MycoKeys 90: | 47-| 62 (2022) ye “." A peer-reviewed open-access journal doi: 10.3897/mycokeys.90.85690 RESEARCH ARTICLE . 03 MycoKkeys research https://mycokeys.pensoft.net Launched to accelerate biodiversity lugisporipsathyra reticulopilea gen. et sp. nov. (Agaricales, Psathyrellaceae) from tropical China produces unique ridge-ornamented spores with an obvious suprahilar plage Sheng-Nan Wang'??, Yu-Guang Fan*°, Jun-Qing Yan'? | Jiangxi Key Laboratory for Conservation and Utilization of Fungal Resources, Jiangxi Agricultural University, Nanchang, Jiangxi 330045, China 2 Key Laboratory of State Forestry Administration on Forest Ecosystem Protection and Restoration of Poyang Lake Watershed, Jiangxi Agricultural University, Nanchang, Jiangxi 330045, China 3 Institute of Edible Mushrooms, Fujian Academy of Agricultural Sciences; National and Local Joint Engineering Research Center for Breeding & Cultivation of Features Edible Mushrooms, Fuzhou 350011, China 4 Key Laboratory of Tropical Translational Medicine of Ministry of Education, Hainan Medical University, Haikou 571199, China § Hainan Provincial Key Laboratory for Research and Development of Tropical Herbs, School of Pharmacy, Hainan Medical University, Haikou 571199, China Corresponding author: Jun-Qing Yan (yanjunging1990@126.com) Academic editor: Kentaro Hosaka | Received 22 April 2022 | Accepted 13 June 2022 | Published 22 June 2022 Citation: Wang S-N, Fan Y-G, Yan J-Q (2022) lugisporipsathyra reticulopilea gen. et sp. nov. (Agaricales, Psathyrellaceae) from tropical China produces unique ridge-ornamented spores with an obvious suprahilar plage. MycoKeys 90: 147— 162. https://doi.org/10.3897/mycokeys.90.85690 Abstract Lugisporipsathyra, a new psathyrelloid genus from tropical red soil of China, is established with /. reticulopilea as the type species. The new genus is characterised by basidiomata psathyrelloid, pileus rugose to appearing reticulate ridged, covered by persistent, but inconspicuous villus, pleurocystidia absent and ridge-ornamented spores with an obvious suprahilar plage. The genus is unique amongst Psathyrellaceae in producing ridge-ornamented spores with an obvious suprahilar plage and forms a distinct lineage within Psathyrellaceae, based on the Maximum Likelihood and Bayesian Inference analyses of a combined three-gene sequence dataset (ITS, LSU and B-tvb). Full descriptions and photographs of the new genus and species are presented. Keywords Basidiomycete, fungal phylogeny, taxonomy Copyright Sheng-Nan Wang et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. 148 Sheng-Nan Wang et al. / MycoKeys 90: 147-162 (2022) Introduction The Psathyrellaceae Vilgalys, Moncalvo & Redhead was established in 2001, based on the type genus Psathyrella (Fr.) Quél. by Vilgalys and Redhead (Redhead et al. 2001). More than 1300 names within the family, including synonyms and subspecies, are listed in Index Fungorum (http://www.indexfungorum.org). Species of Psathyrellaceae are cosmopolitan and often grow on decaying logs, woody debris, humus or soil, in woodlands, lawns or bogs and can have either broad or specific substrate relationships (Kirk et al. 2008). Traditionally, the family included two types of species: psathyrelloid species and coprinoid species. During the classic period of morphological research, Fries (1838) classified the psathyrelloid species to Agaricus L. trib. Psathyrella Fr. Quélet (1872) pro- moted this group to the rank of genus. Psathyrella was finally accepted after the transfer of Drosophila Quél. species and emendations by Singer (1951,1975). Subsequently, Kits van Waveren (1985) removed the species with warty spores from Psathyrella and treated these as the genus Lacrymaria Pat. Although the boundaries of the genus were disputed, most researchers agreed that the psathyrelloid species should be classified in Coprinaceae R.Heim ex Pouzar subfamily Psathyrelloideae (Kuhner) Singer (Hawk- sworth et al. 1983; Kirk et al. 2001). During this same period, the coprinoid spe- cies were classified in Coprinus Pers. (Coprinaceae subfamily Coprinoideae Henn.) (Hawksworth et al. 1995; Kirk et al. 2001). Coprinus was circumscribed by Persoon (1797). However, Fries (1821) did not recognise the genus in his monograph Systema Mycologicum and classified the species in Agaricus. However, in his subsequent mon- ograph Epierisis systematis Mycologici, Fries discarded his previous classification and again placed the coprinoid species in the independent genus Coprinus (Fries, 1838). Although morphological studies provide abundant support for recognition of Psathyrellaceae, morphological data are inadequate to conclusively resolve the systematic relationships amongst the constituent genera and species. When the works of Hopple and Vilgalys (1999) and Redhead et al. (2001) were published, it became apparent that molecular biology techniques would profoundly alter the classical systematics of psathyrelloid species and coprinoid species. Based on these studies, Coprinus was split into four genera (Coprinellus P.Karst., Coprinopsis P.Karst., Coprinus and Parasola Redhead, Vilgalys & Hopple) (Redhead et al. 2001), restraining the generic name Coprinus to a small