683 MycoKeys MycoKeys 107: 189-217 (2024) DOI: 10.3897/mycokeys.107.127907 Research Article New species and new records of Laccaria (Agaricales, Basidiomycota) from Northern Thailand Song-Ming Tang'®, Santhiti Vadthanarat?®©, Bhavesh Raghoonundon*®, Zong-Long Luo™, Xin-Yu Zhu**, Feng-Ming Yu®®, Jun He’®, Shu-Hong Li? an oo FP WO NYP College of Agriculture and Biological Science, Dali University, Dali 671003, China Biotechnology and Germplasm Resources Institute, Yunnan Academy of Agricultural Sciences, Kunming 650205, China Department of Biological Science, Faculty of Science, Ubon Ratchathani University, Ubon Ratchathani, 34190, Thailand School of Science, Mae Fah Luang University, Chiang Rai 57100, Thailand State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China Key Laboratory for Plant Diversity and Biogeography of East Asia, Yunnan Key Laboratory of Fungal Diversity and Green Development, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming 650201, Yunnan, China 7 School of Biotechnology and Bioengineering, West Yunnan University, Lincang, Yunan 677000, China Corresponding author: Shu-Hong Li (shuhongfungi@126.com) OPEN Qaccess This article is part of: Exploring the Hidden Fungal Diversity: Biodiversity, Taxonomy, and Phylogeny of Saprobic Fungi Edited by Samantha C. Karunarathna, Danushka Sandaruwan Tennakoon, Ajay Kumar Gautam Academic editor: Samantha C. Karunarathna Received: 20 May 2024 Accepted: 8 July 2024 Published: 7 August 2024 Citation: Tang S-M, Vadthanarat S, Raghoonundon B, Luo Z-L, Zhu X-Y, Yu F-M, He J, Li S-H (2024) New species and new records of Laccaria (Agaricales, Basidiomycota) from Northern Thailand. MycoKeys 107: 189-217. https://doi.org/10.3897/ mycokeys.107.127907 Copyright: © Song-Ming Tang et al. This is an open access article distributed under terms of the Creative Commons Attribution License (Attribution 4.0 International - CC BY 4.0). Abstract Two new species Laccaria pseudoalba and L. subroseoalbescens are described and il- lustrated, based on morphological characteristics and molecular phylogenetic analysis. Two new records, Laccaria umbilicata and L. yunnanensis from Thailand, are also report- ed. Laccaria subroseoalbescens is characterized by small basidiomata, stipe equal with an enlarged base, and nearly subclavate, pale pink to light orange. Laccaria pseudoalba is characterized by pale orange to orange white pileus, has umbo when young on the pileus, and fistulose stipe of the pale to pastel red color. Phylogenetic analysis based on sequence data from rDNA internal transcribed spacer ITS1-5.8S-ITS2 rDNA (ITS), nuc 28S rDNA (28S), RNA polymerase II subunit 2 (rpb2), and translation elongation factor 1-a (tefl1-a) are provided as further evidence. Molecular analysis confirms the phyloge- netic positions of the two new species and two new records. The differences in charac- teristics of these two new species and closely related species are discussed herein. Key words: 2 new taxa, Hydnangiaceae, phylogeny, taxonomy Introduction The genus Laccaria Berk. & Broome, 1883 is a group of ecologically import- ant ectomycorrhizal fungi that inhabit the soil (He et al. 2019). Laccaria, along with Hydnangium Wallr., Maccagnia Mattir. and Podohydnangium G.W. Beaton, Pegler & T.W.K. Young, belongs to the family Hydnangiaceae within the order Agaricales, phylum Basidiomycota. Species of Laccaria are characterized by collybioid to omphaloid basidi- omata; echinulate, acyanophilous, and inamyloid basidiospores; and a convex, plane, or umbilicate, hygrophanous pileus. Clamps are present in all parts of the basidiomata (Singer 1986; Mueller 1992; Latha and Raj 2019). Approximately 100 species of Laccaria have been described worldwide (according to Index 189 Song-Ming Tang et al.: New species and records of Laccaria species from Northern Thailand Fungorum 2024), known to form symbiotic associations with plants of more than 20 genera (including Abies, Castanea, Fagus, Pinus, Picea, Quercus, Larix, Lithocarpus, and others) (Wilson et al. 2017). These associations benefit plant growth and facilitate effective nutrient acquisition (Smith and Read 2008). Therefore, studying Laccaria diversity is crucial for understanding terrestrial ecosystems and forest management. Laccaria species are globally distributed and have been reported on every continent except Antarctica (Kropp and Mueller 1999). They have been found in association with both angiosperms and gymnosperms worldwide (Wilson et al. 2017) and form ectomycorrhizas (ECM) with many economically important plant species (Kropp and Mueller 1999). However, due to the morphological similarity among Laccaria species, defining species boundaries within the ge- nus is challenging (Sheedy et al. 2013). Since the establishment of Laccaria by Berk. and Broome (1883), many my- cologists have contributed to its taxonomy (Orton 1960; McNabb 1972; Mueller 1984; Wang et al. 2004; Wilson et al. 2013, 2017; Popa et al. 2014; Popa et al. 2016; Luo et al. 2016; Cho et al. 2018; Li 2020). Historically, Laccaria was divided into Russuliopsis by J. Schrot, who only included species with a white spore print in Laccaria (Mueller and Vellinga 1986). To date, seven sectional names have been introduced within Laccaria (http://www.indexfungorum.org/Names/Names.asp; accessed date: 20 June 2024), leading to much controversy in its taxonomy. The number of Laccaria species described from Asia has been increasing, with more studies focusing on Basidiomycetes. Since 2013, twenty-three new species of Laccaria have been described in Asia (Wilson et al. 2013; Popa et al. 2014; Luo et al. 2016; Popa et al. 2016; Cho et al. 2018; Li 2020; Cui et al. 2021; Zhang et al. 2023). Nevertheless, no Laccaria species were reported or described in Thailand during the same period. Thailand is renowned as one of the world’s most important biodiversity hotspots with high fungal diversity (Hyde et al. 2018; Thongbai et al. 2018). During our recent investigation of Laccaria in Thailand, nine Laccaria specimens were col- lected. Based on morphological characteristics and phylogenetic analysis, two un- described species and two new records have been identified. This paper provides detailed descriptions, illustrations, and phylogenetic analyses for these species. Materials and methods Morphological study Specimens were collected from Chiang Mai Province, Thailand. They