Epitypification of Ganoderma sichuanense J.D. Zhao & X.Q. Zhang (Ganodermataceae)

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1 Epitypification of Ganoderma sichuanense J.D. Zhao & X.Q. Zhang (Ganodermataceae) Yi-Jian Yao,1,3 Xin-Cun Wang1,4 & Bo Wang2 1 State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, Beijing , China 2 Soil and Fertilizer Institute, Sichuan Academy of Agricultural Sciences, Chengdu , China 3 Royal Botanic Gardens, Kew, Richmond, Surrey, TW9 3AB, U.K. 4 Current address: Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing , China Author for correspondence: Yi-Jian Yao, yaoyj@im.ac.cn Abstract The widely cultivated Chinese Ganoderma, as Ling-zhi in Chinese and previously named as G. lucidum, is an important fungus with significant medicinal and economic values. The species identity of the fungus was recently clarified as G. sichuanense based on both morphological and molecular data, but an unusual ITS sequence inconsistent with morphological characters was also reported from the holotype of G. sichuanense. The ITS sequence is proved to be of G. weberianum, a species distinctly different from G. sichuanense in morphology. However, repeated amplifications of DNA fragments from the holotype of G. sichuanense were unsuccessful because of the specimen s condition. In order to interpret the species concept of the taxon, both morphologically and molecularly, an epitype of G. sichuanense is designated in this paper from recent collections with living strains from the type locality of the species. Ganoderma lingzhi, a recently proposed new species name for Ling-zhi widely cultivated in China, is determined as a later synonym of G. sichuanense. Keywords China; Ganoderma lingzhi; Ganoderma lucidum; Ling-zhi; polypores Received: 27 Feb. 2013; revision received: 17 June 2013; accepted: 24 July DOI: INTRODUCTION Ling-zhi, the Chinese Ganoderma lucidum, is an important traditional Chinese medicine with significant influence in the Orient. The species is widely cultivated throughout China, Japan, and Korea and now spreading to other parts of the world (Wang & al., 2012). Cultivation of the fungus has become a growing industry in China and the world trade market value of its products reached US$2.5 billion in 2003 (Chang, 2004). Although Ling-zhi was named Ganoderma lucidum (Curtis) P. Karst., a species originally described from London, England (Curtis, 1781: 72, t. 224), and the name has appeared in thousands of publications, the identity of the fungus has been clarified recently. Based on both morphological and molecular data, the identity of the Chinese G. lucidum (Ling-zhi) was re-determined as G. sichuanense J.D. Zhao & X.Q. Zhang, a species originally described from Sichuan Province, China (Wang & Yao, 2009; Wang & al., 2012). The name G. sichuanense was published in 1983 (Zhao & al., 1983) but has not been widely used. Morphological examinations of the holotype, authentic material and topotypes of G. sichuanense, and of cultivated specimens of the Chinese G. lucidum showed that they shared the same microscopic characters such as clavate cutis elements, Bovista-type ligative hyphae and strongly echinulate basidiospores, despite high variability in basidioma morphology (Wang & al., 2012). The results of combined analyses of DNA sequences (ITS + IGS + rpb2) from recent topotypes of G. sichuanense attempts to amplify the fragments from the holotype failed owing to the quality of DNA preserved in the specimen and from cultivated basidiomata and living strains of the Chinese G. lucidum also indicated that they had almost identical sequences. Therefore, both the Chinese G. lucidum (Ling-zhi) and G. sichuanense were considered as conspecific and the latter is the correct name for the fungus (Wang & al., 2012). The original material used in the description