group centred on the type species Coprinu comatus (O.EMiill.) Pers., which is now classified in the Agaricaceae Chevall. The other three genera, together with Psathyrella and Lacrymaria, were incorporated into the newly-established Psathyrellaceae. In 2015, Psathyrella, as a paraphyletic group, was also split, with the establishment of the segregate genera Cystoagaricus Singer emend. Orstadius & E.Larss., Homophron (Britzelm.) Orstadius & E.Larss., Kauffmania Orstadius & E.Larss. and Typhrasa Orstadius & E.Larss. (Orstadius et al. 2015). In 2020, Candolleomyces D.Wacht. & A.Melzer, Britzelmayria D.Wacht. & A.Melzer and Olotia D.Wacht. & A.Melzer were separated from Psathyrella, Punjabia D.Wacht. & Lugisporipsathyra reticulopilea gen. et sp. nov. from Tropical China 149 A.Melzer and Tulosesus D.Wacht. & A.Melzer were separated from Coprinellus, Narcissea D.Wacht. & A.Melzer was segregated from Coprinopsis and Hausknechtia D.Wacht. & A.Melzer was erected for Galerella floriformis Hauskn. (Wachter and Melzer 2020). Heteropsathyrella V.Bau & J.Q.Yan was established in 2021, based on the new species He. macrocystidia T.Bau & J.Q.Yan (Bau and Yan 2021a). Thus, the main systematic framework of Psathyrellaceae has been confirmed. In addition, Ozonium Link and Hormographiella Guarro & Gené, formerly members of the Psathyrellaceae, were established to accommodate the conidial anamorphs of certain species, now classified in Coprinellus (Nagy et al. 2013). Gasteroagaricoides D.A.Reid and Macrometrula Donk & Singer, two genera that, to date, have not been included in phylogenetic analyses, are retained in the Psathyrellaceae. There were 19 genera, in total, in the Psathyrellaceae before the new taxon we discovered was added. From 2015, we initiated a study of Chinese psathyrelloid species and described 15 new taxa (Yan and Bau 2017, 2018a,b; Yan et al. 2019; Bau and Yan 2021a,b; Wang et al. 2021). By chance, we collected a psathyrelloid species with a reticulate-ridged pileus, that was reminiscent of Pluteus thomsonii (Berk. & Broome) Dennis, on the roadside in tropical China. After examining the micromorphology of the specimens, we observed that it produced ridge-ornamented spores with an obvious suprahilar plage. Surprisingly, phylogenetic analysis of molecular data revealed that it belonged to the Psathyrellaceae. Although abundant genera and species are recognised in the Psathyrellaceae, the majority of species have smooth spores. Verrucous spores have been observed only in Lacrymaria. Rough spores have been observed in Coprinopsis, Coprinellus and Psathyrella, but are extremely rare. Thus, the specimens are unique amongst Psathyrellaceae in producing ridge-ornamented spores with an obvious suprahilar plage. On the basis of our morphological and phylogenetic analyses, the specimens are described herein as a new species and a new genus is erected to accommodate the new species. Materials and methods Morphological studies Macroscopic descriptions and habitat details were based on detailed field notes of fresh basidiomata and photos. The location of the collection point is marked on the map (Fig 1). Colour codes follow the Methuen Handbook of Colour (Kornerup and Wan- scher 1978). Microscopic structures were observed and measured from dried speci- mens mounted in water, 5% potassium hydroxide (KOH), 10% ammonium hydrox- ide (NH,OH) or Melzer’s Reagent. Congo red was used as a stain when necessary (Horak 2005). A minimum of 100 basidiospores, basidia and cystidia from seven ba- sidiomata (three collections) were randomly measured using an Olympus BX53 mi- croscope. Detailed observations of spores were made by SEM. The measurements and Q values are recorded as (a)b—c(d), in which “a” is the lowest value, “b—c” covers a 150 Sheng-Nan Wang et al. / MycoKeys 90: 147-162 (2022) FUJIAN Myanmar (Burma) . | Chiang Mail inauiauas| _, Vientiane, idealual aja” 4 af : Poy 43 a we Ai a Paracel | i «& } > “elslands Me (Thailand tye aspen | 7 ° war er F ChinaSéa ~ Bangkok——— . el a ATIUNWUNIUAT : Vietnam | | a | ws | | - Cambodia Figure |. Map showing the location of the collection site of the specimens (red triangle). minimum of 90% of the values and “d” is the highest value. “Q” stands for the ratio of length and width of a spore (Bas 1969; Yu et al. 2020). Specimens were deposited in the Herbarium of Fungi, Jiangxi Agricultural University (HFJAU). DNA extraction and sequencing DNA was extracted from dried specimens with the NuClean Plant Genomic DNA kit (CWBIO, China) (Ge et al. 2021; Na et al. 2022). Three regions (ITS, LSU and B-tud) were selected for the study and were amplified using the primer pairs by ITS1/ITS4 (White et al. 1990), LROR/LR7 (Hopple and Vilgalys 1999) and B36f/B12r (Nagy et al. 2011), respectively. PCR was performed using a touchdown programme for all regions: 5 min at 95 °C; 1 min at 95 °C; 30s at 65 °C (add -1 °C per cycle); 1 min at 72 “Cacyele l5rtimesy Wmintat 95°C; 30-s.ar 50" lminvar7.2. "Ch -cycle 20, times: 10 min at 72 °C (Bau and Yan 2021a). The sequencing was performed by Qing Ke Biotechnology Co. Ltd. (Wuhan City, China). Data analyses A total of 221 nucleotide DNA (ITS, LSU and B-tub) sequences representing 