A phylogenetic overview of the Hydnaceae (Cantharellales, Basidiomycota) with new taxa from China

DNA extraction, PCR, and sequencing

Genomic DNA was extracted from dried herbarium specimens with a Thermo Scientific Phire Plant Direct PCR kit (Thermo Fisher Scientific, Waltham, Massachusetts, USA) according to the manufacturer’s instructions which was also used for the polymerase chain reaction (PCR) (Chen et al. 2016). Nuclear ribosomal RNA markers were used to determine the phylogenetic position of the new species. The internal transcribed spacer (ITS) was amplified with the primers ITS1F/ITS4 (White et al. 1990) and LROR/LR5 (Vilgalys & Hester 1990) for partial nLSU; MS1/MS2 (Matheny 2005) for mtSSU; rpb2-5FCanth/rpb2-7cRCanth (Buyck et al. 2014) for RPB2 of Cantharellus and fRPB2-5F/bRPB2-7.1R (Matheny et al. 2007) for Craterellus and Hydnum; Tef1R/Tef1RF (Morehouse et al. 2003) for TEF1 of Cantharellus and Craterellus, and HEF1F/HEF1R for Hydnum (Feng et al. 2016).

PCR reactions were performed in 30 μL reaction mixtures containing 15 μL of 2 × Phire® Plant PCR buffer, 0.6 μL Phire® Hot Start II DNA Polymerase, 1.5 μL of each PCR primer (10 μM), 10.5 μL double deionised H₂O (ddH₂O), and 0.9 μL template DNA. PCR amplification was confirmed on 1 % agarose electrophoresis gel stained with ethidium bromide (Stöger et al. 2006) and sequenced at the Beijing Genomics Institute (BGI) with the same primers as used in PCR. The newly generated DNA sequences were assembled and manually modified with the software DNAMAN8 (Lynnon Biosoft, Quebec, Canada). The sequence quality control followed the guidelines by Nilsson et al. (2012). All sequences newly obtained were submitted to GenBank (Sayers et al. 2020).

Phylogenetic analyses

Sequences for phylogenetic analysis were found in GenBank (http://www.ncbi.nlm.gov) using the BLAST option and downloaded (Table 1). DNA alignments were performed using the MAFFT v. 7.471 online service (https://mafft.cbrc.jp/alignment/server/index.html; Katoh et al. 2019). Intron regions of RPB2 and TEF1 as well as low-homology regions of ITS1 and ITS2 were removed before phylogenetic analyses, and the two sequence datasets were combined using BioEdit v. 7.2.6 (Hall 2005).

We assembled four datasets for phylogenetic analyses: the Cantharellales dataset based on a five-locus concatenated alignment which included nLSU, ITS, mtSSU, RPB2 and TEF1; the Cantharellus dataset based on a five-locus concatenated alignment which included 5.8S, nLSU, mtSSU, RPB2 and TEF1; the Craterellus and Hydnum datasets both based on a two-locus (nLSU and ITS) concatenated alignment. The four datasets were all partitioned by gene and codon position and the best-fit models were determined by jModelTest v. 2.1.10 (Darriba et al. 2012) based on the Corrected Akaike Information Criterion (AICc). The first dataset (Cantharellales) was divided into nine data partitions and the best-fit models were: GTR + I + G for nLSU, GTR + G for ITS, TrN + G for mtSSU, GTR + I + G for RPB2 1^st, 2^nd, 3^rd and TEF1 1^st, K80 + G for TEF1 2^nd and 3^rd; the second (Cantharellus) was divided into nine: TPM1 + G for 5.8S, TIM1 + I + G for nLSU, F81 + I for mtSSU, TrNef + I + G for PRB2 1^st, TrN + G for PRB2 2^nd and PRB2 3^rd, TIM1ef + I + G for TEF1 1^st, JC for TEF1 2^nd and TPM2 + I + G for TEF1 3^rd; the third (Craterellus) was divided into four: ITM1 + I + G for nLSU, TrN + I for ITS1, JC for 5.8S and TrN + G for ITS2 and the fourth (Hydnum) was divided into four: GIR + I + G for nLSU, JC for ITS1, K80 for 5.8S and TrN + G for ITS2.

Phylogenetic analyses for each dataset were conducted using Bayesian Inference (BI) analysis and Maximum Likelihood (ML) methods. All characters were weighted, and gaps were treated as missing data. BI analysis with MrBayes v. 3.2.7 (Ronquist et al. 2012) implemented the Markov Chain Monte Carlo (MCMC) technique. Four simultaneous Markov chains were run with 15, 10, 5, 5 million generations for the four datasets respectively, starting from random trees and keeping one tree every 100^th generation until the average standard deviation of split frequencies was below 0.01. The value of burn-in was set to discard 25 % of trees when calculating the posterior probabilities. Bayesian Posterior Probabilities (BPP) were obtained from the 50 % majority rule consensus of the trees kept. An ML analysis used the same datasets as the BI analysis and was performed in RAxML v. 8.2.4 (Stamatakis 2014). The best tree was obtained by performing 1 000 rapid bootstrap inferences followed by a thorough search for the most likely tree (Stamatakis et al. 2008). Phylogenetic trees were checked and modified in FigTree v. 1.4 (Rambaut 2012). The alignments and trees were deposited in TreeBASE (No. S28157).

New taxa of Hydnaceae in this study

Cantharellus Adans. ex Fr., Syst. Mycol. (Lundae) 1: 316. 1821. MycoBank MB 17236.



Synonym: Afrocantharellus (Eyssart. & Buyck) Tibuhwa, IMA Fungus 3: 33. 2012. MycoBank MB 518687.

Goossensia Heinem., Bull. Jard. Bot. État Brux 28: 424. 1958. MycoBank MB 17690.



Type species: Cantharellus cibarius Fr., Syst. Mycol. (Lundae) 1: 318. 1821. MycoBank MB 200345.



Notes: Cantharellus was described by Fries (1821), with Cantharellus cibarius selected as the type species by Earle (1909). It is a large ectomycorrhizal genus of the Hydnaceae, comprising many edible species (Moncalvo et al. 2006, Hibbett et al. 2014), and belongs in the core lineage of the cantharelloid clade. Our study reproduced the infrageneric classification of the genus based on a multiple-marker database (including 5.8S, nLSU, mtSSU and two protein coding genes RPB2 and TEF1) (Fig. 2) and the result is similar to Buyck et al. (2014). The phylogenetic status of Cantharellus in the family proposed by Moncalvo et al. (2006) and Hibbett et al. (2014) is also confirmed by our tree (Fig. 1). Cantharellus groups with Craterellus and these two genera as well as Hydnum and several samples of Sistotrema form a strongly supported subclade in Hydnaceae.

 Cantharellus is characterised by fleshy basidiocarps, a colourful pileus, nearly smooth to obvious veined hymenophore, long and stichic basidia, cylindrical hyphal endings in the pileipellis and a solid stipe (Cairney & Chambers 1999, Pine et al. 1999, Buyck 2014, Buyck et al. 2014). The boundary between Cantharellus and its sister group Craterellus had been resolved with molecular data by Dahlman et al. (2000) and Moncalvo et al. (2006). Furthermore, the basidiocarps mostly have a solid stipe which also differentiates Cantharellus from Craterellus (Buyck et al. 2014). Many new taxa of Cantharellus have been published from around the world in the past two decades; for the overview of the detailed references see He et al. (2019). As of now, there are six subgenera, ca. ten sections (Buyck et al. 2014) and up to 300 species recognised in the genus (http://www.indexfungorum.org/). Species of Cantharellus are distributed worldwide but only nine species have been described from China (Chiu 1973, Zang 1980, Shao et al. 2011, 2014, 2016a, b, Tian et al. 2012, An et al. 2017, Jian et al. 2020) and a key to them was provided by Jian et al. (2020).

