Taxonomy and phylogeny of the brown-rot fungi: Fomitopsis and its related genera

Taxonomic and phylogenetic studies on the brown-rot fungi, Fomitopsis and its related genera, are carried out. On the basis of morphological characters and phylogenetic evidence of DNA sequences of multiple loci including the internal transcribed spacer (ITS) regions, the large subunit nuclear ribosomal RNA gene (nLSU), the small subunit nuclear ribosomal RNA gene (nSSU), the small subunit mitochondrial rRNA gene sequences (mtSSU), the translation elongation factor 1-α gene (tef1) and the second subunit of RNA polymerase II (rpb2), six new genera, Fragifomes, Niveoporofomes, Piptoporellus, Rhodofomitopsis, Rubellofomes and Ungulidaedalea are established. Four new species, Buglossoporus eucalypticola, Daedalea allantoidea, Piptoporellus hainanensis and P. triqueter are descibed from China. Illustrated descriptions of the novel species are provided. Identification keys to Fomitopsis and its related genera, as well as keys to the species of each genus are provided.


Introduction
Fomitopsis P. Karst. was established by Karsten and typified by F. pinicola (Sw.) P. Karst. (Karsten 1881a). It is a wellknown cosmopolitan genus of polypores belonging to the antrodia clade (Ortiz-Santana et al. 2013) and includes brown rotting fungi with a dimitic hyphal system (Gilbertson and Ryvarden 1986a;Núñez and Ryvarden 2001;Ryvarden and Melo 2014). The genus is characterized by mostly perennial, sessile to effused-reflexed, tough to woody hard basidiocarps, white to tan or pinkish-colored pore surface with mostly small and regular pores, a dimitic hyphal system with clamped generative hyphae, hyaline, thin-walled, smooth, subglobose to cylindrical basidiospores which are negative in Melzer's reagent, and causing a brown rot (Ryvarden and Johansen 1980;Gilbertson and Ryvarden 1986a;Núñez and Ryvarden 2001;Ryvarden and Melo 2014;. More than 40 species have been accepted in the genus worldwide (Ryvarden and Johansen 1980;Gilbertson and Ryvarden 1986a;Rajchenberg 1995a;Núñez and Ryvarden 2001;Hattori 2003;Kim et al. 2007;Hattori and Sotome 2013;Ryvarden and Melo 2014;, of which 14 species are recorded from China (Dai 2012;Zhou and Wei 2012;Han et al. 2014;. Recently, several studies on the taxonomy and phylogeny of Fomitopsis were carried out, phylogenetic studies showed that Fomitopsis was embed in the antrodia clade and closely related to Daedalea Pers., Piptoporus P. Karst. and some species of Antrodia P. Karst. Kim et al. (2005) sequenced the nLSU regions from 10 species of Fomitopsis and 15 related species, their phylogenetic analysis indicated that Fomitopsis and Piptoporus were phylogenetically heterogenous and members in Fomitopsis were divided into three subgroups, among them, P. betulinus (Bull.) P. Karst. and D. quercina (L.) Pers. were included in Fomitopsis core group Mei-Ling Han and Yuan-Yuan Chen contributed equally to this work. and some Antrodia species were included in F. rosea (Alb. & Schwein.) P. Karst. and F. cajanderi (P. Karst.) Kotl. & Pouzar group. Ortiz-Santana et al. (2013) investigated the phylogenetic relationships among members of the antrodia clade by molecular data from ITS and nLSU regions, in their study, Fomitopsis were divided into five groups, their study supported the polyphyly of Fomitopsis and the transfer of the rosea clade (including F. rosea and F. cajanderi) into the genus Rhodofomes, and confirmed the placement of P. betulinus within Fomitopsis sensu stricto as reported in previous studies (Hibbett and Binder 2002;Binder et al. 2005;Garcia-Sandoval et al. 2011). However, no comprehensive investigation was carried out on a broad phylogenetic overview of Fomitopsis with enough samples from relevant genera, such as Daedalea and Piptoporus, and taxonomic delimitation of Fomitopsis has been controversial and remained insufficiently resolved (Kotlába and Pouzar 1998;Kim et al. 2005Kim et al. , 2007Ortiz-Santana et al. 2013). So, further phylogenetic analyses sampling more species are needed to clarify the relationships of Fomitopsis and its related genera.

