to Jargeat et in Fennoscandia; known from several localities in the northern al. (2010) the same applies for T. argyraceum and T. cingula­ part of Finland, but only from scattered localities in Sweden and tum. Most extreme in this respect is T. roseoacerbum, which Norway. Probably distributed eastwards in Russia, and perhaps according to our data, is present in Finland, Canada, Japan even present in other parts of Europe, e.g. in subalpine forests and Mexico. In Europe it is considered a rarity (Riva 1988, in central European mountain chains. Christensen & Heilmann-Clausen 2013), making the wide distribution particularly intriguing. The above-mentioned spe- Additional material examined. Finland, Outer Ostrobothnia (PeP/Obu), Tervola, Peura, Raemäki, E of the forest road to Syvälampi, between the pond cies with an intercontinental distribution are all associated with Pikku-Ruuntana and Raemäenjänkä, S-sloping, grass-herb spruce forest with widely distributed boreal host tree genera: T. dulciolens and T. spring-fed depressions on calcareous ground, 11 Oct.1997, Ilkka Kytövuori inamoenum are primarily associated with Picea, T. frondosae 97-1187 (H6002040); Koillismaa, Kuusamo, Oulanka National Park, N of the with Populus, T. cingulatum with Salix, while T. aurantium and T. biological field station, E of Puukkosuo, herb rich forest with Picea abies, argyraceum have a broad host selection. The remaining species Pinus sylvestris, Betula, Populus tremula and Salix, eutrophic depression with are associated primarily or exclusively with Pinus. Thus, none Daphne mezereum, Filipendia ulmaria, Goodyera repens, Cirsium helenoides of the species associated exclusively with nemoral deciduous and Elymus caninus, 4 Sept. 2005, Jukka Vauras (23414F, TURA); Koil- lismaa, Kuusamo, Oulanka National Park, Ampumavaara, E of Puukkosuo, hosts, including Fagus and Quercus occurs across continents S of the main road, margin of eutrophic depression with Picea abies, Pinus based on our data. Grubisha et al. (2012), investigated in more sylvestris, Alnus incana, Betula and Salix. 30 Aug. 2007, Emanuele Campo detail the phylogeography of T. populinum and found substan- & Jukka Vauras (25089F, TURA). tial divergence between North American and Fennoscandian populations, pointing to a reproductive isolation established Notes — Tricholoma sulphurescens has long been known 1–1.7 million years ago. Similar studies investigating the phy- as rare but easily identified species characterized by whitish logeography of the apparently circumboreal species mentioned colours and strongly yellowing context. While working with the above would be interesting. volume on Tricholoma in Fungi of Northern Europe (Christensen & Heilmann-Clausen 2013) we realized that collections from Regarding the higher level taxonomy, our study has provided boreal to subalpine Fennoscandia represented a clearly dif- support for several classical sections accepted in Tricholoma, ferent lineage, than collections from southern Europe that are but with some modifications. Most importantly our data showed typically associated with Fagus and Quercus, on warm calcare- T. focale to be a member of sect. Genuina, rather than sect. ous soils. Since T. sulphurescens was originally described from Caligata, while T. sciodes and allied taxa were shown to belong Italy (Bresadola 1905) we here describe the new species as to sect. Tricholoma rather than to sections Terrea or Atrosqua­ T. boreosulphurescens emphasizing its boreal distribution. The mosa. While ITS appears to be a stable marker for species new species is very similar to T. sulphurescens in all important delimitations in Tricholoma (Mouhamadou et al. 2008, Jargeat morphological characters. Our updated phylogeny presented et al. 2010) there are no reasons to believe that the region can here strongly indicates the presence of a third cryptic taxon resolve higher taxonomic relationships at a sufficiently detailed in the group represented by one collection from Italy and one level (e.g. Frøslev et al. 2005). Hence the here suggested in- from a boreonemoral forest with Quercus