mycoscience 58 (2017) 297e301

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Short communication mollissima sp. nov. (, ) from Hainan, southern China

Yu-He Kan a,b, Wen-Min Qin a, Li-Wei Zhou a,* a CAS Key Laboratory of Forest Ecology and Management, Institute of Applied Ecology, Shenyang 110016, PR China b University of Chinese Academy of Sciences, Beijing 100049, PR China article info abstract

Article history: Hyphodontia mollissima is described from Hainan Province, southern China, as a new spe- Received 20 October 2016 cies. This is characterized by resupinate, soft corky basidiocarps, poroid hyme- Received in revised form nophores, a pseudodimitic hyphal system with clamped generative hyphae, presence of 5 April 2017 both bladder-like and apically encrusted cystidia, and broadly ellipsoid, colorless, thin- Accepted 6 April 2017 walled, smooth basidiospores. Morphologically, H. mollissima except for poroid hymeno- Available online 13 May 2017 phores, fits the concept of Palifer. A potential member of Palifer, Hyphodontia erikssonii, re- sembles H. mollissima by similar two kinds of cystidia and also similar shape and size of Keywords: basidiospores, but distinctly differs from the latter by having non-poroid hymenophores and a monomitic hyphal system. Hyphodontia s.l. © 2017 The Mycological Society of Japan. Published by Elsevier B.V. All rights reserved. ITS Palifer Polypores

Hyphodontia J. Erikss., typified by Hyphodontia pallidula (Bres.) J. Hjortstam & Ryvarden, Parmasto, Hastodontia Erikss., was erected by Eriksson (1958) and now belongs to (Parmasto) Hjortstam & Ryvarden, Kneiffiella P. Karst., Lagar- Schizoporaceae, a family of Hymenochaetales (Larsson 2007). obasidium Ju¨ lich, Lyomyces P. Karst., Palifer Stalpers & P.K. In a broad sense, Hyphodontia accommodates 128 valid species Buchanan, Rogersella Liberta & A.J. Navas, Schizopora Velen. (Chen et al. 2016; Yurchenko and Wu 2016), and is character- and (Pers.) Gray (Hjortstam and Ryvarden 2009). ized by white, cream to yellowish, resupinate basidiocarps However, no comprehensive molecular phylogeny fully sup- with poroid, hydnoid, odontioid and smooth hymenophores, a ported these morphology-based taxonomic changes. The lat- monomitic to pseudodimitic hyphal system, various kinds of est phylogenetic study on Hyphodontia s.l. only confirmed cystidia, and ellipsoid, subglobose, cylindrical to allantoid, Hastodontia, Hyphodontia s.s. and as indepen- smooth to finely ornamented, thin- to thick-walled basidio- dent genera, whereas other genera were either nested in spores (Hjortstam and Ryvarden 2009). Recently, some species weakly supported clades or mixed within the same clades of Hyphodontia s.l. were moved to other narrowly defined (Yurchenko and Wu 2014). Moreover, Fibrodontia was excluded genera, viz. Alutaceodontia (Parmasto) Hjortstam & Ryvarden, from Schizoporaceae, and belongs to Trechisporales (Binder Chaetoporellus Bondartsev & Singer, Deviodontia (Parmasto) et al. 2005).

* Corresponding author. Fax: þ86 24 83970348. E-mail address: [email protected] (L.-W. Zhou). http://dx.doi.org/10.1016/j.myc.2017.04.003 1340-3540/© 2017 The Mycological Society of Japan. Published by Elsevier B.V. All rights reserved. 298 mycoscience 58 (2017) 297e301

