Mycologia, 106(5), 2014, pp. 1039–1050. DOI: 10.3852/13-402 # 2014 by The Mycological Society of America, Lawrence, KS 66044-8897

Morphological and molecular evidence for two new species of Laetiporus (Basidiomycota, Polyporales) from southwestern China Jie Song Yuanyuan Chen Baokai Cui1

and Banik 2001, Nu´n ˜ ez and Ryvarden 2001, Tomsˇovsky´ and Jankovsky´ 2008, Ota et al. 2009, Banik et al. 2012), of which three species have been recorded from China (Dai 2012). In North America morphological characters, mating compatibility, RFLPs of the nu-rDNA, ecological data and distribution ranges were used to describe and define species of Laetiporus (Banik et al. 1998, Banik and Burdsall 1999, Banik and Burdsall 2000). Burdsall and Banik (2001) suggested that Laetiporus sulphureus in North America encompassed six morphologically and ecologically distinct species: L. cincinnatus (Morgan) Burds., Banik & T.J. Volk, L. conifericola Burds. & Banik, L. gilbertsonii Burds., L. huroniensis Burds. & Banik, L. persicinus (Berk. & M.A. Curtis) Gilb. and L. sulphureus sensu stricto. These species were grouped into six well supported lineages in the multilocus phylogeny, and except L. persicinus these lineages were considered members of the core clade of Laetiporus (Lindner and Banik 2008). Tomsˇovsky´ and Jankovsky´ (2008) used ITS and nLSU-DNA data to ascertain the taxonomic position ˇ erny´ ex Tomsˇovsky´ & of Laetiporus montanus C Jankovsky´ in Europe. Laetiporus montanus differs from L. huroniensis and L. conifericola in DNA sequences of sampled genetic regions and in basidiospore dimensions, thus L. montanus was recognized as a distinct species restricted to coniferous trees at high altitude. Laetiporus sulphureus var. miniatus (Jungh.) Imazeki, might represent L. montanus in Japan (Tomsˇovsky´ and Jankovsky´ 2008). In eastern Asia, based on ITS and EF1a gene phylogenies, incompatibility and morphological characters, three Laetiporus species have been recognized: L. cremeiporus Y. Ota & T. Hatt., L. montanus and L. versisporus (Lloyd) Imazeki (Ota and Hattori 2008, Ota et al. 2009, Banik et al. 2010). Based on morphological features and ITS phylogeny, Banik et al. (2012) described a new species: Laetiporus caribensis Banik & D.L. Lindner from the Caribbean basin, and 14 Laetiporus lineages had been identified, including four undescribed species. Taxonomy and phylogeny study focusing on brownrot fungi in China have been carried out recently, and some new species have been described (Dai and Niemela¨ 2002, Wang et al. 2004, Cui et al. 2011, Cui and Li 2012, Zhou and Wei 2012, Cui 2013, Cui and Dai 2013, Li and Cui 2013a, Li et al. 2013). In a

Institute of Microbiology, Beijing Forestry University, Beijing 100083, China

Honggao Liu College of Agronomy and Biotechnology, Yunnan Agricultural University, Kunming 650201, China

Yuanzhong Wang Institute of Medicinal Plants, Yunnan Academy of Agricultural Sciences, Kunming 650223, China

Abstract: Two Laetiporus species, L. ailaoshanensis and L. zonatus spp. nov., are described from southwestern China based on morphological and molecular characters. Laetiporus ailaoshanensis is characterized by orange-yellow to reddish orange pileal surface and cream to buff pores when fresh, azonate to faintly zonate pileus, ovoid to ellipsoid basidiospores (5.0–6.2 3 4.0–5.0 mm), and it has been observed only on Lithocarpus. Laetiporus zonatus is characterized by white to cream pileal surface with buff to clay-buff base when fresh, concentrically zonate basidiocarps, ellipsoid to pyriform or dropshaped basidiospores (5.8–7.2 3 4.3–5.5 mm), and it has been found only on Quercus. The phylogenetic relationships of all recognized Laetiporus species were inferred from a combined dataset of ITS and nLSUrDNA sequences, and L. ailaoshanensis and L. zonatus represent two new lineages in this group. Key words: brown-rot fungi, Fomitopsidaceae, phylogeny, polypore, taxonomy INTRODUCTION Laetiporus Murrill, typified by L. sulphureus (Bull.) Murrill, is an important group of wood-inhabiting fungi, which cause a cubical brown rot of hardwoods or conifers, and some Laetiporus species also are known as forest pathogens (Murrill 1904, Dai et al. 2007) and as mushrooms with some medicinal values (Dai et al. 2009). Laetiporus has a worldwide distribution from boreal to tropical zones. Fifteen species have been accepted in the genus worldwide (Gilbertson and Ryvarden 1986, Ryvarden and Gilbertson 1993, Burdsall Submitted 27 Dec 2013; accepted for publication 6 Apr 2014. 1 Corresponding author. E-mail: [email protected]