93 taxa were used in subsequent analyses. Details are presented in Table 1. Some species of Agaricaceae, Mythicomycetaceae Vizzini, Consiglio & M. Marchetti Lugisporipsathyra reticulopilea gen. et sp Table |. Sequences used in this study. Taxon Britzelmayria multipedata B. supernula Candolleomyces eurysporus C. subcacao C. subminutisporus C. subsingeri Coprinellus andreorum C. aureogranulatus C. aureogranulatus C. curtus C. deminutus C. disseminatus C. domesticus C. silvaticus Coprinopsis babosiae C. calospora C. cortinatus C. musae C. musae C. semitalis C. udicola C. villosa Cystoagaricus hirtosquamulosa C. olivaceogrisea C. silvestris C. squarrosiceps C. strobilomyces Hausknechtia floriformis Heteropsathyrellamacrocystidia H. macrocystidia Homophron camptopodum HI. cernuum H. crenulata H. spadiceum Lugisporipsathyra reticulopilea L. reticulopilea L. reticulopilea Kauffmania larga K. larga Lacrymaria glareosa L. hypertropicalis L. lacrymabunda L. pyrotricha L. rigidipes L. subcinnamomea Narcissea cordispora N. cordispora N. patouillardi Olotia codinae Parasola auricoma P. conopilea P. kuebneri Voucher LO237-04 LO250-04 GLM-F126263 Type HMJAU37807 Type HMJAU37801 Type HMJAU37913 Type CS1247 Type CBS973.95 CBS753.96 Isotype NL-2339 NL-0761 NL-2337 NL-1292 LO172-08 NL-4139 Type CBS612.91 Type NL-1621 JV06-179 Type JV06-180 CBS291.77 Type AM1240 Type NL-1758 Type Ramsholm800927 WK8/15/63-5 Type LO191-92 Laessoe44835 E.Nagasawa9740 WU22833 Type HMJAU37803 HMJAU37802 Type 1997/956 LO134-98 W-K8/10/64-5 Type Enderle Epitype HFJAU1352 Type HFJAU3181 HFJAU3182 LO223-90 LAS97-054 LAS06-019 Guzman29585 Type EL70-03 CBS573 LAS00-081 Smith16957 Type SFSUDEH2073 LO41-01 NL-1687 GLM-F112430 Type NL-0087 LO186-02 Neotype Ulje31-V-1987 Type ITS KC992888 KC992867 MT651560 MW301064 MW301066 MG734725 MW621497 GQ249274 MH862611 FM878016 JN159572 FM878017 FN396102 KC992943 FN396128 GQ249275 FN396121 KC992965 KC992966 GQ249278 KC992967 JN943128 KC992945 KC992948 KC992949 KC992950 AY176347 JX968254 MW405101 MW405102 KC992956 DQ389726 KC992957 DQ389729 ON207138 ON207139 ON207140 DQ389694 DQ389695 KC992954 KC992958 DQ389724 GQ249280 KC992953 KC992951 AY461827 DQ389723 FM878009 MG696611 JN943107 DQ389725 KY928608 . nov. from Tropical China LSU KC992888 KC992867 MT651560 MW301092 MW301094 MW301098 MW621007 GQ249283 FM876273 JN159592 FM876274 HQ847132 KC992943 FN396177 GQ249284 FN396171 KC992965 KC992966 GQ249287 KC992967 JQ045877 KC992945 KC992948 KC992949 AY176348 JX968371 MW413358 MW413359 KC992956 DQ389726 DQ389729 ON207137 DQ389694 DQ389695 KC992954 KC992958 DQ389724 GQ249289 KC992953 KC992951 FM876265 MG674714 JQ045871 DQ389725 KY928633 151 B-tub KJ664867 K]664849 MW369460 MW314063 MW314065 MW314068 GQ249258 FN396281 JN159636 FN396282 FN396330 KJ664911 FN396352 GQ249259 FN396346 KJ664920 kJ664921 GQ249262 KJ664922 HQ847173 MW410997 KJ664915 ON210974 ON210975 ON210976 KJ664912 KJ664914 KJ664916 GQ249264 KJ664913 KJ664910 FN396257 FN396252 152 Sheng-Nan Wang et al. / MycoKeys 90: 147-162 (2022) Taxon Voucher ITS LSU B-tub P lactea NL-0466 FM163192 FM160717 FN396254 P. misera NL-0280 Neotype FM163210 FM160699 = P. ochracea NL-3621 Type JN943134 JQ045875 = P parvula CAL1667 Type NR_160509 NG064556 - P plicatilis NL-0295 FM163216 FM160693 FN396253 P plicatilis NL-0075a Epitype NR_171786 NG075167 = P. psathyrelloides CAL1753 Type MK682756 MK682754 - Psathyrella amygdalinospora HMJAU37952 Type MW405104 MW413361 MW410991 Pamygdalinospora HMJAU57044 MW405105 - 7 Pfagetophila L0210-85 (M) Type KC992902 KC992902 K]664879 Pfennoscandica HMJAU37918 MG734723 MW413365 MW410993 Pfennoscandica LO484-05 Type KC992903 KC992903 KJ664881 Pnoli-tangere LO83-03 Neotype DQ389713 DQ389713 KJ664890 Pseminuda Smith34091 (MICH) Type KC992907 KC992907 = Pwarrenensis Smith70162 (MICH) Type KC992906 KC992906 - Punjabia pakistanica MEL2382843 KP012718 KP012718 - P. pakistanica LAH35323 Type MH366736 - - Tulosesus canistri Walleyn877 Isotype HQ846985 - HQ847 142 T. cinereopallidus NL-0177 Type HQ847001 HQ847090 HQ847149 T. fuscocystidiatus NL-2720 Type HQ846977 HQ847064 HQ847152 T. hiascens NL-2536 FM878018 FM876275 FN396284 T. pseudoamphithallus Ulje1288 Type HQ846973 HQ847059 - T. radicellus NL-3168 Type GU227719 HQ847077 GU227737 T. sassii NL-1495 FN396101 FN396155 FN396329 Typhrasa gossypina Schumacher024 KC992946 KC992946 - T. nanispora Barta980706 Type KC992947 KC992947 > T. polycystis HFJAU1454 Type MW466538 MW466544 - T. rugocephala HFJAU1467 Type MW466541 MW466546 - Outgroup Coprinus comatus AFTOL_ID_626 AY854066 AY635772 = Crucibulum laeve REGCrul1/DSH96-02 DQ486696 AF336246 = Cyathus striatus DSH96-028/Cyst1/DSH96-001 DQ486697 AF336247 od Lepiota cristata ZRL20151133 LT716026 KY418841 — Leucocoprinus fragilissimus ZRL20151466 LT716029 KY418844 - Lycoperdon ericaeum ZRL20151498 LT716030 KY418845 - Macrolepiota dolichaula xml2013058 LT716021 KY418836 - Mycocalia denudata AFTOL2018/CBS494.85 DQ911596 DQ911597 - Mythicomyces corneipes AFTOL-ID972 DQ404393 AY745707 Zi M. corneipes KB51 KY648897 = = Nidula niveotomentosa AFTOL1945/CBS250.84 DQ917654 DQ986295 7 Stagnicola perplexa AH25260 Holotype MK351609 MK353793 a S. perplexa AH25282 Paratype MK351610 MK353794 - and