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Species of Psilocybe (Hymenogastraceae) from Yunnan, Southwest China

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Species of Psilocybe (Hymenogastraceae) from Yunnan, Southwest China Phytotaxa 284 (3): 181–193 ISSN 1179-3155 (print edition) http://www.mapress.com/j/pt/ PHYTOTAXA Copyright © 2016 Magnolia Press Article ISSN 1179-3163 (online edition) http://dx.doi.org/10.11646/phytotaxa.284.3.3 Species of Psilocybe (Hymenogastraceae) from Yunnan, southwest China TAO MA1, XIAO-FEI LING1 & KEVIN D. HYDE1, 2* 1 Research Institute of Resource Insects, Chinese Academy of Forestry, the Key Laboratory of Cultivating and Utilization of Resource Insects of State Forestry Administration, Kunming, Yunnan 650224, China 2 Center of Excellence in Fungal Research, Mae Fah Luang University 333 Moo l, Tasud, Muang, Chiang Rai 57100, Thailand Corresponding author: [email protected] Abstract Species of Psilocybe collected in Yunnan Province in southwest China were identified using both morphological characteris- tics and phylogenetic analyses of ITS and nrLSU sequence data. Six phylogenetic species were recognized among the stud- ied collections. In this paper, Psilocybe ruliensis is introduced as a new species, while Psilocybe keralensis is new to China, P. aff. fasciata is new to Yunnan and P. chuxiongensis is recollected in Yunnan Province. The remaining two phylogenetic species are not described or detailed due to the paucity of the material. The four species appear to be endemic to Asia. Key words: basidiomycota, hallucinogenic, taxonomy Introduction The genus Psilocybe sensu lato (sensu Guzmán 1983, 1995) includes the genus Deconica. The name Psilocybe was recently conserved with P. semilanceata (Fr.) P. Kumm. (1871:71) as the conserved type (Redhead et al. 2007), and Psilocybe is now applied to the clade of psychotropic species: Psilocybe sensu stricto (Ramírez-Cruz et al. 2013). However, Psilocybe fuscofulva Peck (1887:7) [formerly known as P. atrobrunnea (Lasch) Gillet (1878:586)] probably does not produce psychotropic alkaloids (Borovička et al. 2015). Eleven species of Psilocybe have been reported from China (Ma et al. 2014, Li et al. 2014, Wang & Tzean 2015) and among these species, P. chuxiongensis T. Ma & K.D. Hyde (2014:213), P. cinnamomea J.F. Liang, Yang K. Li & Ye Yuan (2014:216) and P. angulospora Wang & Tzean (2015:165) were originally described from China. Psilocybe taiwanensis Zhu L. Yang & Guzmán (2010: 185) was described from Taiwan (Guzmán & Yang 2010), but Guzmán et al. (2013) considered it with P. argentipes K. Yokoy (1976: 349) and P. thaizapoteca Guzmán, Karun. & Ram.-Guill. (2012:77) to be a synonym of P. subcaerulipes Hongo (1958:44). Psilocybe chuxiongensis was described from Yunnan Province in southwest China. In this study five phylogenetic species besides P. chuxiongensis were recognized among the studied collections. One of them is described as Psilocybe ruliensis sp. nov. based on evidences from morphology and ITS and LSU sequence data analyses. Psilocybe keralensis K.A. Thomas, Manim. & Guzmán (2002:196) is new to China and P. aff. fasciata Hongo (1957:144) is new to Yunnan Province. The other two phylogenetic species are not described due to inadequate materials and are not detailed further. Materials and Methods Sampling and morphological study Macro-morphological features are described based on detailed field notes made from fresh material and documented by photographs. Materials were dried using an electric food drier and specimens were deposited in the herbarium of the International Fungal Research and Development Centre (IFRDC), Research Institute of Resource Insects, Chinese Academy of Forestry. Facesoffungi and Index Fungorum numbers are registered (Jayasiri et al. 2015, Index Fungorum 2016). Colour designations (e.g. 4A5) are from Kornerup and Wanscher (1981). Microscopic features are described Accepted by Samantha Karunarathna: 25 Oct. 2016; published: 16 Nov. 2016 181 from dried material mounted in 5% KOH, Congo red, or water, measured and illustrated under a compound microscope (Nikon 80i). For terminology and methodology for descriptions see Ma et al. (2014). DNA extraction, PCR, Sequencing and phylogenetic analyses DNA extraction, PCR, sequencing and sequence alignment were followed Ma et al. (2014) and referenced therein. Phylogenetic analyses were conducted based on two nuclear genes (nrLSU and ITS). Sequences newly generated were aligned with selected reference sequences from GenBank. Seven reference taxa from GenBank lacked nrLSU and three lacked ITS sequence data, but were also included in this dataset and omissions were treated as missing data. Most of the reference sequences used are from the studies of Borovička et al. (2011, 2015) and Ramírez-Cruz et al. (2013). Species from the genera Stropharia were used as outgroup taxa, based on former studies (Moncalvo et