1039

1040 TABLE I.

MYCOLOGIA Species, specimens and GenBank accession number of sequences used in this study GenBank accession Nos.

Species

Collection No.

Sample No.

Host

Locality

ITS

nLSU

Laetiporus ailaoshanensis L. ailaoshanensis L. ailaoshanensis L. ailaoshanensis L. caribensis L. caribensis L. caribensis L. cincinnatus L. cincinnatus L. cincinnatus L. conifericola L. conifericola L. conifericola L. cremeiporus

Yuan 3302

IFP 013596

Lithocarpus

Yunnan, China

KF951286a

KF951316a

HKAS 52508 HKAS 53092 Dai 13256 PR914 PR6583 PR6521 DA-37 Dai 12811 JV 0709/168-J CA-8 JAM-1 JV 0709/81-J Cui 10586

HKAS 52508 HKAS 53092 BJFC 016516

BJFC 015824 BJFC 011481

Lithocarpus Lithocarpus Lithocarpus angiosperm Guareaguidonia angiosperm Quercus Quercus Quercus Sequoia Tsuga Picea Lagerstroemia

Yunnan, China Yunnan, China Yunnan, China Puerto Rico Puerto Rico Puerto Rico Wisconsin, USA Connecticut, USA Virginia, USA California, USA Alaska, USA California, USA Sichuan, China

KF951319a KF951318a KF951317a — — — EU402521 KF951304a KF951305a EU402523 EU402524 KF951327a KF951297a

L. cremeiporus L. cremeiporus

Cui 10991 Cui 10988

BJFC 013913 BJFC 013909

Quercus Quercus

Hubei, China Hubei, China

L. L. L. L. L. L. L. L. L.

Dai 10045 Dai 10107 CA-13 TJV2000-101 JV 1109/31 HMC-3 MI-14 Cui 10011 Cui 10015

BJFC 015739 BJFC 015740

BJFC 016042

BJFC 010904 BJFC 010908

Quercus Quercus Quercus Quercus Quercus Tsuga Tsuga gymnosperm gymnosperm

Jilin, China Jilin, China California, USA Florida, USA Texas, USA Michigan, USA Michigan, USA Jilin, China Jilin, China

Cui 1710 Dai 11203 JV 0407/27

IFP 002983 BJFC 015752 BJFC 016517

Larix Larix Abies

Inner Mongolia, China Inner Mongolia, China Czech Republic

Eucalyptus Acacia Unknown Unknown Eucalyptus Eucalyptus Quercus Picea Sorbus Fraxinus Quercus angiosperm Acer Salix angiosperm angiosperm Robinia angiosperm angiosperm Castanea

Hawaii, USA Hawaii, USA Costa Rica Argentina South Africa South Africa Czech Republic Czech Republic Czech Republic Wisconsin, USA Massachusetts, USA Wisconsin, USA Connecticut, USA Wisconsin, USA Hainan, China Jiangxi, China Beijing, China Yunnan, China Fujian, China Yunnan, China

KF951287a KF951288a KF951289a JN684762 JN684766 JN684771 EU402557 KF951291a KF951290a EU402575 EU402577 KF951292a KF951277a KJ480734a KF951279a KF951278a KJ480741a KF951280a KF951281a EU402549 EU402553 KF951293a EU402571 EU402573 KF951274a KF951273a KJ480732a KF951275a KF951276a KF951294a KJ480747a EU402545 EU402546 JN684764 JN684765 EU840662 EU840663 KF951295a KF951296a EU840565 EU402567 EU402568 EU402561 EU402565 EU402566 KF951268a KF951269a KF951270a KF951266a KF951267a KF951271a KJ480736a

cremeiporus cremeiporus gilbertsonii gilbertsonii gilbertsonii huroniensis huroniensis montanus montanus