Morphological studies
The studied specimens are deposited at the herbaria of the Institute of Microbiology, Beijing Forestry University, China (BJFC), the Institute of Applied Ecology, Chinese Academy of Sciences, China (IFP), the private herbarium of Dr. J. Vlasák of Czech Republic (JV), the Botanical Museum of the University of Oslo, Norway (O), Université Claude Bernard, France (LY), Botancal Museum of University of Helsink, Finland (H), Royal Botanic Gardens, Kew, UK (K), Botanic Garden Edinburgh, UK (E), Universidad de Buenos Aires, Argentina (BAFC) and the Pennsylvania State University, USA (PAC). The microscopic routines followed Zhao et al. (2013) and Li et al. (2014). Sections were studied at a magnification up to × 1000 using a Nikon E80i microscope and phase contrast illumination (Nikon, Tokyo, Japan). Drawings were made with the aid of a drawing tube. Microscopic features, measurements and drawings were made from slide preparations stained with Cotton Blue and Melzer's reagent. Spores were measured from sections cut from the tubes. In presenting the variation in the size of the spores, 5 % of measurements were given in parentheses. In the text the following abbreviations were used: IKI = Melzer's reagent, IKI + = amyloid, IKI-= non-dextrinoid and non-amyloid, KOH = 5 % potassium hydroxide, CB = Cotton Blue, CB + = cyanophilous, CB-= acyanophilous, L = mean spore length (arithmetic average of all spores), W = mean spore width (arithmetic average of all spores), Q = variation in the L/W ratios between the specimens studied, n = number of spores measured from given number of specimens. Special color terms followed Petersen (1996).

Phylogenetic analysis
A cetyl trimethylammonium bromide rapid plant genome extraction kit (Aidlab Biotechnologies Co., Ltd, Beijing) was used to extract total genomic DNA from dried specimens, and performed the polymerase chain reaction (PCR) according to the manufacturer's instructions with some modifications Zhao et al. 2015a). The ITS regions were amplified with primer pairs ITS5 and ITS4 (White et al. 1990). The nLSU regions were amplified with primer pairs LR0R and LR7 (http://www.biology.duke.edu/fungi/ mycolab/primers.htm). The nSSU regions were amplified with primer pairs NS1 and NS4 (White et al. 1990). The mtSSU regions were amplified with primer pairs MS1 and MS2 (White et al. 1990). Part of tef1 was amplified with primer pairs EF1-983 F and EF1-1567R (Rehner 2001). rpb2 was amplified with primer pairs bRPB2-6 F and bRPB2-7R (Matheny 2005). The PCR cycling schedule for ITS, mtSSU, and tef1 included an initial denaturation at 95°C for 3 min, followed by 35 cycles at 94°C for 40 s, 54°C for ITS and mtSSU, 54-59°C for tef1 for 45 s, 72°C for 1 min, and a final extension at 72°C for 10 min. The PCR cycling schedule for nLSU and nSSU included an initial denaturation at 94°C for 1 min, followed by 35 cycles at 94°C for 30 s, 50°C for nLSU and 53°C for nSSU for 1 min, 72°C for 1. 5 min, and a final extension at 72°C for 10 min. The PCR cycling schedule for rpb2 followed Kim et al. (2007) with slight modifications: initial denaturation at 95°C for 10 min, followed by 39 cycles at 94°C for 1 min, 56°C for 1 min and 72°C for 1 min + 3 s/cycle, and a final extension at 72°C for 10 min. The PCR products were purified and sequenced at Beijing Genomics Institute (China), with the same primers. All newly generated sequences were deposited at GenBank (Table 1).
Additional sequences were downloaded from GenBank ( Table 1). All sequences were aligned using ClustalX (Thompson et al. 1997) and manually adjusted in BioEdit (Hall 1999). The missing sequences were coded as BN^. Ambiguous nucleotides were coded as BN^. The final concatenated sequence alignment was deposited in TreeBase (http://purl.org/phylo/treebase; submission ID 18345).
