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Exidia Qinghaiensis, a New Species from China

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Exidia Qinghaiensis, a New Species from China Mycoscience: Advance Publication doi: 10.47371/mycosci.2021.03.002 Short Communication (Received December 30, 2020; Accepted March 11, 2021) J-STAGE Advance Published Date: March 27, 2021 Short communication Exidia qinghaiensis, a new species from China Shurong Wanga, R. Greg Thornb, * a College of Food Science and Engineering, Shanxi Agricultural University, Taigu, 030801, China b Department of Biology, University of Western Ontario, London, N6A 5B7, Canada. *Corresponding author. Department of Biology, University of Western Ontario, London, N6A 5B7, Canada. E-mail address: [email protected] (G. Thorn). Text: 12 pages; tables: 1; figures: 3 Advance Publication - 1 - Mycoscience: Advance Publication ABSTRACT A novel, wood-inhabiting jelly fungus from China is described as a new species, Exidia qinghaiensis (Basidiomycota: Auriculariaceae). Phylogenetic analyses were based on sequences of the nuclear ribosomal DNA internal transcribed spacer (nrITS) and large subunit (nrLSU), RNA polymerase II second largest subunit (RPB2), and translation elongation factor 1- (Tef1) regions. Sequences of the new taxon formed a sister group to Exidia thuretiana, a species known from Europe and Asia, and distant to sequences of Exidia repanda from Europe. Fruiting bodies are cushion-shaped to irregularly lobed and yellowish brown, basidiospores are hyaline, allantoid (averaging 12.7 3.4 m; average length/width is 3.7), and the host is Betula. The new species also can be distinguished by nrITS, nrLSU, RPB2, and Tef1 sequences. Our multigene phylogeny supports an Exidia including Exidia japonica, type species of Tremellochaete, but defining generic limits in AuriculariaceaePublication will require more extensive taxon sampling. Keywords: Auriculariaceae, Basidiomycota, one new species, phylogeny, taxonomy Advance - 2 - Exidia qinghaiensis, a new species from China Exidia Fr. is a genus of wood-inhabiting fungi (Basidiomycota: Auriculariaceae) (Hibbett et al., 2014) growing on dead branches and logs, and best known from the temperate regions. Exidia forms a sister group to the much better known “wood ears” of the genus Auricularia (Weiss & Oberwinkler, 2001). The basidia of Exidia are pear-shaped and have longitudinal septa, unlike the tubular and transversely septate basidia of Auricularia Bull. (Weiss & Oberwinkler, 2001). As in Auricularia, basidiocarps (fruiting bodies) are gelatinous and these are diverse in form, ranging from pustular to cup-shaped (Wojewoda, 1977; Moore, 1997); indeed, a few species of Exidia with rather ear-shaped fruiting bodies have regularly been misidentified as Auricularia (Barber, Thorn, & Voitk, 2011). The study of fungal diversity plays an important role in its preservation, not only in China but also on a worldwide scale. The mycota of China, including Exidia, still has not been well investigated because of its great geographical extent, and most Exidia species reported lack supporting molecular data. On the basis of morphological data, a total of 8 species of Exidia, including synonyms and three recently describedPublication species, have been reported in recent years from China (Wu et al., 2020; Ye, Zhang, Wu, & Liu, 2020). Regionally, five species have been reported from Japan (Aoki & Tubaki, 1986; Imazeki, Otani, & Hongo, 1988; Aoki, 1991), three from Korea (Jung, 1993) and ten species from the Russian Far East (Govorova, 1998; Malysheva, 2012; Malysheva & Spirin, 2017). To add to the knowledge of Exidia in China, the first author has undertaken field collection and morphological and molecular studies of Chinese Exidia specimens in herbaria. During our work we detected one additional species-level clade based onAdvance phylogenetic analyses of the nuclear ribosomal DNA internal transcribed spacer region (nrITS) and large subunit (nrLSU), RNA polymerase II second largest subunit (RPB2), - 3 - Mycoscience: Advance Publication and translation elongation factor 1- (Tef1). Herein we describe this new species based on specimens from Qinghai Province, China. For light microscopic observations, freehand sections of a rehydrated portion of specimens were mounted in 2% (w/v) KOH. All measurements of spores and hypobasidia were carried out using oil immersion at 400 and 1000 magnification with differential interference microscopy on a Zeiss AxioImager Z1. All spore dimensions exclude the hilar appendage and are reported as length, width and Q (length/width), given as the 80th percentile range with outliers in parentheses; the average value of Q is reported as Qavg. Color codes (e.g., 8B5) follow Kornerup and Wanscher (1978). The specimens we borrowed and examined are from HMAS (Mycological Herbarium, Institute of Microbiology, Academia