Fungal Diversity DOI 10.1007/s13225-015-0336-7 Porcini mushrooms (Boletus sect. Boletus)fromChina Yang-Yang Cui 1,3 & Bang Feng1 & Gang Wu1 & Jianping Xu2 & Zhu L. Yang1 Received: 3 November 2014 /Accepted: 17 April 2015 # School of Science 2015 Abstract Porcini mushrooms (Boletus sect. Boletus)haveboth importance (Arora 2008; Sitta and Floriani 2008; Sitta and economic and ecological importance. Recent molecular phylo- Davoli 2012). This group of fungi can form ectomyrrizhal genetic study has uncovered rich species diversity of this group symbiosis with plants of several families, such as Pinaceae, of fungi from China. In this study, the Chinese porcini were Fagaceae and Dipterocarpaceae. Meanwhile, porcini mush- characterized by both morphological and molecular phylogenetic rooms are very famous wild edible mushrooms which are evidence. 15 species were recognized, including nine new spe- consumed worldwide (Arora 2008; Sitta and Floriani 2008; cies, namely B. botryoides, B. fagacicola, B. griseiceps, Dentinger et al. 2010;Fengetal.2012; Sitta and Davoli 2012; B. monilifer, B. sinoedulis, B. subviolaceofuscus, Dentinger and Suz 2014). B. tylopilopsis, B. umbrinipileus and B. viscidiceps. Three previ- Since the establishment of the generic name Boletus L. ously described species, viz. B. bainiugan, B. meiweiniuganjun (Linnaeus 1753), many mycologists have contributed to the and B. shiyong, were revised, and B. meiweiniuganjun is treated taxonomic studies of porcini and their allies, either suggesting as a synonym of B. bainiugan.AkeytotheChineseporcini keep the genus Boletus in the broad sense that would represent mushrooms was provided. the currently accepted whole family Boletaceae or split it into small subgenara/sections or different genera (e.g. Gilbert Keywords Boletes . Taxonomy . Morphology . Phylogeny . 1931;Chiu1948, 1957; Singer 1965, 1967, 1986; Watling Wild edible mushrooms 1970; Snell and Dick 1970; Smith and Thiers 1971; Corner 1972;Alessio1985;Zang2006;Horak2005, 2011). Several molecular phylogenetic studies have indicated that the genus Introduction Boletus in the broad sense is polyphyletic (Binder and Hibbett 2006; Dentinger et al. 2010; Feng et al. 2012; Nuhn et al. 2013;Wuetal.2014). However, B. sect. Boletus, the porcini Porcini (Boletus edulis and its allies) are one of the most im- group, typified by B. edulis Bull., is monophyletic (Dentinger portant fungal groups due to their ecological and economic et al. 2010;Fengetal.2012;Nuhnetal.2013;Wuetal.2014). Yang-Yang Cui and Bang Feng contributed equally to this work. This group shares the following common features: the surface of the immature poroid hymenophore is covered with a layer * Zhu L. Yang of tangled white hyphae (referred as Bstuffed pores^, [email protected] Fig. 19a), stipe is more or less reticulated, and the whitish to white flesh is without color change when cut (Coker and Beers 1943; Smith and Thiers 1971; Corner 1972; Singer 1986; 1 Key Laboratory for Plant Diversity and Biogeography of East Asia, Kunming Institute of Botany, Chinese Academy of Sciences, Horak 2005; Halling et al. 2014). Heilongtan, Kunming 650201, China Numerous studies have been conducted to address the tax- 2 Department of Biology, McMaster University, Hamilton, ON L8S onomic issue of porcini mushrooms from different continents. 4K1, Canada In comparison with the taxa from other continents, the porcini 3 University of Chinese Academy of Sciences, No.19A Yuquan Road, from Europe and North America have drawn much more at- Beijing 100049, China tention for a very long time and a more comprehensive Fungal Diversity understanding about them has been reached from both mor- by species epithet. Both macro- and microscopic descriptions phological and molecular phylogenetic views (Singer 1965, are provided for each species. Macroscopic features are based 1967, 1986; Watling 1970; Smith and Thiers 1971;Alessio on detailed field notes and photographs. Microscopic struc- 1985; Bessette et al. 2000; Horak 2005, 2011; Binder and tures were observed on dried material with light microscopy. Hibbett 2006; Beugelsdijk et al. 2008; Dentinger et al. Methods for microscopic studies followed those in Zeng et al. 2010). To date, including stipitate-pileate and sequestrate taxa, (2013), Li et al. (2011, 2014) and Wu et al. (2015). Authors of about 50 species were described and arranged in B. sect. the fungal names listed in Table 1 will not appear in the text. Boletus wordwide (Berkeley 1852; Peck 1888, 1905; Corner 1972;Hongo1973;Horak1980, 2005, 2011;Singer1986; Molecular phylogenetics Both 1993;Nagasawa1994; Zang 2006; Ortiz-Santana et al. 2007; Dentinger 2013;Nuhnetal.2013; Halling et al. 2014; Sequences of three gene makers, the internal transcribed spac- Zeng et al. 2014). Recent molecular phylogenetic analyses er (ITS), the large nuclear ribosomal RNA subunit (nrLSU) indicated that the species diversity of porcini mushrooms and the largest subunit of RNA polymerase II (rpb1), were was heavily under-estimated in East Asia and 15 new phylo- generated for newly