Nidulariaceae Dumort. were chosen as outgroup taxa according to the results of Zhao et al. (2017) and Vizzini et al. (2019). ITS, LSU and B-tud sequence datasets were separately aligned on the MAFFT online server (Katoh et al. 2019). Bayesian Inference (BI) and Maximum Likelihood (ML) phylogenetic analyses of the aligned concatenated dataset were respectively carried out in MrBayes v.3.2.7a and IQTREE v.2.1.2 (Nguyen et al. 2014) via the CIPRES web portal. For the BI analyses, optimal evolutionary models were selected using PartitionFinder2 (Lanfear Lugisporipsathyra reticulopilea gen. et sp. nov. from Tropical China 153 et al. 2017) with the greedy algorithm and the AlCc criterion. Four Monte Carlo Markov chains were run for 2 million generations, sampling every 100" generation, with the first 25% of trees discarded as burn-in (Ronquist et al. 2012). For the ML analysis, models of sequence evolution were assessed in IQ-Tree prior to the analysis. The ML analysis was conducted using the ultrafast bootstrap option with 1,000 replicates and allowing partitions to have different seeds (--p). A nexus file contains alignment sequence and original tree of ML and Bayes is deposited in Suppl. material 1. Results Phylogenetic analysis Based on the BLAST results, the new species were found sharing less than 90.82% (ITS), 97.66% (LSU) and 87.03% (B-tub) similarity with the known species. The aligned concatenated dataset comprised 2,591 characters (ITS 835 bp, LSU 1338 bp and B-tub 418 bp), of which 983 sites were variable and 757 were parsimony in- formative. The best-fit evolutionary models used for the phylogenetic analyses were as follows: for the BI analysis, GTR + 1+ G for ITS and LSU and TIM + 1+ G for B-twb; and for the ML analysis, TIM2 + F + 1+ G4 for ITS, GTR + F + R4 for LSU and HKY + F + 1 + G4 for B-tub. The log-likelihood of the ML consensus tree was —27426.323 and the average standard deviation of split frequencies was less than 0.01 after 1,115,000 generations in the BI analysis. In the resulting trees, clades with a Bayesian posterior probability (BI-PP) > 0.95 and ML bootstrap support (ML-BP) > 75% were considered to be well supported. As shown in the BI tree in Fig. 2, all taxa of Psathyrellaceae formed a well-supported monophyletic lineage (BI-PP = 1; ML-BP = 100%). Within Psathyrellaceae, 18 major supported clades with a high statistical support value (BI-PP = 0.95, ML-BP = 75%) represented a total of 17 (out of 19) known genera and a new genus. /ugisporipsathyra formed a distinct lineage (BI-PP = 1; ML-BP = 100%) clearly separated from currently recognised genera. Taxonomy Iugisporipsathyra J.Q. Yan, Y.G. Fan & S.N. Wang, gen. nov. MycoBank No: 843734 Etymology. lugi-, iugis (Latin), ridge; -spori-, sporis (Latin), spores; lugispori-, refers to its spore ornamentation; -psathyra, one of the synonyms of Psathyrella, refers to its similarity to Psathyrella. Description. Basidiomata psathyrelloid, fragile, non-deliquescent. Pileus hygrophanous, rugose to appearing reticulate ridged, covered by persistent and 154 Sheng-Nan Wang et al. / MycoKeys 90: 147-162 (2022) W190 Parasola conopilea LO186-02 Neotype Parasola psathyrelloides CAL1753 Type woo — Parasola plicatilis NL-0295 1/100 Parasola plicatilis NL-0075a Epitype — Parasola lactea NL-0466 Parasola misera NL-0280 Neotype Parasola W100 Parasola kuehneri Ulje31-V-1987 Type viva 0.5951 Parasola parvula CAL1667 Type , Parasola ochracea NL-3621 Type Parasola auricoma NL-0087 Homophron camptopedum 1997/9536 Homophron spadiceum Enderle Epitype Homophron cernuum LO134-98 H omophr on 09991. Homophron crenulata W-K8/10/64-5 Type Lacrymaria glareosa LAS06-019 Lacrymaria rigidipes LASO0-081 Lacrymaria lacrymabunda EL70-03 . Lacrymaria pyrotricha CBS573 Lac rymaria Lacrymaria subcinnamomea Smith16957 Type Lacrymaria hypertropicalis Guzman29585 Type Coprinopsis babosiae NL-4139 Type Coprinopsis villosa NL-1758 Type Coprinopsis semitalis CBS291,77 Type Coprinopsis calospora CBS612.91 Type ; ; Coprinopsis udicola AM1240 Type Cop rin op Sts Coprinopsis cortinatus NL-1621 Coprinopsis musae JV06-179 Type ‘Kscibn Coprinopsis musae JV06-180 \y Iugisporipsathyra reticulopileus HFJAU1352 Type @ Iugisporipsathyra reticulopileus HFJAU3182 @ Jugisporipsathyra reticulopileus HEJAU3181 @ Punjabia pakistanica MEL2382843 q ® Punjabia pakistanica LAH35323 Type | Punjabia Cystoagaricus hirtosquamulosa Ramsholm800927 Cystoagaricus strobilomyces E.Nagasawa9740 . Cystoagaricus olivaceogrisea WK8/15/63-5 Type Cystoagaricus Cystoagaricus silvestris LO191-92 Cystoagaricus squarrosiceps Laessoe44835 Kauffmania larga LO223-90 , Kauffmania larga Green | Kauffmania Typhrasa gossypina Schumacher024 Typhrasa polycystis HFJAU1454 Type Typhrasa rugocephala HFJAU1467 Type Typhrasa nanispora Barta980706 Type Heteropsathyrella macrocystidia HMJAU37803 Heteropsathyrella macrocystidia WMJAU37802 Type | Heter ops ath ir ella Olotia codinae GLM-F 112430 Type | Olotia Narcissea cordispora SFSUDEH2073 Narcissea cordispora LO41-01 | Narcissea Narcissea patouillardi NL-1687 viooy Psathyrella amygdalinospora HMJAU37952 Type Psathyrella amygdalinospora HMJAUS57044 Psathyrella fagetophila L.O210-85 (M) Type Psathyrella