 As significant ectomycorrhizal (ECM) fungi (Table 2), species of Cantharellus have many host species such as the trees of Fagaceae, Pinaceae, Betulaceae, Salicaceae, Juglandaceae, Polygonaceae, Leguminosae, Phyllanthaceae, Fabaceae etc. (De Kesel et al. 2011, Kumari et al. 2011, Bahram et al. 2012, Tian et al. 2012, Buyck et al. 2012, 2014, 2016a, b, Henkel et al. 2014, Shao et al. 2014, De Kesel et al. 2016, Leacock et al. 2016, Thorn et al. 2017, Das et al. 2018, Parad et al. 2018, Hyde et al. 2019, Buyck et al. 2020). In general, the candidate host of Cantharellus is related to species, regional disparities, and varies with altitude gradient.



Cantharellus subg. Magni T. Cao & H.S. Yuan, subg. nov. MycoBank MB 839393; Fig. 2



Etymology: Magni (Lat.), as the name of the type species.



Type species: Cantharellus magnus T. Cao & H. S. Yuan, MycoBank MB 839407.



Notes: Cantharellus subg. Magni is characterised by a large basidiocarp; smooth, azonate, deep yellow to deep orange pileal surface; always incised pileal margin; decurrent and almost perfectly smooth hymenophore; broadly ellipsoid basidiospores, absence of cystidia, thin- to slightly thick-walled terminal cells of pileipellis hyphae and presence of clamps. The type species, Cantharellus magnus, is distinctly different from the species of the other six subgenera based on morphological characteristics. The subgenus Afrocantharellus Buyck & V. Hofstetter includes small to large species (up to 180 mm wide and 100 mm high, like Cantharellus splendens), and they are differentiated from C. magnus by having four-spored basidia and absence of clamps. C. magnus resembles species of subgenus Cantharellus in having abundant clamps, smooth hymenophore (partly in some species) and yellowish pileus but differs by the extremely large basidiocarps and thin- to slightly thick-walled pileipellis hyphae. The species of subgenus Rubrinus Buyck & V. Hofstetter can be obviously distinguished from Cantharellus magnus by small to medium-sized basidiocarps and absence of clamps. The species of subgenus Cinnabarinus Buyck & V. Hofstetter are similar to C. magnus in having abundant clamps but differ by the thin-walled terminal cells of the pileipellis hyphae and small to medium-sized basidiocarps (except C. afrocibarius). In addition, Cantharellus magnus differs from species of subgenus Parvocantharellus and subgenus Pseudocantharellus Buyck & V. Hofstetter by having large basidiocarps and nearly smooth hymenophore (Buyck 2014).



Cantharellus laevihymeninus T. Cao & H. S. Yuan, sp. nov. MycoBank MB 839405; Fig. 1, Fig. 2, Fig. 5, Fig. 6, Fig. 7



Etymology: Laevihymeninus (Lat.), refers to the almost perfectly smooth hymenophore.



Typus: China, Yunnan Province, Shizong County, Junzishan Mt., on soil in angiosperm and Pinus sp. mixed forest, 8 Aug. 2019, H. S. Yuan & J. R. Yu, holotype, Yuan 13902 (IFP 019441).



Diagnosis: Differs from Cantherallus hainanensis in having larger pilei (30–65 mm vs. 25–55 mm wide in C. hainanensis), discoloured stipes when injured, broader spores (5–6.1 μm vs. 4.5–5 μm wide) and shorter terminal cells (15–38.5 μm vs. 23–82 μm long) of the pileipellis.



Description: Basidiocarps concrescent, medium, fleshy, leathery when fresh, becoming soft corky and light in weight upon drying. Pilei 30–65 mm wide, convex when young, with maturity becoming slightly plano-convex and slightly depressed in the center. Pileal surface dry, subglabrous, smooth to irregularly wrinkled, pale orange to orange (5A3–5A6) when moist, drying light brown (6D6/7D5/7D6). Pileal margin slightly decurved when young, undulate with maturity, involute or irregularly folded, sometimes incised. Pileal context 0.5–3 mm thick, thin towards the pileus margin, light yellow (4A4). Hymenophore decurrent, almost perfectly smooth to a few faint ridges or folds, pale orange to light orange (5A3/5A4). Stipes central, often concrescent, confluent with pilei, 25–45 mm long, 10–18 mm wide, subcylindrical, sometimes hollow; surface glabrous to finely rugulose, orange-white (5A2/6A2) when moist, bruising darker when injured, drying brown to dark brown (6E7–6F8); stipe base slightly enlarged and with a small amount of white basal mycelium. Odour typically of apricots. Taste mild.

 Basidiospores ellipsoid, (6.8–)7.0–8.8(–9.0) × (4.8–)5.0–6.1(–6.2) μm, L_m = 7.87 μm, W_m = 5.52 μm, Q = 1.35–1.42 (n = 60/2), smooth, thin-walled, IKI–, hyaline, some with granular contents; hilar appendix 0.5–1.0 μm long. Basidia subcylindric, subclavate to clavate, 22.5–75 × 6.5–10 μm, sometimes with large guttules or finely granulose contents; sterigmata 4–6, up to 10 μm long, 1–2.5 μm wide at base, slightly curving. Basidioles cylindrical to subclavate, smaller than basidia. Subhymenium trama filamentous, hyphae 2–7 μm wide, thin- to slightly thick-walled, olive yellow in KOH. Cystidia absent. Pileipellis composed of cylindrical hyphae, 3–9 μm wide, thick-walled, interwoven, rarely branched; terminal elements rounded at apex, cells 15–38.5 × 3–10 μm. Stipitipellis composed of cylindrical hyphae, thick-walled, densely interwoven to subparallel, 4.9–7.4 μm wide, terminal elements rounded at apex. Clamp connections present.



Material examined: China. Yunnan Province, Shizong County, Junzishan Mt., on soil in angiosperm and Pinus sp. mixed forest, 8 Aug. 2019, H. S. Yuan & J. R. Yu, paratype, Yuan 13902 (IFP 019442).



Notes: The new species, Cantharellus laevihymeninus, fell in subgenus Cantharellus and together with three other smooth chanterelles viz., C. hainanensis, C. flavolateritius and C. lateritius made up section Sublaeves with a strong support (100 % ML, 1.00 BPP) based on our phylogenetic tree. Members of section Sublaeves usually share the almost smooth hymenophore (apart from C. lateritius which is only partly smooth) (Buyck 2014).

 Cantharellus hainanensis is another Chinese species in section Sublaeves and was described from Hainan Province. Morphologically, Cantharellus hainanensis is like C. laevihymeninus in having a decurrent and almost smooth hymenophore, sometimes hollow stipes, clavate to subcylindrical basidia and 4–6 sterigmata. But Cantharellus laevihymeninus is quite distinct due to its larger pilei (30–65 mm vs. 25–55 mm wide in C. hainanensis), discoloured stipes when injured, broader spores (5–6.1 μm vs. 4.5–5 μm wide) and shorter terminal cells (15–38.5 μm vs. 23–82 μm long) of the pileipellis (An et al. 2017).

