中国蜡伞科云片衣属和地衣小荷叶属担子地衣
刘栋1,3, Bernard Goffinet2, 王欣宇1, 许宰铣3, 石海霞1, 张雁云1, 杨美霞1, 李丽娟1, 银安城1, 王立松1,*
1 中国科学院昆明植物研究所东亚植物多样性与生物地理学重点实验室 云南 昆明 650201
2 康涅狄格大学 美国 康涅狄格 06269-3043
3 国立顺天大学 韩国 顺天市 57922

刘栋

韩国国立顺天大学地衣生物学博士。主要从事地衣分类及分子系统学、淡水生态下地衣多样性研究。工作内容:中国担子地衣的分类学修订,韩国产淡水地衣的多样性调查及黑蜈蚣衣属 Phaeophyscia的分类学修订,及森林地衣监测物种的调查。采集标本约12 000份,覆盖范围主要为中国西南、东北地区及韩国山地、海岸、海岛及淡水环境。发表论文18篇,会议论文若干,参与了《中国药用地衣图鉴》、《中华大典·植物分典》的编写。

摘要

本文报道了中国蜡伞科担子地衣2个属4个种,其中包含1个新种和1个中国新记录种,新种为云南云片衣 Dictyonema yunnanum,新记录种即为亚洲新记录种,灰盖地衣小荷叶 Lichenomphalia velutina。云南云片衣的主要鉴别特征为:地衣体呈蓝绿色纤维状,纤维尖端白色至银白色,直立。本文对所研究的物种进行了详细形态描述并给出了每种的外形特征图,提供了中国蜡伞科担子地衣已知物种检索表。

关键词: 地衣型真菌; 分类; 多样性; 蜡伞科; 中国
Another lineage of basidiolichen in China, the genera Dictyonema and Lichenomphalia (Agaricales: Hygrophoraceae)
LIU Dong1,3, GOFFINET Bernard2, WANG Xin-Yu1, HUR Jae-Seoun3, SHI Hai-Xia1, ZHANG Yan-Yun1, YANG Mei-Xia1, LI Li-Juan1, YIN An-Cheng1, WANG Li-Song1,*
1 Key Laboratory for Plant Diversity & Biogeography of East Asia, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, Yunnan 650201, China;
2 Department of Ecology and Evolutionary Biology, University of Connecticut, 75 North Eagleville Road, Storrs CT, 06269-3043, USA
3 Korean Lichen Research Institute (KoLRI), Sunchon National University, Suncheon 57922, Korea;
Corresponding author. E-mail: wanglisong@mail.kib.ac.cn
Abstract

Several specimens of basidiolichen belonging to the Hygrophoraceae were collected as part of the China lichen mycota. These belong to four species in two genera, Dictyonemaand Lichenomphalia. Dictyonema yunnanumis described as new and Lichenomphalia velutina is newly recorded for Asia based on inferences from morphological, chemical and phylogenetic analysis. Dictyonema yunnanumis characterized by the dark aeruginous blue to black filamentous, ascending to erect, micro-fruticulose thallus. The species of lichenized Hygrophoraceae occurring in China are described and a key to distinguish them is presented.

Key words: lichenized fungi; taxonomy; diversity; Hygrophoraceae; China
Introduction

Lichenization is a successful strategy of establishing a stable mutualistic relationship of fungi with phycobionts. Currently about 19 000 (i.e., 17%) of all known fungal species form lichens and this percentage rises to 27% when only the Ascomycota are considered (Feuerer & Hawksworth 2007; Lü cking et al. 2016). Indeed most lichenized fungi are ascomycetes, whereas only 0.9% of lichen-forming fungi belong to the Basidiomycota (Lawrey et al. 2009; Oberwinkler 2012; Lü cking et al. 2016), the 172 known lichenized basidiomycetes are distributed across five orders (i.e., Agaricales, Atheliales, Lepidostromatales, Cantharellales and Corticiales) with most of the diversity occurring in the Hygrophoraceae (Agaricales), particularly in the genera CoraFr., DictyonemaC. Agardh ex Kunth and LichenomphaliaRedhead, Lutzoni, Moncalvo & Vilgalys (Lü cking et al. 2013a, 2014, 2016, 2017; Lü cking & Timdal 2016). These form either cyphelloid to steroid-corticioid basidiomata lichenized with cyanobacteria or agaricoid-omphalinoid mushrooms associated with green algae (Parmasto 1978; Redhead et al. 2002; Oberwinkler 2012).