of G. sichuanense is still preserved in HMAS, including holotype (HMAS 42798) and paratype (HMAS 43728) (Zhao & al., 1983). Examination of these two specimens revealed morphological differences in the hyphal system, the shape of cutis elements and the size and ornament of basidiospores. The paratype was identified to be G. weberianum (Bres. & Henn. ex Sacc.) Steyaert based on long and subcylindrical cutis elements, colourless skeletal and ligative hyphae, absence of Bovista-type ligative hyphae, small and slightly echinulate basidiospores and presence of chlamydospores in context (Wang & al., 2012). There is an authentic specimen (HMAS 47694) named as G. sichuanense by the late Prof. J.D. Zhao in HMAS, which also shares the microscopic characters with the holotype. Unfortunately, attempts using various methods and kits to amplify DNA fragments from these three specimens holotype, paratype and authentic specimen of G. sichuanense were not successful (Wang & al., 2012). Apparently, DNA has not been well preserved in these specimens compared with that in topotypes of G. sichuanense (HMAS ; Wang & al., 2012). Recently, a new species name, G. lingzhi Sheng H. Wu, Y. Cao & Y.C. Dai, was proposed for the Chinese G. lucidum 1025

2 (Ling-zhi), primarily based on an ITS sequence (GenBank no. JQ781877) from the holotype of G. sichuanense (HMAS 42798; Cao & al., 2012). The ITS sequence is different from that from the Chinese G. lucidum and therefore the latter required a new species name, although some morphological interpretations were added to support the difference between the Chinese G. lucidum and G. sichuanense. However, compared with the dataset of DNA sequences from Ganoderma generated by Wang & al. (2012) and with ITS sequences in GenBank, the ITS sequence claimed by Cao & al. (2012) from the holotype of G. sichuanense is in fact belonging to G. weberianum, a species clearly different from G. sichuanense in morphology, especially in cutis elements, hyphal system, chlamydospores and size and ornamentation of basidiospores as mentioned above (chlamydospores are also formed in the mycelium culture of G. weberianum, unpub. data). Cao & al. (2012) managed to amplify the ITS fragment from the holotype of G. sichuanense and, no matter how unusual the sequence is, they have posted a question about the reliability of the type material. To clarify the morphological variability and the molecular reliability of G. sichuanense, a recent trip to the type locality of the taxon was organized and new collections of the species were obtained. All the collections were subjected to both morphological and molecular studies and their identity was confirmed as G. sichuanense. To fix the application of the name G. sichuanense, which will stabilize the species concept, an epitype is proposed here. The epitype is selected from recent collections made in the type locality of the species based on both morphological and molecular data. The type locality of G. sichuanense was indicated in the protologue (Zhao & al., 1983) as Dokou shi (equivalent to Dokou City in English), which is in the south-west of Sichuan Province, China and is now called Panzhihua City in the new administration system. Panzhihua City is a city including a downtown area of three districts (East, West and Renhe districts) and two outskirt counties (Miyi and Yanbian counties), with an area of 7440 km² and characterized by interleaving distribution of many mountains, forming highlands, valleys and basins. The Chinese G. lucidum (Ling-zhi) has not been cultivated in large scale in this area. Although there is no further information about the type locality in the protologue apart from Dokou shi (Zhao & al., 1983), the type was collected in 1976 from Panzhihua Steel Plant, Dokou City as recorded on the label of type specimen of