 Cantharellus flavolateritius was described from North Carolina in the USA and resembles C. laevihymeninus in having a decurrent and almost smooth hymenophore, stipes bruising darker when injured, absence of cystidia and presence of clamps. However, Cantharellus flavolateritius differs from the new species by having slenderer basidiospores (4.2–5.2 μm), longer basidia (up to 85 μm), 5 sterigmata and longer terminal cells (up to 70 μm long) of the pileipellis (Buyck et al. 2016b). Cantharellus lateritius is similar to C. laevihymeninus in having yellow to orange fruit bodies, sometimes concrescent stipes, ellipsoid spores and absence of cystidia, but differs by larger basidiocarps (up to 9 cm wide and 12 cm high), partly smooth hymenophore, thin-walled hyphae in pileipellis and (3–)4–5 sterigmata (Petersen 1979a, Buyck 2014).



Cantharellus magnus T. Cao & H. S. Yuan, sp. nov. MycoBank MB 839407; Fig. 1, Fig. 2, Fig. 5, Fig. 6, Fig. 8



Etymology: Magnus (Lat.), refers to the large basidiocarps.



Typus: China, Hunan Province, Sangzhi County, Badagong Nature Reserve, on soil in angiosperm forest, 23 Sep. 2020, T. Cao & Y. L. Wei, holotype, Wei 10225 (IFP 019443).



Diagnosis: Differs from Cantherallus miomboensis in having a larger pileus (200 mm vs. 150 mm wide) and smooth hymenophore.



Description: Basidiocarps solitary, fleshy and fragile when fresh, becoming soft corky and light in weight upon drying. Pilei up to 200 mm wide, convex when young, becoming plano-convex and depressed in the center, infundibuliform when mature. Pileal surface dry, subglabrous to velutinate, smooth, azonate, deep yellow to deep orange (4A8–5A8) when moist, drying become light yellow to greyish orange (4A5–5B4). Pileal margin always incised, decurved when young, becoming strongly and irregularly folded and undulate with age. Pileal context 5–15 mm thick, yellowish white (3A2). Hymenophore decurrent, almost perfectly smooth to a few faint ridges or folds, pale yellow to greyish yellow (4A3–4B4) when fresh, pale orange to brownish orange (5A4–5C8) upon drying. Stipes central, confluent with pilei, 30–100 mm long, 6–25 mm wide, subcylindrical, somewhat curved, solid; surface subglabrous to finely rugulose, orange to yellowish white to white (3A1–3A2) when moist, drying pale yellow (4A3); stipe base equal and covered with a small amount of white basal mycelium. Odour typically of apricots. Taste mild.

 Basidiospores broadly ellipsoid, (8.5–)9.0–11.0(–11.5) ×(6.5–)6.8–7.5(–8.0) μm, L_m = 9.73 μm, W_m = 7.26 μm, Q = 1.34–1.37 (n = 60/2), smooth, thin-walled, IKI–, hyaline, some with granular contents; hilar appendix 0.5 μm long. Basidia subcylindric, subclavate to clavate, 85–120 × 10–18 μm, sometimes with large guttules or finely granulose contents; sterigmata 2–6, up to 10 μm long, 1.5–5 μm wide at base, somewhat curving. Basidioles numerous, subcylindrical to subclavate, smaller than basidia, 30–98 × 3–11 μm. Subhymenium trama filamentous, hyphae 3.5–6 μm wide, thin- to slightly thick-walled, olive yellow in KOH. Cystidia absent. Pileipellis composed of cylindrical hyphae, 6–13 μm wide, thin- to slightly thick-walled, interwoven, rarely branched; terminal elements rounded at apex, cells 92–160 × 7–15 μm. Stipitipellis composed of cylindrical hyphae, thick-walled, densely interwoven to subparallel, 7.5–11 μm wide, terminal elements rounded at apex. Clamp connections present.



Material examined: China, Hunan Province, Sangzhi County, Badagong Nature Reserve, on soil in angiosperm forest, 23 Sep. 2020, T. Cao & Y. L. Wei, paratype, Wei 10244 (IFP 019444).



Notes: Cantharellus magnus was collected from a subtropical forest in central China. The phylogenetic analysis shows that it nests in the genus Cantharellus but does not belong to any recognised subgenus. Cantharellus magnus has large basidiocarps, with a deep yellow to deep orange pileal surface, decurrent and smooth hymenophore, broadly ellipsoid spores and large, thin- to slightly thick-walled terminal cells of the pileipellis. Cantharellus magnus resembles C. afrocibarius (up to 180 mm wide) and C. miomboensis (up to 150 mm wide) in having large basidiocarps, but the new species can be differentiated from these two species by having an almost smooth hymenophore (Buyck et al. 2012).



Cantharellus subminor T. Cao & H. S. Yuan, sp. nov. MycoBank MB 839409; Fig. 1, Fig. 2, Fig. 5, Fig. 6, Fig. 9



Etymology: Subminor (Lat.), refers to the affinity with C. minor.



Typus: China, Yunnan Province, Luoping County, Huangnigou Village, on soil in angiosperm and Pinus sp. mixed forest, 9 Aug. 2019, H. S. Yuan, holotype, Yuan 13917 (IFP 019445).



Diagnosis: Differs from Cantharellus minor in the intervenose hymenophore, shorter and broader stipes (15–30 × 2–5 vs. 20–50 × 1–2 mm) and smaller basidiospores (7.8–8.8 × 5.2–5.8 vs. 6–11.5 × 4–6.5 μm).



Description: Basidiocarps solitary, soft and leathery when fresh, becoming brittle upon drying. Pilei thin, 5–15 mm wide, convex to plano-convex and slightly depressed in the center when young, becoming broadly uplifted and infundibuliform with age. Pileal surface dry, subglabrous, smooth to irregularly wrinkled, vivid yellow to light yellow (3A8/3A5) when moist, drying become brownish orange to light brown (6C6–6D7). Pileal margin entire and decurved when young, becoming plane, uplifted, sometimes incised with age. Pileal context thin, ca. 0.2 mm thick. Hymenophore decurrent, composed of low forking veins, sometimes intervenose at margin, concolorous with pileal surface. Stipes central, confluent with pilei, 15–30 mm long, 2–5 mm wide, subcylindrical, solid; surface glabrous to finely rugulose, orange to golden yellow (5B7–5B8) when moist, drying brownish orange to light brown (6C5–7D7), deeper than the pileal surface; leathery or fleshy when fresh, become hard upon drying; stipe base slightly enlarged and covered with a small amount of white basal mycelium. Odour faintly fruity. Taste mild.

 Basidiospores ellipsoid, (7.5–)7.8–8.8(–9.0) × (4.8–) 5.2–5.8 (–6.0) μm, L_m = 8.38 μm, W_m = 5.57 μm, Q = 1.47–1.50 (n = 60/2), smooth and thin-walled, IKI–, hyaline, some with granular contents; hilar appendix 0.3–0.8 μm long. Basidia subcylindric, subclavate to clavate, 43–96.5 × 6.5–10 μm, sometimes with large guttules or finely granulose contents; sterigmata 4–5, up to 10 μm long, 1–3 μm wide at base, somewhat curving. Basidioles numerous, cylindrical to subclavate, smaller than basidia, 10–62.5 × 2–10 μm. Subhymenium trama filamentous, hyphae 3–5.5 μm wide, thin- to slightly thick-walled, olive yellow in KOH. Cystidia absent. Pileipellis composed of cylindrical hyphae, 8.5–12 μm wide, thin- to slightly thick-walled, interwoven, rarely branched; terminal elements rounded at apex, cells 22–96 × 8.5–15 μm. Stipitipellis composed of cylindrical hyphae, thick-walled, densely interwoven to subparallel, 5–13.5 μm wide, terminal elements rounded at apex. Clamp connections present.