The genus Dictyonema contained five species when it was first monographed by Parmasto (1978), but recent inferences from DNA sequences suggest that the diversity is vastly underestimated (Chaves et al. 2004; Lawrey et al. 2009; Lü cking et al. 2014), and that at least 144 species may be recognized and segregated into five genera: the filamentous Cyphellostereum D.A. Reid and Dictyonema s. str., the squamulose Acantholichen P.M. Jø rg. and the foliose Cora and Corella Vain. The distribution of the species is mainly pantropical, and parts of temperate areas (Dal-Forno et al. 2013; Lü cking et al. 2013a, 2013b, 2014, 2015, 2016; Vargas et al. 2014; Lü cking & Timdal 2016).

Agaricoid-omphalinoid fungi were originally accommodated in a single genus, Omphalina Qué l., which hence included lichenized and non-lichenized species. The lichenized species formed either a globular (Botrydina-type) or squamulose (Coriscium-type) thallus. Redhead et al. (2002) segregated the lichenized species to the new genus Lichenomphaliaon morphological and phylogenetic grounds. Lichenomphaliacurrently comprises 15 species distributed across artic and alpine areas of the Northern Hemisphere, Australia and Chile (Bigelow 1970; Redhead & Kuyper 1987; Barrasa 2001; Lü cking et al. 2017; Sandoval-Leiva et al. 2017).

Lichenized basidiomycetes occurring in China belong primarily to the Lepidostromatales, following the study by Liu et al. (2017) who transferred four species of MulticlavulaR.H. Petersen [Cantharellales. (Petersen & Zang 1986; Zang et al. 1996; Chou 2000; Jia et al. 2008)], to Sulzbacheromyces B.P. Hodk. & Lü cking, placed one species of LepidostromaMä gd. & S. Winkl. reported from China (He et al. 2016), in synonymy with S. sinensis(R.H. Petersen & M. Zang)

D. Liu & Li S. Wang and described two new species in this genus. Furthermore, three other species belonging to Lichenomphalia(Zang et al.1996; Xiao et al. 2005), and three more to Dictyonema(Wei 1991) were reported in last two dacades. Here we report on the Hygrophoraceae collected in southwestern China, combining a detailed morphological and anatomical study with molecular phylogenetic analyses of the internal transcribed spacer regions (ITS) and 28S rDNA to diagnose species and assess their ecological and geographic distribution in China.

1 MATERIALS AND METHODS
1.1 Materials and morphological observation

All the specimens used in this study are deposited in the Lichen Herbarium of the Kunming Institute of Botany (KUN-L). Morphological characters were examined with Nikon SMZ 745T dissecting microscope. Anatomical characters were investigated by Nikon Eclipse 50i stereomicroscope. Microscopic measurements were made in water, and the following notations are used for spore: mean values (in italics), and minimum-maximum values are given for all microstructures and derived parameters [Q= length/width ratios, Q (m) = mean values of Q]. The number (N) of basidiospores that were measured is given. Photographs were taken using a Nikon digital camera head DS-Fi2. Chemical spot tests were conducted following Orange et al. (2010).

1.2 DNA isolation, PCR and phylogenetic analysis

Total genomic DNA was extracted from freshly collected and frozen herbarium specimens using the Axyprep Multisource Genomic DNA Miniprep Kit following the manufacturer’ s instructions (Axygen Biosciences). The internal transcribed spacer (ITS) region and a 5’ portion of the 28S were amplified and sequenced using primers pairs ITS1F (Gardes & Bruns 1993) / ITS4 (White et al.1990) and LR0R (Rehner & Samuels 1994) / LR5 (Vilgalys & Hester 1990) respectively, following the protocols presented in Liuet al. (2014).