Most parsimonious phylogenies were inferred from the combined 3-gene dataset (ITS + nLSU + rpb2) and 6-gene dataset (ITS + nLSU + nSSU + mtSSU + tef1 + rpb2), and their congruences were evaluated with the incongruence length difference (ILD) test (Farris et al. 1994) implemented in PAUP* 4.0b10 (Swofford 2002), under heuristic search and 1000 homogeneity replicates. Phylogenetic analysis approaches followed Zhao et al. (2014Zhao et al. ( , 2015b. Sequences of Trametes suaveolens (L.) Fr. and Coriolopsis polyzona (Pers.) Ryvarden                obtained from GenBank were used as outgroups to root trees following Binder et al. (2013). Maximum parsimony analysis was applied to the combined multiple genes datasets and the tree construction procedure was performed in PAUP* version 4.0b10. 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 1000 random sequence additions. Max-trees were set to 5000, branches of zero length were collapsed and all parsimonious trees were saved. Clade robustness was assessed using a bootstrap (BT) analysis with 1000 replicates (Felsenstein 1985). Descriptive tree statistics tree length (TL), consistency index (CI), retention index (RI), rescaled consistency index (RC), and homoplasy index (HI) were calculated for each Most Parsimonious Tree (MPT) generated. RAxML v.7.2.8 was used to construct a maximum likelihood (ML) tree with GTR + G + I model of site substitution including estimation of Gamma-distributed rate heterogeneity and a proportion of invariant sites (Stamatakis 2006). The branch support was evaluated with bootstrapping method of 1000 replicates (Hillis and Bull 1993). Phylogenetic trees were visualized using Treeview (Page 1996).

Rhodofomes cajanderi
MrModeltest 2. 3 (Posada and Crandall 1998;Nylander 2004) was used to determine the best-fit evolution model for the combined multi-gene dataset for Bayesian inference (BI). Bayesian inference was calculated with MrBayes 3.1.2 with a general time reversible (GTR) model of DNA substitution and a gamma distribution rate variation across sites (Ronquist and Huelsenbeck 2003). Four Markov chains were run for 2 runs from random starting trees for 8.2 million generations (ITS + nLSU + rpb2), for 10 million generations (ITS + nLSU + nSSU + mtSSU + tef1 + rpb2) and trees were sampled every 100 generations. The first one-fourth generations were discarded as burn-in. A majority rule consensus tree of all remaining trees was calculated. Branches that received bootstrap support for maximum parsimony (MP), maximum likelihood (BS) and Bayesian posterior probabilities (BPP) greater than or equal to 75 % (MP and BS) and 0.95 (BPP) were considered as significantly supported, respectively.
The combined six gene (ITS, nLSU, nSSU, mtSSU, tef1, rpb2) sequences dataset had an aligned length of 4789 characters, of which 2874 characters were constant, 257 were variable and parsimony-uninformative, and 1658 were parsimony-informative. Maximum parsimony analysis yielded 14 equally parsimonious trees (TL = 11,070, CI = 0.305, RI = 0.740, RC = 0.225, HI = 0.695). Best model for the combined ITS + nLSU + nSSU + mtSSU + tef1 + rpb2 sequence dataset estimated and applied in the Bayesian analysis was: GTR + I + G with equal frequency of nucleotides. ML analysis resulted in a similar topology as MP and Bayesian analysis, and the ML topology was shown in Fig. 2.
Remarks. Buglossoporus was proposed by Kotlába and Pouzar (1966), it was treated as a synonym of Piptoporus (Ryvarden 1991;Hattori 2000). In our study, Piptoporus is treated as a synonym of Fomitopsis, and Buglossoporus is an independant genus. Phylogenetically, B. quercinus and B. eucalypticola formed a well-supported lineage (Figs. 1 and 2), which was distant from Fomitopsis betulina (= P. betulinus), and closely related to Neolentiporus Rajchenb. However, Neolentiporus has a dimitic hyphal system in trama with irregularly thick-walled generative hyphae and metachromatic skeletal hyphae (Rajchenberg 1995b Diagnosis. Differing from other Buglossoporus species by its clay-pink to cinnamon pileal surface usually with a pellicle, pinkish buff or clay-buff to dark brown pore surface, and cylindrical to fusiform basidiospores (4.5-6.8 × 2-2.8 μm), and exclusively growing on Eucalyptus.