Material examined: China, Yunnan Province, Qujing City, Luoping County, Huangnigou Village, on soil in angiosperm and Pinus sp. mixed forest, 9 Aug. 2019, H. S. Yuan, paratype, Yuan 13925 (IFP 019446); Yuan 13926 (IFP 019447).



Notes: Cantharellus subminor is the smallest species of Cantharellus described from China. In the phylogenetic tree, C. subminor is embedded in subg. Parvocantharellus and forms a clade with two other small chanterelles, C. minor and C. romagnesianus.

 Morphologically, Cantharellus subminor is closely related to C. minor by having a smooth and yellowish pileal surface, small pileus (less than 15 mm wide) and 4–5 sterigmata, ellipsoid basidiospores and presence of clamps, but C. minor differs by the non-intervenose hymenophore, longer and slenderer stipes (20–50 × 1–2 mm) and larger basidiospores (6–11.5 × 4–6.5 μm) (Peck 1873, Lao et al. 2019). Cantharellus romagnesianus is like C. subminor in having subcylindrical stipes, subcylindric basidia, absence of cystidia and presence of clamps, however, it is differentiated by having a larger pileus (up to 25 mm wide), discoloured context when injured, slenderer basidiospores (4.5–5 μm wide) and 5–6 sterigmata (Eyssartier & Buyck 1999). Cantharellus albus is another Chinese species embeded in subg. Parvocantharellus, but it can be distinctly differentiated from C. subminor in having a larger pileus (up to 50 mm wide and 70 mm high) and snow white basidiocarps (Jian et al. 2020). The Mexican Cantharellus parvoflavus M. Herrera, Bandala & Montoya is also a member of subg. Parvocantharellus and has small size basidiocarps as C. subminor, however, it differs from C. subminor by having orangish pilei, slenderer basidiospores (Q = 1.52–1.57 vs. 1.47–1.50), shorter basidia (50–89 vs. 43–96.5 μm long) and smaller terminal elements (23–80 × 3.5–8 vs. 22–96 × 8.5–15 μm) of pileipellis. Besides, the similarity of TEF1 sequences between C. parvoflavus and C. subminor is 94.69 %.



Cantharellus yunnanensis W.F. Chiu, Acta Microbiol. Sin. 13(2): 129. 1973. MycoBank MB 310378.



Synonym: Cantharellus tuberculosporus M. Zang, Acta Microbiol. Sin. 20(1): 31. 1980. MycoBank MB 118474.

Cantharellus anzutake W. Ogawa, N. Endo, M. Fukuda and A. Yamada, Mycoscience 59: 158. 2017. MycoBank MB 813057.



Typus: China, Yunnan Province, Kunming City, Xishan Forest Park, on soil in angiosperm and Pinus sp. mixed forest, 11 Aug. 1942, S. J. Shen, holotype, Tsinghua 8090 (HMAS 4090).



Materials examined: China, Yunnan Province, Kunming City, Xishan Forest Park, on soil in angiosperm and Pinus sp. mixed forest, 15 Aug. 2019, T. Cao, Yuan 13983 (IFP 019448), 13985 (IFP 019449); Liaoning Province, Fushun City, Xinbin County, Gangshan Mt., on soil in angiosperm and Pinus sp. mixed forest, 12 Aug. 2020, H. S. Yuan, Yuan 14539 (IFP 019450); 14636 (IFP 019451).



Notes: Cantharellus yunnanensis is the first species of the genus to be described from China (Chiu 1973). Shao et al. (2021) have selected the epitype (Herrera 263C) and redescribed the species. In this study, we collected several samples (Yuan 13983 and Yuan 13985) from Xishan Forest Park of Kunming, where the type material of C. yunnanensis (HMAS 4090, Tsinghua 8090) and epitype (Herrera 263C) were collected and we recognised them as the same taxa as C. yunnanensis based on phylogenetical and morphological evidences. Phylogenetic analyses shows that four samples (Yuan 13983, 13985, 14539, 14363) which were collected from Liaoning Province, group with C. yunnanensis (XieXD 174), C. tuberculosporus (HKAS58195 and HKAS58196) and C. anzutake (TNS-F-61925). The eight samples form a strongly supported (84 % in ML, 0.99 BPP) isolated lineage (Fig. 2). Furthermore, Yuan 13983 showed a high similarity of TEF1 sequences to KU720337 (XieXD 174), KM893834 (HKAS58195), KM893835 (HKAS58196) and LC179800 (TNS-F-61925) with 99.01 %, 99.12 %, 99.01 % and 98.76 % respectively.

 According to the original description, Cantharellus yunnanensis possesses small basidiocarps with a white to pale salmon hymenophore and small basidiospores (4–5 × 2–3.5 μm) (Chiu 1973). The verification of the epitype specimen (Herrera 263C) shows that the basidiospore size, 6.5–8.5 × 5–6.5 μm overlaps the spore size of the newly collected specimens Additionally, according to the observations of the specimens (epitype, Yuan 13983 and Yuan 13985), the size of the basidiocarps of C. yunnanensis is also larger than the original description, and the hymenophore can be white, pale salmon or even yellow in some individuals.

 Cantharellus anzutake, described from Japan, was collected from a forest of Japanese red pine and is characterised by a pale yellow to orange-yellow pileal surface, white to pale yellow hymenophore, 4–6 spored basidia and ellipsoid basidiospores. The spore size of the C. yunnanensis epitype (Herrera 263C, 6.5–8.5 × 5–6.5 μm), Yuan 13983 and Yuan 13985 (7.5–9 × 5–6.5 μm) overlaps with those of C. anzutake (5.8–9.2 × 4–6.3 μm), and the pileal and hymenophore colour of C. anzutake often change with weather conditions; having a pale-yellow pileus and white hymenophore in drier conditions or yellow in wet is very similar to C. yunnanensis (Ogawa et al. 2018). Phylogenetic analyses (Fig. 2) suggest the type Cantharellus anzutake (TNS-F-61925) and several samples of C. yunnanensis belong to a separate lineage which confirms C. anzutake is the later synonym of C. yunnanensis.

 Cantharellus tuberculosporus was described from Xizang, China. According to the original description, the species is characterised by a bright yellow pileal surface and hymenophore as well as tuberculate basidiospores (Zang 1980). Shao (2011) corrected the type specimen number (HKAS5412) to HKAS 28930 and redescribed the species as the spores of HKAS 28930 are smooth, 7–8 × 5–6 μm, and obviously different from the tuberculate spores in the original description. The tuberculate spores may be due to the shrinkage caused by dehydration (Shao 2011). The anatomical features of C. tuberculosporus are closely related to C. anzutake and C. yunnanensis. In addition, the macroscopic morphology of the fruitbody and molecular evidence in our study both strongly support that C. tuberculosporus is the same species as C. anzutake and C. yunnanensis. Cantharellus yunnanensis has priority, therefore, C. tuberculosporus and C. anzutake are later synonyms of C. yunnanensis.