In China, about 60 species of Hyphodontia s.l. have been trees were used to construct a 50% majority consensus tree reported (Chen et al. 2016). A poroid species collected from and for calculating Bayesian posterior probabilities (BPPs). Hainan, southern China is presented herein as new to science, Tracer 1.5 (http://tree.bio.ed.ac.uk/software/tracer/) was used based on morphological and phylogenetic evidence. to confirm the convergence of chains. The studied specimens are deposited at IFP (acronym ac- The current ITS dataset had 71 sequences of Hyphodontia cording to the Index Herbariorum; http://sweetgum.nybg.org/ s.l. and one outgroup sequence, and resulted in an align- science/ih/). Special color terms for macroscopic characters of ment of 846 characters. BS searches in the ML analysis basidiocarps follow Petersen (1996). The microscopic proced- stopped after 200 replicates. BI converged after 10,000,000 ure follows Zhou et al. (2016). Sections were prepared in Cot- generations, which was indicated by the effective sample ton Blue (CB), Melzer's reagent and 5% (w/v) potassium sizes of all parameters over 3000 and the potential scale hydroxide (KOH), and then studied at magnifications of up to reduction factors close to 1.000. Both ML and BI algorithms 1000 using a Nikon Eclipse 80i microscope (Nikon, Tokyo, constructed almost congruent topologies, and thus only the Japan). Microscopic measurements were made from sections topology from ML algorithm is presented with statistical stained in CB. When presenting variation in basidiospore size, values at the nodes where BS and BPP over 50% and 0.8, the top and bottom 5% of measurements were excluded and respectively (Fig. 1). The ITS-based phylogeny (Fig. 1)shows are given in parentheses. Microscopic structures were drawn that the four newly sequenced specimens formed a fully from sections stained in CB with the aid of a light tube. The supported terminal lineage, which was clearly separated abbreviations used in the text are as follows: L ¼ mean from all other sampled species of Hyphodontia s.l. This basidiospore length (arithmetic average of all basidiospores), lineage was nested within the Xylodon-Schizopora-Palifer W ¼ mean basidiospore width (arithmetic average of all ba- clade (92%/1) and mostly related to Hyphodontia dimitica Jia J. sidiospores), Q ¼ variation in the L/W ratios among studied Chen & L.W. Zhou (90%/0.99). Combined with morpho- specimens and n ¼ number of basidiospores measured from logical characters, this lineage was described as a new the given number of specimens. species below. Dried herbarium specimens were subjected to DNA extraction using CTAB Rapid Plant Genome Extraction Kit- DN14 (Aidlab Biotechnologies Co., Ltd., China) according to the manufacturer's instruction. The resulted DNA was used as Hyphodontia mollissima L.W. Zhou, sp. nov. Figs. 2, 3. template for amplifying internal transcribed spacer (ITS) re- MycoBank no.: MB 818722. gion with primers ITS5 and ITS4 (White et al. 1990) using 2 ® EasyTaq PCR SuperMix (TransGen biotech, China). The PCR Type: CHINA, Hainan Province, Baoting County, Qixianling procedure was as follows: initial denaturation at 95 C for National Forest Park, approximate altitude 800 m, on fallen 3 min, followed by 34 cycles of 94 C for 40 s, 54 C for 45 s and angiosperm branch, 18 Mar 2016, LWZ 20160318-3 (holotype, 72 C for 1 min, and a final extension at 72 C for 10 min. The IFP 019149). PCR products were sequenced at Beijing Genomics Institute, rRNA gene sequences ex holotype: KY007517 (ITS). China with primers ITS5 and ITS4 (White et al. 1990). The Etymology: mollissima (Lat.): referring to the soft corky newly generated sequences were deposited at GenBank basidiocarps. (http://www.ncbi.nlm.nih.gov/genbank/; Fig. 1). Basidiocarps: annual, resupinate, adnate, inseparable, soft The four newly generated ITS sequences were incorpo- corky, without odor or taste when fresh, up to 10 cm long, rated with the datasets of previous studies on Hyphodontia s.l. 4.5 cm wide and 2.5 mm thick at center. Pore surface white to (Yurchenko and Wu 2014; Zhao et al. 2014; Riebesehl et al. cream when fresh, cream to buff when dry; pores angular, 5e7 2015; Chen et al. 2016), which comprise all reliable ITS se- per mm; dissepiments thin to thick, entire; sterile margin quences for species of Hyphodontia s.l. populinus white, up to 2 mm wide. Subiculum cream, up to 0.5 mm thick. (Schumach.) Donk was used as the outgroup taxon (Larsson Tube buff, up to 2 mm long. et al. 2006). The new ITS dataset was aligned using MAFFT 7 Hyphal structure: hyphal system pseudodimitic; gener- (Katoh and Standley 2013) with the G-INS-i option (Katoh et al. ative hyphae with clamp connections; tissue unchanged in 2005). The resulting alignment was deposited at TreeBASE KOH. Subiculum: generative hyphae dominant, colorless, (http://www.treebase.org; accession number S20770). The thin- to slightly thick-walled, frequently branched, loosely best-fit evolutionary model of the resulting alignment was interwoven, 3e5 mmdiam;skeletal-likehyphaerare,color- estimated as GTR þ I þ G using jModelTest 2.1.4 (Guindon and less, thick-walled with a wide to narrow lumen, un- Gascuel 2003; Darriba et al. 2012). Following this model, branched, 2.5e4.5 mm diam. Tubes: generative hyphae maximum likelihood (ML) and Bayesian inference (BI) algo- frequent, colorless, thin- to slightly thick-walled, frequently rithms were used to perform phylogenetic analysis. raxmlGUI branched, interwoven, 2e5 mm diam; skeletal-like hyphae 1.2 (Stamatakis 2006; Silvestro and Michalak 2012) with the occasional, colorless, thick-walled with a wide to narrow auto FC option (Pattengale et al. 2010) in bootstrap (BS) repli- lumen, unbranched, rarely clamped, 2.5e4 mmdiam. cates was used to conduct ML algorithm. BI algorithm Bladder-like cystidia colorless, thin- to slightly thick- employing two independent runs was carried out using walled, 20e35 7e10 mm. Apically encrusted cystidia cy- MrBayes 3.2 (Ronquist and Huelsenbeck 2003). Each run had lindrical, colorless, slightly thick-walled, 10e30 3e8 mm. 10,000,000 generations and started from random trees. Trees Basidia suburniform to clavate, with four sterigmata, were sampled every 1000th generation; the first 25% of sam- 10e17 3.5e5 mm. Basidiospores broadly ellipsoid, color- pling trees were discarded as burn-in, whereas all remaining less, thin-walled, usually bearing one small guttule, mycoscience 58 (2017) 297e301 299