Sinica, Beijing, China). Genomic DNA was extracted from dried materials using the E.Z.N.A. Forensic DNA extraction kit (Omega Bio-Tek, Norcross, Georgia, USA). PCR amplification was performed with primers ITS8F and ITS6R (Dentinger, Margaritescu,Publication & Moncalvo, 2010) or ITS8F/5.8S and 5.8SR/LS1R (Vilgalys & Hester, 1990; Hausner, Reid, & Klassen, 1993) for the ITS region, primers LS1 and LR3 (Vilgalys & Hester, 1990) for the 5'-LSU region, primers b-6F and b-7.1R (Matheny, 2005) or b-6F/f-7cR and b-6.9F/b-7.1R (Raja, Miller, Pearce, & Oberlies, 2017) for the RPB2 region, and ef1-983-F and ef1-1567-R (Rehner & Buckley, 2005) for Tef1. Successfully amplified products were cleaned using an EZ-10 Spin Column PCR Products Purification Kit (BioBasic Canada, Markham, Ontario) and sequenced using dye- terminatorAdvance sequencing at Robarts Research Institute (London, Canada) or Sangon Biotech Co., Ltd. (Shanghai, China). Following DNA sequencing, chromatograms of partial sequences were cleaned and assembled using SeqEd v1.0.3 (Applied Biosystems, Foster City, California, USA). - 4 - Exidia qinghaiensis, a new species from China Newly acquired sequences have been deposited in GenBank as MW353408–MW353409 and MW358923–MW358926. Few Exidia sequences available from GenBank contained all of the chosen gene regions but, after preliminary analyses based solely on the ITS region that found no matches to our putative new species (data not shown), we chose to focus on a small dataset that had coverage of as many of these regions as possible (Table 1). For Exidia glandulosa (Bull.) Fr. (HHB 12029) and the outgroup Auricularia heimuer F. Wu, B.K. Cui & Y.C. Dai (Dai 13782), draft genome sequences are available and were queried using BLASTn to obtain sequences of RPB2 or Tef1. Sequences of the different gene regions were separately aligned using MAFFT v7 online (Katoh & Standley, 2013) with the G-INS-i strategy and “leave gappy regions” option invoked, then the rough ends of alignments trimmed using MEGA X (Kumar, Stecher, & Tamura, 2016) before concatenating to yield a combined matrix of 2,572 aligned bases. Prior to concatenation, evolutionary models were compared using MEGA X, and since the GTR+G+I model received the best ln(L) score in all cases, Publicationthis model was used in maximum likelihood (ML) analyses implemented in MEGA X for the combined dataset. Bayesian analyses were conducted in MrBayes 3.2.6 (MB; Ronquist et al., 2012) with 5 000 000 generations, 4 chains, and a burn-in of 25% (when the average standard deviation of split frequencies between chains had stabilized below 0.001). Node support was determined as posterior probabilities in MrBayes, and as bootstrap support in ML analyses using 100 replicates. The alignments and trees have been deposited to TreeBase (http://www.treebase.org) as S27878. AdvancePhylogenetic analyses of the combined nrITS, nrLSU, RPB2 and Tef1 data (Fig. 1) supports the segregation of Exidia qinghaiensis as a sister species to Exidia thuretiana (Lév.) Fr. A sample representing the type species of Tremellochaete Raitv., Exidia japonica Yasuda - 5 - Mycoscience: Advance Publication [syn. Tremellochaete japonica (Yasuda) Raitv.], was placed with strong support in the genus Exidia, as the sister to Exidia candida Lloyd. However, a more inclusive taxon sample will be required to define generic limits in the Auriculariaceae. Taxonomy Exidia qinghaiensis S.R. Wang & Thorn, sp. nov. Figs. 2, 3. MycoBank no.: MB 838343. Diagnosis: Fruiting bodies are cushion-shaped to irregularly lobed and adpressed, becoming confluent, yellowish brown (drying fuscous), with paler flesh, basidiospores are hyaline, allantoid, averaging 12.7 3.4 m, with Qavg = 3.7, and the host is Betula. Basidiospores of Exidia saccharina Fr., on conifers, are slightly wider (10–14 3.5–4.0 m), and those of E. thuretiana, also on angiosperms,Publication are both longer and broader (13–19 4.5–6.0 m), but similar in shape to those of E. qinghaiensis. These species can be distinguished by their nrITS, nrLSU, RPB2, and Tef1sequences. Holotype: CHINA, Qinghai Province, Menyuan County, Xianmi wood farm, approx. 37°17' N, 101°57' E, 2,850 m above sea level (a.s.l.), on a fallen branch of Betula, 19 Oct 2004, leg. Zhang Xiaoqing, HMAS 156328 (Mycological Herbarium, Institute of Microbiology, Academia Sinica). AdvanceGene sequences ex-holotype: MW353409 (nrITS, nrLSU), MW358924 (RPB2) and MW358926 (Tef1). Etymology: qinghaiensis (Latin), referring to Qinghai Province, where the holotype was collected. - 6 - Exidia qinghaiensis, a new species from China Basidiomata gelatinous, cushion-shaped to irregularly lobed, adpressed, orbicular, typically growing separately and adhering to the substrate, sometimes fusing together and becoming confluent, up to 3 cm in widest dimension and 0.5 cm thick; when dried
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