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Porpomyces Submucidus (Hydnodontaceae, Basidiomycota), a New Species from Tropical China Based on Morphological and Molecular Evidence

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Porpomyces Submucidus (Hydnodontaceae, Basidiomycota), a New Species from Tropical China Based on Morphological and Molecular Evidence See discussions, stats, and author profiles for this publication at: https://www.researchgate.net/publication/283034971 Porpomyces submucidus (Hydnodontaceae, Basidiomycota), a new species from tropical China based on morphological and molecular evidence Article in Phytotaxa · October 2015 DOI: 10.11646/phytotaxa.230.1.5 CITATIONS READS 2 186 2 authors, including: Fang Wu Beijing Forestry University 73 PUBLICATIONS 838 CITATIONS SEE PROFILE All content following this page was uploaded by Fang Wu on 30 May 2018. The user has requested enhancement of the downloaded file. Phytotaxa 230 (1): 061–068 ISSN 1179-3155 (print edition) www.mapress.com/phytotaxa/ PHYTOTAXA Copyright © 2015 Magnolia Press Article ISSN 1179-3163 (online edition) http://dx.doi.org/10.11646/phytotaxa.230.1.5 Porpomyces submucidus (Hydnodontaceae, Basidiomycota), a new species from tropical China based on morphological and molecular evidence FANG WU*, YUAN YUAN & CHANG-LIN ZHAO 1Institute of Microbiology, PO Box 61, Beijing Forestry University, Beijing 100083, China * Corresponding authors’ e-mails: [email protected] (FW) Abstract A new species is described from tropical China as Porpomyces submucidus on the basis of both morphological characters and molecular evidence. Phylogenetic analyse based on the ITS sequence, LSU sequence and the ITS+LSU sequence show that the new species belongs to Porpomyces and forms a distict clade as a sister group of Porpomyces mucidus. The fungus is characterized by thin, white to cream, resupinate basidiome with cottony to rhizomorphic margin, small pores (7–9 per mm), a monomitic hyphal structure with clamped generative hyphae, usually ampullated at most septa, and small, hyaline, thin-walled, ellipsoid basidiospores measuring 2.2–2.8 × 1.2–1.8 μm. It is closely related to Porpomyces mucidus, but the latter has larger pores (4–5 per mm) and larger basidiospores (2.8–3.9 × 2–2.8 μm). Key words: phylogeny, taxonomy, Trechisporales Introduction Porpomyces Jülich (Hydnodontaceae, Trechisporales), typified with Porpomyces mucidus (Pers. : Fr.) Jülich, was erected by Jülich (1982) and characterized by a combination of monomitic hyphal structure, clamped generative hyphae and a negative reaction in Melzer’s reagent (Jülich 1982, Tomšovský et al. 2010). The genus was treated as a synonym of Ceriporiopsis Domański by many mycologists (Gilbertson & Ryvarden 1986, Núñez & Ryvarden 2001, Ryvarden & Melo 2014). However, Rajchenberg (2003) found that the species formed ampulliform septa in culture. Additionally, molecular phylogeny demonstrated that it formed a distinct clade as a sister group of Trechispora P. Karst., and is not related to Ceriporiopsis (Larsson 2001). So, Porpomyces was revived as a valid genus based on morphological and molecular evidence by Rajchenberg (2003), Niemelä (2005), Tomšovský et al. (2010) and Yurchenko et al. (2014). Morphologically, Porpomyces differs from Ceriporiopsis by having cottony to rhizomorphic margin and ampullate clamp connections. China has a high diversity of polypores, especially in Hainan Province which is an island located between 18°10‘– 20°10‘N and 108°37‘–111°05‘E in Southern China, and 260 polypore species have been recorded from the island (Dai et al. 2011, 2012, Tian et al. 2013, Li et al. 2014). During a repeated field survey for wood-decaying fungi of Hainan Province in 2014, a nice resupinate polypore with white pores and rhizomorphic margin was found, and it was tentatively identified as Porpomyces mucidus. However, its smaller pores and basidiospores than those of P. mucidus suggest that it might be an undescribed species. Phylogenetic analysis confirms that it is new species of Porpomyces. The illustrated description of the new species is provided in the present paper. Materials and methods Morphological studies.—Studied specimens were deposited in the herbarium of the Institute of Microbiology, Beijing Forestry University (BJFC). The microscopic procedure followed He & Dai (2012). Sections were studied at magnifications up to × 1000 using a Nikon Eclipse 80i microscope and phase contrast illumination (Nikon, Tokyo, Japan). Drawings were made with a drawing tube. Microscopic features, measurements and drawings were made from Accepted by Samantha Karunarathna: 14 Sept. 2015; published: 6 Oct. 2015 61 slides stained with Cotton Blue and Melzer’s reagent. Spores were measured from sections cut from the tubes. Five percent of measurements were excluded from each end of the range, and were given in parentheses. In the text the following abbreviations were used: KOH = 5% potassium hydroxide, IKI = Melzer’s reagent, IKI– = neither amyloid nor dextrinoid, CB = Cotton Blue, CB+ = cyanophilous, L = mean spore length (arithmetic average of all spores), W = mean spore width (arithmetic average of all spores), Q = variation in the ratios of L/W between specimens studied, n = number of spores measured from given number of specimens (Zhao et al. 2013, Li et al. 2014, Zhou et al. 2015). Special color terms followed Petersen (1996). DNA extraction, amplification, sequencing and phylogenetic analyses.