G. sichuanense (HMAS 42798; Fig. 1). The Panzhihua Steel Plant (now called Panzhihua Iron and Steel (Group) Company Limited ) is much surrounded by downtown area nowadays. Eight collections of G. sichuanense were made in the recent field trip, one from Renhe District and seven from Miyi County. In addition to G. sichuanense, two collections of G. multipileum D. Hou and one of G. gibbosum (Blume & T. Nees) Pat. were also made at the base of dead trees on a hill next to workshops of the Panzhihua Iron and Steel (Group) Company Limited on the edge of downtown area of Panzhihua City and on a tree stump in a street within the city, respectively. Among the G. sichuanense collections, one of the specimens from a mountainous area within the administration of Caochang Township, Miyi County, displaying a range of growth stages and various pileus shapes, is selected to serve as the epitype of G. sichuanense. Isolations of living strains were successfully performed from the fresh material including the proposed epitype. Morphological observation, molecular work and data analyses followed those described in Wang & al. (2012). ITS, IGS and rpb2 sequences from the proposed epitype (HMAS ) were submitted to GenBank with the accession numbers of KC All the ITS sequences named G. lingzhi or G. sichuanense in Cao & al. (2012), as well as the one derived from the proposed epitype in this study, were added to the dataset generated in Wang & al. (2012) to constitute a compiled dataset for phylogenetic analyses. The morphological description of the epitype of G. sichuanense is provided below. DESCRIPTION OF THE EPITYPE OF GANODERMA SICHUANENSE Fig. 1. The holotype of Ganoderma sichuanense (HMAS 42798). Scale bar = 1.5 cm. Ganoderma sichuanense J.D. Zhao & X.Q. Zhang in Acta Mycol. Sin. 2: Epitype (designated here): CHINA. Sichuan Province, Panzhihua City, Miyi County, Caochang Township, Huangqiao Village, N, E, alt m, on or near the base of a dead tree of Quercus sp., 12 Oct. 2012, Y.J. Yao, B. Wang & X.C. Wang 003 (HMAS ). The living strain (no. 2187) from the epitype was preserved at the China General Microbiological Culture Collection with the accession number of CGMCC Figure 2 Basidiomata annual, substipitate to stipitate, corky to woody. Pileus cm, cm thick, flabellate to reniform, concave or convex; upper surface orange brown to bay, laccate, with radial furrows, sometimes covered by a layer of rust brown basidiospores; margin cream-coloured to concolorous with the pileus, acute to obtuse. Pores 5 6 per mm, 1026

3 suborbicular, rarely angular; surface yellowish brown, darker when bruised; tubes mm long, cinnamon. Stipe cm long, cm thick, lateral, cylindrical or flattened, bay, laccate. Context mm thick, cream-coloured to buff, darker near the tube layer, without black, horn-like layer in the context; hyphal system trimitic; generative hyphae hyaline, thin-walled, rare; skeletal hyphae hyaline in 5% KOH, thickwalled to solid, aciculiform or arboriform, μm diam.; ligative hyphae hyaline, thick-walled, branched, μm diam., typically Bovista-type. Basidiospores ovoid, truncate at the apex; yellowish brown, with a dark brown eusporium bearing thick echinulae, overlaid by a hyaline myxosporium, (9.0 ) ( 11.0) μm (average length/width ratio of basidiospores: Q = 1.40) cum myxosp. and ( 6.5) μm (Q = 1.45) sine myxosp. Basidia not seen. Cutis hymeniodermiformic, yellowish brown, elements clavate, about 20 30( 45) μm long and (7.0 ) μm thick at the top. The above description is solely based on the epitype proposed. However, some variations in macroscopic morphology in basidiomata, pileus, pores, stipe, context thickness, spore size, cutis elements, etc., were found in the other collections made in the same trip, especially in the collection from Xuefang Village, Zhongba Township, Renhe District, which