Newly generated sequences of Dictyonema and Lichenomphaliawere separately assembled using SeqMan (DNAstar packages) and edited by BioEdit 7.09 (Hall 1999) into two matrices: Dictyonema- matrix and Lichenomphalia-matrix. To assess the affinities of the Dictyonemaspecimen, which was phenotypically distinct from the known species, we sequenced two loci (i.e., ITS and 28S) and added these to a matrix of available ITS, 28S and also RPB2 sequences for 34 taxa plus one outgroup [i.e., Eonema pyriforme (M.P. Christ.) Redhead, Lü cking & Lawrey; Table 1]. Sequences for each locus were automatically aligned with MAFFT v7.273 (Katoh & Standley 2016), then screened for ambiguously aligned regions using the GUIDANCE webserver (Penn et al. 2010a, 2010b). Regions aligned with low confidence (ITS below 0.558, 28S below 0.90) were removed; no RBP2 sites were excluded. The three aligned matrices were combined by Phyutility 2.2.6 (Smith & Dunn 2008). The Lichenomphalia-matrix comprised only ITS sequences, sampled for 36 accessions representing seven taxa. Sixteen sequences are newly generated sequences and 20 are retrieved from GenBank. Ambiguous regions were selected using GUIDNCE (below 0.9), and excluded from further analyses.

Table 1 Voucher specimens information of ITS, LSU or RPB2 sequence data used in phylogenetic analysis

Both matrices were analyzed under the criterion of maximum likelihood (ML) using RAxML v7.2.6 (Stamatakis 2014) with the GTR+G+I model and branch support estimated via nonparametric bootstrapping based on 1 000 replicates. For the Bayesian inference (BI) performed with MrBayes v3.1.2 (Huelsenbeck & Ronquist 2001), the best-fitted substitution model was selected based on the Akaike Information Criterion (AIC) using jModelTest 3.7 (Posada 2008): TIMef+I+G for ITS, TrN+I+G for 28S and TrNef+G for RBP2 in Dictyonema-matrix, and GTR+I+G for ITS in Lichenomphalia-matrix. Bayesian inference was conducted using four chains and run for 2 million generations. Trees were sampled every 100th generations. Eonema pyriforme(M.P. Christ.) Redhead, Lü cking & Lawrey was selected as outgroup for the reconstruction of relationships within Dictyonema, and L. umbellifera (L.) Redhead, Lutzoni, Moncalvo & Vilgalys was used to root the tree comprising the remaining Lichenomphaliaaccessions in order to determine the position and closet relative of Chinese specimens based on the recent phylogenetic study (Lodge et al. 2014; Lü cking et al. 2017; Sandoval-Leiva et al. 2017).

2 RESULTS AND DISCUSSION

After excluding the ambiguous sites, the Dictyonema- matrix comprises 3 145 sites [667 for ITS, 1 451 for 28S and 1 027 for RBP2 (Dictyonema-matrix)] and the Lichenomphalina-matrix comprises 509 sites for ITS.

The most-likely phylogenetic tree (Fig. 1) inferred for the relationships within the Dictyonema s. str.

suggests that Cyphellostereumis monophyletic and sister to the remaining species, and that Dictyonemaforms a grade leading to the reciprocally monophyletic Acantholichen and Cora as previously proposed by Dal-Forno et al. (2013). A collection of Dictyonemafrom Yunnan was resolved sister to D. irpicinumalthough with no support. Three species and one variety of Dictyonema [currently D. scaridum, D. sericeum and D. thelephora, (Lü cking et al. 2013a)] were recorded in China (Wei 1991), which were treated as two species and four varieties by Parmasto (1978). We have not seen any populations of Dictyonema other than the one described here as D. yunnanum.

Fig. 1 Phylogenetic trees inferred by maximum likelihood (RAxML) using three loci (ITS, 28S, RPB2). Nodes supported by ML bootstrap values > 70% and Bayesian posterior probabilities > 0.95 are identified by thickened branches.

Inferences from ITS sequences resolve the Chinese specimens of Lichenomphalia as belonging to three species (bold in Fig. 2), which all occur in the Hengduan Mountains and Himalayan region. Zang et al. (1996) reported L. luteovitellina from the same overall area but we have not been able to locate populations of L. luteovitellina and also didn’ t find voucher specimens in his publication, hence the species could be included in our phylogenetic analysis. The genus Lichenomphalia primarily contained eight lichenized species when it was firstly revised by Redhead et al. (2002) from genus Omphalina. However, differences between these two genera are ambiguous, and some lichenized species may hide if the thallus is not obvious or collected in some historical specimens. Recently more species in Lichenomphalia were discovered or revised from Omphalina based on the phylogenetic analysis, such as L. altoandinafrom Chile (Sandoval-Leiva et al.2017) and L. oreades (=O. oreades) from North America and Europe (Lü cking et al. 2017). Molecular study is necessary to clarify species constitution in genera Lichenomphlia and Omphalina, nevertheless, type species and type location species of them need further studies in order to make clear the real nature relationship and evolution among species in Lichenomphalia.