were pho- tographed in the field, then separately wrapped in aluminium foil or kept in a plastic collection box. The fresh basidiomata were macro-morphologically de- scribed on the same day of collection. Colour codes were determined following Kornerup and Wanscher (1978). After being thoroughly dried at 50 °C (Hu et al. 2022) in a food drier, the specimens were stored in sealed plastic bags and deposited in Mae Fah Luang University Herbarium (MFLU) and Herbarium of Cryptogams Kunming Institute of Botany, Academia Sinica (KUN-HKAS). Dried materials were sectioned under a stereo microscope, transferred onto slides, and mounted in a 5% KOH solution. For microscopic characteristics, anatomical and cytological characteristics including basidia, basidiospores, and cystidia, MycoKeys 107: 189-217 (2024), DOI: 10.3897/mycokeys.107.127907 190 Song-Ming Tang et al.: New species and records of Laccaria species from Northern Thailand were observed and photographed using a Nikon Eclipse 80i microscope at mag- nifications up to x 1000. For SEM studies, fragments of the lamellae of the dried material were taken, sputter coated with gold, and analysis with a Hitachi $520 (Hitachi Japan). The notation [x/y/z] specifies that measurements were made on x basidiospores measured from y basidiomata of z collections. At least 50 ba- sidiospores and 20 basidia were measured from one basidioma. Basidiospores dimensions are given as (a—) b—c (-d). Where “a” and “d” refer to the minimum and maximum values of all measurements, respectively, b—c presents the range of 95% of the measured values, and Q is the length/width ratio of basidiospores, Q_is the average Q of all basidiospores and is given as Q_ + standard deviation. DNA extraction, PCR amplification, and sequencing Genomic DNA was extracted from dried specimens using Ezup Column Fungi Ge- nomic DNA extraction kit (Sangon China) following the manufacturer's protocol. Primer pairs for PCR were respectively ITS1/ITS4 (White et al. 1990), LR5/LROR (Vilgalys and Hester 1990), rpb2-5F/rpb2-7cR (Liu et al. 1999), and tef1-983F/tef1- 2218R (Rehner and Buckley 2005). ITS, LSU, rpb2, and tef1 were amplified in 25 uL reactions containing 12.5 uL 2x Taq Plus Master Mix II (Vazyme Biotech Co., Ltd China), 9.5 pL ddH,0, 1 pL 10 uM of forward and reverse primers, 1 uL DNA. PCR conditions were carried out as follows in Table 1 using a C1000 thermal cycler (Bio- Rad China). The PCR amplicons were sent to Sangon Biotech (China) for Sanger sequencing. Sequence reads were assembled in SeqMan II (DNA STAR Inc.). Sequence alignment and phylogenetic analysis The newly generated sequences were checked using BioEdit Sequence Align- ment Editor version 7.0.4 and assembled using SeqMan (DNAstar, Madison, WI, USA). The sequences were then blasted using the Basic Local Alignment Search Tool (BLAST) against the GenBank database (Nilsson et al. 2006) to check the most closely related sequences. Reference sequences for a total of 103 specimens representing 55 species were retrieved (Table 2) and minimally adjusted by hand in BioEdit v.7.0.4 (Hall 2007) first, and then aligned using Tri- mAIl (Salvador et al. 2009). Maximum likelihood (ML) analysis was performed separately for each locus and the concatenated dataset using RAxML-HPC2 v. 8.2.12 (Stamatakis 2014) as implemented on the CIPRES portal (Miller et al. 2010), with the GTR+G mod- el for both genes and 1,000 rapid bootstrap (BS) replicates. For Bayesian Infer- ence (BI), the best substitution model for each character set was determined Table 1. PCR primers and cycling conditions used in the study. Locus Primers (Reference) PCR conditions? ITS ITS1, ITS4 (White et al. 1990) 94 °C: 30 s, 48 °C: 30 s, 72 °C: 1.00 min. (35 cycles) LSU LROR, LR5 (Vilgalys and Hester 1990) 94 °C: 30 s, 48 °C: 30s, 72 °C: 1.30 min. (35 cycles) rpb2 f RPB2-5F, b RPB2-7cR (Liu et al. 1999) 95 °C: 30 s, 55 °C: 1 min, 72 °C: 1.30 min. (35 cycles) tef 983F, 2218R (Rehner and Buckley 2005) 95 °C: 30 s, 55 °C: 1 min, 72 °C: 1.30 min. (35 cycles) @ The three steps given for each primer pair were repeated for 35 cycles, preceded by an initial denaturation step of 5 min at 94 °C, and followed by a final elongation step of 10 min at 72 °C and a final hold at 4 °C. MycoKeys 107: 189-217 (2024), DOI: 10.3897/mycokeys.107.127907 19] Song-Ming Tang et al.: New species and records of Laccaria species from Northern Thailand Table 2. Laccaria taxa and sample IDs with geographic location and GenBank ID numbers for ITS, LSU, rpb2, and tef1 sequences used in phylogenetic analysis. Sequences produced in this study are marked in bold. “*” following a species name indicates that the specimen is the holotype of that species. GenBank accession Species name Sample no. Location Laccaria acanthospora HKAS45998 KU685719 KU685870 KU686069 z ASIS18039 MG519546 | - | MG551620 MG551652 TPML20120807-69 MG519542 MG519583 MG551616 MG551649 Te wL2012000769 Kepacr | America| paar? |= | paver 2a20s canada | Hasso7e2 | ~waraoorr09 | dapan | ugenme | S| : ewe | usa) —kwos7ass |S «SCs Heastorrs1 | china | Mwsaoeo7 | ~Heasios7s1 China| MNSBSSSQ | ~wuwrorori0s China| tsar ~wiwasaszzss [chine | opzaaeor |) : JX504152 JX504120 JX504226 L. negrimarginata GMM7631 France BAP360* China ews. France wore | america | pavasas2 | ~Wkas 108s _|—china—~|~—=ONSS7a79_—~«»~=CNSSGAB4 owen | France | uxsoanse | xs0azze papacor [chine | ansoanzo |= | MycoKeys 107: 189-217 (2024), DOI: 10.3897/mycokeys.107.127907 192 Song-Ming Tang et al.: New species and records of Laccaria species from Northern Thailand GenBank accession Species name Sample no. Location L. neovinaceoavellanea GDGM52852* OR689447 OR785479 pe GDGM53063 OR689448 OR785480 Fe GDGM89621 China OR689449 OR785481 L. nobilis F1091206 America KU685636 KU685779 L. oblongospora ObiFr France GQ406466 L. ochropurpurea PRL4777 America KU685733 KU685883 KU686025 L. ohiensis GMM7539 New Zealand KU685712 KU685853 KU685994 L. prava A3394 Japan JN942788 JN939770 JN993522 ASIS19814 South Korea MG519531 MG519575 MG551606 SFC2012091940* South Korea MG519525 MG551600 L. prava HKAS106742* China MN585660 HKAS106745 China MN585661 L. proxima F1081079 Argentina KU685633 KU685777 KU685928 GMM7584 Russia KU685717 KU685858 KU685999 L. pseudoalba MFLU 22-0106* Thailand ON557377 ON556492 ON598886 HKAS 110664 Thailand ON557376 ON556491 ON598887 L. pseudomontana psel625* America