—Rapid extraction of genomic DNA kit CTAB (Aidlab Biotechnologies Co., Ltd, Beijing, China) was used to obtain DNA from dried specimens and cultural strains, according to the manufacturer’s instructions. ITS regions were amplified with primers ITS4 and ITS5 (White et al. 1990), and the LSU with primers LR0R and LR7 (http://www.biology.duke.edu/fungi/mycolab/primers.htm). PCR conditions were as follows: for ITS, initial denaturation at 95°C for 3 min, followed by 34 cycles at 94°C for 40 s, 54°C for 45 s, 72°C for 1 min; for LSU, initial denaturation at 94°C for 1 min, followed by 34 cycles at 94°C for 30 s, 50°C for 1 min, 72°C for 1.5 min, and a final extension of 72°C for 10 min. DNA sequencing was performed at Beijing Genomics Institute, China, with the same primers. All newly generated sequences were submitted to GenBank (Table 1). Sequences generated for this study were aligned with additional sequences downloaded from GenBank (Table 1) using BioEdit (Hall 1999) and ClustalX (Thompson et al. 1997). Sequence alignments were deposited at TreeBase (submission ID 17710). TABLE 1. List of species, specimens and GenBank accession number of sequences used in this study. GenBank accessions Species name Sample no. ITS LSU Brevicellicium olivascens (Bres.) K.H. Larss. & Hjortstam KHL 8571 - JN649327 Fibrodontia alba Yurchenko & Sheng H. Wu TNM F25503 JQ612713 JQ612714 F. alba TNM F24944 KC928274 KC928275 F. brevidens (Pat.) Hjortstam & Ryvarden Wu 9807-16 KC928276 KC928277 F. gossypina Parmasto AFTOL-ID 599 DQ249274 AY646100 Hyphodontia floccosa (Bourdot & Galzin) J. Erikss. Berglund 150-02 DQ873618 DQ873617 H. subalutacea (P. Karst.) J. Erikss. GEL2196 DQ340341 DQ340362 Porpomyces mucidus BRNM 710171 FJ496660 FJ496696 P. mucidus Cui 6919 KT157831a KT157836a P. mucidus JV 040725 KT157832a KT157837a P. mucidus Dai 12692 KT157833a KT157838a P. mucidus Dai 10726 KT157834a KT157839a P. mucidus Dai 175 KT157835a - P. submucidus F. Wu & C.L. Zhao Cui 5183 KT152143a KT152145a P. submucidus Dai 13708 KT152144a KT152146a Sistotremastrum suecicum Litsch. ex J. Erikss. KHL 11849 EU118666 EU118667 Subulicystidium longisporum (Pat.) Parmasto ZJ 4 JQ905612 - Trechispora alnicola (Bourdot & Galzin) Liberta AFTOL-ID 665 DQ411529 - T. farinacea (Pers. : Fr.) Liberta TUB 011825 EU909231 EU909231 T. hymenocystis (Berk. & Broome) K.H. Larss. KHL 8795 AF347090 - T. incisa K.H. Larss. EH 24/98 AF347085 - T. mollusca (Pers. Fr.) Liberta CFMR: DLL2011-186 KJ140681 - T. mollusca DLL2010-077 JQ673209 - T. nivea (Pers. : Fr.) K.H. Larss. G. Kristiansen - AY586720 T. regularis (Murrill) Liberta KHL 10881 AF347087 - Trechispora sp. 4Bart146S HQ022084 - Trechispora sp. G9-10 KF527456 - Trechispora sp. CFMR: DLL2011-162 KJ140660 - Trechispora sp. CFMR: DLL2011-213 KJ140703 - Trechispora sp. CFMR: DLL2011-008 KJ140534 - T. thelephora (Lév.) Ryvarden 1820 AMV KF937369 - Tubulicium vermiferum (Bourdot) Oberw. KHL 8714 - AY463477 a Newly generated sequences for this study 62 • Phytotaxa 230 (1) © 2015 Magnolia Press WU ET AL. Maximum parsimony analysis was applied to ITS, LSU and a concatenated dataset of ITS and LSU sequences. Sequences of Hyphodontia floccosa (Bourdot & Galzin) J. Erikss. and H. subalutacea (P. Karst.) J. Erikss. obtained from GenBank were used as outgroups (Yurchenko & Wu 2014). MP analysis was performed with PAUP* version 4.0b10 (Swofford 2002). All characters were equally weighted and gaps were treated as missing data. Trees were inferred using the heuristic search option with TBR branch swapping and 1,000 random sequence additions. Max-trees were set to 5,000, branches of zero length were collapsed and all parsimonious trees were saved. Clade robustness was assessed using bootstrap (BT) with 1,000 replicates (Felsenstein 1985). MrMODELTEST2.3 (Posada & Crandall 1998, Nylander 2004) was used to determine the best-fit evolution model for each data set for Bayesian inference (BI). Bayesian inference was calculated with MrBayes3.1.2 with a general time reversible (GTR) model of DNA substitution and a gamma distribution rate variation across sites (Ronquist & Huelsenbeck 2003). Four Markov chains were run for two runs from random starting trees for 0.5 million generations (ITS), for 0.2 million generations (LSU), for 0.5 million generations (ITS+LSU), and trees were sampled every 100 generations. The first one-fourth generations were discarded as burn-in (Tomšovský et al. 2010, Zhao et al. 2013, Li et al. 2014).
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