Fig. 2 Phylogenetic trees inferred by maximum likelihood (RAxML) using ITS. Nodes supported by ML bootstrap values > 70% and Bayesian posterior probabilities > 0.95 are identified by thickened branches. Chinese accessions are in bold.

3 TAXONOMY

Dictyonema yunnanumD. Liu, X.Y. Wang & Li S. Wang, sp. nov. Fig. 3A-E

Fungal Name FN570516

Fig. 3 Dictyonema yunnanum(Li S. Wang & M.X. Yang 15-49922, holotype).A: Habitat; B: Erect fibroid thallus; C: Micro-structure of fibril; D: Filaments with heterocytes in microscopic view; E: Associated hyphae in the fibril tip, cyanobacterial cell absent. Bars: A=5cm; B=1cm; C, D, E=50µ m.

Etymology: The epithet refers to the type locality.

Holotype - CHINA: Yunnan Province, Pu’ er City, Ximeng Wa Nationality Autonomous County, Mengsuo Dragon Pond Park (Mengsuolongtan), 1 085m, on moss over bark, mixed with moss, 15 Nov 2015, Li S. Wang & M.X. Yang 15-49922 (KUN-L!).

Diagnosis: differs from D. thelephora by the erect fibrils with silvery or white tips.

Description. Thallus filamentous, ascending or erect, micro-fruticose, tightly interwoven, irregularly dispersed to confluent patches, entire thallus 5-10cm in diam.; prothallus absent; fibrils blue to dark blue, even black, apex white or silvery due to lack of cyanobacteria, 1-8mm tall, 180-248µ m thick. Photobiont cyanobacteria, not forming distinct medulla, filaments numerous, cells dark aeruginous blue, 9.9-12.1µ m wide, 4.7-5.5µ m high, wrapped in a hyphal sheath of paraplectenchymatous or jigsaw puzzle shaped cells; heterocysts sparse, pale yellow, 6-9.9µ m wide and 4-6µ m high; hyphae associated with hyphal sheath straight, 6.1-9.0µ m wide, lacking clamp connections.

Hymenophore not observed.

Chemistry. All spot tests (K, C, P) negative.

Habitat and distribution. Grow on mosses over bark, together with species of Cladonia, Graphis and Sticta. Known from a single collection from the tropical area of the Yunnan Province in Southwestern China.

Notes. This species is characterized by the dark aeruginous blue to black filaments, the ascending to erect, micro-fruticulose thallus, with white to silvery fibril tips, the absence of a prothallus and the pale yellow heterocysts. Dictyonema yunnanumdiffers from Acantholichen, Cora and Corella spp. by the filamentous versus squamulose or foliose thallus. It resembles Cyphellostereum spp. by its filamentous thallus, but differs in the hyphal sheath surrounding the photobiont filaments composed of paraplectenchymatous, jigsaw-puzzle-shaped cells. The new species is most similar with D. thelephoraby having dark blue fibrils but differs in the erect fibrils with silvery or white tips. Dictyonema yunnanum resembles members of the D. sericeumgroup in the blue green fibrils but differs in not forming lobes. It can be distinguished from D. pectinatum and D. schenckianum by fibrils not all pointing in a single direction, from D. metallicumand D. phyllophilum by absence of a silvery prothallus, from D. caespitosum and D. irrigatum by the absence of clamp connections in the hyphae and from D. irpicinumby the thallus surface lacking coarse, finger-like outgrowths, and the hyphae lacking clamps.

Lichenomphalia hudsoniana(H.S. Jenn.) Redhead, Lutzoni, Moncalvo & Vilgalys, Mycotaxon 83: 38 (2002). Fig. 4A, D

Fig. 4 Species of the genus Lichenomphalia in China. A: Habitat of L. hudsoniana (Li S. Wang etc. 12-34740); B: Habitat of L. umbellifera (Li S. Wang etc. 17-56025); C: Habitat of L. velutina(Li S. Wang etc. 12-35156); D: Botrydina-type thallus (Li S. Wang etc. 12-35137); E:Coriscium-type thallus (Li S. Wang etc. 12-34740). Bars: A=3cm; B=2cm; C=1cm; D, E=0.5mm.