DQ149871 L. pumila pum1252 America DQ149864 L. roseoalbescens LM5099* Mexico KJ874328 KJ874331 L. rubroalba MsS15 China KX449358 MS20 China KX449357 L. rufobrunnea GDGM82878* China OR689443 OR785482 OR835197 GDGM89627 China OR689444 OR785483 L. salmonicolor GMM7596* China JX504143 JX504218 KU686045 GMM7602 China JX504145 L. squarrosa DM63* Mexico MF669958 MF669965 SYC109 Panama KP877340 L. subroseoalbescens MFLU23-0339* Thailand PP785397 PP789598 MFLU23-0340 Thailand PP785398 PP789599 L. tetraspora F1080957 Germany KU685631 KU685775 L. torosa SFC2015090217* South Korea MG519561 MG519598 MG551631 KA12-1306 South Korea MG519562 L. tortilis ASIS22273* South Korea MG519533 MG519576 MG551608 GMM7635 F1111951 GMM7733 PRL7587 GDGM82883 GDGM82911* MFLU 22-0105 HKAS 110652 KNU2012100803 SFC20120926-01* A2986 A0559 SFC20150810-10 GMM7520 CAL1389* KUNF78558* MFLU 22-0107 HKAS 110636 HKAS 110638 ES11.10.2.A AFTOLID972 JX504155 KU685640 JX504157 JX504170 OR689445 OR689446 ON557372 ON557371 MG519560 MG519556 JN942810 JN942803 MG519539 KU685707 MK141034 NR154115 ON557374 ON557373 ON557375 KC964108 DQ404393 KU685906 KU685784 KU686053 KU686063 KU686013 KU686047 OR835194 OR835192 ON598888 France L. trichodermophora Costa Rica America China China China Thailand Thailand South Korea JX504247 OR785485 OR785486 ONS56490 ON556489 MG519597 MG519594 JN939738 JN939756 MG519580 KU685848 L. trullisata L. umbilicata MG551630 MG551627 JN993520 JN993512 MG551614 KU685990 L. versiforma South Korea L. vinaceoavellanea Japan Japan South Korea L. violaceonigra New Zealand L. violaceotincta China Thailand Thailand Thailand Germany L. yunnanensis ON556488 ON556487 ON556486 ON598889 ON598890 Mythicomyces corneipes AY745707 Germany DQ408110 MycoKeys 107: 189-217 (2024), DOI: 10.3897/mycokeys.107.127907 tef1 KU686119 MG551642 KU686120 ON598894 OR826272 KU686151 MG551664 MG551644 KU686156 KU686153 OR826270 OR826268 ON598896 ON598895 MG551663 MG551660 MG551646 ON598892 ON598891 DQ029197 193 Song-Ming Tang et al.: New species and records of Laccaria species from Northern Thailand with MrModeltest 2.2 (Nylander 2004) on CIPRES, using the Akaike information criterion. Bayesian analysis was performed using MrBayes ver. 3.2.7a (Ron- quist et al. 2011) as implemented on CIPRES (Miller et al. 2010). Results Phylogenetic analyses Thirty-three new sequences (11 of ITS, 11 of LSU, 5 of rpb2, and 6 of tef1) were generated for Laccaria species and deposited in GenBank (Table 2). The ITS dataset included 103 specimens representing 55 species, while the ITS-LSU- rpb2-tef1 dataset included 71 specimens representing 42 species. Two phy- logenetic analyses were conducted: one for the 5.8S, ITS1+ITS2 dataset, and the other with concatenated matrix of 5.8S+LSU, ITS1+ITS2, rob2 codon, rpb2 intronsttef1 introns and tefl codons (Vaidya et al. 2011). The ITS final aligned matrix contained 687 positions (170 for 5.8S, 517 for ITS1+ITS2), while the concatenated matrix contained 3,509 positions (1,054 for 5.8S+LSU, 430 for ITS1+ITS2, 1,026 for rpb2 exons, 163 for tef1 introns+rpb2 introns, 836 for tef1 exons). Based on previous phylogenies (Wilson et al. 2013, 2017; Popa et al. 2014, 2016; Luo et al. 2016; Cho et al. 2018; Li 2020; Cui et al. 2021; Zhang et al. 2023), species of the Mythicomyces corneipes (Fr.) Redhead & A.H. Sm. were selected as the outgroup. In the 5.8S-ITS1-ITS2 dataset, the following models were selected by mrModelTest: SYM for 5.8S and GTR+I+G for ITS1+ITS2. In the ITS, LSU, rpb2, and tefl datasets, the models selected by mrModelTest were: GTR+I+G for 5.8S+LSU and tefl codon, GTR+G for ITS1+ITS2 and rpb2 codon, GTR+G for rpb2 introns+tef1 introns. In MrBayes analysis, two runs of five chains each were run for 2,000,000 gen- erations and sampled every 200 generations. Convergence was further evalu- ated by checking that the potential scale reduction factor (PSRF) statistic was close to 1 for all parameters. Moreover, the effective sample size (ESS) was much higher than 200 for all parameters. A clade was considered to be sup- ported if showing a bootstrap support value (BS) =>75% and/or a posterior prob- ability (PP) 0.90. Trees were edited in FigTree version 1.4.0 and PowerPoint. Fig. 2 presents the phylogeny from the combined datasets. Nine specimens collected in northern Thailand formed three monophyletic clades, here de- scribed as L. pseudoalba, L. subroseoalbescens, L. umbilicata, and L. yunnan- ensis, respectively. Each clade was well supported by both ML and BI in the concatenated trees (Fig. 2). In our phylogenetic analysis, the four species clus- tered as separate clades with high support. Thus, these species are formally described in this paper. Taxonomy Laccaria pseudoalba S.M Tang & S.H. Li, sp. nov. MycoBank No: 844144 Figs 3, 4, 5, 12 Etymology. The epithet “pseudoalba’” refers to its similarity to L. alba in their small basidiomata and orange-white to pale orange pileus. MycoKeys 107: 189-217 (2024), DOI: 10.3897/mycokeys.107.127907 194 Song-Ming Tang et al.: New species and records of Laccaria species from Northern Thailand Holotype. THAILAND. Chiang Mai Province: Mae On district, Huay Keaw sub- district, Pox village, 18°43'55.6'N, 99°17'50.1"E, elevation 789 m., 6 September 2020, S. M. Tang, 2020090608 (MFLU 22-0106). Description. Basidiomata small. Pileus 9-15 mm in diam., convex to appla- nate, hemispherical, applanate to plano-concave, pale orange (5A2-3, 6A2-3), orange-white (5A2-3, 6A2-3), when dry moisture loss of moisture or with age becoming whitish, clearly striate on the surface; umbo when young, becoming papilla to abrupt papilla with age; margin inflexed, sometime reflexed; context thin, 1-2 mm, pale orange (5A2-3), unchanging. Lamellae distant, arcuate, ad- nate with decurrent tooth, orange white (542-3, 6A2—3) when young, become pale orange with age, 3-4 mm in height; lamella edge even or entire, some- time undate; lamellulae in 3-4 tiers. Stipe 28.0-—41.1 x 1.8-2.7 mm, cylindrical, central, equal with an enlarged base and nearly subclavate, pale (7—8A6) to pastel red (7A4—5, 8A4—5), smooth, basal mycelium white (1A1); stipe context stuffed, pastel red. Odor and taste not observed. Basidia 29-38 x 9-13 um, (mean length = 32 + 2.5, mean width = 11 + 1.2), clavate, mostly 4-spored, rarely 2-spored, sterigmata 5-8 um x 2-3 um, (mean length = 6.0 + 1.22, mean width = 2.4 + 0.45). Basidiospores (exclud- ing ornamentation) [150/3/2] (6.0-) 7.1-11.0 (-12.0) x (6.5-) 7.0-10.4 (-10.9) um, (mean length = 8.9 + 0.83, mean width = 8.4 + 0.71), Q = 1.00-1.36, Q_, = 1.08 + 0.07, globose to subglobose, hyaline, echinulate, spines 2-3 ym long, ca. 1-2 um wide at the base, crowded. Cheilocystidia 20-31 x 6-9 um, (mean length = 25 + 3.5, mean width = 7 + 1.0), narrowly clavate, thin-walled, colorless and hyaline, abundant. Pleurocystidia 15-31 x 6-8 um, (mean length = 21 + 4.2, mean width = 7 + 0.8), narrowly clavate to subclavate, flex- uose or mucronate, thin-walled, hyaline, abundant. Lamellar trama 50-70 um thick, regular, composed of slightly thick-walled, filamentous hyphae 2-8 um wide. Lamellar edge more in number of sterile basidia. Subhymenium 7-10 um thick, tightly interwoven, fusiform or irregular cells, 5-8 x 3-4 um, (mean length = 7 + 0.8, mean width = 3.6 + 0.5). Pileipellis 70-100 um thick, orange hyaline in KOH, composed of appressed, parallel, simply septate, thin-walled, cylindrical, filamentous hyphae 4-6 um wide, colorless and hyaline. Stipitipel- lis composed of appressed, parallel, simply septate, thick-walled, hyphae 3-7 um wide; stipe tama composed of longitudinally arranged, pastel red in KOH, clavate terminal cells, infrequently branching, septate, thick-walled, hyphae hy- aline 3-10 um wide. Caulocystidia not seen. Clamp present at some septa in pileipellis, lamellae and stipitipellis. Habitat and phenology. Scattered, gregarious, or caespitose on the ground in the Fagus and Dipterocarpus. Additional specimens examined. THAILAND. Chiang Mai Province: Mae On District, Huay Keaw Sub-district, elevation 799 m. 6 September 2020, S. M. Tang, HKAS110664; ibid., 6 September 2020, S. M. Tang, HKAS110663. Notes. In our single gene (Fig. 1) phylogenetic analysis, the phylogenetic po- sition of L. fengkaiensis, L. prava, L. vinaceoavellanea, L. violaceotincta, L. umbil- icata and L. yunnanensis, within L. pseudoalba is well supported (100/1.00) as monophyletic clades. However, L. yunnanensis has bigger basidiomata (pileus 60-100 mm wide), brownish to flesh-colored pileus, and relatively bigger basid- ia (45-50 x 9-10 pm) (Popa et al. 2014). Laccaria vinaceoavellanea has vina- ceous-buff pileus, and rare pileocystidia (Li 2020). Laccaria violaceotincta has MycoKeys 107: 189-217 (2024), DOI: 10.3897/mycokeys.107.127907 195 Song-Ming Tang et al.: New species and records of Laccaria species from Northern Thailand of tichodermopiera F1111951 Costa Rica forlilis type ASIS22273 South Korea Q0)7. versiforma KNU2012100803 South Korea oot a monicolor GMM7602 China jcolor type GMM7596 China »L proxina F1081079 Argentina 99) L. proxima GMM7584 Russia L_ negrimarginate GMM7631 France -L. oblongospora ObiFr France L. tana type pse1623 America gl. montana TWOS91 America sgl cam "L, amethystina GMM Russia J trultisata PRLIS8? China poJl torosa type SFC2015090217 South Korea E. torasa KA12-1306 South Korea ‘parva type SFC2012091940 South Korea L. punsila puml252 America a cident [alba TEMES ROPES CARN? Mexico 09 f- aide: ASIS18039 South Korea alba F1121461 China DOF 1. fengkatensis HKASIOSTAI China pow! nsendoalba HKAS 110664 Mpprusdoatia SUING? (96 Zns Thailand OO) Thsubretseoalbescens MELU23-0339 Type Thailand IL. versiforma type SFC20120926-01 South Korea L. trichodermophora F1111951 Costa Rica L. trichodermophora GMM7733 America L. nobilis F1091206 America L. bicolor GM7712 USA 100) L.bicolor HKAS44062 China L. bicolor KA130253 South Korea $6 L. bicolor GMM7620 France L. longipes F1092175 America D8) L. araneosa type SFC2013091721 South Korea L. araneosa TPML20120912-40 South Korea 100 |Z. tortilis GMM7635 France L. tortilis type ASIS22273 South Korea 100 |Z. versiforma KNU2012100803 South Korea 87 L. versiforma type SFC20120926-01 South Korea 98 -—L. salmonicolor GMM7602 China L. salmonicolor type GMM7596 China L. proxima F1081079 Argentina 97] L. proxima GMM7584 Russia 5 L. negrimarginata GMM7631 France L. oblongospora ObiFr France L. pseudomontana type pse1625 America O0|L. montana TWOS91 America L. montana TWO319 America LOOQL. macrocystidia GMM7612 France L. macrocystidia GMM7616 France 99 | L. himalayensis AWW463 China L. himalayensis type AWW484 China L. bullipellis type AWW465 China gg Ll. amethystina GMM7621 France L. amethystina GMM7041 Russia L. trullisata PRL7587 China 100. |£. torosa type SFC2015090217 South Korea L. torosa KA12-1306 South Korea L. parva type SFC2012091940 South Korea 100 7 parva ASIS19814 South Korea L. parva A3394 Japan L. pumila pum1252 America L. amethysteo-occidentalis AWW556 America 2N'L. amethysteo-occidentalis DAVFP 28205 Canada L. amethysteo-occidentalis type KGP40 America 9] pL. nanlingensis type GDGM 84954 China L. nanlingensis GDGM 84949 China L. laccata GMM7615 France L, moshujun HKAS110661 China 91) 7. moshujun HKAS110653 China L. moshujun type HKAS 93732 China il L. japonica type F64167 Japan L. japonica SFC2012072212 South Korea L. squarrosa type DM63 Mexico 100) L. murina ASIS24249 South Korea L. murina ASIS216 South Korea L. ochropurpurea PRL4777 America L. roseoalbescens type LM5099 Mexico L. alba TPML20120807-69 South Korea _100_|7. aiba ASIS18039 South Korea L. alba F1121461 China L. stellata SYC109 Panama 100 | L. rubroalba MS20 China L. rubroalba MS15 China 90.L. aurantia type KUNF78557 China L. aurantia MBFBO001109 Japan 92) L. fagacicola HKAS107731 China L. fagacicola type HKAS90435 China L. negrimarginata type BAP360 China Figure 1. Maximum likelihood tree based on ITS1-5.8S-ITS2. Bootstrap support values => 70%. The new sequences are highlighted in red, and the holotype of each species is in bold. MycoKeys 107: 189-217 (2024), DOI: 10.3897/mycokeys.107.127907 196 Song-Ming Tang et al.: New species and records of Laccaria species from Northern Thailand 80 .L. trichodermophora F1111951 Costa Rica “mi 0 A 93| L. guizhouensis type HMAS352265 China I tr ata seu L. guizhouensis HMAS352266 China re 100_|L. neovinaceoavellanea GDGM89621 China 72 L. neovinaceoavellanea type GDGM52852 China 92 'L. neovinaceoavellanea GDGM53063 China L. violaceotincta type CAL1389 India L. yunnanensis HKAS110636 L. yunnanensis 2020091004 a me ne Pm ; 85 100} 7. yunnanensis HKAS110638 y nn op sro Ss th Kore L. yunnanensis type KUNF78558 China 06 South Kor ze ete sab Sx uth Korea OORT. parva A ASH U)4 So South Ki parva A3: L. vinaceoavellanea A0559 Japan