Craterellus Pers., Mycol. Eur. (Erlanga) 2: 4. 1825. MycoBank MB 17398.



Synonym: Pseudocraterellus Corner, Beih. Sydowia 1: 268. 1958. MycoBank MB 18388.

Pterygellus Corner, Monogr. Cantharelloid Fungi: 166. 1966. MycoBank MB 18424.



Type species: Craterellus cornucopioides (L.) Pers., Mycol. Eur. (Erlanga) 2: 5. 1825. MycoBank MB 153130.



Notes: Craterellus was described in 1825, with C. cornucopioides as the type species (Persoon 1825). The genus is traditionally characterised by funnel-shaped basidiocarps with a hollow stipe that may also be much reduced (Petersen 1979a). The genus belongs to Hydnaceae according to Hibbett et al. (2014) which is also supported by our analysis based on dataset 4 (this study provides four markers for genus Craterellus: nLSU, ITS, RPB2 and TEF1) (Fig. 1). Craterellus with the sister genus Cantharellus form a fully supported lineage which is closely related to the genera Hydnum and Sistotrema. Pseudocraterellus and Pterygellus Corner, two other genera in Hydnaceae, have been recognised as synonyms of Craterellus. Although in the tree (Fig. 3) three samples of Pseudocraterellus have been appropriately embeded in Craterellus, the validity of the recombined species and research on typification in the two genera are needed to confirm their status (Feibelman et al. 1997, Yomyart et al. 2012, Henkel et al. 2014, Hembrom et al. 2017). Approximately 140 taxa names are recorded for Craterellus, and up to 70 species are currently accepted (http://www.indexfungorum.org). The species in the genus often possess an ectomycorrhizal nutritional mode and are distributed worldwide. Most of the known species are edible and show diversity in colour (Dahlman et al. 2000, Dunham et al. 2003, Porter et al. 2008, Wright et al. 2009, Matheny et al. 2010, Wilson et al. 2012, Osmundson et al. 2013, Henkel et al. 2014, Raja et al. 2017, Bijeesh et al. 2018, Zhong et al. 2018). Up to now, there are only four species described from China (Berkeley & Curtis 1860, Zhong et al. 2018, Zhang et al. 2020, Cao et al. 2021).

 Although the molecular evidence has been widely used in taxonomy of Craterellus since Dahlman et al. (2000), most studies do a phylogenetic analysis using only nLSU or ITS sequences but our study is based on the combined nLSU and ITS dataset (dataset 2). There are six distinct clades with high support have been recognised in the phylogenetic tree (Fig. 3). The species in the same clade often share several similar morphological characteristics, thus we propose them as six subgenera in the genus Craterellus.



Craterellus subg. Cariosi T. Cao & H. S. Yuan, subgen. nov. MycoBank MB 839396; Fig. 3



Etymology: Cariosi (Lat.), refers to the habit of fruiting on decayed wood.



Type species: Craterellus pleurotoides (T.W. Henkel, Aime & S.L. Mill.) A.W. Wilson, Mycologia 104: 1475. 2012. MycoBank MB 510239.



Notes: The subgenus consists of two South American species, Craterellus pleurotoides and C. olivaceoluteus. Although Craterellus is an ECM genus, the two species in subgen. Cariosi, C. pleurotoides and C. olivaceoluteus both fruit on very decayed wood and they also share the characteristics of small-sized basidiocarps, smooth hymenophore and presence of clamps (Henkel et al. 2006, 2014).



Craterellus subg. Craterellus MycoBank MB 839401; Fig. 3



Etymology: Craterellus (lat.), refers to the subgenus in which the type species of the genus is located.



Type species: Craterellus cornucopioides (L.) Pers., Mycol. Eur. (Erlanga) 2: 5. 1825. MycoBank MB 153130.



Notes: This subgenus includes the genus type Craterellus cornucopioides as well as five other ‘black trumpet’ species viz., C. badiogriseus, C. caeruleofuscus, C. croceialbus, C. macrosporus and C. squamatus, and they all have tuberiform to infundibuliform blackish brown basidiocarps. However, the yellow individuals (e.g., C. konradii Bourdot & Maire) rule out blackish brown basidiocarps as an iconic feature of this group. The species in clade Craterellus often have a fully perforated pileus with smooth or wrinked hymenophore and lack distinct stipes. Although lacking molecular evidence, we suspect other species in the C. cornucopioides complex (e.g., C. cornucopioides var. cornucopioides (L.) Pers., C. cornucopioides var. crispus Sacc., C. cornucopioides var. flavicans Sacc., C. cornucopioides var. mediosporus Corner, C. cornucopioides var. parvisporus Heinem., C. cornucopioides var. roseus R. Heim, C. philippinensis Bres. and C. verrucosus Massee) also belong to subgenus Craterellus. Craterellus cornucopioides, C. fallax, C. macrosporus and C. squamatus form a well-supported subclade in Clade III and they share the large basidiospore size (up to 14 μm long) (Smith 1968, Gulden & Høiland 1989, Matheny et al. 2010).



Craterellus subg. Imperforati T. Cao & H. S. Yuan, subgen. nov. MycoBank MB 839397; Fig. 3



Etymology: Imperforati (Lat.), refers to the non-perforated pileus.



Type species: Craterellus carolinensis R.H. Petersen, Persoonia 5(2): 217. 1968. MycoBank MB 329222.



Notes: This subgenus is comprised of our new species Craterellus badiogriseus and eight other species viz., C. albidus, C. albostrigosus, C. carolinensis, C. hesleri, C. indicus, C. inusitatus, C. parvogriseus, C. shoreae and Pseudocraterellus sinuosus (Fig. 3). They often have variably coloured pilei but share the small basidiocarps (less than 40 mm wide and high), non-perforated pilei, smooth or slightly wrinked hymenophore, broadly ellipsoid basidiospores and absence of clamps (Reid 1962, Petersen 1969, Petersen 1975, Deepika et al. 2012, Henkel et al. 2014, Das et al. 2017, Hembrom et al. 2017, Bijeesh et al. 2018, Zhang et al. 2020).



Craterellus subg. Lamelles T. Cao & H. S. Yuan, subgen. nov. MycoBank MB 839398; Fig. 3



Etymology: Lamelles (Lat.), refers to the hymenophore with folds or well-developed false gills.



Type species: Craterellus cinereus R.H. Petersen, Mycol. Eur. (Erlanga) 2: 6. 1825. MycoBank MB 357303.



Notes: There are five species in subgenus Lamelles viz., Craterellus cinereus, C. ignicolor, C. lutescens, C. melanoxeros and C. tubaeformis. They often have medium to large basidiocarps, perforated or occasionally perforated pilei and presence of clamps. Most species in this clade have a hymenophore with folds or well-developed false gills, but C. lutescens has a smooth hymenophore (Dahlman et al. 2000, Redhead et al. 2002, Contu et al. 2009).



Craterellus subg. Longibasidiosi T. Cao & H. S. Yuan, subgen. nov. MycoBank MB 839399; Fig. 3



Etymology: Longibasidiosi (Lat.), refers to the long basidia.



Type species: Craterellus excelsus T.W. Henkel & Aime, Mycotaxon 107: 202. 2009. MycoBank MB 510899.