and Tilia in Estonia. frageneric classification should be viewed as preliminary, and It remains to be determined which of the two non-boreal line- we encourage further studies using multiple molecular mark- ages corresponds to the type specimen of T. sulphurescens ers to investigate the infrageneric phylogeny of the genus. As originally described by Bresadola, and the degree to which they the majority of known species in Tricholoma occurs in North are separable based on morphological or ecological characters. America it is obvious that a careful sampling of North American taxa should be part of such a study, but even Asia, Australia, New Zealand and southern South America host Tricholoma Discussion species that are highly relevant to include in future attempts to unravel the biodiversity, evolution and phylogeography of this With the present study we have provided a first comprehensive important ectomycorrhizal genus. phylogenetically supported taxonomic overview of the genus Tricholoma in northern Europe. Based on this we consider sec- tions Caligata, Atrosquamosa and Terrea as rather well evalu- Acknowledgements Irene Andersson, Niclas Bergius, Tor Erik Brandrud, ated taxonomically on the European continental scale. All three Gro Gulden, Liz Holden, Claes Ingvert, Thomas S. Jeppesen, Kuulo Kala- mees, Tommy Knutsson, Michael Krikorev, Lasse Kosonen, Ilkka Kytövuori, sections have been sampled intensively in this or other stud- Christian Lange, Perry Larsen, Thomas Læssøe, Jens Maarbjerg, Michal ies, and we would be surprised if future studies will change Mikšik, Pierre-Arthur Moreau, Siw Muskos, Johan Nitare, Machiel Noordeloos, fundamentally with the species delimitations presented here Clark Ovrebo, Scott Redhead, Alfredo Riva, Svengunnar Ryman, Sigvard and elaborated in more detail by Christensen & Heilmann- Svensson, Jukka Vauras and Jan Vesterholt† are thanked for valuable discus- Clausen (2013). For all other sections our sampling is limited sions, for giving us the opportunity to study their interesting collections and for allowing us to use their sequence data. The curators of the herbaria C, and additional European species are likely to occur, not least E, H, K, L, LIP, LUG, M, O, OULU, S, TAA and UPS are thanked for arrang- in southern Europe. The sections Genuina, Contextocutis, Seri- ing loans. We want to thank the J. E. Lange and the Flora Agaricina Danica cella and Tricholoma in particular are in need of further phylo- Foundations for supporting collection trips outside Denmark, while UNITE genetic studies with T. equestre s.lat., T. sulphureum s.lat. and is thanked for supporting the sequencing of collections. T. viridilutescens/sejunctum representing species complexes with considerable cryptic diversity. These are all represented References across the northern hemisphere and future studies address- ing these two groups should apply a comprehensive sampling Basso MT, Candusso M. 1997. Tricholoma bonii. Documents Mycologiques strategy and apply multiple genetic markers to unravel the 27, 107: 61–71. complex phylogeography of both groups. Bessette AE, Bessette AR, Roody WC, et al. 2013. Tricholomas of North America. University of Texas Press, US. Many Tricholoma species appear to have a circumboreal Bidaud A, Thévenard G. 2003. Tricholoma cingulatum var. alboflavescens distribution based on the data presented in our study. At least var. nov. Documents Mycologiques 32, 127-128: 69–74. J. Heilmann-Clausen et al.: Tricholoma in northern Europe 57