PNT. vinaceoavellanea A2986 J apan L. vinaceoavellanea SFC20150810-10 South Korea a a L. umbilicata. HKAS110652 Thailand i - Ae umbilicata type GDGM82911 China cnr 100_|7. umbilicata MFLU22-0105 Thailand cine Th OT SRB PES LSI? ene A teria che L. umbilicata GDGM82883 China | eco 100 |Z. prava HKAS106745 China eam es L. prava type HKAS106742 China S 100 | L. rufobrunnea GDGM89627 China 73 L. rufobrunnea type GDGM82878 China — Sane as «i100 F-L. fengkaiensis type HKAS106739 China : aa — 2 L. fengkaiensis HKAS106741 China O) JL. pseudoalba HKAS110664 Thailand L. pseudoalba MFLU22-0106 Type Thailand sce meet - On) | L. subroseoalbescens MFLU23-0340 Thailand L. subroseoalbescens MFLU23-0339 Type Thailand 89/—L. fulvogrisea type KUN-F78556 China se L. fulvogrisea KUN-FB-101105 China i cNBD Nov L. acanthospora HKAS45998 China pay fa mA L. canaliculata GMM7267 Australia 74) L. tetraspora F1080957 Germany L. violaceonigra GMM7520 New Zealand 99 L. miniata type GDGM76043 China 100 L. glabripes GMM7521 New Zealand L. ohiensis GMM7539 New Zealand L. ambigua type PDD89696 New Zealand 100 Mythicomyces corneipes AFTOLID972 Germany — 7104 Mythicomyces corneipes ES11-10/2 A Germany Figure 1. Continued. dark brown to reddish brown pileus and pleurocystidia absent (Latha and Raj 2019). Laccaria fengkaiensis has relatively larger basidiomata (pileus 50-90 mm), more obvious striate, stipitipellis hyphal ends are either ascending or ag- gregating into scattered clusters, smaller basidiospores (5.2—6.3 x 5.1-6.3 um) and narrower basidia (30-45 x 6—-8.5 um) (Li 2020). Laccaria prava has larger basidiomata (pileus 30-75 mm), presence of caulocystidia, and absence of pleurocystidia (Li 2020). Laccaria pseudoalba can be confused with Laccaria alba Zhu L. Yang & Lan Wang due to their similar orange-white to whitish basidiomata. However, L. alba has white to whitish stipe while L. pseudoalba has pale to pastel red stipe, relatively thicker pileipellis (30-75 um), absent pleurocystidia, narrower cheilocystidia (4-6 um), and present clavate, hyaline caulocystidia (Wang et al. 2004). MycoKeys 107: 189-217 (2024), DOI: 10.3897/mycokeys.107.127907 197 Song-Ming Tang et al.: New species and records of Laccaria species from Northern Thailand -/0,95 __97/L.00 B6/1.0U 100/L.O0f L. moshujun HKAS110661 L. moshujun HKAS110653 L. japonica SFC2012072212 South Korea L. squarrosa type DM63 Mexico L. roseoalbescens type LM5099 Mexico L.ochropurpurea PRL4777 America 100/1.00 | — L, amethysteo-occidentalis AWW556 America PL. amethystina GMM7621 France L. amethystina GMM7041 Russia L. parva A3394 Japan L. parva type SFC2012091940 South Korea L. parva ASIS19814 South Korea L. proxima F1081079 Argentina 100/1.00 | 7 proxima GMM7584 Russia L. negrimarginata GMM7631 France L. himalayensis AWW463 China 91/0.99 is L. himalayensis type AWW484 China 100/1.00 IN L. bullipellis type AWW465 China [| ~F L. tortilis type ASIS22273 South Korea L. tortilis GMM7635 France L. trullisata PRL7587 China L. murina ASIS24249 South Korea 100/1.00 100/1.00 90/1.00 L. bicolor HKAS44062 China 99/1.00 L. bicolor KA130253 South Korea 79/1.00| * ; L. bicolor GMM7620 France L. bicolor GM7712 USA 84/1.00 ; L. trichodermophora F1111951 Costa Rica 96/1.00 L. trichodermophora GMM7733 America LE. nobilis F1091206 America L. longipes F1092175 America PS L. araneosa TPML20120912-40 South Korea L. araneosa type SFC2013091721 South Korea 100/1.00¢ LZ. versiforma type SFC20120926-01 South Korea L. versiforma KNU2012100803 South Korea L. salmonicolor type GMM7596 China L. macrocystidia GMM7616 France L. macrocystidia GMM7612 France L. alba TPML20120807-69 South Korea 00/1.00 L. alba ASIS18039 South Korea L. nanlingensis GDGM 84949 China L. nanlingensis type GDGM 84954 China L. torosa type SFC2015090217 South Korea L. neovinaceoavellanea GDGM53063 China L. neovinaceoavellanea type GDGM52852 China L. neovinaceoavellanea GDGM89621 China L. yunnanensis HKAS110636 Thailand P—! /. yunnanensis HKAS110638 Thailand L. yunnanensis MFLU-0107 Thailand L. vinaceoavellanea AQ559 Japan 100/ L. vinaceoavellanea SFC20150810 10 South Korea L.OOLL. vinaceoavellanea A2986 Japan L. umbilicata type GDGM82911 China 100/1.004 L- “mbilicata HKAS110652 Thailand NL. wumbilicata MFLU 22-0105 Thailand 82/1.00} | gg 100/1.00 TP 100/1.00 100/1.00 100/1.00 91/1.00 100/1.00 72/1.00 100/1.00. 99/1.00 85/1.00 94/1.00 L. umbilicata GDGM82883 China 1.001 00/-L. rufobrunnea type GDGM82878 China 1.00°— L. rufobrunnea GDGM89627 China 100/1.00 | L. psedoalba MFLU220106 type Thailand L. psedoalba HKAS110664 Thailand L. fengkaiensis type HKAS106739 China 100/1.00_|L. subroseoalbescens MFLU23-0339 type Thailand L. subroseoalbescens MFLU23-0340 Thailand L. acanthospora HKAS45998 China L. tetraspora F1080957 Germany L. violaceonigra GMM7520 New Zealand 92/ L. miniata type GDGM76043 China -/1.00 84/1.00 0.04 L. ohiensis GMM7539 New Zealand L. glabripes GMM7521 New Zealand L. canaliculata GMM7267 Australia L. ambigua type PDD89696 New Zealand M. corneipes AFTOLID972 Germany Figure 2. Maximum likelihood phylogeny using ITS1-5.8S-ITS2, LSU, rpb2, and tef1 sequence data to identify species of Laccaria growing on roots of Mythicomyces corneipes. ML bootstrap (270%) and posterior probabilities (20.90) are indi- cated above branches or in front of the branch leading to each node. The new species and a new record are highlighted in red; the holotype of each species is in bold. MycoKeys 107: 189-217 (2024), DOI: 10.3897/mycokeys.107.127907 198 Song-Ming Tang et al.: New species and records of Laccaria species from Northern Thailand Figure 3. Fresh basidiomata of Laccaria pseudoalba (a holotype, MFLU 22-0106 b, e HKAS 110664 c, d HKAS 110663). Scale bars: 5mm. Photographs by Song-Ming Tang. Laccaria subroseoalbescens S.M. Tang & S.H. Li, sp. nov. MycoBank No: 853964 Figs 6-8, 15 Etymology. The epithet “subroseoalbescens” refers to its similarity to L. ro- seoalbescens in their pale orange to greyish orange and clearly striate on the pileus surface. Holotype. THAILAND. Chiang Rai Province: Thasud, Muang District, Mae Fah Luang University Park, elevation 488 m, dominated by Dipterocarpus sp., 10 August 2020, OR1663 (MFLU23-0339). Basidiomata small. Pileus 2-8 mm in diam., plano-concave to concave, gla- brous, pale yellow (4A3), light yellow (4A4), pale orange (5A3) to greyish orange (5A4), light orange at center, becoming paler towards