Fruiting body. Basidiocarps annual, solitary, pileate, usually with a central to lateral stipe. Pileus applanate to slightly convex, flabelliform or semicircular with a series of inwardpointing notches, projecting up to 10 cm, 6.5 cm wide, 7 mm thick at base. Pileal surface peach to brownish orange when fresh, becoming clay-pink to cinnamon when dry, usually with a pellicle, glabrous, azonate, rugose; margin flesh-pink when fresh, becoming cinnamon to dark brown when dry, acute. Pore surface white when fresh, turning pinkish buff or claybuff to dark brown upon drying, sterile margin indistinct; pores round to angular, 2-6 per mm; dissepiments thin, entire. Context distinctly thicker than tubes, cream to pinkish buff, corky, up to 6.5 mm. Tubes concolourous with pore surface, fragile, very short, up to 0.5 mm long. Stipe glabrous, often with a pellicle, cream to dark brown, up to 16 cm long and 2.7 cm thick, fleshy and flexible when fresh, fragile and light in weight when dry.
Buglossoporus malesianus Corner, Beihefte zur Nova Hedwigia 78: 165, 1984. (Fig. 3b Remarks. Hattori (2000) studied the type specimens of Buglossoporus malesianus, B. matangensis and B. rufescens described by Corner and considered the type specimens of B. matangensis and B. rufescens represent B. malesianus, and the former two were treated as synonyms of Piptoporus malesianus. We also examined the types of these three species, and comfirmed the identifications of B. matangensis and B. rufescens as Hattori (2000). We tried to extract DNA from materials of Buglossoporus malesianus, B. matangensis and B. rufescens, but did not success. However, Piptoporus malesianus has annual, pileate, substipitate basidiocarps, a dimitic hyphal system in context and monomitic in trama, and cylindrical to fusiform basidiospores, which are similar to Buglossoporus quercinus. Therefore, we keep this species in Buglossoporus although without phylogenetic support. Hattori (2000) Kotlába and Pouzar (1966) established Buglossoporus with B. quercinus as the type species. However, Buglossoporus was treated as a synonym of Piptoporus (Ryvarden 1991;Hattori 2000), and B. quercinus was accepted as P. quercinus. In our study, Piptoporus is treated as a synonym of Fomitopsis, B. quercinus was formed in the Buglossoporus lineage ( Figs. 1 and 2), For a detailed description, see Ryvarden and Melo (2014)  Basidiocarps mostly perennial, effuse-reflexed or most often pileate, broadly sessile, coriaceous to corky or hard corky when dry. Pileal surface smooth to velutinate, often concentrically zonate and sulcate. Hymenophore surface ochraceous to dark-brown or grey, hymenophores irregular, labyrinthine/daedaleoid to lamellate, hydnoid or poroid. Context more or less brownish, sometimes with a cuticle or crust at the upper surface. Tubes coriaceous to corky or hard corky. Hyphal system dimitic with more or less branched skeletal hyphae, generative hyphae with clamp connections, skeletal hyphae colorless to pale yellow or pale brown, thick-walled, IKI-, CB-. Catahymenium present or not. Cystidia occasionally present, thin-walled cystidioles usually present. Basidiospores cylindrical to ellipsoid, hyaline, thinwalled, smooth, IKI-, CB-. Usually grows on angiosperm wood and causes a brown rot.
Remarks. Daedalea, typified by D. quercina, was originally established by Persoon (1801), then it was treated as a Fig. 1 Maximum likelihood tree illustrating the phylogeny of Fomitopsis and its related genera in the antrodia clade based on the combined sequences dataset of ITS + nLSU + rpb2. Branches are labeled with maximum likelihood bootstrap higher than 50 %, parsimony bootstrap proportions higher than 50 % and Bayesian posterior probabilities more than 0.95 collective genus for all species with a daedaleoid to labyrinthine hymenophore (Fries 1821). During the last century, more microscopic and chemical characters were applied in taxonomy, many Daedalea species have been transferred to other genera (Singer 1944;Donk 1966;Ryvarden 1984).