warrenensis Smith70162 (MICH) Type Psathyrella seminuda Smith34091 (MICH) Type P. sathyr ella Psathyrella noli-tangere LO83-03 Neotype Psathyrella fennoscandica HMJAU37918 Psathyrella fennoscandica LO484-05 Type Hausknechtia floriformis WU22833 Type | - Coprinellus curtus NL-2339 : Coprinellus disseminatus NL-2337 Coprinellus domesticus NL-1292 Coprinellus andreorum CS1247 Type ‘ Coprinellus aureogranulatus CBS973,95 Cop. rinellus Coprinellus aureogranulatus CBS753.96 Isotype Coprinellus silvaticus LO172-08 Coprinellus deminutus NL-0761 Tulosesus canistri Walleyn877 Isotype ai ps Tulosesus cinereopallidus NL-0177 Type Tulosesus hiascens NL-2536 0.98/- YO thiosesus Suscocystidiatus NL-2720 Type Tu lo. SESUS V/100 0.99/94 1/95 Tugisporipsathyrae Typhrasa AVAOV TITHaAH.LVSd 0.99/76 Tulosesus sassii NL-1495 Tulosesus radicellus NL-3168 Type Tulosesus pseudoamphithallus Ulje1288 Type Candolleomyces eurysporus GLM-F 126263 Type Candolleomyces subminutisporus HMJAU37801 Type ' Candolleomyces subcacao HMJAU37807 Type Can d ol leo my ces Candolleomyces subsingeri HMJAU37913 Type Britzelmayria multipedata LO237-04 r ; Britzelmayria supernula L.O250-04 | Britzelmay. ria woor Mythicomyces corneipes AFTOL-ID972 1/100 Mythicomyces corneipes KB51 4 v0) Stagnicola perplexa AH25260 Holotype Myth icomycetaceae 0997) Stagnicola perpleca AH25282 Paratype Lepiota cristata 2RL20151133 Lycoperdon ericaeum £RL20151498 V/100 Leucocoprinus fragilissimus ZRL20151466 I sits Macrolepiota dolichaula xml2013058 Aga ricaceae Coprinus comatus AFTOL_ID_626 Crucibulum laeve REGCrull/DSH96-02 Mycocalia denudata AFTOL2018/CBS494,85 . . Nidula niveotomentosa AFYOL1943/CBS250.34 Nidulariaceae Cyathus striatus DSH96-028/Cyst1/DSH96-001 1/100) 0.96/80 0.95/- dNOUDLNO 0.08 Figure 2. Phylogeny generated by Bayesian Inference, based on a concatenated sequence dataset for three nuclear DNA regions (ITS, LSU and B-tub). The tree was rooted with Agaricaceae spp., Mythicomyc- etaceae spp. and Nidulariaceae spp. Bayesian Inference posterior probabilities (BI-PP) > 0.95 and Maxi- mum Likelihood bootstrap percentages (ML-BP) = 75% are shown as PP/BP at relevant nodes. (black circle) indicates newly-described taxa. Lugisporipsathyra reticulopilea gen. et sp. nov. from Tropical China 155 inconspicuous villus. Lamellae adnexed, brown. Stipe white, central, hollow. Spores amygdaliform in profile view, ovoid to elongate in face view, inamyloid, brown, fades in concentrated sulphuric acid, ridged and rarely verrucose ornamentation, suprahilar plage obvious. Basidia monomorphic. Pseudoparaphyses abundant. Pleurocystidia absent. Cheilocystidia present. Pileipellis hymeniderm, pyriform cell mixed with simple hairs. Type species. /ugisporipsathyra reticulopilea J.Q. Yan, Y.G. Fan & S.N. Wang Notes. The combination of veil absent, pleurocystidia absent and spores orna- mented with ridges or rarely verrucose, with an obvious suprahilar plage is unique in Psathyrellaceae. Iugisporipsathyra reticulopilea J.Q. Yan, Y.G. Fan & S.N. Wang, sp. nov. MycoBank No: 843801 Fig. 3 Etymology. reticulo-, reticular; reticulopilea, referring to the surface characteristic of the pileus. Description. Pileus 30-90 mm broad, oblate when young, expanding to plane, surface dry, rugose to appearing reticulate ridged, hygrophanous, pale yellow to grey- ish-yellow (4A3—4B2), becoming yellowish-white (4A2) as pileus dries, centre and ridged area darker, brown to dark brown (7D6-—7F6), becoming greyish-yellow (4B2) as pileus dries. Pileus surface covered by inconspicuous villus. Villus very short, white (4A2), persistent. Veil absent. Context 3.0—4.0 mm broad, fragile, dirty white (7A1— 7B2). Lamellae 3.5-10 mm broad, crowded, adnexed, 2—3 tiers of lamellulae, dirty white (7A1—7B2), becoming brown (7E6—7E8) as spores mature, edge white (7A1— 7B1) and saw-toothed under 20x magnification. Stipe 50-80 mm long, 3.0-10 mm thick, fragile to fibrous, white to dirty white (7A1—7B1), cylindrical, hollow, gradually thickening towards base, 8.0—17 mm thick at base. Stipe surface covered with small, white, evanescent fibrils. Spores (7.5—)8.0—9.7(-10.5) x (4.0—-)4.5-6.0 um, Q = 1.5—2.0, amygdaliform in profile view, (4.5—)4.8-6.0(-6.3) pm broad, ovoid to elongate in face view, inamyloid, red-brown in water, brown in alkaline solution, fades in concentrated sulphuric acid, ornamentation up to 1.0 um high, composed of irregular ridges and rarely verrucose, variable in length, partly connected, sometimes forming a zebroid pattern or closed meshes, suprahilar plage obvious, germ pore absent. Basidia (19- )22—29 x 9.5-12.0 um, clavate, hyaline, 4- or 2-spored. Pseudoparaphyses abundant. Pleurocystidia absent. Cheilocystidia (37—)40-61(-68) x (9.5—-)12-18(—22) um, hyaline, utriform with obtuse to broadly obtuse apex, base tapering to a short or long stipe. Caulocystidia 50-90 x 6.0—14 um, scattered or caespitose, various, mostly narrow clavate, hyaline. Trama of gills subparallel. Pileipellis hymeniderm, composed of a 1-cell-deep layer of pyriform cells, mixed with sparsely simple hairs, pyriform cells (35—)38—60 (—62) x (12—)14—23 um, hairs hyphae, separate, 7.0—10 um broad. Clamps present. Sheng-Nan Wang et al. / MycoKeys 