the margin, without umbo, when loss of moisture or with age becoming whitish, clearly striate on the surface; MycoKeys 107: 189-217 (2024), DOI: 10.3897/mycokeys.107.127907 199 Song-Ming Tang et al.: New species and records of Laccaria species from Northern Thailand Figure 4. Laccaria pseudoalba a basidiospores b basidium and basidioles ¢ basidia d cheilocystidia e pleurocystidia. Scale bars: 10 um. Photographs by Song-Ming Tang. MycoKeys 107: 189-217 (2024), DOI: 10.3897/mycokeys.107.127907 Song-Ming Tang et al.: New species and records of Laccaria species from Northern Thailand Figure 5. Laccaria pseudoalba A stipitipellis B pileipellis. Scale bars: 10 um. Photographs by Song-Ming Tang. MycoKeys 107: 189-217 (2024), DOI: 10.3897/mycokeys.107.127907 Song-Ming Tang et al.: New species and records of Laccaria species from Northern Thailand &. | | RS | 3 nas SSS ae Oy | Figure 6. Fresh basidiomata of Laccaria subroseoalbescens (holotype a, c OR 1663, MFLU23-0339 b, d OR 1664, MFLU23- 0340). Scale bars: 1 cm. context thin, below 1 mm, pale orange (5A2-3), unchanging. Lamellae distant, arcuate, adnate with decurrent tooth, pale pink (6-7A2), 1-2 mm in height; lamella edge even or entire, sometime undate; lamellulae in 2-3 tiers. Lamellae pale pink (6—-7A2) to bright flesh-pink, 1.2 mm diam., subdecurrent or decur- rent, thick, regular, close. Stipe 5.0-13.0 x 0.8-1.7 mm, cylindrical, central or eccentric, equal with an enlarged base and nearly subclavate, pale pink (7A2) to light orange (6A5), concolorous with pileus, becoming whitish after loss of moisture or with age, smooth; stipe context stuffed, pale orange. Odor and taste not observed. Basidia 30-46 x 8-14 um, av. 38 + 4.1 x 13.8 + 1.3 um, clavate, mostly 4- spored, rarely 2—spored; sterigmata6-14umx 2-4 um, av. 8.5+2.9x3.3+0.8um. Basidiospores [78/2/2] 7.0-8.9 x 6.8-9.0 ym, av. 8.3 + 0.6 x 7.8 + 0.6 um, Q. = 1-1.3, Q,. = 1.1 + 0.08, globose, hyaline; echinulate spines 2-3 x 1-2 um, crowded. Cheilocystidia 23-37 x 4-8 um, av. 34 + 8.5 x 6.5 + 1.5 um, narrowly clavate, thin-walled, colorless and hyaline, abundant. Pleurocystidia 36-59 x 5-8 um, av. 48 + 7.6 x 6.5 + 1.3 um, subclavate, narrowly clavate, flexuose or mucronate, thin-walled, hyaline hyphae. Lamellar edge more in number of ster- MycoKeys 107: 189-217 (2024), DOI: 10.3897/mycokeys.107.127907 202 Song-Ming Tang et al.: New species and records of Laccaria species from Northern Thailand Figure 7. Laccaria subroseoalbescens (OR1663, MFLU23-0339) a basidia b cheilocystidia ¢ pleurocystidia d basidio- spores. Scale bars: 10 um. ile basidia, composed of clavate, cylindrical inflated cells 11-23 x 8-15 um, thin-walled, colorless, similar to basidioles in shape. Subhymenium 10-24 um thick, tightly interwoven, fusiform or irregular cells, 4-7 x 5-6 um. Lamellar trama 74-90 um thick, regular, composed of slightly thick-walled, filamentous hyphae 2-5 um wide. Pileipellis 60-90 um thick, colorless hyaline in KOH, MycoKeys 107: 189-217 (2024), DOI: 10.3897/mycokeys.107.127907 203 Song-Ming Tang et al.: New species and records of Laccaria species from Northern Thailand B Figure 8. Laccaria subroseoalbescens (OR1663, MFLU23-0339) A pileipellis B stipitipellis. Scale bars: 10 um. Photo- graphs by Song-Ming Tang. MycoKeys 107: 189-217 (2024), DOI: 10.3897/mycokeys.107.127907 Song-Ming Tang et al.: New species and records of Laccaria species from Northern Thailand composed of appressed, parallel, simply septate, thin-walled, cylindrical, fila- mentous hyphae 7-11 um wide, colorless and hyaline. Stipitipellis composed of appressed, parallel, simply septate, thick-walled, hyphae 3-5 um wide; stipe trama composed of longitudinally arranged, pastel red in KOH, clavate terminal cells, infrequently branching, septate. Caulocystidia abundant, flexuose, thin- walled, hyaline hyphae, 4-5 um wide. Clamp present at some septa in pileipel- lis, lamellae and stipitipellis. Habitat and phenology. Scattered on the ground in subtropical forests of Dipterocarpus. Additional specimens examined. THAILAND. Chiang Rai Province: Thasud, Muang District, Mae Fah Luang University, 10 August 2020, elev. 489 m, OR1664 (MFLU23-0340). Notes. In single gene (Fig. 1) phylogenetic analysis, L. subroseoalbescens is closely related to L. pseudoalba. However, L. pseudoalba has a pale orange to orange white pileus, and larger basidiospores 7.1-11.0 x 7.0-10.4 um, shorter basidia sterigmata (5-8 um x 2-3 um). Laccaria acanthospora A.W. Wilson & G.M. Muell., L. ambigua K. Hosaka, A.W. Wilson & G.M. Mueller, and L. negrimarginata A.W. Wilson & G.M. Muel- ler have similar small basidiomata (pileus < 15 mm) as L. subroseoalbescens. However, L. acanthospora has orange pileus, relatively longer spines (2-6 um) on the basidiospores, and longer basidia (40-56 x 10-14 um) (Wilson et al. 2013). Laccaria ambigua has orange-brown basidiomata, without the striates on the pileus margin, and stipe orange-brown to ochraceous buff (Wilson et al. 2017). Laccaria negrimarginata has dark blackish brown to dark brown pileus and stipe, fibrillose to appressed squamulose on the pileus surface (Wilson et al. 2013). Laccaria indohimalayana K. Das, |. Bera & Vizzini and L. roseoalbescens T.J. Baroni, Montoya & Bandala are similar to L. subroseoalbescens in their sharing a light yellow basidiomata. However, L. indohimalayana doesn't have cheilo- cystidia and pleurocystidia, and is clearly separated in the phylogeny (Wang et al. 2019). Laccaria roseoalbescens has larger pileus (7-~29 mm), and shorter echinae (1-2.5 um) (Montoya et al. 2015). Laccaria umbilicata Ming Zhang, in Zhang, Gao, Mu & Deng, Journal of Fungi 9(12, no. 1179): 16 (2023) Figs 9-11,15 Description. Basidiomata small. Pileus 7-11 mm in diam., applanate to pla- no-concave, depressed to subumbilicate shape of center, light yellow (1B4), when loss of moisture or with age becoming whitish, clearly striate towards the margin on the surface, without umbo; margin straight, eroded of margin; context thin, 0.5-1 mm, light yellow (1B4). Lamellae reddish brown (8E5-8), 3-5 mm wide; lamellulae subdecurrent to decurrent, thick, regular, distant, 3-4 mm in height; lamella edge even or entire, sometime undate, lamellulae in 3-4 tiers. Stipe 11.0-18.0 x 1.4-—2.0 mm, cylindrical, fistulose, central or eccen- tric, equal with an enlarged base and nearly subclavate, white (1A1), sometime pale orange, basal