added samples following Feng et al. genetic species (fourteen from China) were uncovered (Feng (2012). ITS sequences were also generated for the Bstufffed et al. 2012). Four of them were subsequently described as pores^ of two specimens from B. bainiugan and B. shiyong, B. shiyong, B. bainiugan, B. meiweiniuganjun and respectively. Detailed information about these newly generat- B. orientialbus (Dentinger 2013;Zengetal.2014), while the ed sequences is provided in Table 2. These sequences were remaining species have not been documented yet. combined with the sequences of the representatives of porcini Overall, eight species, viz. B. bainiugan, B. edulis, mushrooms which were used in Dentinger et al. (2010), Feng B. hiratsukae, B. meiweiniuganjun, B. orientialbus, et al. (2012), Nuhn et al. (2013) and Wu et al. (2014) for B. reticuloceps, B. shiyong and B. violaceofuscus, were phylogenetic analyses. Two data matrices, the combined reported from East Asia (Chiu 1948, 1957;Zangetal.1993; dataset of nrLSU and rpb1, and the ITS dataset, were analyzed Nagasawa 1994; Li and Song 2003; Wang and Yao 2005; for different purposes following Feng et al. (2012). However, Dentinger 2013;Zengetal.2014). different from those in the Feng et al. (2012), the current B ^ In porcini, the feature stuffed pores (Fig. 19a; Coker and analyses included the introns of rpb1, while the ambiguously Beers 1943; Smith and Thiers 1971;Singer1986) has been aligned sequences of nrLSU and rpb1 were deleted by using used as one of the key characters to distinguish porcini mush- Gblocks0.91b with the default setting. The procedures for rooms from other boletes. Sitta et al. (2007) and Sitta and constructing the phylogenetic trees were the same as those Davoli (2012), however, pointed out that the white mycelia used in Feng et al. (2012), except that the 3.0 version of would be the result of infection by fungi of the genus MrBayes was used in this study. Hypomyces based on their morphological observations. This strongly argues the validity to use the Bstuffed pores^ as one of the key diagnostic characteristics for porcini mushrooms. To verify the nature of the white mycelia, molecular techniques Results may provide useful evidence. The aim of this study are: 1) to characterize the all known Phylogenetic analyses species of porcini from China; 2) to document the new species of porcini in China using morphological features, molecular Fourteen, eleven and eighteen new sequences were generated evidence and ecological data; and 3) to illustrate the nature of for ITS, nrLSU and rpb1, respectively. Phylogenetic tree gen- the Bstuffed pores^ using molecular data. erated from ITS database (Fig. 1) clustered all newly added samples into the phylogenetic species recognized by Feng et al. (2012). Furthermore, phylogenetic analyses based on Material and methods the combined nrLSU-rpb1 dataset identified one additional new phylogenetic species (B. tylopilopsis in Fig. 2). These Morphological studies undecribed species were compared with known species using macro- and micro- morphological characteristics, geographi- The examined materials were collected from many subtropical cal distribution patterns and ecological preferences. and temperate parts of China and are deposited in the Cryptogamic Herbarium of Kunming Institute of Botany, Taxonomy Chinese Academy of Sciences (HKAS). Color codes indicat- ed in the descriptions are from Kornerup and Wanscher 1. Boletus bainiugan Dentinger, Index Fungorum 29: 1 (1981). The descriptions of species are in alphabetical order (2013) (Figs. 1, 2, 3 and 4) Fungal Diversity Table 1 Currently known Chinese porcini species identified Species Type location Corresponding species in by molecular data Feng et al. (2012) B. bainiugan Dentinger (including Yunnan, China B.sp.6–7 B. meiweiniuganjun Dentinger) B. botryoides B. Feng et al. Hunan, China B.sp.2 B. edulis Bull. Europe B. edulis B. fagacicola B. Feng et al. Hunan, China B.sp.9 B. griseiceps B. Feng et al. Fujian, China B. sp. HKAS 71346 B. monilifer B. Feng et al. Yunnan, China B.sp.1 B. orientialbus N.K. Zeng & Zhu L. Yang Fujian, China B.sp.14 B. subviolaceofuscus B. Feng et al. Yunnan, China B. violaceofuscus-2 B. reticuloceps (M. Zang et al.) Q.B.Wang & Sichuan, China B. reticuloceps Y. J. Yao B. shiyong Dentinger Yunnan, China B.sp.5 B. sinoedulis B. Feng et al. Sichuan, China B.sp.10 B. tylopilopsis B. Feng et al. Yunnan, China This study B. umbrinipileus B. Feng et al. Yunnan, China B.sp.3 B. violaceofuscus W.F. Chiu Yunnan, China B. violaceofuscus-1 B. viscidiceps B. Feng et al. Yunnan, China B. sp. 4-1 B. sp. 4-2 Yunnan, China B. sp. 4-2, immature B.sp.8 Hunan,China B. sp. 8, only a single collection available B. sp. 11 Shandong, China B. sp. 11, only a single collection available B. sp. 12 Jiangsu, China B. sp. 12, only a single collection available Synonym: Boletus meiweiniuganjun Dentinger, Index 5 μmlong.Hymenophoral trama boletoid and bilateral-diver- Fungorum 29: 1 (2013) gent, composed of hyphae 4–16 μmindiam.Pleurocystidia Basidioma medium-sized to large.