comprises 26 basidiomata. Among the collections, sessile basidiomata, orbicular and yellowish pileus (up to cm) ornamented with concentrically sulcate zones, whitish to yellowish brown pore surface, eccentric to centric stipe up to 12.5 cm long and 6.0 cm thick, and context up to 10.0 mm thick were present, and basidiospores measured up to μm (Q = ) cum myxosp. and 8.5( 9.0) 6.0( 6.5) μm (Q = ) sine myxosp., and cutis elements μm thick at the top. Additional specimens examined: CHINA. Sichuan Province, Panzhihua City, Miyi County, Caochang Township, Huang qiao Village, around the stump of Quercus sp., 12 Oct. 2012, Fig. 2. The epitype of Ganoderma sichuanense (HMAS No ). A, the habitat of the collection; B, specimens of the collection; C, reversed side view of the specimens in B; D, tissue of cutis; E, Bovista-type ligative hyphae; F, basidiospores. Scale bars: B, C = 4 cm; D, E = 20 µm; F = 10 µm. 1027

4 Y.J. Yao, B. Wang & X.C. Wang 005 (HMAS ); ibid., Y.J. Yao, B. Wang & X.C. Wang 006 (HMAS ); ibid., N, E, alt m, close to the stump of Quercus sp., 12 Oct. 2012, Y.J. Yao, B. Wang & X.C. Wang 004 (HMAS ); ibid., Y.J. Yao, B. Wang & X.C. Wang 009 (HMAS ); ibid., N, E, alt m, on the base of the stump of Quercus sp., Y.J. Yao, B. Wang & X.C. Wang 007 (HMAS ); ibid., close to the stump of Quercus sp., Y.J. Yao, B. Wang & X.C. Wang 008 (HMAS ); Panzhihua City, Renhe District, Zhongba Township, Xuefang Village, N, E, alt m, purchased from a villager who gathered the specimens from mountainous areas surrounding the village, 14 Oct. 2012, Y.J. Yao, B. Wang & X.C. Wang 019 (HMAS ). Compared with the description in the previous study (Wang & al., 2012), the morphology of G. sichuanense is further extended as follows (Italics indicate emendations): Basidiomata annual, sessile, substipitate to stipitate, corky to woody. Pileus cm, cm thick, flabellate, dimidiate, reniform to orbicular, concave or convex; upper surface yellowish brown to bay, laccate, sometimes with radial furrows or concentrically sulcate zones, sometimes covered by a layer of rust brown basidiospores; margin cream-coloured to concolorous with the pileus, acute to obtuse. Pores 4 6 per mm, suborbicular, rarely angular; surface whitish to yellowish brown, darker when bruised; tubes mm long, cinnamon. Stipe cm long, cm thick, lateral, eccentric to centric, cylindrical or flattened, bay to black, laccate. Context mm thick, cream-coloured to buff, darker near the tube layer, with or without black, hornlike layer in the context; hyphal system trimitic; generative hyphae hyaline, thin-walled, sometimes rare, μm diam., with clamp connections; skeletal hyphae hyaline to yellowish brown in 5% KOH, thick-walled to solid, aciculiform or arboriform, μm diam.; ligative hyphae hyaline to yellowish brown, thick-walled, branched, μm diam., typically Bovista-type. Basidiospores ovoid, truncate or not at the apex; yellowish brown, with a dark brown eusporium bearing thick echinulae, overlaid by a hyaline myxosporium, (8.0 ) ( 12.5) (5.5 ) ( 8.5) μm (Q = ) cum myxosp. and (5.0 ) ( 9.5) ( 7.0) μm (Q = ) sine myxosp. Basidia not seen. Cutis hymeniodermiformic, yellowish brown, elements clavate, 20 40( 45) μm long and μm thick at the top. A total of 60 ITS sequences (Appendix 1) were included in the compiled dataset for phylogenetic analyses. The dataset consisted of 563 bp, of which 93 were parsimony informative. Maximum-parsimony (MP) analyses of the dataset yielded 681 equally parsimonious trees (length = 145, consistency index = , retention index = ). The HKY model of DNA substitution with gamma-distributed rate variation across invariant sites was determined as best-fit for Bayesian analyses. There was no prominent difference between the topology of MP analyses and the Bayesian consensus tree, and one of the 681 equally parsimonious