Hygrophorus hudsonianusH.S. Jenn., Mem. Carnegie Mus.: 2 (1936) [MB#252625].

Omphalina hudsoniana(H.S. Jenn.) H.E. Bigelow, Mycologia 62: 15 (1970) [MB#357716].

Description. See Bigelow (1970).

Chemistry. All spot tests (K, C, P) negative in the thallus and basidiomata.

Habitat and distribution. Grow on moss, soil, bark and rotten wood. It’ s worldwide in arctic and alpine regions of the Northern hemisphere (Bigelow 1970; Redhead & Kuyper 1987; Lutzoni & Vilgalys 1995; Barrasa 2001; Zoller & Lutzoni 2003); in China known from the Himalayan region among Sichuan, Xizang and Yunnan Provinces, typically above 2 500m elevation.

Notes. This species is characterized by the squamulose (Coriscium-type) thallus, and the milky white to yellow orange basidiomata. The fruiting bodies of Chinese specimens exhibit a great variation of the pileus surface morphology and size (up to 4cm in diam.). Lichenomphalia hudsonianaoccurs in the same habitat as various Cladoniaspecies, when sterile its squamules can be distinguished from those of Cladonia by being more appressed and white patch consisting of numerous interwoven hyphae on the substrate.

Specimens examined. CHINA. SICHUAN PROVINCE: Muli Co., Ninglangshan Mountains, 3 900m, on soil, 24 Jun 1982, Y. Xuan 82-42; Kangding Co., Dujuanshan Mountains, 29° 54'10.8"N, 101° 59'59.34"E, 3 880m, on soil, 7 Jul 2006, Li S. Wang 06-26099; Yanyuan Co.: Bailinggongshe, the Fourth Production Brigade, 3 780m, on stump, Li S. Wang 83-1431; Dalingongshe, Huolushan Mountain, 2 750m, on Abiesspp., 21 Jul 1983, Li S. Wang 83-1153; 3 750m, on Abiesspp., 23 Jul 1983, Li S. Wang 83-1178(a); Shuazhisi Temper, Yakeshan Mountains, 3 600m, on soil under Abiesforest, 13 Jul 1858, X.J. Li 1561. Xizang Province: Linzhi Co., Lulang Town, nek of Sejilashan Mountains, 29° 37'15.96"N, 94° 40'4.2"E, 4 260m, on Sabina sp., 24 Aug 2007, Li S. Wang etc.