L. montanus L. montanus L. montanus L. L. L. L. L. L. L. L. L. L. L. L. L. L. L. L. L. L. L. L.

sp. sp. sp. sp. sp. sp. sulphureusb sulphureusb sulphureusb sulphureusc sulphureusc sulphureusd sulphureusd sulphureusd versisporus versisporus versisporus versisporus versisporus versisporus

EUC-1 KOA-1 Munez 207 Robledo 1122 RV4A RV5A Dai 12154 JV 1106/15 L13-706696 TJV99-150 MAS-2 GR-12 CT-1 DA-41 Cui 5488 Cui 7882 Cui 9154 Dai 13160 Dai 7268 Dai 13052

BJFC 013116 BJFC 016049

BJFC 012671 BJFC 015826

BJFC BJFC BJFC BJFC BJFC BJFC

003529 006371 008092 013371 001160 013275

KF951298a KF951299a KF951300a KF951301a EU402527 EU402528 KF951306a EU402540 EU402539 KF951315a KF951311a KF951314a KF951313a KF951312a EU402541 EU402542 — — — — KF951302a KF951303a EU884420 EU402530 EU402531 EU402534 EU402532 EU402533 KF951321a KF951323a KF951322a KF951320a KF951326a KF951324a

SONG ET AL.: TWO NEW SPECIES OF LAETIPORUS TABLE I.

1041

Continued GenBank accession Nos.

Species

Collection No.

Sample No.

Host

ITS

Locality

L. versisporus

Cui 10992

BJFC 013914

Quercus

Yunnan, China

L. zonatus

Cui 10403

BJFC 011298

Quercus

Yunnan, China

L. zonatus L. zonatus L. zonatus Wolfiporia dilatohypha

Cui 10404 HKAS 54701 HKAS 71806 FP72162

BJFC 011299 HKAS 54701 HKAS 71806

Quercus angiosperm angiosperm Quercus

Yunnan, China Yunnan, China Yunnan, China Tennessee, USA

nLSU a

KF951272 KJ480746a KF951282a KJ480733a KF951283a KF951285a KF951284a EU402556

KF951325a KF951307a KF951308a KF951309a KF951310a EU402517

a

Newly generated sequences for this study. Species distributed in Europe only. c Species distributed in North America; white-pored. d Species distributed in North America, South America and Europe; yellow-pored. b

continuation of these surveys two Laetiporus species were found to be undescribed. Phylogenetic analysis of ITS and nLSU regions verified their affinity within the genus. MATERIALS AND METHODS Morphological studies.—The studied specimens were deposited at the herbaria of the Institute of Microbiology, Beijing Forestry University (BJFC), the Institute of Applied Ecology, Chinese Academy of Sciences (IFP) and the Kunming Institute of Botany, Chinese Academy of Sciences (HKAS). Microscopic examinations followed Zhao and Cui (2013). Sections were studied at a magnification up to 10003 with Nikon E 80i 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 excluded from each end of the range and are given in parentheses. In the text the following abbreviations are used: IKI 5 Melzer’s reagent, IKI2 5 negative in Melzer’s reagent, KOH 5 5% potassium hydroxide, CB 5 cotton blue, CB2 5 acyanophilous, L 5 mean spore length (arithmetic average of all spores), W 5 mean spore width (arithmetic average of all spores), R 5 mean of L/W ratios, Q 5 variation in the L/W ratios between the specimens studied, n 5 number of spores measured from given number of specimens. Color terms followed Petersen (1996). DNA extraction and sequencing.—The fungal specimens used in this study are listed (TABLE I). A Phire plant direct PCR kit (Thermo, Vilnius, Lithuania) procedure was used to extract total genomic DNA from the fruit bodies and for polymerase chain reaction (PCR). The DNA was amplified with primers ITS4 and ITS5 for ITS (White et al. 1990) and LR0R and LR7 for nLSU (http://www.biology.duke.edu/ fungi/mycolab/primers.htm). DNA sequencing was per-