Fig. 1 e Phylogenetic position of Hyphodontia mollissima within Hyphodontia s.l. inferred from ITS sequences. The topology is constructed by maximum likelihood algorithm, and bootstrap values and Bayesian posterior probabilities are calculated, respectively, from maximum likelihood and Bayesian inference algorithms. GenBank accession numbers and voucher numbers of sequences are given after each species name. The newly sequenced specimens are in boldface. The clade names are adapted from Yurchenko and Wu (2014). smooth, inamyloid, indextrinoid, acyanophilous, (3.9e) approximate altitude 500 m, on fallen angiosperm branch, 14 4e4.8(e5) (3e)3.3e4(e4.2) mm, L ¼ 4.37 mm, W ¼ 3.64 mm, Nov 2007, Yuan 4391 (paratype, IFP 018864); Ledong County, Q ¼ 1.18e1.23 (n ¼ 90/3) (Table 1). Jianfengling National Natural Reserve, approximate altitude Additional specimens examined: CHINA, Hainan Province, 850 m, on fallen angiosperm branch, 16 Nov 2007, Yuan 4562 Changjiang County, Bawangling National Natural Reserve, (paratype, IFP 019013), Yuan 4568 (paratype, IFP 019015). 300 mycoscience 58 (2017) 297e301

recently described as the first poroid species in Hyphodontia s.s. from phylogenetic and morphological perspectives by Riebesehl et al. (2015), who also emended the concept of Hyphodontia s.s. accordingly. However, in the current case, H. mollissima was separated from the only sampled species of Palifer, P. verecundus (G. Cunn.) Stalpers & P.K. Buchanan in the ITS-based phylogeny (Fig. 1), and more importantly, no comprehensive phylogenetic framework of Hyphodontia s.l. is available. Therefore, we tentatively place the new species in Hyphodontia s.l., not in Palifer. Further studies on the phylogeny of Hyphodontia s.l. via sampling more taxa and more importantly employing multiloci are needed to clarify the exact phylogenetic positions of H. mollissima and also other members of Hyphodontia.  Fig. 2 e Basidiocarps of Hyphodontia mollissima (holotype, Hyphodontia erikssonii (M. Galan & J.E. Wright) Hjortstam & LWZ 20160318-3, IFP 019149). Bar: 1 cm. Ryvarden, a member of Palifer as circumscribed by Yurchenko and Wu (2016), has similar two kinds of cystidia to H. mollis- sima (Galan et al. 1993). Moreover, both species have similar Hyphodontia mollissima is characterized by resupinate, basidiospores in shape and size (Galan et al. 1993). However, soft corky basidiocarps, poroid hymenophores, a pseudodi- H. erikssonii distinctly differs in its monomitic hyphal system mitic hyphal system with clamped generative hyphae, besides non-poroid hymenophores (Galan et al. 1993). presence of both bladder-like and apically encrusted cys- Hyphodontia mollissima is closely related to H. dimitica in the tidia, and broadly ellipsoid, colorless, thin-walled, smooth current phylogeny (Fig. 1). The latter species was recently basidiospores. The combination of these morphological described as new (Chen et al. 2016), but is considered to be characters but poroid hymenophores fits well with the conspecific with H. nongravis (Lloyd) Sheng H. Wu (Chen et al. circumscription of Palifer (Yurchenko and Wu 2016). This 2017). H. mollissima differs mainly in its smaller pores, which new species thus might be a poroid member of Palifer. are 2e4 per mm in H. dimitica (Chen et al. 2016) and 3e4 per Likewise, H. borbonica Riebesehl, E. Langer & Barniske was mm in H. nongravis (Wu 2000).

Fig. 3 e Microscopic structures of Hyphodontia mollissima (holotype, LWZ 20160318-3, IFP 019149). A: Basidiospores. B: A part of section of trama. C: Hyphae from subiculum. Bars:A5mm; B, C 10 mm.

Table 1 e Basidiospore measurements of each specimen of Hyphodontia mollissima. Specimen Range (mm) L (mm) W (mm) Q n LWZ 20160318-3 (IFP 019149) 4e4.8(e5) (3e)3.2e4(e4.2) 4.39 3.57 1.23 30 Yuan 4391 (IFP 018864) (3.9e)4e4.6(e4.7) (3.2e)3.3e3.9(e4) 4.24 3.58 1.18 30 Yuan 4568 (IFP 019015) (4e)4.2e4.7(e4.8) (3.3e)3.5e4 4.49 3.75 1.2 30 mycoscience 58 (2017) 297e301 301

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