See discussions, stats, and author profiles for this publication at: https://www.researchgate.net/publication/309467764 Three New Species of Rhizopogon from Southwest China Article in Phytotaxa · October 2016 DOI: 10.11646/phytotaxa.282.2.7 CITATIONS READS 0 31 11 authors, including: Yongchang Zhao Fu-Qiang Yu Yunnan Academy of Agricultural Sciences Chinese Academy of Sciences 21 PUBLICATIONS 90 CITATIONS 21 PUBLICATIONS 63 CITATIONS SEE PROFILE SEE PROFILE Some of the authors of this publication are also working on these related projects: CHEMICAL COMPOSITION AND FUNCTIONS OF EDIBLE MUSHROOMS— View project HIMALAYAN HYPOGENOUS FUNGI—Diversity and distribution of sequestrate fungi, based on the high throughput sequencing of mycophagous animal feces View project All content following this page was uploaded by Fu-Qiang Yu on 09 March 2017. The user has requested enhancement of the downloaded file. 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Phytotaxa 282 (2): 151–163 ISSN 1179-3155 (print edition) http://www.mapress.com/j/pt/ PHYTOTAXA Copyright © 2016 Magnolia Press Article ISSN 1179-3163 (online edition) http://dx.doi.org/10.11646/phytotaxa.282.2.7 Three New Species of Rhizopogon from Southwest China LIN LIa,b,g, YONGCHANG ZHAOa, DEQUN ZHOUb*, FUQIANG YUc, LINYONG ZHENGd, YUN WANGe, XIAOLEI ZHANGa, ZHONGJIAN DUANf, XINGYUN ZHAOf, ZHIHONG HEf & SHUHONG LIa,b* aKey Laboratory for Agricultural Biotechnology of Yunnan, Biotechnology and Germplasm Resources Institute, Yunnan Academy of Agricultural Sciences, Kunming 650223 Yunnan, China bFaculty of Environmental Sciences and Engineering, Kunming University of Science and Technology, Kunming 650500 Yunnan, China cKunming Institute of Botany, Chinese Academy of Sciences, Kunming 650201 Yunnan, China dSichuan Academy of Agricultural Sciences, Chengdu 610066, Sichuan, China eNew Zealand Institute for Plant and Food Research Limited, Christchurch 8140, New Zealand fJianchuan County Forestry Bureau, Jianchuan 671300, Yunnan, China gThe College of Agriculture and Biological Science, Dali University, Dali 671000, Yunnan, China *Correspondence author: Shuhong Li Email: [email protected] Dequn Zhou Email: [email protected] Abstract Rhizopogon fruiting bodies were collected from Pinus forests in Yunnan and Sichuan Provinces in southwest China. Based on morphological and molecular phylogenetic analyses, three new species are described as Rhizopogon jiyaozi, Rhizopogon flavidus and Rhizopogon sinoalbidus. Their phylogenetic relationships to each other and previously described species are discussed. Key words: Hypogeous fungi, ITS, morphology, phylogeny, taxonomy Introduction The genus Rhizopogon Fr. & Nordholm (Basidiomycota, Boletales, Rhizopogonaceae) contains approximately 150 species of hypogeous fungi (Kirk et al. 2008). They are associated mainly with Pinaceae and are distributed worldwide in natural and exotic forests (Molina et al. 1999). Despite this cosmopolitan range, most known species are found in pine (Pinus) and Douglas-fir [Pseudotsuga menziesii (Mirb.) Franco] forests of the Pacific Northwestern United States, Europe and New Zealand (Smith 1964; Smith and Zeller 1966; Martín 1996; Visnovsky et al. 2010; Walbert et al. 2010; Cooper 2012). Rhizopogon are common ectomycorrhizal fungi in these coniferous forests and play an important role in the ecology of these forest ecosystems. The first Rhizopogon species was described in Europe by Elias Magnus Fries in 1817. Smith and Zeller published their North American monograph in 1966 and divided the genus into two subgenera, Rhizopogonella and Rhizopogon. Species in the subgenus Rhizopogonella were moved into Alpova by Trappe in 1975. Based on macroscopic and microscopic characteristics of the sporocarp and color changes on the peridium from chemical reactions, the subgenus Rhizopogon was divided into four sections, Amylopogon, Fulviglebae, Rhizopogon and Villosuli (Smith and Zeller 1966). The Morden taxonomy divided the genus Rhizopogon into five subgenera, Versicolores, Rhizopogon, Roseoli, Villosuli and Amylopogon, based on morphological and DNA molecular phylogenetic analyses (Grubisha 2001; Grubisha et al. 2002). To date six Rhizopogon species have been reported from China: Rhizopogon roseolus (Corda) Th. Fr. Rhizopogon shanxiensis B. Liu, Rhizopogon nigrescens Coker & Couch, Rhizopogon piceus Berk. & M.A. Curtis, Rhizopogon cylindriosporus A.H. Sm., Rhizopogon luteolus Fr. and Rhizopogon superiorensis A.H. Sm. (Liu B 1985; Tao and Chang 1988; Yu FQ and Liu PG 2005; Dai YC and Yang ZL 2008; Dai YC et al. 2010; Shao DH et al. 2013). In 2012 and 2013, a few Rhizopogon specimens were collected in Yunnan and Sichuan in southwest China (Plate.1). Morphological