mycelium white (1A1); stipe context fistulose, white, some- time pale orange. Odor and taste not observed. MycoKeys 107: 189-217 (2024), DOI: 10.3897/mycokeys.107.127907 205 Song-Ming Tang et al.: New species and records of Laccaria species from Northern Thailand Figure 9. Fresh basidiomata of Laccaria umbilicata (a-d MFLU 22-0105, e, f HKAS 110652, g HKAS 110651). Scale bars: 5 mm. Photographs by Song-Ming Tang. Basidia 30-49 x 9-15 um, (mean length = 39 + 6.3, mean width = 12 + 1.9), clavate, hyaline, 4-spored; sterigmata 5-8 x 2-3 um (mean length = 6 + 0.7, mean width = 2.5 + 0.23). Basidiospores (excluding ornamentation) [150/3/2] (6.4-) 7.9-11.0 (-12.0) x (5.7-) 7.4-9.6 (-10.8) ym, (mean length = 9.4 + 0.76, mean width = 8.9 + 0.73), Q = 1.00-1.34, Q_ = 1.07 + 0.06, globose to MycoKeys 107: 189-217 (2024), DOI: 10.3897/mycokeys.107.127907 206 Song-Ming Tang et al.: New species and records of Laccaria species from Northern Thailand Figure 10. Laccaria umbilicata (MFLU 22-0105) a basidiospores b-c basidia d cheilocystidia e pleurocystidia. Scale bars: 10 um. Photographs by Song-Ming Tang. subglobose, hyaline, echinulate; spines 0.2-0.5 um long, ca. 0.5-0.8 um wide at the base, crowded. Cheilocystidia 15-20 x 3-5 um, (mean length = 17 + 1.7, mean width = 4 + 0.8), subclavate, narrowly clavate, hyphae-like, flexuose or mucronate, thin-walled, hyaline, abundant. Pleurocystidia 17-25 x 4-6 um, (mean length = 20 + 2.6, mean width = 5 + 0.7), subclavate, narrowly clavate, hy- Mycokeys 107: 189-217 (2024), DOI: 10.3897/mycokeys.107.1 27907 207 Song-Ming Tang et al.: New species and records of Laccaria species from Northern Thailand Figure 11. Laccaria umbilicata A pileipellis B stipitipellis. Scale bars: 10 um. Photographs by Song-Ming Tang. MycoKeys 107: 189-217 (2024), DOI: 10.3897/mycokeys.107.127907 Song-Ming Tang et al.: New species and records of Laccaria species from Northern Thailand phae-like, flexuose or mucronate, thin-walled, hyaline, abundant. Lamellar trama regular, 50-70 um wide, composed of slightly thick-walled, filamentous hyphae 2-8 um wide. Subhymenium 7-11 um thick, tightly interwoven, fusiform or ir- regular cells, 5-10 x 2-3 um, (mean length = 8 + 1.2, mean width = 2.3 + 0.3). Lamellar edge heteromorphous, more in number of cheilocystidia. Pileipellis 60-100 um thick, composed of interwoven radiating, thin-walled, cylindrical, fil- amentous hyphae 3-8 um wide. Stipitipellis composed of appressed, parallel, simply septate, thick-walled, hyphae 2-8 um wide; stipe trama composed of longitudinally arranged, pale orange in KOH, clavate terminal cells, infrequent- ly branching, septate, thick-walled, hyphae 8-20 um wide. Caulocystidia not seen. Clamp present at some septa in pileipellis, lamellae and stipitipellis. Habitat and phenology. Gregarious or caespitose on the ground associated with the Fagus and Dipterocarpus. Additional specimens examined. THAILAND, Chiang Mai Province: Mae On District, Huay Keaw, Pox Village, elevation 795 m., 6 September 2020, S. M. Tang, HKAS110652; ibid., 6 September 2020, S. M. Tang, HKAS110651. Chiang Mai Province, Mae On District, Huay Keaw, elevation 812 m, 6 September 2020, S. M. Tang, 2020090626 (MFLU 22-0105) Notes. Following BLASTn searches of NCBI GenBank, the closest matches of the ITS and LSU sequences of our new collection (HKAS110652) is L. umbil- icata (specimen GDGM82911 (holotype) ITS 99.67% shared identity; specimen GDGM82883 LUS 99.54% shared identity). The morphology of Thai collections was fit to the original description of L. umbilicata from Southwest China (Yun- nan) by Zhang et al. (2023), including small basidiomata (10-28 mm), pale yel- low, pale orange to light orange pileus, and clavate to ellipsoid pleurocystidia. Phylogenetically, our specimens grouped with L. umbilicata GDGM82911 (holo- type) have high support values (Fig. 1, 100). Thus, we identified this specimen as anew record from Thailand. Laccaria yunnanensis F. Popa, Rexer, G. Kost, Mycol. Progress 13(4): 1113 (2014) Figs 12-15 Description. Basidiomata large. Pileus 50-110 mm in diam., plano-con- cave, concave to hemisphaericus, glabrous, without umbo, yellowish-brown (5D4—5D8), brown (6E5-6E8), dark brown (7-8F5-8), yellowish brown when young, becoming dark brown with age, clearly striate on the surface; margin inflexed, sometimes reflexed; context thin, 2-3 mm, yellowish brown (5D4). Lamellae adnate, distant, yellowish-brown (5D4—5D8), brown (6E5-6E8), dark brown (7-8F5-8), 5-8 mm in height; lamella edge even or entire, sometime undate; lamellulae subdecurrent or decurrent, thick, regular, close; lamellulae in 2-3 tiers. Stipe 24.2-77.8 x 2.1-5.6 mm, cylindrical, central or eccentric, equal, smooth, same color as the pileus, yellowish-brown (5D4-8), brown (6E5-8), dark brown (7F 5-8, 8F5-8), to whitish at the base, basal mycelium white (1A1); stipe context stuffed, yellowish brown. Odor and taste not observed. Basidia 41-53 x 7-15 um, (mean length = 45 + 8.3, mean width = 10 + 2.3), clavate, mostly 4-spored, rarely 2—spored, sterigmata 6-9 um, 2-3 um wide at base. Basidiospores (excluding ornamentation) [250/5/2] 7.9-10.9 MycoKeys 107: 189-217 (2024), DOI: 10.3897/mycokeys.107.127907 209 Song-Ming Tang et al.: New species and records of Laccaria species from Northern Thailand “ge iN ig Bs: Ar Be ‘ : , Figure 12. Laccaria yunnanensis a—d basidiomata (a, b HKAS 110638 c HKAS 110631 d HKAS 110630). Scale bars: 1 cm. Photographs by Song-Ming Tang. x 8.0-10.9 um, (mean length = 9.5 + 0.81, mean width = 9.4 + 0.73), Q = 1.00-1.21, Q. = 1.12, globose, hyaline, echinulate, spines 1-2 ym long, ca. 0.5-1.0 um wide at the base, crowded. Pleurocystidia 50-70 x 10-25 um, (mean length = 59 + 5.4, mean width = 18 + 2.4), clavate to ellipsoid, thin- walled, hyaline hyphae. Cheilocystidia abundant, 25-50 x 4-8 um, (mean length = 38 + 3.5, mean width = 6 + 0.8), subclavate, narrowly clavate to cylindrical, flexuose or mucronate, thin-walled, hyaline hyphae. Lamellar tra- ma 60-100 mm thick regular, composed of slightly thick-walled, filamentous hyphae 2-12 um wide. Lamellar edge more in number of cheilocystidia. Sub- hymenium 7-10 um thick, tightly interwoven, fusiform or irregular cells, 2-4 x 4—5 um, (mean length = 3 + 0.2, mean width = 4.3 + 0.3). Pileipellis 40-80 um thick, yellowish brown in KOH, composed of radiating interwoven, thin- walled, cylindrical, filamentous