07-28373. Yunnan Province: Dali Bai Nationality Autonomous Prefecture: Dali City, Cangshan Mountains, 25° 41'05.32"N, 100° 06'14.37"E, 3 500m, on soil, 8 Jul 2012, Li S. Wang etc. 12-35050; Jianchuan Co., Laojunshan Mountains, 26° 37'53.53"N, 99° 43'36.79"E, 3 900m, on soil, 30 Jul. 2013, Li S. Wang etc. 13-38763; 3 800m, on bark, 30 Jul 2013, Li S. Wang etc. 13-38934, 13-38935, 13-38936; 26° 37'58.24"N, 99° 44'01.22"E, 4 050m, on bark, 30 Jul 2013, Li S. Wang etc. 13-38781; Gongshan Dulong Nationality Autonomous Co., on the way from Xishaofang to Dongshaofang, 27° 41'47.02"N, 98° 27'13.8"E, 3 510m, on rock, 3 Jun 2000, Li S. Wang 00-19099; 27° 42'15.18"N, 98° 26'40.62"E, 2 500m, on stump, 5 Jun 2000, Li S. Wang etc. 00-19276; on the way from Qiqi to Xishaofang, 27° 42'9.84"N, 98° 29'67"E, 2 800m, on rock, 2 Jun 2000, Li S. Wang 00-18777; Diqing Zang Nationality Autonomous Prefecture: Deqin Co.: nek of Baimaxueshan Mountains, 28° 22′ 05.90"N, 99° 00′ 20.00"E, 4 300m, on soil, 21 Jun 2013, Li S. Wang & X.Y. Wang 13-38411; on the way from Meilishi Village to nek of Suola Mountains, 28° 38′ 04.54"N, 98° 40′ 10.03"E, 3 336m, on soil, 9 Sep 2012, D. Liu etc. 12-35551; 28° 38′ 25.75"N, 98° 38′ 47.47"E, 3 950m, on rotten wood, 9 Sep 2012, D. Liu etc. 12-35682; on the way from God Waterfall (Shenpu) to Xiayubeng Village, 28° 23′ 3"N, 98° 47′ 46.2"E, 3 120m, on rotten wood, 13 Sep 2012, D. Liu etc. 12-36872; on the way from Yubeng Village to Xidan Co., nek of Nanzonglashan Mountains, 28° 23′ 57.48"N, 98° 46′ 20.04"E, 3 210m, on rotten wood, 14 Sep 2012, D. Liu etc. 12-36892; on the way from Yubeng Village to Xiaonong Camp, 28° 24′ 13.2"N, 98° 45′ 55.2"E, 3 620m, on rotten wood, D. Liu etc. 12-36868, 12-36876, 12-36877; Lijiang City, Jiuhexiang Town, Laojunshan Mountains, 26° 39'N, 99° 46'E, 3 690m, on soil, 3 Sep 2005, Li S. Wang 05-25056; 26° 39'N, 99° 46'E, 3 750m, on the soil under Rhododendronspp., 28 Aug 2005, Li S. Wang 05-25035; 26° 37'52.68"N, 99° 43'29.46"E, 3 850-3 895m, on soil and bark, 24-25 Aug 2006, Li S. Wang 06-26500, 06-26504, 06-26579, 06-26583; 26° 38'32.4"N, 99° 45'59.52"E, 3 900m, 16 Jul 2010, Li S. Wang 10-31510; 26° 37'51.66"N, 99° 42'57.84"E, 4 020m, on stump, 22 May 2011, Li S. Wang & M.M. Liang 11-32112, 11-32116; 26° 37'21.36"N, 99° 43'30.12"E, 3 900m, on moss, 13 Jun 2013, Li S. Wang & X.Y. Wang 13-38213; Shangeru-la City: Baishuitai, 27° 39'N, 100° 01'E, 3 400m, 19 Sep 1994, Li S. Wang 94-14675; Shuogenhu Lake, 3 500m, on moss, 26 May 1988, Li S. Wang 98-18138; nek of Daxueshan Mountains, 28° 34'N, 99° 49'E, 4 500m, on rock, 11 Oct 2001, Li S. Wang 01-20754; Tianbaoshan Mountains, 27° 36'16.93"N, 99° 53'06.61"E, 3 678m, on rotten wood, 6 Jul 2012, Li S. Wang etc. 12-34739, 12-34740, 12-35140, 12-35141, 12-35142, 12-35143; Habacun Village, Habaxueshan Mountains, 27° 20'21.36"N, 100° 04'46.56"E, 3 700m, on rock, 26 Oct 2002, Li S. Wang 02-22183; 3 900m, 12 Dec 2006, Li S. Wang etc. 06-21900; Xiaozhongdian Co., Tianchi Lake, 27° 37'N, 99° 33'E, 3 700m, on soil, 13 Jun 2004, Li S. Wang 04-23292.

Lichenomphalia umbellifera(L.) Redhead, Lutzoni, Moncalvo & Vilgalys, Mycotaxon 83: 38 (2002). Fig. 4B, E

Agaricus umbelliferL., Species Plantarum: 1175 (1753) [MB#516829].

Description. See (Bigelow 1970).

Chemistry. All spot tests (K, C, P) negative in the thallus and basidiomata.

Habitat and distribution. Grow on mosses, soil, bark and rotten wood. It widely distributes in arctic and alpine regions of the Northern Hemisphere (Bigelow 1970; Redhead & Kuyper 1987; Lutzoni & Vilgalys 1995; Barrasa 2001; Geml et al. 2012; Lü cking et al. 2017), and occurring between 1 500 and 4 200m in China.

Notes. This species is characterized by the globose (Botrydina-type) thallus and brownish fruitbody.