trees was presented in Fig. 3. Six clades in Ganoderma were recognized in the analysis, i.e., G. lucidum, G. multipileum, G. resinaceum Boud., G. sichuanense, G. tropicum (Jungh.) Bres. and G. weberianum, with Tomophagus colossus (Fr.) Murrill as an outgroup (Fig. 3). All the ITS sequences named G. lingzhi in Cao & al. (2013) and KC from the proposed epitype (HMAS ) of G. sichuanense were nested with those of G. sichuanense used in Wang & al. (2012) with strong support (BP = 100%, PP = 1.00). The sequences purported from the holotype of G. sichuanense (HMAS 42798, JQ781877), and from another specimen (Cui 7691, JQ781878) named G. sichuanense in Cao & al. (2012) were both clustered with those of G. weberianum in the same clade with strong support (BP = 100%, PP = 1.00). Apparently, the ITS sequences named as G. lingzhi in Cao & al. (2012) are conspecific with those from G. sichuanense in Wang & al. (2012), including topotypes and the epitype (obtained in this study) of G. sichuanense, and the sequences from both the holotype of G. sichuanense and Cui 7691 in Cao & al. (2012) are of G. weberianum. As illustrated in Wang & al. (2012: figs. 3-A, 3-C 3-E for G. sichuanense, including the Chinese G. lucidum [Lingzhi], and figs. 3-B and 4-C for G. weberianum), G. sichuanense has clavate and comparatively short (20 40 µm) cutis elements, Bovista-type ligative hyphae, strongly echinulate basidiospores of µm long cum myxosp.; whilst G. weberianum has subcylindrical and comparatively long (30 60 µm) cutis elements, slightly echinulate basidiospores of μm long cum myxosp. and also chlamydospores in context (illustrated in Fig. 4 of this paper), but lacks Bovista-type ligative hyphae. It is worthy of note that although cells clavate were described for the cutis of Ling-zhi (G. lingzhi) in Cao & al. (2012: 54), the illustration of apical cells from the cuticle in fig. 3c in that paper did not show the typical clavate shape. It may be necessary to check the illustration with the specimen used again, although no specimen was indicated for fig. 3 in Cao & al. (2012). Further, the size range of cutis elements given as µm (for G. lingzhi) in Cao & al. (2012: 54), is generally in overlap with µm in Wang & al. (2012: 6), 20 30( 45) (7.0 ) µm (for the epitype) and 20 40( 45) µm (overall for G. sichuanense) in the present study. These differences between measurements may be regarded as variations of specimens or standard deviation of measurement, but the size in Cao & al. (2012) tends to be slightly longer and slenderer than that recorded by this research group. The morphology of G. weberianum was based on HMAS (cultivated specimen from a strain isolated from a basidioma originally collected in Hainan Province, China), from which DNA sequences of ITS (JF915411), IGS (JF915425) and rpb2 (JF915434) were obtained, and HMAS (the paratype of G. sichuanense collected in Sichuan Province, China) in Wang & al. (2012). The specimen Cui 7691, from which the ITS sequence (JQ781878) derived and identified as G. sichuanense, was collected in Guandong Province, China (Cao & al. 2012), a province next to Hainan but separated by Hainan Strait. JQ is very similar to JF from HMAS and grouped in the same clade of phylogenetic analyses (Fig. 3). Although Cui 7691 was named as G. sichuanense in Cao & al. (2012), it is likely that the specimen is morphologically similar to HMAS (G. weberianum), but different from the 1028