Diagnosis. Differing from other Daedalea species by its annual, pileate basidiocarps, pinkish buff to cinnamon buff or pale mouse-grey pileal surface, light clay-buff to fawn pore surface with large pores (1-3 per mm), a catahymenium formed by skeletal hyphae and allantoid basidiospores.
Fruiting body. Basidiocarps annual, pileate, imbricate, corky, without odor or taste when fresh, hard corky and light in weight upon drying. Pileus conchate or triquetrous, projecting up to 4.4 cm, 2.5 cm wide, 9 mm thick at base. Pileal surface pinkish buff to cinnamon buff or pale mousegrey, glabrous to tuberculate, slightly zonate and radially streaked; margin pinkish buff to clay-buff, acute. Pore surface light clay-buff to fawn; pores round to angular or elongated, 1-3 per mm; dissepiments thin, entire. Context cream, hard corky, up to 5 mm thick. Tubes pinkish buff to salmon, corky, up to 4 mm long.
Remarks. Daedalea dickinsii produce pileate basidiocarps, similar colored pileal surface and pore surface, similar sized pores and basidiospores with D. allantoidea, but D. dickinsii differs by a perennial growth habit, broadly concentrically sulcate pileal surface and cylindrical basidiospores, and it grows mostly on wood of Quercus in temperate areas (Núñez and Ryvarden 2001).
Type species. Fomitopsis pinicola (Sw.) P. Karst. Basidiocarps annual to perennial, mostly sessile, occasionally effused-reflexed or substipitate, soft, corky, tough to woody. Pileal surface white to greyish, yellowish or brown, velutinate to glabrous, concentrically sulcate or not. Pore Section of hymenium; e. Hyphae from trama; f. Hyphae from context. Bars: a = 5 μm; bf = 10 μm surface white, cream to greyish or tan; pores mostly small, round to angular. Context white to greyish or straw, fibrous to corky, sometimes with a thin crust or cuticle at the upper surface. Hyphal system mostly dimitic with more or less branched skeletal hyphae, generative hyphae with clamp connections, skeletal hyphae colorless to pale grey, thick-walled with a narrow lumen to subsolid, IKI-, CB-. Cystidia occasionally present, thin-walled cystidioles usually present. Basidiospores cylindrical to ellipsoid, hyaline, thin-walled, smooth, IKI-, CB-. Grows on angiosperm wood or gymnosperm wood, and causes a brown rot.
Remarks. In our six-loci phylogenetic study (Fig. 2 Four samples of Piptoporus betulinus from China and Finland formed a highly supported subgroup (100 % BS, 100 % MP, 1.00 BPP), and then grouped with Fomitopsis pinicola. The above species subsequently clustered together forming a monophyletic lineage, namely Fomitopsis s. s., with high support (91 % BS, 96 % MP, 1.00 BPP). These ten species share similar morphological characters and form the core group of Fomitopsis. We did not get any sample of F. ostreiformis (Berk.) T. Hatt., but F. ostreiformis has annual, sessile or effuse-reflexed basidiocarps, greyish pileal surface, white or greyish white pore surface, white to brownish and fibrouscorky context, a dimitic hyphal system with more or less branched skeletal hyphae, and cylindrical basidiospores (De 1981;Hattori 2003). In addition, F. ostreiformis clustered with F. durescens, F. iberica, F. nivosa, F. hemitephra and F. palustris in Fomitopsis s. s. based on phylogeny of ITS + nLSU + rpb2 sequences (Fig. 1) generative hyphae and more or less branched skeletal hyphae, cylindric to slightly allantoid, hyaline and smooth basidiospores which are negative in Melzer's reagent (Gilbertson and Ryvarden 1986b;Ryvarden and Melo 2014). In our study, it is closely related to Fomitopsis pinicola and grouped into the Fomitopsis s. s. clade. Therefore, Piptoporus betulinus is transferred to Fomitopsis. For a detailed description of P. betulinus, see Ryvarden and Melo (2014). Etymology. Fragifomes (Lat.): refers to the fragile and layered basidiocarps.
Diagnosis. Differing from Fomitopsis s. s. by its soft corky to fragile basidiocarps.