90: 147-162 (2022) q ny — 4 Figure 3. Macroscopic and microscopic structures of lugisporipsathyra reticulopilea a-d Basidiomata e, f spores viewed by scanning electron microscopy g spores in Melzer’s Reagent h spores in water i hyme- nophore j, k cheilocystidia I, m pileipellis and hairs hyphae n, 0 caulocystidia. Scale bars: 20 mm (a=d); 20 pm (g-0). Structures of imo were observed in 5% KOH solution and Congo red was used as the stain. Lugisporipsathyra reticulopilea gen. et sp. nov. from Tropical China 157 Known distribution. Tropical China (Hainan Province). Habit and habitat. Scattered or 2—3 caespitose on red soil of roadside under broadleaf tree. Specimens examined. Cuina. Hainan Province, Ding’an County, Longhu Town, 2 Jan 2019, Yu-Guang Fan, Jun-Qing Yan HFJAU 1352 (holotype); 4 Jan 2019, Jun- Qing Yan, Sheng-Nan Wang, HFJAU 3181, HFJAU 3182. DNA sequence of type. ON207138 (ITS), ON207137 (LSU), ON210974 (B-tub). Notes. Differs from other species in Psathyrellaceae by having ridge-ornamented spores with an obvious suprahilar plage. Discussion The discovery of 1. reticulopilea has transformed our traditional understanding of Psathyrellaceae. The species is unique amongst Psathyrellaceae in producing ridge- ornamented spores with an obvious suprahilar plage. This feature is so unusual that it seems difficult to associate it with Psathyrellaceae. However, the characteristic of the spores of fading in concentrated sulphuric acid is in common with other species in this family (Singer 1986; Kirk et al. 2008; Padamsee et al. 2008; Nagy et al. 2013; Orsta- dius et al. 2015; Wachter and Melzer 2020). Macroscopically, the psathyrelloid basidiomata of L reticulopilea enables ready discrimination from the coprinoid taxa of Psathyrellaceae. Gasteroagaricoides spp. have a densely granular-warty pileus and Macrometrula spp. have a volva (Singer 1948; Reid 1986). Lugisporipsathyra reticulopilea can be distinguished from these species by the smooth pileus and absence of a volva. Amongst the abundant psathyrelloid taxa of Psathyrellaceae, only the species of Typhrasa have slight to distinct ridge-like folds on the pileus. However, no species has a reticulate-ridged pileus similar to that of I. reticulopilea. \n addition, the pileus surface of /. reticulopilea is covered by a white, inconspicuous, but persistent villus. This feature also readily distinguishes /. reticulopilea from known species of Typhrasa (Orstadius et al. 2015; Wang et al. 2021). Microscopically, almost all species of Psathyrellaceae have smooth spores. Granu- lose spores are observed only in Coprinopsis, Coprinellus and Psathyrella, but are ex- tremely rare. Verrucose spores are known only in Lacrymaria. No species has an obvi- ous suprahilar plage as in L. reticulopilea (Guzman et al. 1990; Orstadius and Knudsen 2012; Orstadius et al. 2015). In the classification system of Smith (Smith 1972), some species with ornamented spores were classified in Psathyrella subg. Panaeolina (Maire) A.H. Smith. Those species are now excluded from the Psathyrellaceae and are classified in Panaeolina Maire, based on phylogenetic relationships and spores that do not fade in concentrated sulphuric acid (Kirk et al. 2013; Zhao et al. 2017). Detailed morpho- logical comparison of Lugisporipsathyra and psathyrelloid genera of Psathyrellaceae is presented in Table 2. Sheng-Nan Wang et al. / MycoKeys 90: 147-162 (2022) 158 quasoid quasqe Apeses AI9A quasqe quasqe quasqe yuasqe quasqe quasqe quasqe quasqe quasaid erpnsAd02]1 quasaid quasaid quasaid quasaid quasaid yuasaid quasoid quasaid quasoid quasoid quasaid erpnsA0]1947) parejnorpod sa[nqoys Jo sdosp | payem-yprya_-| AjSuons pue Afio repnyooennur | ysis Ajerer | padeys-emnyeds qm ‘paypem-urua | Jo paypea-ury | Apueurwopard | payyem-urgi | paype-ury? yuosqe poyem-yorya poyyea-urya poyyea-urga quasqe poyyess-urga erposAs0mo, gq quasaid oeyddy Y soummouros | peurpiiod jo s04e| sro Ayores ajduns Ajosseds | ssrey o;durs doasp ]J2° [ B Aq uopesed ‘wopered uuopesed 03 ursopered 03 Ypis saxruy ‘wapesed 0} | pazoaoo ‘unopered uopesed 01 uaprusudy | ‘woprucwdy | uopruswAy | wuopruswAy | wuioprusudy | ‘wepruowAPE | wopruowdy | 0} WopruowiAy uopesed 0) woprusurAy uapered sypedrpg quasqe quasaid Ayares quasqe quasqe quasqe yuasard quasqe quasaid quasqe quasqe quasqe saskydesedopnasg orydrounp orydiowrouowr | srydiourouour | stydiowrouow | 01 -ouour = |orydiowouour| srydsourouowr |otydrowouour| s1ydsourouowr orydrourouowr | stydiowouour | stydiowrouow PIpiseg urmiodsoxdur aSeyd sepryesrdns qu Jo SNOLAGO Y)TAA asonuess uOonrUsWIeUIO qoours Ajeres ‘qoours qoours Ayre ua1yJo qoours sa3 pri qioours qioours qioours qioours sovzpans a10dg¢ quaosanbyap | yuaosonbypp | iuassonbyap | auaosanbrjap quaosanbryap yuaosanbryap yuaosanbryap qusosanbryap-uou -uou -uou -uou -uou yuaosonbrjap-uou -uou quaosanbyap-uou | yuadsanbyyap-uou -uou -uou aeypurey to dey sy]eo asoqojsqns Ajarer ovyddy oeyddy oeyddéy oeyddy oeydday aeyddy ‘oqqvadim | Saqeadrim ‘aqqvodim quasqe soqquodim yuasqe quesqe ovyd dy ‘oqeadim quasqe soqqeadim ‘oqquadim TPA oeyddy ‘asorsrenbs Jo ‘urds SPOF 2yL]-OSpra oeyddy seyd Ay fsnqqra ‘gsopnurenbs JOUNISIP 03 YSIS qloours qioous s9so]uau0} yoours