Bon M. 1975. Tricholomes de France (3’me tome: sections Atrosquamosa Kirby G. 2012. The genus Tricholoma in Britain. Private print, UK. et Equetria, ss. sect. Albata). Documents Mycologiques 5, 18: 111–164. Knudsen H, Vesterholt J (eds). 2008. Funga Nordica. Nordsvamp, Copen- Bon M. 1984a. Les Tricholomes de France et d’Europe occidentale. Ency- hagen, Denmark. clopedie Mycologique 36. Paris. Kornerup A, Wanscher JH. 1974. Farver i Farver. Politikens Forlag, Denmark. Bon M. 1984b. Novitates – Validations, nouvelles combinaisons et espèces. Kost G. 1984. Megatricholoma nov. gen. Eine neue agaricoide Gattung mit Documents Mycologiques 14, 56: 22. verwandtschaftlichen Beziehungen zu Arten anderer Organisationsstufen Bon M. 1991. Flore Mycologique d’Europe, vol. 2, Les Tricholomes et res- der Homobasidiomyzeten. Sydowia 37: 53–74. semblants. St Valerysur-Somme, France. Krieglsteiner GJ. 1982. Über einige neue, seltene, kritische Makromyzeten Bougher NL. 1996. Diversity of ectomycorrhizal fungi associated with in der Bundesrepublik Deutschland. III. Zeitschrift für Mykologie 48: 44. eucalypts in Australia. In: Brundrett M, Dell B, Malajczuk N, et al. (eds), Krombholz JV. 1836. Naturgetreue Abbildungen und Beschreibungen der Mycorrhizas for plantation forestry in Asia: 8–15. Australian Centre for Essbare, Schadlichen und Verdachtigen Schwämme, vol. 4. Prague. International Agricultural Research, Australia. Kytövuori I. 1988. The Tricholoma caligatum group in Europe and North Bresadola G. 1905. Hymenomycetes novi vel minus cogniti. Annales Myco­ Africa. Karstenia 28: 65–77. logici 3: 159–164. Leake JR, McKendrick SL, Bidartondo M, et al. 2004. Symbiotic germination Bresadola G. 1927. Iconographia Mycologica, 2: pl. 51–100. Mediolani, Italy. and development of the myco-heterotroph Monotropa hypopitys in nature Chapela IH, Garbelotto M. 2004. Phylogeography and evolution in matsutake and its requirement for locally distributed Tricholoma spp. New Phytologist and close allies inferred by analyses of ITS sequences and AFLPs. My- 163: 405–423. cologia 96: 730–41. Ludwig E. 2012. Pilzkompendium, band 3. Fungicon-verlag, Germany. Chapon P. 2011. Les Tricholomes du groupe ‘Fucatum’. Bulletin Mycologique Moncalvo JM, Vilgalys R, Redhead SA, et al. 2002. One hundred seventeen et Botanique Dauphiné-Savoie 200: 79–90. clades of euagarics. Molecular Phylogenetics and Evolution 23: 357–400. Christensen M, Heilmann-Clausen J. 2009. Two new boreal species of Tri­ Moreau PA. 2011. Deux tricholomes peu connus retrouvés en Savoie: choloma from Fennoscandia. Mycotaxon 107: 431–440. Tricholoma inodermeum et T. subfusipes. Bulletin Mycologique et Botani­ Christensen M, Heilmann-Clausen J. 2012. Tricholoma (Fr.) P. Kumm. In: que Dauphiné-Savoie 200: 21–34. Vesterholt J, Knudsen H (eds), Funga Nordica. 2nd edn.: 494–510. Nords­ Mouhamadou B, Carriconde F, Gryta H, et al. 2008. Molecular evolution of vamp, Copenhagen, Denmark. mitochondrial ribosomal DNA in the fungal genus Tricholoma: barcoding Christensen M, Heilmann-Clausen J. 2013. The genus Tricholoma. Fungi of implications. Fungal Genetics and Biology 45: 1219–1226. Northern Europe, vol. 4. Svampetryk, Denmark. Moukha S, Férandon C, Beroard E, et al. 2013. A molecular contribution Christensen M, Noordeloos ME. 1999. Notulae ad floram agaricinam neer- to the assessment of the Tricholoma equestre species complex. Fungal landicam – Tricholoma. Persoonia 17: 295–317. Biology 117: 145–155. Clémençon H. 1983. Die Erdritterlinge und ihre nächst verwandten Arten Murata H, Ota Y, Yamaguchi M, et al. 2013. Mobile DNA distributions refine aus der Gattung Tricholoma, Sektion Tricholoma. Mycologica Helvetica the phylogeny of ‘matsutake’ mushrooms, Tricholoma sect. Caligata. 1: 17–30. Mycorrhiza 23: 447–461. Comandini O, Haug I, Rinaldi AC, et al. 2004. Uniting Tricholoma sulphureum Musumeci E, Contu M. 2008. Tricholoma clavocystis (Agaricomycetes, and T. bufonium. Mycological Research 108: 1162–1171. Basidiomycota), una nuova specie della sezione Tricholoma osservata in Ferrarese GG, Zaffalon C. 2010. A new Tricholoma of the section Atrosqua- Svizzera. Bollettino dell’Associazione Micologica ed Ecologica Romana mosa: Tricholoma urbicum sp. nov. Micologia e Vegetazione Mediterranea 73–74: 47–55. 25: 119–128. Noordeloos ME, Christensen M. 1999. Tricholoma. In: Bas C, Kuyper TW, Fries E. 1821. Systema Mycologicum. Lund, Sweden. Noordeloos ME, et al. (eds), Flora Agaricina Neerlandica, Vol. 4: 107–148. Frøslev TG, Matheny PB, Hibbett D. 2005. Lower level relationships in the Balkema, The Netherlands. mushroom genus Cortinarius (Basidiomycota, Agaricales): A comparison Orlovich DA, Cairney JWG. 2004. Ectomycorrhizal fungi in New Zealand, of RPB1, RPB2, and ITS phylogenies. Molecular Phylogenetics and Evolu- current perspectives and future directions. New Zealand Journal of Botany tion 37: 602–618. 