Additional specimens examined. CHINA. Sichuan Province: Jiulong Co., Wuxuhai, 29° 09'7.5"N, 101° 24'23.7"E, 3 600m, on stump, 24 Sep 2007, Li S. Wang 07-29193; Kangding Co, Yajiageng, 29° 47'56.4"N, 102° 03'42.24"E, 2 962m, on stump, 29 Sep 2010, Li S. Wang 10-31833; Muli Co., 3# District, 3 850m, on soil under the Abies forest, 8 Sep 1983, K.K. Chen 932. Xizang Province: Bomi Co, on the way from Daxing Village to Pangzhu Village, 2 900m, on soil, 1 Sep 1982, Y.G. Su 721; Mangkang Co., Honglashan Mountains, 23° 16′ 14.28″N, 98° 40′ 34.92″E, 4 135m, 16 Aug 2007, Li S. Wang etc. 07-27900; Motuo Co., Jiankaigongshe Gangrikabuqu South side, 2 350m, on soil, 23 Apr 1982, Y.G. Su 1337. Yunnan Province: Dali Bai Nationality Autonomous Prefecture, Dali City, Cangshan Mountains: 27° 36'16.93"N, 99° 53'06.61"E, 3 678m, on rotten wood, 6 Jul 2012, Li S. Wang etc. 12-34741; 25° 40'28.5"N, 100° 06'1.68"E, 3 570m, on rock, 28 Jul 2006, Li S. Wang 06-26213; Diqing Zang Nationality Autonomous Prefecture: Deqin Co.: on the way from Yubeng Village to Xiaonong Camp, 28° 24′ 13.2"N, 98° 45′ 55.2"E, 3 620m, on rotten wood, 14 Sep 2012, D. Liu etc. 12-36869; on the way from God Waterfall (Shenpu) to Xiayubeng Village, 28° 23′ 3.1"N, 98° 47′ 4.62"E, 3 120m, on rotten wood, 13 Sep 2012, D. Liu etc. 12-36873; on the way from Yubeng Village to Xidan Co., nek of Nanzonglashan Mountain, 28° 23′ 57.48"N, 98° 46′ 20.04"E, 3 210m, on rotten wood, 14 Sep 2012, D. Liu etc. 12-36881, 12-36882, 12-36883, 12-36884, 12-36885, 12-36886, 12-36887, 12-36888, 12-36889, 12-36890, 12-36891, 12-36893, 12-36894; Lijiang Co.: Jiuhexiang Town, Laojunshan Moutains, 26° 37'52.68"N, 99° 43'29.46"E, 3 850-3 895m, on soil, 24 Aug 2006, Li S. Wang etc. 06-26501, 06-26502, 06-26503; Yulongxueshan Moutains, Sandaowan, 3 000m, on soil, 15 Sep 1993, M. Zang 11948; 3 600m, on soil under Abies forest, 3 Jul 1985, Li S. Wang 85-0238; Weixi Co., Lidipingxiang Town, 27° 13'15.72"N, 99° 24'36.54"E, 3 430m, on stump, 18 Oct 2007, Li S. Wang 07-28817; Shangri-la City, Tianbaoshan Moutains: 27° 36'16.93"N, 99° 53'06.61"E, 3 678m, on rotten wood, 6 Jul 2012, Li S. Wang etc. 12-35145, 12-35147, 12-35150, 12-35151, 12-35152, 12-35153, 12-35154; 27° 36'17.81"N, 99° 53'08.44"E, 3 662m, on rotten wood, 8 Jul 2017, Li S. Wang etc. 17-56025; Nek of Xiaoxueshan Mountains, 28° 17'08.35"N, 99° 45'58.20"E, 3 461m, on rotten wood, 5 Jul 2012, Li S. Wang etc. 12-35137, 12-35138, 12-35139; Hongshan Mountains, 3 700m, on rotten wood, 29 Jul 1986, M. Zang 10551.

Lichenomphalia velutina(Qué l.) Redhead, Lutzoni, Moncalvo & Vilgalys, Mycotaxon 83: 43 (2002). Fig. 4C

Omphalia velutinaQué l., Comptes Rendus de l´ Association Franç aise pour l´ Avancement des Sciences 14(2): 445, t. 12 (1886) [MB#184594].

= Botrydina velutina(Qué l.) Redhead & Kuyper, Arctic and Alpine Mycology 2: 333 (1987) [MB#134045].

Description. Thallus not obvious, Botrydina-type, globular, with globules scattered or clustered, hygrophanous when wet, yellow-green to dark green, 35-90µ m in diam., diffuse; margin indistinct; globules consist of clusters that the unicellular green algae aggregated and enveloped by the hyaline hyphae, hyphae 1-5µ m wide, lacking clamps. Photobiont Coccomyxa, 4-8µ m in diam.