formed at Beijing Genomics Institute, China. Some of the sequences possessed ambiguous base calls, possibly as a result of heterozygosity; all PCR products exhibiting this phenomenon were cloned into a pMDTM 19-T Vector (Takara, Tokoyo, Japan) and the DNA of at least eight of the resulting transformed clones from each sample were amplified and sequenced (Ota et al. 2009). All newly generated sequences were submitted to GenBank and are listed (TABLE I). Phylogenetic analysis.—Sequences generated for this study were aligned with additional sequences downloaded from GenBank (TABLE I) with BioEdit (Hall 1999) and Clustal X (Thompson et al. 1997). Before phylogenetic analysis, ambiguous sequences were trimmed and gaps were added to optimize local alignments. Sequence alignment was deposited at TreeBASE (http://purl.org/phylo/treebase; submission ID 15441). Polymorphic sequences derived from one isolate or one specimen were labeled in the analyses. The best-fit model of nucleotide evolution was selected by hierarchical likelihood ratio tests (hLRT) in MrModeltest 2.3 (Posada and Crandall 1998, Nylander 2004). Phylogenetic analysis used in this study followed Li and Cui (2013b). Maximum parsimony (MP) analysis and Bayesian inference were applied to the combined dataset of ITS and nLSU sequences. Wolfiporia dilatohypha Ryvarden & Gilb. was used as outgroup to root trees (Lindner and Banik 2008, Ota et al. 2009, Banik et al. 2010, Banik et al. 2012). Four Markov chains were run twice from random starting trees for 2.3 million generations, and trees were sampled every 100 generations. The first 25% trees sampled was discarded as burn-in. A majority rule consensus tree of all remaining trees was calculated. Maximum parsimony (MP) bootstrap proportions higher than 75% and Bayesian posterior probabilities (BPP) more than 0.95 were considered as significant support.

RESULTS Molecular phylogeny.—The combined ITS+nLSU dataset included sequences from 53 fungal specimens or

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MYCOLOGIA

isolates representing 16 taxa. The dataset had an aligned length of 1514 characters including gaps, of which 1305 are constant, 55 are variable and parsimony uninformative and 154 are parsimony informative. Maximum parsimony analysis yielded 10 equally parsimonious tree (length 5 359, CI 5 0.694, RI 5 0.894, RC 5 0.620, HI 5 0.306), and we illustrated (FIG. 1) a strict consensus tree of these trees. The best model for the combined ITS+nLSU partition was a GTR+I+G model. The Bayesian inference analysis resulted in a similar topology. In the phylogenetic inferences, our collections from southwestern China are distributed into two new lineages; collections from subtropical areas on Lithocarpus formed one lineage (MP 5 91%, BPP 5 0.96), while collections from high mountains of temperate areas formed the second lineage (MP 5 99%, BPP 5 1.00). They are considered distinct phylogenetic species. TAXONOMY Laetiporus ailaoshanensis B.K. Cui & J. Song, sp. nov. FIGS. 2a, b, 3 MycoBank MB808203 Laetiporus ailaoshanensis is characterized by orange-yellow to reddish orange pileal surface and cream to buff pores when fresh, azonate to indistinctly zonate pileus, ovoid to ellipsoid basidiospores (5.0– 6.2 3 4.0–5.0 mm). Etymology: ailaoshanensis (Lat.), referring to the locality (Ailaoshan) of the type specimens.

Holotype: CHINA, YUNNAN PROVINCE: Jingdong County, Ailaoshan Nature Reserve, on dead tree of Lithocarpus, 12 Jul 2013, Dai 13256 (BJFC 016516). Basidiocarps annual, pileate; pileus sessile to laterally substipitate, imbricate, fleshy, crumbly or chalky and light when dry, applanate, flabelliform to dimidiate, projecting up to 8 cm, 10 cm wide and 1.3 cm thick at base; pileal surface orange-yellow to reddish orange when fresh, becoming pinkish buff to orange-brown on drying, glabrous, azonate to faintly zonate; margin cream to buff when juvenile, becoming orange-yellow to reddish orange with age, acute; pore surface cream to buff when fresh, becoming buff to cinnamon buff when dry; sterile margin buff to clay-buff, up to 1 mm wide; pores angular, 3–5 per mm; dissepiments thin, entire to lacerate; context white to cream when fresh, becoming cream to buff on drying, fleshy when fresh, crumbly or chalky when dry, up to 1 cm thick; tubes concolorous with pore surface, crumbly or chalky, up to 3 mm long. Odor not distinctive, flavor acidic. Hyphal structure: hyphal system dimitic; generative hyphae simple-septate; skeletal hyphae IKI2, CB2, dissolving in KOH.