42: 721–738. Gardes M, Bruns TD. 1993. ITS primers with enhanced specificity for ba- Ota Y, Yamanaka T, Murata H, et al. 2012. Phylogenetic relationship and sidiomycetes – application to the identification of mycorrhizae and rusts. species delimitation of matsutake and allied species based on multilocus Molecular Ecology 2: 113–118. phylogeny and haplotype analyses. Mycologia 104: 1369–1380. Grubisha LC, Levsen N, Olson MS, et al. 2012. Intercontinental divergence Ovrebo CL. 1986. Three new species of Tricholoma with a description of in the Populus-associated ectomycorrhizal fungus, Tricholoma populinum. Tricholoma luteomaculosum. Mycologia 78: 418–425. New Phytologist 194: 548–560. Rambaut A. 1996 Se-Al: Sequence Alignment Editor. http://tree.bio.ed.ac. Gulden G. 1969. Musseronflora. Universitetsforlaget, Norway. uk/software/seal/ Gulden G. 1992. Tricholoma (Fr.) Staude. In Hansen L, Knudsen H (eds), Nordic Macromycetes, vol. 2: 183–191. Nordsvamp, Copenhagen, Den- Riva A. 1988. Tricholoma (Fr.) Staude. Fungi Europaei, vol. 3, Libreria Gio­ mark. vanni Biella, Italy. Gulden G, Trudell S, Frøslev TG, et al. 2014. Species of Tricholoma section Ronquist F, Teslenko M, Van der Mark P, et al. 2012. MrBayes v. 3.2: Efficient Caligatum in Newfoundland and Labrador. Omphalina 5, 6: 5–9. Bayesian phylogenetic inference and model choice across a large model Hermosilla CE, Sánchez J. 1994. Aportaciones a un posible catálogo de space. Systematic Biology 61: 539–542. Tricholoma Fr. Belarra 10–11: 71–78. Ryberg M, Matheny PB. 2011. Asynchronous origins of ectomycorrhizal Hongo T. 1988. On the genus Tricholoma of Japan. Transactions of the clades of Agaricales. Proceedings of the Royal Society B 279: 2013–2011. Mycological Society of Japan 29: 441–447. Sánchez-García M, Matheny PB, Palfner G, et al. 2014. Deconstructing the Horton TR. 2002. Molecular approaches to ectomycorrhizal diversity studies: Tricholomataceae (Agaricales) and introduction of the new genera Albo- variation in ITS at a local scale. Plant and Soil 244: 29–39. magister, Corneriella, Pogonoloma and Pseudotricholoma. Taxon 63: 993– Huijsman HSC. 1968. Observations sur les Tricholomataceae 1. Schweizer­ 1007. ische Zeitschrift für Pilzkunde 46: 143–153. Singer R. 1986. The Agaricales in modern taxonomy. 4th ed. Koeltz Scientific Intini M, Dogan HH, Riva A. 2003. Tricholoma anatolicum spec. nov.: a new Books, Germany. member of the matsutake group. Micologia e Vegetatione Mediterranea Smith AH. 1942. New and unusual Agarics from Michigan - III. Papers of the 18: 135–142. Michigan Academy of Sciences. 27: 57–74. Jacobsson S, Muskos S, Larsson E. 2006. Tricholoma borgsjoeënse, a Stamatakis A, Hoover P, Rougemont J. 2008. A rapid bootstrap algorithm for new species from a boreal coniferous forest in Fennoscandia. Mycotaxon the RAxML web servers. Systematic Biology 57, 5: 758–771. 95: 195–200. Staude F. 1857. Die Schwämme Mitteldeutschlands, in besondere des Her­ Jargeat P, Martos F, Carriconde F, et al. 2010. Phylogenetic species delimi- zogthums. Coburg, Germany. tation in ectomycorrhizal fungi and implications for barcoding: the case of Tedersoo L, May TW, Smith ME. 2010. Ectomycorrhizal lifestyle in fungi: the Tricholoma scalpturatum complex. Molecular Ecology 19: 5216–5230. global diversity, distribution, and evolution of phylogenetic lineages. Mycor­ Kalamees K. 2001. Taxonomy and ecology of the species of the Tricholoma rhiza 20: 217–263. equestre group in the Nordic and Baltic countries. Folia Cryptogamica White TJ, Bruns T, Lee S, et al. 1990. Amplification and direct sequencing Estonica 38: 13–23. of fungal ribosomal RNA genes for phylogenetics. In: Innis MA, Gelfand Katoh K, Kuma K, Toh H, et al. 2005. MAFFT version 5: improvement in accu- DH, Sninsky JJ, et al. (eds), PCR protocols: A guide to methods and ap- racy of multiple sequence alignment. Nucleic Acids Research 33: 511–518. plications: 315–322. Academic Press, Inc., US.