Remarks. In our study, two samples of F. niveomarginata formed a single lineage (Figs. 1 and 2), which was distant from Fomitopsis s. s. Morphologically, Fragifomes differs from Fomitopsis s. s. by its soft corky to fragile basidiocarps. For a detailed description of Fomitopsis niveomarginata, see Zhou and Wei (2012). Etymology. Niveoporofomes (Lat.): refers to the white pore surface and layered basidiocarps.
Diagnosis. Differing from Fomitopsis s. s. by its annual basidiocarps, ovoid to broadly ellipsoid basidiospores.
Remarks. Three samples of Niveoporofomes spraguei from France, USA and China formed a single lineage with a high support ( Figs. 1 and 2), and was distinct from Fomitopsis s. s. Morphologically, Niveoporofomes differs from Fomitopsis s. s. in its annual growth habit and ovoid to broadly ellipsoid basidiospores.
Remarks. In our study, Piptoporus soloniensis (Dubois) Pilát was clustered with two new species from China and formed a highly supported lineage (Figs. 1 and 2) in the antrodia clade. Piptoporus is treated as a synonym of Fomitopsis, therefore, Piptoporellus gen. nov. is proposed to accommodate P. soloniensis and the two new species from China, described below.
Diagnosis. Differing from other Piptoporellus species by its cream to buff pileal surface and lacking cystidia or other sterile hymenial elements.
Fruiting body. Basidiocarps annual, pileate, with a lateral base, solitary, corky, without odor or taste when fresh, hard corky and light in weight upon drying. Pileus dimidiate, flabelliform or semicircular, applanate to convex, projecting up to 9 cm, 7.8 cm wide, 1.7 cm thick at base. Pileal surface cream to buff, glabrous, azonate, radially streaked; margin cream, acute, incurved. Pore surface cream to buff or yellow, shinning; sterile margin indistinct; pores round to angular or irregular, 4-5 per mm; dissepiments thin, entire. Context cream, hard corky, up to 1.1 cm thick. Tubes paler than or concolourous with pore surface, fragile, up to 6 mm long.
Fruiting body. Basidiocarps annual, pileate, sessile, solitary, corky, without odor or taste when fresh, fragile and light in weight upon drying. Pileus triquetrous, projecting up to 3.5 cm, 2.3 cm wide, 1.5 cm thick at base. Pileal surface Fig. 6 Microscopic structures of Piptoporellus hainanensis (drawn from the holotype). a. Basidiospores; b. Basidia and basidioles; c. Hyphae from trama; d. Hyphae from context. Bars: a = 5 μm; b-d = 10 μm buff-yellow or salmon to brownish orange, glabrous, azonate; margin salmon to brownish orange, acute. Pore surface cream or buff to light brown; sterile margin distinct, up to 7 mm wide, cream to brownish orange; pores round to angular, 3-4 per mm; dissepiments thin, entire. Context distinctly thicker than tubes, cream to pinkish buff, corky, up to 1.45 cm thick. Tubes concolourous with pore surface, fragile, up to 0.5 mm long.
Remarks. According to Nobles (1971), Fomitopsis cajanderi was closely related to the species of Daedalea s. s. sharing several important morphological characters. After Nobles' view, Donk (1974) suggested the removal of F. cajanderi from Fomitopsis and its inclusion into Daedalea or any other related group. Subsequently, a new genus, Rhodofomes Kotl. & Pouzar, was proposed by Pouzar (1990, 1998) with R. rosea (Alb. & Schwein.) Kotl. & Pouzar as type species. But the concept of the genus has been controversial and remained insufficiently resolved (Kim eat al. 2005(Kim eat al. , 2007Ortiz-Santana et al. 2013;Han et al. 2014;Ryvarden and Melo 2014). In our study, species of Rhodofomes formed a well-supported lineage ( Figs. 1 and 2), which was distant from Fomitopsis s. s., and closely related to species of Antrodia serialis (Fr.) Donk group Rubellofomes and Niveoporofomes. However, Antrodia serialis group has mostly resupinate to effused-reflexed basidiocarps, white, cream to wood-colored pore surface and a dimitic hyphal system with mostly unbranched skeletal hyphae (Núñez and Ryvarden 2001;Ryvarden and Melo 2014); Rubellofomes has white to cream or purple pink pore surface when fresh, which becoming straw yellow to cinnamon brown when dry; Niveoporofomes has soft coky to fragile basidiocarps, white pore surface when fresh, which becoming yellowish brown upon drying and cream context. Thus, Rhodofomes is recognized as an independent genus, and five species including four new combinations are proposed in the genus.