Context: generative hyphae rare, difficult to observe; binding hyphae dominant, hyaline, frequently branched, occasionally simple-septate, thick-walled with a wide lumen, interwoven, 6–14 mm diam. Tubes: generative hyphae hyaline, thin-walled, rarely branched, 3.8–5 mm diam; skeletal hyphae slightly thick-walled with a wide lumen, occasionally branched and simple-septate, loosely interwoven to subparallel along the tubes, 3–5 mm diam. Cystidia absent, but fusoid cystidioles occasionally present, hyaline, thin-walled, 10–13 3 4–6 mm. Basidia clavate, bearing four sterigmata and a basal simple-septum at the base, 12–15 3 5–8 mm; basidioles dominant, in shape similar to basidia but smaller. Basidiospores ovoid to ellipsoid, hyaline, smooth, usually bearing a large guttule, IKI2, CB2, (4.5–)5.0– 6.2 3 4.0–5.0(–5.2) mm, L 5 5.29 mm, W 5 4.18 mm, Q 5 1.22–1.31 (n 5 60/2). Type of rot: brown. Additional specimens (paratypes) examined: CHINA, YUNNAN PROVINCE: Jingdong County, Ailaoshan Nature Reserve, on living tree of Lithocarpus, 14 Jul 2007, HKAS 52508 & 53092 (HKAS); 3 Aug 2007, Yuan 3302 (IFP 013596); on fallen trunk of Lithocarpus, 15 Oct 2013, Dai 13566 (BJFC 015028), Dai 13567 (BJFC 015029) & Dai 13574 (BJFC 015036).

Laetiporus zonatus B.K. Cui & J. Song, sp. nov. FIGS. 2c, d; 4 MycoBank MB808204 Laetiporus zonatus is characterized by white to cream pileal surface with buff to clay-buff base when fresh, white to cream pores, concentrically zonate basidiocarps, ellipsoid to pyriform or drop-shaped basidiospores (5.8–7.2 3 4.3–5.5 mm). Etymology: zonatus (Lat.), referring to the concentrically zonate pileal surface.

Holotype: CHINA, YUNNAN PROVINCE: Lanping County, Tongdian, Laojun Mountains, Luoguqing, on fallen trunk of Quercus, 20 Sep 2011, Cui 10404 (BJFC 011299). Basidiocarps annual; pileus sessile to laterally substipitate, imbricate, fleshy, crumbly or chalky and light when dry, applanate, flabelliform to dimidiate, projecting up to 10 cm, 17 cm wide and 3 cm thick at base; pileal surface white to cream and buff to claybuff at base when fresh, becoming pale buff-yellow to orange brown on drying, glabrous, concentrically zonate with buff-yellow to cinnamon-buff zones; margin buff-yellow to yellowish brown when juvenile, becoming cinnamon-buff to cinnamon-brown when dry, waved, acute; pore surface white to cream with buff-yellow to yellowish brown zones when fresh, becoming cream buff to yellowish brown when dry; sterile margin distinct, clay-buff to cinnamon brown,

SONG ET AL.: TWO NEW SPECIES OF LAETIPORUS

1043

FIG. 1. Strict consensus tree illustrating the phylogeny of two new species and related species generated by maximum parsimony based on ITS+nLSU sequence data. Branches are labeled with parsimony bootstrap proportions (before the slash markers) higher than 50% and Bayesian posterior probabilities (after the slash markers) more than 0.95. Lowercase letter after isolates number, such as JV 0407/27a and JV 0407/27b, show clone sequences.

up to 2 mm wide; pores angular, 2–5 per mm; dissepiments thin, entire to lacerate; context white to cream when fresh, becoming cream to buff on drying, fleshy when fresh, crumbly or chalky when dry, up to 2.5 cm thick; tubes concolorous with pore surface, crumbly or chalky, up to 5 mm long. Odor not distinctive; flavor mild.

Hyphal structure: hyphal system dimitic; generative hyphae simple-septate; skeletal hyphae IKI2, CB2, dissolving in KOH. Context: generative hyphae rare, difficult to observe; binding hyphae dominant, hyaline, frequently branched, occasionally simple-septate, thick-walled with a wide lumen, interwoven, 6–15 mm diam.

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FIG. 2. 2.5 cm.