snorAqo-uou qioours qloours ‘QsoqPLIqu qioours qoours soups snot gq vsvayddy uosqdowmopy | vyashgvdsosagazy | snatsvsvo4shy sastmoayopuvy ViADUaZILAg ‘avaoeTpaidyiesg ay} ur vious proyjardyresd ayeurwtsdsip 03 pasn sonstioieseyp yes1sojoydiow jo Arewuing *7 ajqe Lugisporipsathyra reticulopilea gen. et sp. nov. from Tropical China 159 Acknowledgements This work was financed by the National Natural Science Foundation of China (31960008, 31860009), The Project of FAAS (XTCXGC2021007) and Jiangxi Pro- vincial Natural Science Foundation (20202BABL213041). Sincere thanks to the anonymous reviewers of the manuscript. References Bas C (1969) Morphology and subdivision of Amanita and a monograph of its section Lepidella. Persoonia 5: 285-573. Bau T, Yan J-Q (2021a) A new genus and four new species in the/Psathyrella s.l. clade from China. MycoKeys 80: 115-131. https://doi.org/10.3897/mycokeys.80.65123 Bau T, Yan J-Q (2021b) Two new rare species of Candolleomyces with pale spores from China. MycoKeys 80: 149-161. https://doi.org/10.3897/mycokeys.80.67 166 Fries EM (1821) Systema mycologicum. Gryphiswaldiae, Sumtibus Ernesti Mauritii, 1-510. Fries E (1838) Epicrisis Systematis Mycologici. seu synopsis Hymenomycetum. Typographia Academica, Uppsala, Sweden, 1-610. Ge Y-P, Liu Z-W, Zeng H, Cheng X-H, Na Q (2021) Updated description of Atheniella (Mycenaceae, Agaricales), including three new species with brightly coloured pilei from Yunnan Province, southwest China. MycoKeys 81: 139-164. https://doi.org/10.3897/ mycokeys.8 1.67773 Guzman G, Bandala VM, Montoya L (1990) Observaciones taxondémicas sobre el género Psathyrella subgénero Lacrymaria en México y descripcidn de nuevos taxa (Basidiomyco- tina, Agaricales). Scientia Fungorum: 105-123. Hawksworth DL, Sutton BC, Ainsworth GC (1983) Ainsworth and Bisby’s Dictionary of the Fungi. Commonwealth mycological institute, Surrey, USA, 444 pp. Hawksworth DL, Kirk PM, Sutton BC, Pegler DN (1995) Ainsworth & Bisby‘s Dictionary of the Fungi. The Cambridge University Press, Cambridge, UK, 616 pp. Hopple Jr JJ, Vilgalys R (1999) Phylogenetic relationships in the mushroom genus Coprinus and dark-spored allies based on sequence data from the nuclear gene coding for the large ribosomal subunit RNA: Divergent domains, outgroups, and monophyly. Molecular Phy- logenetics and Evolution 13(1): 1-19. https://doi.org/10.1006/mpev.1999.0634 Horak E (2005) Rohrlinge und Blatterpilze in Europa: Bestimmungsschliissel ftir Polyporales (pp), Boletales, Agaricales, Russulales. Elsevier, Munich, Germany, 1-555. Katoh K, Rozewicki J, Yamada KD (2019) MAFFT online service: Multiple sequence alignment, interactive sequence choice and visualization. Briefings in Bioinformatics 20(4): 1160-1166. https://doi.org/10.1093/bib/bbx108 Kirk PM, Cannon PF, David J, Stalpers JA (2001) Ainsworth and Bisby’s dictionary of the fungi. CABI publishing, 665 pp. Kirk PM, Cannon PE, Minter DW, Stalpers JA (2008) Dictionary of the Fungi. CABI International, Wallingford, USA, 771 pp. 160 Sheng-Nan Wang et al. / MycoKeys 90: 147-162 (2022) Kirk PM, Stalpers JA, Braun U, Crous PW, Hansen K, Hawksworth DL, Hyde KD, Liicking R, Lumbsch TH, Rossman AY, Seifert KA, Stadler M (2013) A without-prejudice list of generic names of fungi for protection under the International Code of Nomenclature for algae, fungi, and plants. IMA Fungus 4(2): 381-443. https://doi.org/10.5598/imafun- gus.2013.04.02.17 Kits van Waveren E (1985) The Dutch. French and British species of Psathyrella. Persoonia 2: 1-284. Kornerup A, Wanscher JHK (1978) The methuen handbook of colour 3"! Edn. Eyre Methuen Ltd. Reprint., London, UK, 1-252. Lanfear R, Frandsen PB, Wright AM, Senfeld T, Calcott B (2017) PartitionFinder 2: New methods for selecting partitioned models of evolution for molecular and morphologi- cal phylogenetic analyses. Molecular Biology and Evolution 34: 772-773. https://doi. org/10.1093/molbev/msw260 Na Q, Liu Z-W, Zeng H, Cheng X-H, Ge Y-P (2022) Crepidotus yuanchui sp. nov. and C. caspari found in subalpine areas of China. Mycoscience 63(1): 1-11. https://doi.org/10.47371/ mycosci.2021.10.004 Nagy LG, Walther G, Hazi J, Vagvolgyi C, Papp T (2011) Understanding the Evolutionary Pro- cesses of Fungal Fruiting Bodies: Correlated Evolution and Divergence Times in the Psath- yrellaceae. Systematic Biology 60(3): 303-317. https://doi.org/10.1093/sysbio/syr005 Nagy LG, Vagvélgyi C, Papp T (2013) Morphological characterization of clades of the Psath- yrellaceae (Agaricales) inferred from a multigene phylogeny. Mycological Progress 12(3): 505-517. https://doi.org/10.1007/s11557-012-0857-3 Nguyen LT, Schmidt HA, von Haeseler A, Minh BQ (2014) IQ-TREE: A fast and effective stochastic algorithm for estimating maximum-likelihood phylogenies. Molecular Biology and Evolution 32(1): 268-274. https://doi.org/10.1093/molbev/msu300 Orstadius L, Knudsen H (2012) Psathyrella (Fr.) Quél. In: Knudsen H, Vesterholt J (Eds) Funga Nordica Agaricoid, boletoid, cyphelloid and gasteroid genera. Nordsvamp, Copen- hagen, Denmark, 586-623. Orstadius L, Ryberg M, Larsson E (2015) Molecular phylogenetics and taxonomy in Psathyrellace- ae (Agaricales) with focus on psathyrelloid species: Introduction of three new genera and 18 new species. Mycological Progress 14(5): 1-42. https://doi-org/10.1007/s11557-015-1047-x Padamsee M, Matheny PB, Dentinger BT, McLaughlin DJ (2008) The mushroom family Psathyrellaceae: Evidence for large-scale polyphyly of the genus Psathyrella. Molecular Phy- logenetics and Evolution 46(2): 415-429. https://doi.org/10.1016/j.ympev.2007.11.004 Persoon CH (1797) Tentamen dispositionis methodicae fungorum in classes, ordines, genera et familias. Cum supplemento adjecto. Arcum Caeci, Leipzig, Germany, 1-76. https://doi. org/10.5962/bhI.title.42674 Quelet L (1872) Les champignons du Jura et des vosges. Mémoires de la Société d’ Emulation de Montbéliard 5: 43-332. Redhead SA, Vilgalys R, Moncalvo JM, Johnson J, Hopple Jr JS (2001) Coprinus Pers. and the disposition of Coprinus species sensu lato. Taxon 50(1): 203-241. https://doi. org/10.2307/1224525 Lugisporipsathyra reticulopilea gen. et sp. nov. from Tropical China 161 Reid DA (1986) New or interesting records of Australasian basidiomycetes: VI. Transactions of the British Mycological Society 86(3): 429-440. https://doi.org/10.1016/S0007- 1536(86)80186-3 Ronquist F, Teslenko M, Van Der Mark P, Ayres DL, Darling A, Héhna S, Larget B, Liu L, Suchard MA, Huelsenbeck JP (2012) MrBayes 3.2: Efficient Bayesian phylogenetic infer- ence and model choice across a large model space. Systematic Biology 61(3): 539-542. https://doi.org/10.1093/sysbio/sys029 Singer R (1948) New Genera of Fungi-IV. Mycologia 40(2): 262—264. https://doi.org/10.108 0/00275514.1948.12017704 Singer R (1951) Type study on basidiomyceteV. Sydowia 5: 445-475. Singer R (1975) The Agaricales in modern taxonomy. J. Cramer, Vaduz, Liechtenstein, 912 pp. Singer R (1986) The Agaricales in Modern Taxonomy. 4 ed. Koeltz Scientific Books, Koenig- stein, Federal Republic of Germany, 981 pp. Smith AH (1972) The North American species of Psathyrella. The New York Botanical Garden 24 = 633% Vizzini A, Consiglio G, Marchetti M (2019) Mythicomycetaceae fam. nov. (Agaricineae, Agaricales) for accommodating the genera Mythicomyces and Stagnicola, and Simocybe parvispora reconsidered. Fungal Systematics and Evolution 3(1): 41-56. https://doi. org/10.3114/fuse.2019.03.05 Wachter D, Melzer A (2020) Proposal for a subdivision of the family Psathyrellaceae based on a taxon-rich phylogenetic analysis with iterative multigene guide tree. Mycological Progress 19(11): 1151-1265. https://doi.org/10.1007/s11557-020-01606-3 Wang S-N, Hu Y-P, Chen J-L, Qi L-L, Zeng H, Ding H, Huo G-H, Zhang L-P, Chen F-S, Yan J-Q (2021) First record of the rare genus Typhrasa (Psathyrellaceae, Agaricales) from China with description of two new species. MycoKeys 79: 119-128. https://doi.org/10.3897/ mycokeys.79.63700 White TJ, Bruns TD, Lee SB, Taylor JW, Innis MA, Gelfand DH, Sninsky JJ (1990) Amplifica- tion and direct sequencing of Fungal Ribosomal RNA Genes for phylogenetics. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ (Eds) PCR protocols: a guide to methods and applications. New York, USA, Academic Press, 315-322. https://doi.org/10.1016/B978- 0-12-372180-8.50042-1 Yan JQ, Bau T (2017) New and newly recorded species of Psathyrella (Psathyrellaceae, Agaricales) from Northeast China. Phytotaxa 321(1): 139-150. https://doi.org/10.11646/ phytotaxa.321.1.7 Yan JQ, Bau T (2018a) The Northeast Chinese species of Psathyrella (Agaricales, Psathyrel- laceae). MycoKeys 33: 85-102. https://doi.org/10.3897/mycokeys.33.24704 Yan JQ, Bau T (2018b) Psathyrella alpina sp. nov. (Psathyrellaceae, Agaricales), a new species from China. Phytotaxa 349(1): 85-91. https://doi.org/10.11646/phytotaxa.349.1.11 Yan J-Q, Ge Y-P, Hu D-M, Zhou J-P, Huo G-H (2019) Psathyrella tintinnabula sp. nov. (Psathyrellaceae, Agaricales), a new species from southwest China. Phytotaxa 400(2): 64-70. https://doi.org/10.11646/phytotaxa.400.2.2 162 Sheng-Nan Wang et al. / MycoKeys 90: 147-162 (2022) Yu W-J, Chang C, Qin L-W, Zeng N-K, Fan Y-G (2020) Pseudosperma citrinostipes (Inocy- baceae), a new species associated with Keteleeria from southwestern China. Phytotaxa 450: 8-16. https://doi.org/10.11646/phytotaxa.450.1.2 Zhao R-L, Li G-J, Sanchez-Ramirez S, Stata M, Yang Z-L, Wu G, Dai Y-C, He S-H, Cui B-K, Zhou J-L, Wu EK He M-Q, Moncalvo J-M, Hyde KD (2017) A six-gene phylogenetic over- view of Basidiomycota and allied phyla with estimated divergence times of higher taxa and a phyloproteomics perspective. Fungal Diversity 84(1): 43-74. https://doi.org/10.1007/ s13225-017-0381-5 Supplementary material | Iugisporipsathyra reticulopilea gen. et sp. nov. (Agaricales, Psathyrellaceae) from Tropical China Produces Unique Ridge-ornamented Spores with an Obvious Suprahilar Plage Authors: Jun-Qing Yan, Yu-Guang Fan, Sheng-Nan Wang Data type: phylogenetic Explanation note: A nexus file contains alignment sequence and original tree of ML and Bayes. Copyright notice: This dataset is made available under the Open Database License (http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License (ODDbL) is a license agreement intended to allow users to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited. Link: https://doi.org/10.3897/mycokeys.90.85690.suppl1