hyphae 3-5 um wide. Stipitipellis composed of appressed, parallel, simply septate, thick-walled, hyphae 5-20 um wide; stipe trama composed of longitudinally arranged, pale yellowish in KOH, clav- ate terminal cells, infrequently branching, septate, thick-walled, hyphae 6-18 MycoKeys 107: 189-217 (2024), DOI: 10.3897/mycokeys.107.127907 210 Song-Ming Tang et al.: New species and records of Laccaria species from Northern Thailand Figure 13. Laccaria yunnanensis A pileipellis B stipitipellis. Scale bars: 20 um. Photographs by Song-Ming Tang. MycoKeys 107: 189-217 (2024), DOI: 10.3897/mycokeys.107.127907 Song-Ming Tang et al.: New species and records of Laccaria species from Northern Thailand Figure 14. Laccaria yunnanensis a basidiospores b basidia ¢ cheilocystidia d pleurocystidia. Scale bars: 10 um. Photo- graphs by Song-Ming Tang. um wide. Caulocystidia not seen. Clamp present at some septa in pileipellis, lamellae and stipitipellis. Habitat and phenology. Scattered, gregarious, or caespitose on the ground in Dipterocarpus and Fagus. Material examined. THAILAND. Chiang Mai Province: Mare Taeng District, Pha Deng Village, 14 July 2020, S. M. Tang, HKAS 110638; ibid., 14 July 2020 S. M. Tang, HKAS 110636; ibid., 11 August 2020 S. M. Tang, MFLU 22-0107; ibid., 10 September 2020 F. M. Yu, HKAS 110630. Notes. The morphology of Thai collections fit the original description of L. yunnanensis from Southwest China (Yunnan) by Popa et al. (2014) including large basidiomata (pileus 50-110 mm in diam.), yellowish brown, brown, dark MycoKeys 107: 189-217 (2024), DOI: 10.3897/mycokeys.107.127907 212 Song-Ming Tang et al.: New species and records of Laccaria species from Northern Thailand Figure 15. Characteristics of basidiospores ornamentations a, b Laccaria pseudoalba c, d Laccaria subroseoalbescens e-g Laccaria umbilicata h-i Laccaria yunnanensis. Scale bars: 2 um. Photographs by Song-Ming Tang. brown, yellowish brown or dark brown pileus, basidia clavate, and clavate to ellipsoid pleurocystidia. Our molecular analysis also indicated that four Thai collections belong to the same species. Discussion With the development of molecular phylogenetic analysis, many new Lac- caria species have been rapidly described (Wilson et al. 2013, 2017; Popa et al. 2014, 2016; Luo et al. 2016; Cho et al. 2018; Li 2020; Cui et al. 2021; Zhang et al. 2023). Morphological characteristics and systematically infor- mative traits are few in Laccaria; hence, molecular analyses are important for classification and species identification. In this study, we used the molecular MycoKeys 107: 189-217 (2024), DOI: 10.3897/mycokeys.107.127907 213 Song-Ming Tang et al.: New species and records of Laccaria species from Northern Thailand phylogenetic analysis (single gene ITS1+ITS2 and 5.8 S Fig. 1), and multi-lo- cus phylogenetic analysis (ITS, LSU, RPB2 and TEF1 sequences Fig. 2) to evaluate the taxonomy of Laccaria in Thailand. We identified 2 new species, namely L. pseudoalba, L. subroseoalbescens, and two new records, L. umbil- icata and L. yunnanensis. Before this study, five Laccaria species, namely L. amethystina Cooke, L. lac- cata, L. ohiensis (Mont.) Singer, L. proxima (Boud.) Pat., and L. vinaceoavellanea Hongo were reported to occur in Thailand based on morphological charac- teristics, but the specimens lacked detailed descriptions (Chandrasrikul et al. 2011). In the future, more extensive specimen collection is needed in Thailand to determine whether these species are indeed distributed there. So far, only Fagus and Dipterocarpus have been found to host L. pseudo- alba, L. umbilicata and L. subroseoalbescens. Species in Laccaria are similar in morphology characters, so habitat and host trees can provide important in- formation for species identification. It is clear that several Laccaria species have a wide range of host trees while other species of Laccaria associate with a limited group or single host (Mueller 1992). For example, L. laccata (Scop.) Cooke (hosts: Castanea, Quercus, Pinus) and L. himalayensis A.W. Wilson & G.M. Muell. (hosts: Abies, Pinus, Picea) have been reported with a variety of hosts in forests; whereas L. trichodermophora G.M. Muell. (host: Quercus) and L. masoniae G. Stev. (host: Nothofagus) have only been found with a single host tree species (Mueller 1984 ). To date, 42 species of Laccaria have been reported in Asia (Wilson et al. 2013, 2017; Popa et al. 2014, 2016; Luo et al. 2016; Cho et al. 2018; Li 2020; Zhang et al. 2023). These species are described in China (26 species), South Korea (12 species), Japan (seven species), India (Seven species), and Thai- land (four species, this study). The taxonomy of Laccaria species in Thailand is still poorly understood and unclear. As a result of their very similar morpho- logical characteristics, many Laccaria species are misidentified as the same species. Thus, for a better understanding of the species diversity of Laccaria in Thailand and their relationships within the genus, additional studies and data are required. Acknowledgement We thank Dr. Olivier Raspé for his comments on the phylogenetic analysis and for providing specimens. Additional information Conflict of interest The authors have declared that no competing interests exist. Ethical statement No ethical statement was reported. Funding This study was financially supported by the earmarked fund for CARS (Project ID: CARS- 20) and the National Natural Science Foundation of China (Project ID: 32060006). MycoKeys 107: 189-217 (2024), DOI: 10.3897/mycokeys.107.127907 214 Song-Ming Tang et al.: New species and records of Laccaria species from Northern Thailand Author contributions Investigation: XYZ, JH. Software: FMY. Supervision: SHL. Writing - original draft: SMT. Writing - review and editing: ZLL, SV, BR. Author ORCIDs Song-Ming Tang ® https://orcid.org/0000-0002-61 74-7314 Santhiti Vadthanarat © https://orcid.org/0000-0002-9035-0375 Bhavesh Raghoonundon ® hitps://orcid.org/0000-0001-6671-2404 Zong-Long Luo © https://orcid.org/0000-0001-7307-4885 Feng-Ming Yu ® https://orcid.org/0000-0001-9133-8645 Jun He © https://orcid.org/0000-0001-7027-7206 Data availability All of the data that support the findings of this study are available in the main text. Reference Berkeley MJ, Broome CE (1883) Notices of British fungi (1989-2027). 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