MYCOLOGIA

Basidiomata of Laetiporus species. a, b. L. ailaoshanensis. c, d. L. zonatus. Bars: a 5 2 cm, b 5 2.5 cm, c 5 3 cm, d 5

Tubes: generative hyphae hyaline, thin-walled, occasionally branched, 4–5.5 mm diam; skeletal hyphae thick-walled with a wide lumen, occasionally branched and simple-septate, loosely interwoven to subparallel along tubes, 4–5.5 mm diam. Cystidia absent, but fusoid cystidioles occasionally present, hyaline, thin-walled, 9–17 3 5.5–7 mm. Basidia clavate, bearing four sterigmata and a basal simple-septum at the base, 18–26 3 5–8 mm; basidioles dominant, clavate to pyriform, smaller than basidia. Basidiospores ellipsoid to pyriform or drop-shaped, hyaline, smooth, IKI2, CB2, (5.5–)5.8–7.2 3 (4.0–)4.3– 5.5 mm, L 5 6.46 mm, W 5 4.92 mm, Q 5 1.29–1.34 (n 5 90/3). Type of rot: brown. Additional specimens (paratypes) examined: CHINA, YUNNAN PROVINCE: Lanping County, Tongdian, Laojun Mountains, Luoguqing, on fallen trunk of Quercus, 20 Sep 2011, Cui 10403 (BJFC 011298); Dali, Jianchuan County, Laojun Mountains, on fallen trunk of Quercus, 16 Sep 2009, HKAS 71806 (HKAS); Lijiang, Yulongxueshan Nature

Reserve, Heibaishui, on fallen trunk of Quercus, 24 Jul 2008, HKAS 54701 (HKAS).

DISCUSSION Laetiporus sulphureus is common in the northern hemisphere (Gilbertson and Ryvarden 1986, Ryvarden and Gilbertson 1993, Nu´n ˜ ez and Ryvarden 2001), and it was recorded as a tree pathogen in China (Dai et al. 2007). Recent studies suggested that Laetiporus sulphureus is a species complex, which include morphologically and/or ecologically distinct species (Burdsall and Banik 2001, Ota et al. 2009). Two additional species, L. cremeiporus and L. montanus, were reported in northeastern China (Ota et al. 2009), but many collections were identified as L. sulphureus sensu lato. In the present study two Laetiporus species, L. ailaoshanensis and L. zonatus, from southwestern China can be distinguished from L. sulphureus sensu stricto and other Laetiporus species with morphological

SONG ET AL.: TWO NEW SPECIES OF LAETIPORUS

1045

FIG. 3. Microscopic structures of Laetiporus ailaoshanensis (drawn from the holotype). a. Basidiospores. b. Basidia and basidioles. c. Cystidioles. d. Hyphae from trama. e. Hyphae from context. Bars: a 5 5 mm, b–e 5 10 mm.

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MYCOLOGIA

FIG. 4. Microscopic structures of Laetiporus zonatus (drawn from the holotype). a. Basidiospores. b. Basidia and basidioles. c. Cystidioles. d. Hyphae from trama. e. Hyphae from context. Bars: a 5 5 mm, b–e 5 10 mm.

SONG ET AL.: TWO NEW SPECIES OF LAETIPORUS characters and molecular data. The combined dataset of ITS and nLSU sequences supported that L. ailaoshanensis and L. zonatus are new species, and no close relationships between the two new Laetiporus species and among them and other Laetiporus species were found. Laetiporus conifericola, L. huroniensis and L. montanus are morphologically similar to L. ailaoshanensis and L. zonatus; they all produce imbricate basidiocarps with similar pileal surface; however, the former three species produce slightly larger pores and longer basidiospores than L. ailaoshanensis and produce basidiospores that different from those of L. zonatus in size and shape (TABLE II); in addition the former three species occur on conifers in boreal or cool temperate areas (Burdsall and Banik 2001, Tomsˇovsky´ and Jankovsky´ 2008, Ota et al. 2009), while L. ailaoshanensis was found so far exclusively on Lithocarpus in subtropical China and L. zonatus was found only on Quercus at high altitude in temperate areas in southwestern China (TABLE II). Laetiporus caribensis might be confused with L. ailaoshanensis and L. zonatus, with a similar pileal surface and a hardwoods habitat; however L. caribensis produces smaller pores and smaller basidiospores (TABLE II); moreover L. caribensis lives in tropical areas of the Caribbean and Central America (Banik et al. 2012). Laetiporus cincinnatus and L. gilbertsonii resemble L. ailaoshanensis and L. zonatus in similar pileal surface and pore surface and growth on hardwoods, but they produce smaller and/or more flat basidiospores and slightly larger pores than L. ailaoshanensis, smaller and difform basidiospores than L. zonatus (TABLE II; Burdsall and Banik 2001). Laetiporus cremeiporus is similar to L. ailaoshanensis and L. zonatus by having imbricate basidiocarps with light orange to reddish orange pileal surface, yellowish white to cream pore surface and growth on hardwoods, but it produces slightly larger pores and longer basidiospores than L. ailaoshanensis, more flat and difform basidiospors than L. zonatus; moreover it is in cool temperate to boreal areas (TABLE II; Ota et al. 2009). So far five species of Laetiporus have been recognized in China: L. ailaoshanensis, L. cremeiporus, L. versisporus, L. zonatus and L. montanus. The first four species usually occur on hardwoods, and L. montanus is found only on conifers. Laetiporus montanus is found in boreal zones in northeastern China; L. cremeiporus is found in boreal to warm temperate zones from northeastern to central China; L. versisporus has wide distribution in cool temperate to tropical areas from central to southern China including Hainan island; L. ailaoshanensis has been found exclusively on Lithocarpus in subtropical zones