5 holotype of G. sichuanense (HMAS 42798). However, the specimen Cui 7691 is not available for examination after a request for a loan of the specimen and, therefore, the morphological confirmation cannot be made at this time. Ganoderma lingzhi was believed to be morphologically different from G. sichuanense in the presence or absence of a stipe and in basidiospore sizes, i.e., stipe present and basidiospores (8.0 ) ( 12.0) (5.2 ) ( 7.5) μm in the former while stipe absent and basidiospores (7.0 ) ( 9.3) (4.6 ) ( 6.8) μm in the latter (Cao & al., 2012). However, the presence or absence of a stipe is not a stable character in Ganoderma, as indicated by Moncalvo & Ryvarden (1997) and also as displayed in recently collected specimens (HMAS and ; Fig. 5). The basidiospore size for G. sichuanense given by Cao & al. (2012) was similar to that in the protologue (Zhao & al., 1983; as μm), which was apparently with the inclusion of the paratype (HMAS 43728, a specimen of G. weberianum; Wang & al., 2012). The spore measurement of the holotype of G. sichuanense is μm cum myxosp. (Wang & al., 2012), similar to the measurement from Ganoderma sichuanense HMAS60537 Ganoderma sichuanense HMAS62503 Ganoderma sichuanense HMAS76566 Ganoderma sichuanense HMAS Ganoderma lingzhi Dai12443 Ganoderma lingzhi IFP01021 Ganoderma lingzhi Dai12479 Ganoderma lingzhi Li245 Ganoderma sichuanense HMAS99391 Ganoderma sichuanense HMAS Ganoderma sichuanense HMAS Ganoderma sichuanense CGMCC5.75 Ganoderma sichuanense CGMCC5.425 Ganoderma lingzhi Wu Ganoderma lingzhi Cui9164 Ganoderma sichuanense HMAS Ganoderma sichuanense HMAS Ganoderma sichuanense HMAS Ganoderma lingzhi Dai12425 Ganoderma lingzhi Dai12426 Ganoderma lingzhi Dai12441 Ganoderma lingzhi Dai12447 Ganoderma lingzhi Dai12449 Ganoderma sichuanense HMAS Ganoderma sichuanense HMAS Ganoderma sichuanense HMAS Ganoderma sichuanense HMAS Ganoderma sichuanense CGMCC5.533 Ganoderma lingzhi Dai12438 Ganoderma lingzhi Cui6982 Ganoderma sichuanense HMAS25066 Ganoderma sichuanense HMAS25067 Ganoderma sichuanense HMAS42605 Ganoderma sichuanense HMAS42745 Ganoderma sichuanense HMAS47337 Ganoderma sichuanense HMAS /1.00 Ganoderma sichuanense HMAS Ganoderma lingzhi Dai3583 Ganoderma lingzhi Dai12374 Ganoderma sichuanense HMAS Ganoderma sichuanense HMAS Epitype Ganoderma lingzhi Dai10631 Ganoderma lingzhi Dai12573 Ganoderma lingzhi Cui /1.00 Ganoderma lingzhi Cui / Ganoderma sichuanense HMAS Ganoderma multipileum HMAS Ganoderma multipileum BCRC /1.00 Ganoderma tropicum HMAS /1.00 Ganoderma tropicum BCRC37122 Ganoderma weberianum HMAS /1.00 Ganoderma sichuanense Cui7691 Ganoderma sichuanense HMAS42798 Holotype 67/ / Ganoderma weberianum SUT-H Ganoderma resinaceum HMAS86599 Ganoderma resinaceum GrTO96 100/1.00 Ganoderma lucidum HMAS86597 Ganoderma lucidum G1TO99 Tomophagus colossus CGMCC5.763 Tomophagus colossus CBS G. sichuanense G. multipileum G. tropicum G. weberianum G. resinaceum G. lucidum Fig. 3. One of the 681 most parsimonious trees derived from the ITS dataset. Numbers on branches denote the percentage of bootstrap replicates/bayesian posterior probabilities. Only bootstrap values higher than 50% in MP or posterior probabilities higher than 0.90 in Bayesian were shown. 1029

6 Fig. 4. Chlamydospores observed in the contexts of Ganoderma weberianum specimens. A, chlamydospores in HMAS (the paratype of G. sichuanense), collected from Panzhihua City, Sichuan Province; B, chlamydospores in HMAS 97365, cultivated specimen in Huairou District, Beijing from the strain CGMCC 5.71, which was isolated from a basidioma originally collected from Hainan Province. Scale bars = 10 µm. Fig. 5. Sessile basidiomata of Ganoderma sichuanense. A, top view of basidioma (HMAS ); B, reversed side view of the basidioma in A; C, top view of basidioma (HMAS ); D, reversed side view of the basidioma in C. Scale bars = 1 cm. the epitype described above. In fact, there is no substantial difference in morphology between G. sichuanense and G. lingzhi, especially in short and clavate cutis elements, Bovista-type ligative hyphae, strongly echinulate basidiospores and their measurement (Wang & al., 2012). Apart from the ITS sequence from the holotype of G. sichuanense (JQ781877) by Cao & al. (2012), G. lingzhi is, based on all other molecular data analyzed, the same as G. sichuanense as defined by the epitype proposed here. Therefore, G. lingzhi is conspecific with G. sichuanense and is a later synonym. ACKNOWLEDGEMENTS The authors are grateful to Dr. Paul Kirk for critical reading the manuscript and valuable suggestions. This work is supported by the National Natural Science Foundation of China ( ) LITERATURE CITED Cao, Y., Wu, S.