and molecular phylogenetic analyses determined that the Yunnan and Sichuan collections contained three new species. Here, the new species Rhizopogon jiyaozi belonging to subgen. Roseoli, Rhizopogon flavidus belonging to subgen. Rhizopogon and Rhizopogon sinoalbidus belonging to subgen. Amylopogon are described, and their relationships with other species of Rhizopogon are discussed. Accepted by Samantha Karunarathna: 2 Oct. 2016; published: 27 Oct. 2016 151 Material & methods Morphological characteristics examination Twenty-seven specimens were collected in Yunnan and Sichuan and deposited in the YAAS Herbarium, Yunnan Academy of Agricultural Sciences, Kunming, China. Macroscopic characteristics were described from fresh specimens. Sections were cut with a razor blade by hand, mounted in 5% KOH solution or water, and then stained with Melzer’s reagent. The sections were examined under an Olympus BH-2 microscope. DNA extraction, PCR amplification, sequencing and phylogenetic analyses The total genomic DNA of specimens was extracted from a piece of a fruiting body tissue using the Zomanbio Plant Genomic DNA Kit. The internal transcribed spacer (ITS) rDNA region was amplified with PCR primers ITS4 and ITS5. The PCR reactions were run on a BIO-RAD C1000TM Thermal Cycler with the following settings: initial denaturation for 5 min at 94°C, followed by 32 cycles of 40 s, denaturation at 94°C, annealing at 56°C for 40 s, extension for 1 min at 72°C and final extension at 72°C for 10 min. The purifying and sequencing of PCR products were conducted by the Beijing Genomics Institute. The other sequences data of ITS rDNA used in this study were downloaded from GenBank, as shown in Table 1. TABLE 1. Database of materials used for molecular analyses. Species name Voucher Origin GenBank No. of References ITS R. abietis A.H.Smith OSC41455 USA KC346843 From GenBank R. abietis A.H.Smith Trappe 7603 USA EU837243 From GenBank R. albidus A.H. Smith AHS 69642 USA AM085519 From GenBank R. alkalivirens A.H. Smith AHS69037 USA AF377154 Bidartondo & Bruns (2002) (holotype) R. arctostaphyli A.H. Smith JPT5705 USA AF377167 Bidartondo & Bruns (2002) (holotype) R. arctostaphyli A.H. Smith - USA EU726304 Hynes et al.(2010) R. atroviolaceus A.H. Smith AHS68263 USA AF377131 Bidartondo & Bruns (2002) R. bacillisporus A.H. Smith OSC63507 USA EU837230 From GenBank R. bacillisporus A.H. Smith SOC1307 USA JN022517 From GenBank R. brunsii Grubisha & Trappe LG1296 USA AY971814 Grubisha et al. (2005) R. brunsii Grubisha & Trappe LG701 USA AY971823 Grubisha et al. (2005) (Holotype) R. burlinghamii A.H.Smith JMT17882 USA AF058303 Grubisha et al. (2002) R. corsicus Demoulin & Moy. V. Demoulin s.n17-IX- Belgium AM085521 From GenBank 1989 R. ellenae A.H.Smith AHS66137 USA AF071445 Grubisha et al. (2002) R. ellenae A.H.Smith JMT17476 USA AF058311 Grubisha et al. (2002) R. ellenae A.H.Smith - USA JF695019 From GenBank R. ellipsosporus Trappe, Castellano OSC138981 USA JX310372 From GenBank & Amar. R. ellipsosporus Trappe, Castellano OSC138996 USA JX310373 From GenBank & Amar. R. evadens A.H.Smith AHS65484 USA AF062927 Grubisha et al. (2002) ...Continued on next page 152 • Phytotaxa 282 (2) © 2016 Magnolia Press LI ET AL. TABLE 1. (Continued) Species name Voucher Origin GenBank No. of References ITS R. evadens A.H.Smith JMT16402 USA AF058312 Grubisha et al. (2002) R. evadens A.H.Smith JMT12321 USA AF062932 Grubisha et al. (2002) R. fallax A.H. Smith AHS66116 USA AF377143 Bidartondo & Bruns (2002) R. fallax A.H. Smith AHS65762 USA AF377144 Bidartondo & Bruns (2002) R. flavidus YAASL2957(holotype) China KP893813 This study R. flavidus YAASL2956 China KP893814 This study R. flavidus YAASL2959 China KP893815 This study R. fuscorubens A.H.Smith JMT17446 USA AF058313 Grubisha et al. (2002) R. fuscorubens A.H.Smith Ash58b - AY880943 Ashkannejhad & Horton (2006) R. graveolens Tul. & C. Tul. PRM619028 Czech AJ810037 Martín & García (2009) R. idahoensis A.H. Smith - - AF224472 Kretzer et al. (2000) R. idahoensis A.H. Smith AHS70715 USA AF377123 Bidartondo & Bruns (2002) R. jiyaozi YAASL2929(holotype) China KP893830 This study R. jiyaozi YAASL2335 China KP893823 This study R. jiyaozi YAASL2336 China KP893824 This study R. jiyaozi YAASL2911 China KP893825 This study R. jiyaozi YAASL2916 China KP893826 This study R. jiyaozi YAASL2918 China KP893827 This study R. jiyaozi YAASL2925 China KP893828 This study R. jiyaozi YAASL2927 China KP893829 This study R. jiyaozi YAASL2930 China KP893831 This study R. jiyaozi YAASL2931 China KP893832 This study R. jiyaozi YAASL2932 China KP893833 This study R. jiyaozi YAASL2950 China KP893834 This study R. jiyaozi YAASL2395 China KP893835 This study R. jiyaozi YAASL2399 China KP893836