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of southwestern China; while L. zonatus was reported only on Quercus in high mountains of temperate southwestern China. In the present study (FIG. 1) the ITS and nLSU sequences of Laetiporus sensu stricto were analyzed and the results were consistent with Lindner and Banik (2008), Ota et al. (2009) and Banik et al. (2010). The two new species, Laetiporus ailaoshanensis (Clade N) and L. zonatus (Clade O), are monophyletic and are distant from other known clades of Laetiporus. Laetiporus montanus (Clade A2, Banik et al. 2010) from eastern Asia and Europe, L. huroniensis (Clade A1, Banik et al. 2010) and L. conifericola (Clade B, Banik et al. 2010) grouped together in a weakly supported ‘‘Conifericola clade’’. Collections identified as Laetiporus sulphureus were positioned in three clades: C (restricted to Europe and occurs on angiosperms and gymnosperms, Banik et al. 2010), E1 (white-pored species distributed in North America and grows on angiosperms) and E2 (yellow-pored species distributed in Europe, North America and South America and grows on angiosperms). Further study is needed to resolve the phylogeny and taxonomy of this species complex. Laetiporus cremeiporus is a well supported clade (Clade D, Banik et al. 2010), and it has been widely reported in eastern Asia (China, Korea, Japan). Laetiporus gilbertsonii is common in South and North America, and isolates of this species formed a well supported clade (F, Banik et al. 2010), which is sister of the white-pored L. sulphureus. Laetiporus versisporus (Clade G, Banik et al. 2010) has been found throughout eastern Asia, and the sampled specimens of this fungus were split into two well supported subclades. The unidentified Laetiporus taxa from South Africa and Hawaii were named Clade H and Clade I respectively after Banik et al. (2010, 2012). Laetiporus caribensis (Clade J, Banik et al. 2010) clusters with some unidentified Laetiporus taxa from Costa Rica (Clade L, Banik et al. 2012) and Argentina (Clade M, Banik et al. 2012). Laetiporus cincinnatus formed a strongly supported clade (Clade K, Banik et al. 2010). The key morphological characters, host types and distribution of Laetiporus sensu stricto clade (species in Clade A-O except the unidentified taxa) were listed (TABLE II). Our research expanded the number of Laetiporus species in China. For the time being taxonomy and phylogeny of the Laetiporus sulphureus species complex in the northern hemisphere has been relatively well studied, however there remain undescribed Laetiporus diversity worldwide and the relationships among Laetiporus species remain unresolved (Lindner and Banik 2008, Vasaitis et al. 2009,

2–4/mm 2–4/mm

bright sulphureous yellow

sulfur yellow

white lemon yellow

light orange to reddish orange pale salmon orange or pale pinkish orange

bright orange

light orange to reddish orange

citric yellow to orange

bright salmon orange bright salmon orange

L. cremeiporus (Clade D) L. gilbertsonii (Clade F)

L. huroniensis (Clade A1) L. montanus (Clade A2)

L. sulphureus (Clade C)

L. sulphureus (Clade E1) L. sulphureus (Clade E2)

2–5/mm

1–4/mm

2–4/mm

2–4/mm

2–4/mm

2–4/mm

lemon yellow to bright creamy yellow yellowish white to cream lemon yellow to pale lemon yellow (in West USA) or isabelline to nearly white (in Southeast USA) lemon yellow