Notes: There are two species in this subgenus characterised by medium to large basidiocarps (up to 65 mm wide in Craterellus cinereofimbriatus and 150 mm in C. excelsus), greyish brown and often perforated pilei, smooth hymenophore, long basidia (up to 100 μm long), broadly ellipsoid basidiospores and absence of clamps. Besides, both species grow on soil under Dicymbe corymbosa (Henkel et al. 2009, 2014).



Craterellus subg. Ovoidei T. Cao & H. S. Yuan, subgen. nov. MycoBank MB 839400; Fig. 3



Etymology: Ovoidei (Lat.), refers to the narrowly ellipsoid to ovoid basidiospores.



Type species: Craterellus odoratus (Schwein.) Fr., Epicr. Syst. Mycol. (Upsaliae): 532. 1838. MycoBank MB 190345.



Notes: Craterellus luteus and C. odoratus constitute sugenus Ovoidei and the perforated pilei, smooth hymenophore, narrowly ellipsoid to ovoid basidiospores (8.5–12.5 × 5.7–7.8 μm in C. luteus and 8.9–11.8 × 4.4–6.3 μm in C. odoratus) and absence of clamps are the common features. In addition, the two species both have brightly coloured basidiocarps (light yellow and bright orange) (Petersen 1979b, Zhong et al. 2018).



Craterellus badiogriseus T. Cao & H. S. Yuan, sp. nov. MycoBank MB 839410; Fig. 1, Fig. 3, Fig. 5, Fig. 6, Fig. 10



Etymology: Badiogriseus (Lat.), refers to the brownish grey pileal surface.



Typus: China, Liaoning Province, Qingyuan County, on soil in angiosperm and Pinus sp. mixed forest, 7 Sep. 2019, H. S. Yuan, holotype, Yuan 14776 (IFP 019452).



Diagnosis: Differs from Craterellus indicus in having a brownish grey to black pileal surface, longer basidia (55–106 vs. 48–85 μm long) and broader terminal cells of pileipellis hyphae (5–10 vs. 2.5–8 μm long).



Description: Basidiocarps concrescent, infundibuliform, leathery when fresh, becoming brittle and light in weight upon drying. Pilei 10–15 mm wide, convex-expanded towards the margin, slightly or deeply depressed in the center but not perforated. Pileal surface dry, velutinate, brownish grey (8F2) to black when moist, drying greyish brown (7D3–7F3). Pileal margin thin, slightly involute or irregularly involute, orange-grey (5B2), lighter than pileal surface. Pileal context 0.3–0.5 mm thick, orange-grey to brownish orange (5B2–5C3). Hymenophore decurrent, smooth, brownish grey to grey (4C1–4C2) when moist drying yellowish grey to orange-grey (4B2–5B2). Stipes 10–15 mm long, 3–5 mm wide, subcylindrical, hollow; surface glabrous, concolorous with hymenophore; stipe base equal or slightly enlarged and rarely covered with basal mycelium. Odour fruity.

 Basidiospores broadly ellipsoid, (7.5–)8.0–10.5(–11.0) × (6.5–)6.8–7.5(–8.0) μm, L_m = 9.35 μm, W_m = 7.33 μm, Q = 1.25–1.28 (n = 60/2), smooth, thin-walled, IKI–, hyaline, some with granular contents, inamyloid; hilar appendix 0.3–0.8 μm long. Basidia subcylindric, subclavate to clavate, 55–106 × 8–12 μm, sometimes with large guttules or finely granulose contents; sterigmata 2–4, up to 10 μm long, 2–5 μm wide at base, slightly curving. Basidioles numerous, subcylindrical to subclavate, smaller than basidia, 15–95 × 5–10 μm. Subhymenium trama filamentous, hyphae 3–5 μm wide, thick-walled, olive yellow in KOH; pileal trama hyphae 3–6 μm wide, thick-walled, secondary septation absent. Cystidia absent. Pileipellis composed of cylindrical hyphae, 5–15 μm wide, thick-walled, interwoven to subparallel, rarely branched; terminal elements rounded at apex, cells 20–65 × 6–15 μm. Clamp connections absent.



Material examined:China, Liaoning Province, Qingyuan County, on soil in angiosperm and Pinus sp. mixed forest, H. S. Yuan, 7 Sep. 2019, paratype, Yuan 14779 (IFP 019453).



Notes: The new species, Craterellus badiogriseus, was discovered from a temperate forest in northeast China. In the phylogenetic tree, Craterellus badiogriseus and three samples of Pseudocraterellus Corner form a lineage with strong support (100 % ML and 0.99 BPP). Although Pseudocraterellus was regarded as a later synonym of Craterellus by Feibelman et al. (1997), the molecular and morphological evidence of the type material in the genus are unconvincing. P. sinuosus is similar to C. badiogriseus in having dark greyish brown and non-perforated pileus, a smooth hymenophore and absence of clamps. However, it can be distinctly differentiated from the new species in having a larger pileus (12–25 mm wide), longer stipes (up to 37 mm long) and presence of secondary hyphal septation (Henkel et al. 2014).

 Craterellus badiogriseus and Pseudocraterellus group with two other Asiatic species C. indicus and C. parvogriseus and form a large clade with support, 87 % in ML and 0.95 BPP. Craterellus badiogriseus resembles C. indicus in having a non-perforated pileus, smooth hymenophore, broadly ellipsoid basidiospores, 2–4 sterigmata and absence of clamps, but the latter differs from the new species by the light brownish pileal surface, shorter basidia (48–85 μm long) and slenderer terminal cells (2.5–8 μm wide) of the pileipellis hyphae. Craterellus parvogriseus is related to C. badiogriseus in having a brownish grey pileal sueface, broadly ellipsoid basidiospores and absence of clamps, however, the former can be distinguished from C. badiogriseus by the irregularly folded hymenophore, longer stipes (up to 27 mm long), 2–6 sterigmata and smaller basidia (47–78 × 8–9 μm) (Das et al. 2017).



Craterellus croceialbus T. Cao & H. S. Yuan, sp. nov. MycoBank MB 839411; Fig. 1, Fig. 3, Fig. 5, Fig. 6, Fig. 11



Etymology: Croceialbus (Lat.), refers to the orange-white pileal margin.



Typus: China, Liaoning Province, Xinbin County, Gangshan Mt., on soil in angiosperm and Pinus sp. mixed forest, 26 Aug. 2020, H. S. Yuan, Yuan 14623 (holotype IFP 019454).



Diagnosis: Differs from Craterellus cornucopioides in having smaller basidiocarps (up to 2 cm vs. 15 cm wide), 2–4 sterigmata and smaller basidiospores (up to 12.5 μm vs. 14 μm long).



Description: Basidiocarps solitary to concrescent, infundibuliform, leathery when fresh, becoming brittle upon drying. Pilei 10–20 mm wide, convex-expanded towards the margin, broadly and deeply depressed in the center and perforation continuous with hollow stipe. Pileal surface dry, glabrous to velutinate, smooth, brownish grey to greyish brown (6D2–6D3) when moist, drying greyish brown (6F3–7E3). Pileal margin thin, slightly revolute and undulate, orange-white (5A2), distinctly lighter than pileal surface. Pileal context 0.3–1 mm thick, orange-white (5A2). Hymenophore decurrent, almost perfectly smooth to having a few faint ridges or folds, moist pale grey to grey (1B1–3B1), drying yellowish grey to orange-grey (4B2–5B2). Stipes indistinct, confluent with pileus, 20–35 mm long and 3–8 mm wide, subconic, hollow; surface glabrous, concolorous with the hymenophore; stipe base equal or slightly enlarged and rarely covered with basal mycelium. Odour fruity.