Basidiomata agaricoid, pileus 4-12mm diam., hygrophanous, grey brown to dark brown, convex, usually depressed in center, slightly undulate and streaked near margin, glabrous or slightly subfibrillose, gill line pale to dark black, obvious; context concolorous with pileus, thick and fragile, hyphae interleave, hyphae 4-8µ m wide with ‘ zebroid’ brown stripe. Odor and taste not distinctive. Lamellae distant, ventricose, decurrent to deeply decurrent, concolorous with or little paler than pileus consist of long and short lamellulae. Lamellar trama irregular, 4-8µ m wide with ‘ zebroid’ brown stripe. Stipe 0.5-1.5cm high, cylindric, solid, concolorous with pileus or little paler toward base, tomentose, white mycelial patch present at the base. Cystidia and clamps absent.

Basidia 14-25× 4-8.5µ m, clavate or subcylindrical, 4-spored, or 2-spored sometimes, sterigmata 2-6µ m long. Basidiospores 6-7.6-9× 4-5.1-6µ m [Q (m)= 1.54, N=20], elliptical, oval, obovate or subround, hyaline, non-amyloid, with oil droplets sometimes, hilar appendix present.

Chemistry. All spot tests (K, C, P) negative in the thallus and basidiomata.

Distribution and habitat. Grow on mosses and soil. Widespread in the lowland and alpine areas of Europe and North America (Lutzoni & Vilgalys 1995; Barrasa 2001; Lü cking et al. 2017), and known in China only from the Yunnan Province, above 3 000m elevation; on mosses and soil.

Notes. This species is new to China and Asia. The diagnostic morphological characters of this taxon are the globose (Botrydina-type) thallus, the dark grey brownish small fruiting body and the brown zebroid hyphae in the pileus. This species is known from few collections in Europe and North America, and only published sequences identified as Omphalina velutina(= L. velutina) in GenBank, and several other species (i.e. L. grisella), the morphological differences were not rather distinct, but they mixed with L. velutina, L. oreades or L. grisella. Although these species are morphologically indistinguishable, and our specimen forms a clade with the accession of L. velutina from Greenland, and together they compose a sister clade to L. oreades(incl. L. grisella; Fig. 2). The populations of L. velutinafrom China occurred over a narrower and higher altitudinal distribution (over 3 000m) than those in Europe, which occur below 1 500m elevation (Barrasa 2001).

Specimens examined. CHINA. Yunnan Province, Dali Bai Nationality Autonomous Prefecture, Dali City, Cangshan Mountains: 25° 41'09.81"N, 100° 06'30.12"E, 3 244m, on soil, 8 Jul 2012, Li S. Wang etc. 12-35149, 12-35156, 12-35157, 12-35159; 25° 41'05.32"N, 100° 06'14.37"E, 3 500m, on soil, 8 Jul 2012, Li S. Wang etc. 12-35160, 12-35161, 12-35162, 12-35163, 12-35164, 12-35165, 12-35166, 12-35167, 12-35168; 25° 41'6.6"N, 100° 06'14.4"E, 3 160m, on soil, 14 Aug. 2011, Li S. Wang etc. 11-32289.

Key to species of lichenized Hygrophoraceae in China

1. Photobiont cyanobacteria, thallus usually filamentous

Dictyonema 2

1. Photobiont green algae, thallus granules or squamulose

Lichenomphalia 3

2. Thallus forming semicircular lobes4

2. Thallus appressed filamentous 5

4. Fibrils narrow, densely arranged, giving the lobes an almost smooth appearance; thallus surface with coarse, finger-like outgrowths, lobe surface intensely blue-green; clamp connections present D. scabridum

4. Fibrils broad, loosely and irregularly arranged, giving the lobes a rough appearance; lobe surface mottled white and blue-green; clamp connections absent D. sericeum

5. Fibrils irregularly erect, apical silvery D. yunnanum

5. Fibrils irregularly appressed, apical ochraceous

D. thelephora

3. Thallus squamuloseL. hudsoniana

3. Thallus granulose 6

6. Hyphae zebroid brown L. velutina

6. Hyphae hyaline 7

7. Stipe bright yellow L. luteovitellina

7. Stipe pale grey to brownL.umbellifera

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