For a detailed description of Fomitopsis cajanderi, see Carranza-Morse and  and Ryvarden and Melo (2014).
Specimens examined. CHINA. Heilongjiang Prov., Yichun, Fenglin Nature Reserve, on fallen trunk of Picea, 2 s. s. by annual, pileate, corky or soft-fibrous basidiocarps, cream to pinkish buff context without a pellicle or crust at the upper surfac, moreover, its skeletal hyphae are thickwalled with a distinctly wide lumen and mostly dissolved in KOH. Piptoporus roseovinaceus Choeyklin, T. Hatt. & E.B.G. Jones, living on dead bamboo culms and collected from Thailand, has annual, pileate, sessile basidiocarps, light orange to greyish orange pileus surface, sordid white to greyish orange pore surface, white to pale orange context, a dimitic hyphal system in context and monomitic in trama with clamped generative hyphae, and thin-walled, short ellipsoid to ellipsoid basidiospores (Choeyklin et al. 2009). These characters fit well with Buglossoporus. But unfortunately, no sequences data were available for P. roseovinaceus, therefore, the taxonomic position of P. roseovinaceus remained uncertain. Kotlába and Pouzar (1990) suggested a narrow concept of Fomitopsis and proposed a new genus, Pilatoporus Kotl. & Pouzar, and transferred F. palustris into it as a type species based on the presence of pseudoskeletal hyphae with conspicuous clamp connections. However, the main morphological characters of F. palustris fit well with Fomitopsis s. s. Moreover, in our phylogenetic tree ( Figs. 1 and 2), F. palustris was strongly clustered within Fomitopsis s. s. Therefore, we prefer to keep F. palustris in Fomitopsis, which coincides with Kim et al. (2005Kim et al. ( , 2007.
23 species of Fomitopsis s. l. were divided into seven genera based on morphological characters and phylogenetic evidence in our study. However, another idea to use of a single generic name (the Bnew^Fomitopsis) for Fomitopsis s. l. and related genera altogether is not accepted. Because in our phylogenetic analysis ( Figs. 1 and 2), Fomitopsis s. l. together with its related genera are polyphyletic, which was also confirmed by Kim et al. (2005Kim et al. ( , 2007 and Ortiz-Santana et al. (2013). If a single generic name (Fomitopsis) for Fomitopsis s. l., Daedalea, Piptoporus and Antrodia was used, the morphological concept of the BnewF omitopsis needs to be expanded to allow species with irregular and daedaleoid hymenophores (Daedalea, Rhodofomitopsis), a catahymenium formed by skeletal hyphae and / or cystidia (Daedalea), mostly respinate to effused-reflexed basidiocarps (Antrodia) and substipitate or stipitate and fragile basidiocarps (Piptoporus); and many new combinations should be proposed. This solution is straightforward, but it is not totally free of problems. Species of Antrodia embed into the antrodia clade forming at least three clades, and Buglossoporus and Neolentiporus were also included in this Bnew^Fomitopsis, however, Buglossoporus has a monomitic hyphal system in trama and Neolentiporus has irregularly thick-walled generative hyphae and metachromatic, unbranched skeletal hyphae, these characters were quite different from this Bnew^Fomitopsis and cannot be suitable for the inclusion of Buglossoporus and Neolentiporus in this BnewF omitopsis. Therefore, we prefer to set up new genera for different monophyletic clades.
In summary, we performed a comprehensive study on Fomitopsis s. l. and its related genera. On the basis of morphological characters and phylogenetic evidence, six new genera, Fragifomes, Niveoporofomes, Rhodofomitopsis, Rubellofomes, Ungulidaedalea and Piptoporellus are proposed, four new species, Buglossoporus eucalypticola, Daedalea allantoidea, Piptoporellus hainanensis and P. triqueter are described, and 16 new combinations are proposed. However, the DNA sequences data of ca. 20 species of