 Basidiospores ellipsoid, (9.0–)10.0–12.0(–12.5) × (6.5–)6.8–8.0(–8.2) μm, L_m = 10.31 μm, W_m = 7.33 μm, Q = 1.41–1.44 (n = 60/2), smooth, thin-walled, IKI–, hyaline, some with granular contents, inamyloid; hilar appendix 0.5 μm long. Basidia subcylindric, subclavate to clavate, 60–85 × 10–12 μm, sometimes with large guttules or finely granulose contents; sterigmata 2–4, up to 10 μm long, 1–4 μm wide at base, somewhat curving. Basidioles numerous, subcylindrical to subclavate, smaller than basidia, 16–75 × 5–10 μm. Subhymenium trama filamentous, hyphae 3–5 μm wide, thick-walled, brownish yellow in KOH; pileal trama hyphae 3–5.5 μm wide, thick-walled, secondary septation absent. Cystidia absent. Pileipellis composed of cylindrical hyphae, 9–20 μm wide, thin- to slightly thick-walled, subparallel, rarely branched; terminal elements rounded at apex, cells 35–75 × 10–23 μm. Clamp connections absent.



Material examined: China, Liaoning Province, Fushun City, Xinbin County, Gangshan Mt., on soil in angiosperm and Pinus sp. mixed forest, 26 Aug. 2020, H. S. Yuan, paratype, Yuan 14647 (IFP 019455).



Notes: Craterellus croceialbus resembles C. cornucopioides in the brownish grey pileus, hollow stipes, ellipsoid basidiospores and absence of clamps, but the latter species differs from C. croceialbus in having larger basidiocarps (pileus up to 15 cm wide), basidia with 2 sterigmata and larger basidiospores (up to 14 μm long) (Smith 1968, Smith et al. 1979, Gulden & Høiland 1989, Hansen & Knudsen 1997, Matheny et al. 2010, Kumari et al. 2011, Tibuhwa 2018).



Craterellus macrosporus T. Cao & H. S. Yuan, sp. nov. MycoBank MB 839412; Fig. 3, Fig. 5, Fig. 6, Fig. 12



Etymology: Macrosporus (Lat.), refers to the large basidiospores.



Typus: China, Liaoning Province, Qingyuan County, on soil in angiosperm and Pinus sp. mixed forest, 7 Sep. 2019, T. Cao, holotype, Yuan 14782 (IFP 019456).



Diagnosis: Differs from Craterellus cornucopioides in having smaller basidiocarps (less than 35 mm wide and 60 mm high), velutinate and smooth pileal surface and broader basidiospores (up to 11.5 μm wide).



Description: Basidiocarps solitary, infundibuliform, leathery when fresh, becoming brittle and light in weight upon drying. Pilei 30–35 mm wide, convex-expanded towards the margin, broadly and deeply depressed in the center and perforation continuous with hollow stipe. Pileal surface dry, glabrous to velutinate, smooth, brownish grey to greyish brown (5C2–5D3) when moist, drying brown (6E4–6E5). Pileal margin thin, slightly revolute and undulate, brownish grey (7F2–8F2), deeper than pileal surface. Pileal context 1–2 mm thick, brownish grey to greyish brown (8E2–8E3). Hymenophore decurrent, almost perfectly smooth or forming a few faint ridges or folds, grey (3B1) when moist, drying greyish brown to brownish grey (6D3–7C2). Stipes indistinct, confluent with pilei, 55–60 mm long and 8–15 mm wide, subconic, slightly curved, hollow; surface glabrous, concolorous with the hymenophore; stipe base equal or slightly enlarged and rarely with basal mycelium. Odour fruity.

 Basidiospores broadly ellipsoid, (12.5–)12.8–14.5(–15.0) × (8.8–)9.0–11.0(–11.5) μm, L_m = 13.46 μm, W_m = 10.27 μm, Q = 1.31–1.37 (n = 30/1), smooth, thin-walled, IKI–, hyaline, some with granular contents, inamyloid; hilar appendix 0.5 μm long. Basidia subcylindric, subclavate to clavate, 80–105 × 9–13.5 μm, sometimes with large guttules or finely granulose contents; sterigmata 2, up to 10 μm long, 3–5 μm wide at base, slightly curving. Basidioles numerous, subcylindrical to subclavate, smaller than basidia, 13–85 × 5–10 μm. Subhymenium trama filamentous, hyphae 3–5 μm wide, thick-walled, pale yellow in KOH; pileal trama hyphae 4–5 μm wide, thick-walled, secondary septation absent. Cystidia absent. Pileipellis composed of cylindrical hyphae, 10–23 μm wide, thin-walled, subparallel, rarely branched; terminal elements rounded at apex, cells 35–98 × 10–25 μm. Clamp connections absent.



Notes: Craterellus macrosporus resembles C. cornucopioides in the infundibuliform basidiocarps, dark brownish pileus with almost smooth hymenophore, hollow stipes, absence of clamps, and basidia with 2 sterigmata, but C. cornucopioides differs in having scaly pileal surfaces, larger basidiocarps (up to 15 cm wide), shorter basidia (less than 96 μm long) and narrower basidiospores (less than 9 μm wide) (Smith 1968, Smith et al. 1979, Gulden & Høiland 1989, Hansen & Knudsen 1997, Matheny et al. 2010, Kumari et al. 2011, Tibuhwa 2018). The large (up to 15 μm long) spores of Craterellus macrosporus are like those of C. konradii. The European Craterellus konradii has the habit of C. cornucopioides and may be a member of the C. cornucopioides complex. However, Craterellus konradii has yellowish basidiocarps (Gulden & Høiland 1989). Craterellus cornucopioides var. parvisporus Heinem. is similar to C. macrosporus in the blackish brown pileal surface but differs by the smaller basidiospores (6.8–8.5 × 4.3–6 μm) (Heinemann 1958). Another new species from a temperate forest in northeast China, Craterellus croceialbus, is similar to C. macrosporus in having greyish brown basidiocarps, smooth pileal surface, grey hymenophore and absence of clamps, but C. macrosporus differs from C. croceialbus in having a larger pileus (30–35 mm wide), longer and wider stipes (55–60 mm long and 8–15 mm wide), larger basidiospores (12.8–14.5 × 9–11 μm), and longer basidia (80–105 × 9–13.5 μm) with 2 sterigmata.



Craterellus squamatus T. Cao & H. S. Yuan, sp. nov. MycoBank MB 839413; Fig. 1, Fig. 3, Fig. 5, Fig. 6, Fig. 13



Etymology: Squamatus (Lat.), refers to the scaly pileus.



Typus: China, Liaoning Province, Kuandian County, Baishilazi Nature Reserve, ground in angiosperm forest, 8 Aug. 2020, H.S. Yuan, holotype, Yuan 14520 (IFP 019457).



Diagnosis: Differs from Craterellus cornucopioides in having smaller basidiocarps (pileus less than 13 mm wide and stipes less than 35 mm long), broader basidiospores (up to 10 μm long) and basidia with 2–4 sterigmata.



