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Exidia Qinghaiensis, a New Species from China Shurong Wanga, R

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Exidia Qinghaiensis, a New Species from China Shurong Wanga, R Mycoscience VOL.62 (2021) 212-216 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. 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 (nrL- SU), RNA polymerase II second largest subunit (RPB2), and translation elongation factor 1-a (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 μm; 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 Auriculariaceae will require more extensive taxon sampling. Keywords: Auriculariaceae, Basidiomycota, one new species, phylogeny, taxonomy Article history: Received: 30 December 2020, Revised: 11 March 2021, Accepted: 11 March 2021, Available online 20 May 2021 Exidia Fr. is a genus of wood-inhabiting fungi (Basidiomycota: field collection and morphological and molecular studies of Chi- Auriculariaceae) (Hibbett et al., 2014) growing on dead branches nese Exidia specimens in herbaria. During our work we detected and logs, and best known from the temperate regions. Exidia forms one additional species-level clade based on phylogenetic analyses a sister group to the much better known “wood ears” of the genus of the nuclear ribosomal DNA internal transcribed spacer region Auricularia (Weiss & Oberwinkler, 2001). The basidia of Exidia are (nrITS) and large subunit (nrLSU), RNA polymerase II second pear-shaped and have longitudinal septa, unlike the tubular and largest subunit (RPB2), and translation elongation factor 1-a (Tef1). transversely septate basidia of Auricularia Bull. (Weiss & Oberwin- Herein we describe this new species based on specimens from Qin- kler, 2001). As in Auricularia, basidiocarps (fruiting bodies) are ghai Province, China. gelatinous and these are diverse in form, ranging from pustular to For light microscopic observations, freehand sections of a rehy- cup-shaped (Wojewoda, 1977; Moore, 1997); indeed, a few species drated portion of specimens were mounted in 2% (w/v) KOH. All of Exidia with rather ear-shaped fruiting bodies have regularly measurements of spores and hypobasidia were carried out using oil been misidentified as Auricularia (Barber, Thorn, & Voitk, 2011). immersion at 400 × and 1000 × magnification with differential -in The study of fungal diversity plays an important role in its pres- terference microscopy on a Zeiss AxioImager Z1. All spore dimen- ervation, not only in China but also on a worldwide scale. The my- sions exclude the hilar appendage and are reported as length, width cota of China, including Exidia, still has not been well investigated and Q (length/width), given as the 80th percentile range with out- because of its great geographical extent, and most Exidia species liers in parentheses; the average value of Q is reported as Qavg. Color reported lack supporting molecular data. On the basis of morpho- codes (e.g., 8B5) follow Kornerup and Wanscher (1978). The speci- logical data, a total of 8 species of Exidia, including synonyms and mens we borrowed and examined are from HMAS (Mycological three recently described species, have been reported in recent years Herbarium, Institute of Microbiology, Academia Sinica, Beijing, from China (Wu et al., 2020; Ye, Zhang, Wu, & Liu, 2020). Region- China). ally, five species have been reported from Japan (Aoki & Tubaki, Genomic DNA was extracted from dried materials using the 1986; Imazeki, Otani, & Hongo, 1988; Aoki, 1991), three from Ko- E.Z.N.A. Forensic DNA extraction kit (Omega Bio-Tek, Norcross, rea (Jung, 1993) and ten species from the Russian Far East (Govor- Georgia, USA). PCR amplification was performed with primers ova, 1998; Malysheva, 2012; Malysheva & Spirin, 2017). To add to ITS8F and ITS6R (Dentinger, Margaritescu, & Moncalvo, 2010) or the knowledge of Exidia in China, the first author has undertaken 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 * Corresponding author. Department of Biology, University of Western Ontario, London, N6A 5B7, Canada. b-7.1R (Matheny, 2005) or b-6F/f-7cR and b-6.9F/b-7.1R (Raja, E-mail address: [email protected] (G. Thorn). Miller, Pearce, & Oberlies, 2017) for the RPB2 region, and ef1-983-F This is an open-access paper distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivative 4.0 international license (CC BY-NC-ND 4.0: https://creativecommons.org/licenses/by-nc-nd/4.0/). doi:org/10.47371/mycosci.2021.03.002 ― 212 ― S. Wang, R.G. Thorn / Mycoscience VOL.62 (2021) 212-216 and ef1-1567-R (Rehner & Buckley, 2005) for Tef1. Successfully da [syn. Tremellochaete japonica (Yasuda) Raitv.], was placed with amplified products were cleaned using an EZ-10 Spin Column PCR strong support in the genus Exidia, as the sister to Exidia candida Products Purification Kit (BioBasic Canada, Markham, Ontario) Lloyd. However, a more inclusive taxon sample will be required to and sequenced using dye-terminator sequencing at Robarts Re- define generic limits in the Auriculariaceae. search Institute (London, Canada) or Sangon Biotech Co., Ltd. (Shanghai, China). Following DNA sequencing, chromatograms of Taxonomy partial sequences were cleaned and assembled using SeqEd v1.0.3 (Applied Biosystems, Foster City, California, USA). Newly acquired Exidia qinghaiensis S.R. Wang & Thorn, sp. nov. Figs. 2, 3. sequences have been deposited in GenBank as MW353408– MycoBank no.: MB 838343. MW353409 and MW358923–MW358926. Few Exidia sequences available from GenBank contained all of Diagnosis: Fruiting bodies are cushion-shaped to irregularly the chosen gene regions but, after preliminary analyses based sole- lobed and adpressed, becoming confluent, yellowish brown (drying ly on the ITS region that found no matches to our putative new fuscous), with paler flesh, basidiospores are hyaline, allantoid, av- species (data not shown), we chose to focus on a small dataset that eraging 12.7 × 3.4 μm, with Qavg = 3.7, and the host is Betula. Basid- had coverage of as many of these regions as possible (Table 1). For iospores of Exidia saccharina Fr., on conifers, are slightly wider Exidia glandulosa (Bull.) Fr. (HHB 12029) and the outgroup Auric- (10–14 × 3.5–4.0 μm), and those of E. thuretiana, also on angio- ularia heimuer F. Wu, B.K. Cui & Y.C. Dai (Dai 13782), draft ge- sperms, are both longer and broader (13–19 × 4.5–6.0 μm), but nome sequences are available and were queried using BLASTn to similar in shape to those of E. qinghaiensis. These species can be obtain sequences of RPB2 or Tef1. Sequences of the different gene distinguished by their nrITS, nrLSU, RPB2, and Tef1sequences. regions were separately aligned using MAFFT v7 online (Katoh & Holotype: CHINA, Qinghai Province, Menyuan County, Xianmi Standley, 2013) with the G-INS-i strategy and “leave gappy regions” wood farm, approx. 37°17’ N, 101°57’ E, 2,850 m above sea level option invoked, then the rough ends of alignments trimmed using (a.s.l.), on a fallen branch of Betula, 19 Oct 2004, leg. Zhang Xiao- MEGA X (Kumar, Stecher, & Tamura, 2016) before concatenating qing, HMAS 156328 (Mycological Herbarium, Institute of Microbi- to yield a combined matrix of 2,572 aligned bases. Prior to concate- ology, Academia Sinica). nation, evolutionary models were compared using MEGA X, and Gene sequences ex-holotype: MW353409 (nrITS, nrLSU), since the GTR+G+I model received the best ln(L) score in all cases, MW358924 (RPB2) and MW358926 (Tef1). this model was used in maximum likelihood (ML) analyses imple- Etymology: qinghaiensis (Latin), referring to Qinghai Province, mented in MEGA X for the combined dataset. Bayesian analyses where the holotype was collected. were conducted in MrBayes 3.2.6 (MB; Ronquist et al., 2012) with 5 Basidiomata gelatinous, cushion-shaped to irregularly lobed, 000 000 generations, 4 chains, and a burn-in of 25% (when the av- adpressed, orbicular, typically growing separately and adhering to erage standard deviation of split frequencies between chains had the substrate, sometimes fusing together and becoming confluent, stabilized below 0.001). Node support was determined as posterior up to 3 cm in widest dimension and 0.5 cm thick; when dried uni- probabilities in MrBayes, and as bootstrap support in ML analyses formly greyish or fuscous brown (7F3) on both hymenial and abhy- using 100 replicates. The alignments and trees have been deposited menial surfaces, rehydrating to yellowish brown or caramel (5DE7 to TreeBase (http://www.treebase.org) as S27878. to 6E7), with paler flesh; the hymenial, downward-facing surface Phylogenetic analyses of the combined nrITS, nrLSU, RPB2 and smooth, glabrous and a little shining, becoming slightly eroded in Tef1 data (Fig. 1) supports the segregation of Exidia qinghaiensis as age; abymenial surface slightly granular, usually without wrinkles a sister species to Exidia thuretiana (Lév.) Fr. A sample represent- and alveolate-venose. Flesh thin and pliant-gelatinous; odor and ing the type species of Tremellochaete Raitv., Exidia japonica Yasu- taste not noted. Table 1. Sequences of Auricularia and Exidia used in phylogenetic analyses. New sequences derived for this study are in bold font. Name Voucher Country ITS LSU RPB2 Tef1 A.
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