al. 2002, Matheny et al. 2006). The combined nuclear dataset (LSU and ITS) was analysed using RAxML 7.2.6 (Stamatakis 2006) and MrBayes v3.1.2 (Huelsenbeck & Ronquist 2005), respectively. ML trees were inferred by running 1000 replicates combined with a ML+Rapid bootstrap. Bayesian inference (BI) best-fit models were determined by Akaike Information Criterion (AIC) in Modeltest Version 2.3 (Posada & Crandall 1998), and GTR+I+G model was suggested as the substitution model for each of the two nuclear regions and the combined dataset. BI analyses using six chains were run for 1 000 000 generations and trees sampled every 100th generation. The first 25% (2500) of samples were discarded as burn-in. Node support was estimated as posterior probabilities (pp) from the remaining trees. Clades receiving ML Bootstrap values (BS)>70% and BI pp>0.95 were considered to be significantly supported. Tree visualization and drawing were carried out with TreeView version 1.5.2 (Page 1996). Results Molecular studies Twenty-four new sequences (12 for ITS and LSU) were generated from twelve specimens of the seven species (Table 1) and are deposited in GenBank. The combined LSU+ITS dataset consisted of 69 taxa representing 44 species and 1375 nucleotide sites were included in the final alignment matrix. For this dataset, RAxML and BI analysis yielded nearly identical tree topologies and only the tree inferred from BI analysis is shown for the datasets. Phylogenetic trees generated showed that the samples collected from Yunnan grouped into six lineages which including P. chuxiongensis. Two lineages corresponded to P. fasciata and P. keralensis. Psilocybe aff. fasciata from Yunnan and P. fasciata from Japan clustered together with 83% RAxML BS support, but with a relatively low BI pp value. Four collections of P. keralensis from three sites in Yunnan clustered together although with relatively low support. Psilocybe ruiliensis, a novel species described in the present paper, with three samples from the same site, formed an independent clade with strong support (RAxML BS=100%, PP=1.00), and had a sister relationship with P. samuiensis (RAxML BS=79%, PP=0.80). The remaining species had limited material and are tentatively named Psilocybe spp. 1 and 2. Psilocybe sp. 1 formed a monophyletic clade, but with little support, and is related with P. keralensis, P. antioquiensis, P. zapotecorum, P. thaizapoteca and P. argentipes. Psilocybe sp. 2 and P. stuntzii formed a group with 78% RAxML BS and 0.92 Bayesian PP support (Fig. 1). TABLE 1. New sequences generated in this study GenBank accession Species name Voucher Coll. No. Locality ITS nrLSU Psilocybe aff. fasciata IFRD415224 MS007 Mangshi, Yunnan, ChinaKX357868 KX357869 P. keralensis IFRD414040 CX063 Chuxiong, Yunnan, China KX357870 KX357871 P. keralensis IFRD415173 ZY039 Zhaotong, Yunnan, China KX357872 KX357873 P. keralensis IFRD415174 ZY040 Zhaotong, Yunnan, China KX357874 KX357875 P. keralensis IFRD414213 ZD040 Shangri-La, Yunnan, China KX357876 KX357877 P. ruiliensis IFRD415240 RLC004 Ruili, Yunnan, China KX357878 KX357879 P. ruiliensis IFRD415241 holotype RLC005 Ruili, Yunnan, China KX357880 KX357881 P. ruiliensis IFRD415243 RLC007 Ruili, Yunnan, China KX357882 KX357883 Psilocybe sp.1 IFRD415205 SB143 Shuangbai county, Yunnan, China KX357884 KX357885 Psilocybe sp.2 IFRD414050 CX025 Chuxiong, Yunnan, China KX357886 KX357887 P. thaiduplicatocystidiata MFLU10-0850 NTS-120 Chiang Mai, Thailand KX357888 KX357889 P. thaiaerugineomaculans MFLU10-0851 NTS-121 Chiang Mai, Thailand KX357890 KX357891 182 • Phytotaxa 284 (3) © 2016 Magnolia Press MA ET AL. FIGURE 1. Phylogram inferred from analyses of a combined LSU and ITS dataset using MrBayes. Values above and below branches are posterior probabilities (pp≥0.80) and RAxML bootstrap (BS≥50%) values. For node with little difference between RAxML and BI analyses, only the pp value is displayed and BS value is not shown but indicated by ‘/’. The terminal taxa represent individual specimens with GenBank accession numbers. nrLSU and ITS numbers are indicated as LSU/ITS and the sequences that are lacking are not shown, and only one number is represented the sequence that is including nrLSU and ITS data. Species from Yunnan province and new sequences generated in this study are in bold. PSILOCYBE (HYMENOGASTRACEAE) Phytotaxa 284 (3) © 2016 Magnolia Press • 183 Taxonomy Psilocybe ruiliensis T. Ma, X.F. Ling & K.D. Hyde, sp. nov. (Figs. 2, 3) Index Fungorum number: IF552166, Facesoffungi number: FoF 02246 Etymology:—the species epithet ‘ruiliensis’ refers to the location Rili where the type collections were found. Holotype:—CHINA, Yunnan Province: Dehong Dai-Jingpo Autonomous Prefecture, Ruili, Longchuan County, on grassland near a reservoir, E097°48´03.6”, N24°15´48.9”, 940m, 24 September 2011, Tao Ma, Xiao-Fei Ling RLC005 (IFRD 415241). Diagnosis:―the
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