bright orange to salmon orange

L. conifericola (Clade B)

2–4/mm

pale cream

4–5/mm

lemon yellow

bright salmon orange

3–5/mm

Pores

cream to buff

Pore surface

L. cincinnatus (Clade K)

L. ailaoshanensis orange-yellow to (Clade N) reddish orange L. caribensis orange to pale (Clade J) orange

Pileal surface

Distribution

ovoid to ellipsoid North America (Wisconsin, 5.0–6.8 3 4.0–5.0 mm Massachusetts) ovoid to ellipsoid North America, Europe and 5.0–6.8 3 4.0–5.0 mm South America

broadly ovoid eastern North America 5.0–7.0 3 4.2–5.0 mm and Great Lakes areas pyriform or ovoid to boreal zones in northeastern China and in mountain ellipsoid areas of Japan and central 6.0–8.0 3 4.0–5.5 mm Europe ovoid to ellipsoid most of Europe 5.0–7.0 3 3.5–5.0 mm

ovoid to ellipsoid cool temperate to boreal 5.6–7.0 3 3.9–4.7 mm areas of eastern Asia broadly ovoid North America, Central 5.0–6.5 3 3.5–4.5 mm and South America

ovoid to ellipsoid subtropical areas of 5.0–6.2 3 4.0–5.0 mm southwestern China ellipsoid tropical zones of the 4.0–4.5 3 2.7–3.6 mm Caribbean basin and Central America broadly ovoid throughout the eastern 4.5–5.5 3 3.5–4.0 mm USA except for in the states along the Gulf of Mexico, common in the Great Lakes regions broadly ovoid western North America 6.5–8.0 3 4.0–5.0 mm from California to Alaska

Basidiospores

Main morphological characters, host trees and distribution areas of species in the Laetiporus sensu stricto clade

Species

TABLE II.

Burdsall and Banik 2001 Tomsˇovsky´ and Jankovsky´ 2008, Ota et al. 2009 Ota et al. 2009

Burdsall and Banik 2001, Vasaitis et al. 2009, Banik et al. 2012

Ota et al. 2009

Burdsall and Banik 2001

Burdsall and Banik 2001

in the present study Banik et al. 2012

References

Quercus, Sorbus Populus, Taxus, Prunus, Picea, Castanea Fraxinus, Quercus Burdsall and Banik 2001 Acer, Salix, Burdsall and Gleditisa, Banik 2001 Quercus, Fraxinus, Castanea, Salix

Picea, Larix, Abies

Tsuga

Quercus, Pyrus, Prunus Eucalyptus Quercus, Prunus

Tsuga, Picea, Abies, Pinus

arising from the soil (Quercus)

Guareaguidonia, Dacroydes

Lithocarpus

Host

1048 MYCOLOGIA

ellipsoid to pyriform or drop-shaped 5.8–7.2 3 4.3–5.5 mm 2–5/mm

Robinia, Castanea, Nu´n ˜ ez and Ryvarden Quercus, 2001, Ota et Elaeocarpus, al. 2009 Castanopsis high mountains of temperate Quercus in the present areas of southwestern China study

ovoid to short cool temperate to tropical ellipsoid areas of eastern Asia 4.0–6.8 3 3.0–5.5 mm 2–6/mm

L. zonatus (Clade O)

whitish to sulphur usually yellow, yellow sometimes pale yellow to nearly white white to cream white to cream and buff to clay-buff at base L. versisporus (Clade G)

Species

TABLE II.

Continued

Pileal surface

Pore surface

Pores

Basidiospores

Distribution

Host

References

SONG ET AL.: TWO NEW SPECIES OF LAETIPORUS

1049

Ortiz-Santana et al. 2013). More comprehensive studies on Laetiporus depend on additional collections and data from poorly sampled areas. ACKNOWLEDGMENTS The authors thank Prof Yu-Cheng Dai (BJFC, China) for collecting specimens and improving the text. Dr Zheng Wang (Yale University) is deeply thanked for improving the English. Drs Shuang-Hui He and Chang-Lin Zhao (BJFC, China) are acknowledged for companionship during field collections. Drs Karl-Henrik Larsson (O, Norway), Yan-Chun Li (HKAS, China), Hai-Sheng Yuan (IFP, China) and Josef ˇ eske´ Budeˇjovice, Czech Republic) are deeply Vlasa´k (C thanked for loan of specimens. The research was financed by the Fundamental Research Funds for the Central Universities (Project No. JC2013-1) and the National Natural Science Foundation of China (Project No. 31170018).

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