-H. & Dai, Y.-C Species clarification of the prize medicinal Ganoderma mushroom Lingzhi. Fungal Diversity 56: Chang, S.-T Ganoderma lucidum A leader of edible and medicinal mushrooms. Int. Agric. Trade 90: Curtis, W Flora londinensis. London: printed for the author. Guglielmo, F., Bergemann, S.E., Gonthier, P., Nicolotti, G. & Garbelotto, M A multiplex PCR-based method for the detection and early identification of wood rotting fungi in standing trees. J. Appl. Microbiol. 103: Guglielmo, F., Gonthier, P., Garbelotto, M. & Nicolotti, G A PCR-based method for the identification of important wood rotting fungal taxa within Ganoderma, Inonotus s.l. and Phellinus s.l. F. E. M. S. Microbiol. Lett. 282:

7 Moncalvo, J.-M. & Ryvarden, L A nomenclatural study of the Ganodermatacea Donk. Oslo: Fungiflora. Moncalvo, J.-M., Wang, H.-F., Wang, H.-H. & Hseu, R.-S The use of ribosomal DNA sequence data for species identification and phylogeny in the Ganodermataceae. Pp in: Buchanan, P.K., Hseu, R.-S. & Moncalvo, J.-M. (eds.), Ganoderma: Systematics, phytopathology and pharmacology. Proceedings of Contributed Symposia 59A, B, 5th International Mycological Congress, Vancouver, August 14 21, Taipei: [s.n.]. Roberts, L.M Australian Ganoderma: Identification, growth & antibacterial properties. Dissertation, Swinburne University of Technology, Melbourne, Australia. Wang, D.-M., Wu, S.-H., Su, C.-H., Peng, J.-T., Shih, Y.-H. & Chen, L.-C Ganoderma multipileum, the correct name for G. lucidum in tropical Asia. Bot. Stud. (Teipei) 50: Wang, X.-C. & Yao, Y.-J Taxonomic studies on Ganoderma lucidum in China. Pp in: Abstracts of the 5th International Medicinal Mushroom Conference, Nantong, September 5 8, Jiangsu, China: [s.n.]. Wang, X.-C., Xi, R.-J., Li, Y., Wang, D.-M. & Yao, Y.-J The species identity of the widely cultivated Ganoderma, G. lucidum (Ling-zhi), in China. PLoS ONE 7: e Zhao, J.-D., Xu, L.-W. & Zhang, X.-Q Taxonomic studies on the family Ganodermataceae of China II. Acta Mycol. Sin. 2: Appendix 1. ITS sequences with their voucher materials used in this study. Taxon name, country and/or locality, collector(s) and collection number or herbarium (or living collection) accession number, GenBank accession number of ITS, and reference. A newly generated sequence is indicated by an asterisk (*). Ganoderma lingzhi Sheng H. Wu, Y. Cao & Y.C. Dai, China, Hubei, Wu (holotype), JQ781858, Cao & al., 2012; G. lingzhi, China, Jiangsu, Cui 4018, JQ781856, Cao & al., 2012; G. lingzhi, China, Tianjin, Cui 6982, JQ781862, Cao & al., 2012; G. lingzhi, China, Shandong, Cui 9164, JQ781859, Cao & al., 2012; G. lingzhi, China, Zhejiang, Cui 10165, JQ781857, Cao & al., 2012; G. lingzhi, China, Hunan, Dai 3583, JQ781868, Cao & al., 2012; G. lingzhi, China, Anhui, Dai 10631, JQ781860, Cao & al., 2012; G. lingzhi, China, Yunan, Dai 12374, JQ781867, Cao & al., 2012; G. lingzhi, China, Shandong, cultivated, Dai 12425, JQ781871, Cao & al., 2012; G. lingzhi, China, Sichuan, cultivated, Dai 12426, JQ781870, Cao & al., 2012; G. lingzhi, China, Henan, cultivated, Dai 12438, JQ781861, Cao & al., 2012; G. lingzhi, China, Zhejiang, cultivated, Dai 12441, JQ781869, Cao & al., 2012; G. lingzhi, China, Guangdong, cultivated, Dai 12443, JQ781866, Cao & al., 2012; G. lingzhi, China, Fujian, cultivated, Dai 12447, JQ781872, Cao & al., 2012; G. lingzhi, China, Jiangsu, cultivated, Dai 12449, JQ781873, Cao & al., 2012; G. lingzhi, China, Anhui, cultivated, Dai 12479, JQ781864, Cao & al., 2012; G. lingzhi, China, Liaoning, cultivated, Dai 12573, JQ781855, Cao & al., 2012; G. lingzhi, China, Hubei, cultivated, IFP 01021, JQ781865, Cao & al., 2012; G. lingzhi, China, Henan, Li 245, JQ781863, Cao & al., 2012; G. lucidum (Curtis) P. Karst., U.K., HMAS 86597, AY884176, Wang & al., 2012; G. lucidum, Italy, G1T 099, AM269773, Guglielmo & al., 2007; G. multipileum D. Hou, China, Sichuan, HMAS , JF915409, Wang & al., 2012; G. multipileum, China, Taiwan, BCRC 37033, EU021462, Wang & al., 2009; G. resinaceum Boud., U.K., HMAS 86599, AY884177, Wang & al., 2012; G. resinaceum, Italy, GrT 096, AM906065, Guglielmo & al., 2008; G. sichuanense J.D. Zhao & X.Q. Zhang, China, Sichuan, HMAS (holotype), JQ781877, Cao & al., 2012; G. sichuanense, China, Yunnan, HMAS 25066, JN197275, Wang & al., 2012; G. sichuanense, China, Yunnan, HMAS 25067, JN197276, Wang & al., 2012; G. sichuanense, China, Yunnan, HMAS 42605, JN197277, Wang & al., 2012; G. sichuanense, China, Shanghai, cultivated, HMAS 42745, JN197278, Wang & al., 2012; G. sichuanense, China, Hubei, HMAS 47337, JN197279, Wang & al., 2012; G. sichuanense, China, Henan, HMAS 59482, JN197280, Wang & al., 2012; G. sichuanense, China, Shandong, cultivated, HMAS 60537, JN197281, Wang & al., 2012; G. sichuanense, China, Jiangxi, cultivated, HMAS 62503, JF915405, Wang & al., 2012; G. sichuanense, China, Shanghai, cultivated, HMAS 76566, JF915406, Wang & al., 2012; G. sichuanense, China, Beijing, cultivated, HMAS 99391, JF915407, Wang & al., 2012; G. sichuanense, China, Sichuan, HMAS , JF915404, Wang & al., 2012; G. sichuanense, China, Sichuan, cultivated, HMAS , JF915408, Wang & al., 2012; G. sichuanense, China, Henan, cultivated, HMAS , JF915393, Wang & al., 2012; G. sichuanense, China, Zhejiang, cultivated, HMAS , JF915394, Wang & al., 2012; G. sichuanense, China, Fujian, cultivated, HMAS , JF915395, Wang & al., 2012; G. sichuanense, China, Shandong, cultivated, HMAS , JF915396, Wang & al., 2012; G. sichuanense, China, Zhejiang, cultivated, HMAS , JF915397, Wang & al., 2012; G. sichuanense, China, Henan, cultivated, HMAS , JF915398, Wang & al., 2012; G. sichuanense, China, Jiangsu, cultivated, HMAS , JF915399, Wang & al., 2012; G. sichuanense, China, Sichuan, HMAS , JF915400, Wang & al., 2012; G. sichuanense, China, Sichuan, HMAS , JF915401, Wang & al., 2012; G. sichuanense, China, Sichuan, HMAS , JF915402, Wang & al., 2012; G. sichuanense, China, Sichuan, HMAS , JF915403, Wang & al., 2012; G. sichuanense, China, Sichuan, Y.J. Yao, B. Wang & X.C. Wang 003 (HMAS , epitype), *KC662402, this study; G. sichuanense, China, CGMCC 5.75, JN197282, Wang & al., 2012; G. sichuanense, Japan, CGMCC 5.425, JN197283, Wang & al., 2012; G. sichuanense, Japan, CGMCC 5.533, JN197284, Wang & al., 2012; G. sichuanense, China, Guangdong, Cui 7691, JQ781878, Cao & al., 2012; G. tropicum (Jungh.) Bres., China, Hainan, HMAS , JF915410, Wang & al., 2012; G. tropicum, China, Taiwan, BCRC 37122, EU021457, Wang & al., 2009; G. weberianum (Sacc.) Steyaert, China, Hainan, HMAS 97365, JF915411, Wang & al., 2012; G. weberianum, Australia, SUT H2, AY569451, Roberts, 2004; Tomophagus colossus (Fr.) Murrill, Philippines, CGMCC 5.763, JQ081068, Wang & al., 2012; T. colossus, Philippines, CBS , Z37071, Moncalvo & al., 1995; T. colossus, Philippines, CBS , Z37091, Moncalvo & al., Abbreviations and acronyms: BCRC, Bioresources Collection and Research Center, Taiwan, China; CBS, Centraalbureau voor Schimmelcultures; CGMCC, China General Microbiological Culture Collection Center, Beijing, China; GlT, Ganoderma lucidum; GrT, Ganoderma resinaceum; HMAS, Fungarium, Institute of Microbiology, Chinese Academy of Sciences; SUT, Swinburne University of Technology, Australia. 1031