Description: Basidiocarps solitary to concrescent, tuberiform to infundibuliform, leathery when fresh, becoming brittle and light in weight upon drying. Pilei 5–15 mm wide, broadly and deeply depressed in the center and perforation continuous with hollow stipe. Pileal surface dry, glabrous, grey, light brown to dark brown (6B1/7D4/7F2) when moist, drying become dark brown (7F3–7F4); scabrous, often with darker brown raised scales. Pileal margin thin, slightly undulate, decurved or involute. Pileal context 1–3 mm thick, orange-white to orange-grey (5A2–5B2). Hymenophore decurrent, almost perfectly smooth or forming a few faint ridges or folds, orange-grey (5B1–6B2). Stipes indistinct, confluent with pilei, 15–35 mm long, 2–5 mm wide, subcylindrical, somewhat curved and irregular, hollow; surface glabrous, concolorous with the hymenophore; stipe base enlarged and covered with white basal mycelium. Odour fruity.

 Basidiospores ellipsoid, (11.5–)12.0–13.8(–14.0) × (8.2–)8.5–9.5(–10.0) μm, L_m = 13.03 μm, W_m = 8.89 μm, Q = 1.46–1.51 (n = 60/2), smooth, thin-walled, IKI–, hyaline, some with granular contents, inamyloid; hilar appendix 0.5–0.8 μm long. Basidia subcylindric, subclavate to clavate, 35–62 × 5–7.5 μm, sometimes with large guttules or finely granulose contents; sterigmata 2–4, up to 5 μm long, 1.5–3 μm wide at base, slightly curving. Basidioles numerous, subcylindrical to subclavate, smaller than basidia, 13.5–45 × 3–6.2 μm. Subhymenium trama filamentous, hyphae 3–8 μm wide, thick-walled, olive yellow in KOH; pileal trama hyphae 4–8 μm wide, thick-walled, secondary septation absent. Cystidia absent. Pileipellis composed of cylindrical hyphae, 6–15 μm wide, thick-walled, interwoven to subparallel, frequently branched; terminal elements rounded at apex, cells 15–53.5 × 9–20 μm. Clamp connections absent.



Material examined: China, Liaoning Province, Kuandian County, Baishilazi Nature Reserve, on soil in angiosperm forest, 3 Sep. 2020, H. S. Yuan, paratype, Yuan 14721 (IFP 019458).



Notes: Morphologically, Craterellus squamatus is reminiscent of the C. cornucopioides complex clade (including C. cornucopioides and C. fallax A.H. Sm.) (Dahlman et al. 2000, Moncalvo et al. 2006) which is also confirmed by the molecular evidence. In the phylogenetic tree (Fig. 3), C. squamatus and C. macrosporus clustered together with the C. cornucopioides complex and formed a group with moderate support (77 % ML and 0.96 BPP). Craterellus squamatus is similar to the genus type species, C. cornucopioides, in having a dark-coloured pileus (except for the yellow individuals from Europe) (Dahlman et al. 2000), scaly pileal surface, almost perfectly smooth hymenophore, hollow stipes and absence of clamp connections. However, the latter species differs from C. squamatus by the larger basidiocarps (up to 15 cm wide), longer basidia (up to 96 μm long) and basidia with 2 sterigmata (Smith 1968, Smith et al. 1979, Gulden & Høiland 1989, Hansen & Knudsen 1997, Matheny et al. 2010, Kumari et al. 2011, Tibuhwa 2018). Additionally, there is the molecular similarity of ITS sequences between C. squamatus (Yuan 14520, holotype) and the two sequences of C. cornucopioides (KT693262 and UDB000053) with 95.43 % and 94.95 %, respectively. Craterellus fallax is regarded as a more recent synonym of C. cornucopioides. The morphological features of C. fallax are very similar to the latter, expect for the yellowish to orangy spore print of C. fallax (which in C. cornucopioides is white) and the presence of secondary septation (which in C. cornucopioides is absent) (Smith 1968, Petersen 1975, Bigelow 1978). However, phylogenetic analysis supports it as a species distinct from C. cornucopioides (Matheny et al. 2010). The presence of the secondary septation in C. fallax distinctly distinguishes it from C. squamatus and three new species of Craterellus in this study. Among the described varieties of Craterellus cornucopioides, several species also have black pilei, but the European C. cornucopioides var. cornucopioides (L.) Pers. can be distinguished by its regularly bi-sterigmate and long basidia (up to 100 μm long) (Corner 1966), the Malaysian C. cornucopioides var. mediosporus Corner differs by the 6-sterigmate basidia and blackish brown hymenophore (Corner 1966) and the Congolese C. cornucopioides var. parvisporus has smaller basidiospores (6.8–8.5 × 4.3–6 μm) than C. squamatus (Heinemann 1958).

 Craterellus croceialbus, C. squamatus, C. macrosporus, and the C. cornucopioides complex clustered together and formed a large subclade with strong support (96 % ML and 0.95 BPP). Craterellus croceialbus is closely related to C. squamatus in having an almost perfectly smooth hymenophore, hollow stipes, basidia with 2–4 sterigmata, and absence of cystidia. However, Craterellus croceialbus can be distinctly differentiated from C. squamatus by the larger pilei (10–20 mm), velutinate and smooth pileal surface, smaller spores (10–12 × 6.8–8 μm), larger basidia (60–85 × 10–12 μm) and larger terminal cells (35–75 × 10–23 μm) of the pileipellis hyphae. Craterellus macrosporus and C. squamatus form a well-supported lineage (Fig. 3). The similarity of the ITS sequences between C. squamatus and C. macrosporus is 97.70 % and they share several morphological characteristics including the infundibuliform basidiocarps, almost perfectly smooth hymenophore, absence of cystidia and clamp connections. But C. macrosporus differs C. squamatus by having a larger plieus (30–35 mm wide), longer and wider stipes (55–60 mm long and 8–15 mm wide), smooth pileal surface, larger basidia (80–105 × 9–13.5 μm) with 2 sterigmata, broader basidiospores (Q = 1.31–1.37) and larger terminal cells (35–98 × 10–25 μm) of pileipellis hyphae. Craterellus macrosporus is a separate species distinctly different from C. squamatus based on the phylogenetic and morphological analyses.



Hydnum L., Sp. pl. 2: 1178. 1753. MycoBank MB 17797.



Synonym: Malacodon Bataille, Bull. Soc. Mycol. Fr. 39: 203. 1923. MycoBank MB 22227.



Type species: Hydnum repandum L., Sp. Pl. 2: 1178. 1753. MycoBank MB 225014.



Notes: Hydnum typified by H. repandum and traditionally characterised by stipitate and pileate basidiocarps with aculeate hymenophore, stichic basidia, fleshy and azoned monomitic context, smooth and subglobose to obovoid-elliptic basidiospores and usually the absence of cystidia (Vizzini et al. 2013, Niskanen et al. 2018, Swenie et al. 2018). Hydnum form ECM associations with a variety of host species including members of Betulaceae, Dipterocarpaceae, Fagales, Magnoliaceae, Malvaceae, Myrtaceae, Pinaceae, Salicaceae and Ulmaceae (McNabb 1971, Agerer et al. 1996, Lee et al. 2002, Feng et al. 2016, Niskanen et al. 2018, Swenie et al. 2018).

 The genus was placed in Cantharellales by Kreisel (1969) based on its stichic basidia and this was confirmed by several subsequent molecular analyses (Pine et al. 1999, Moncalvo et al. 2006, Matheny et al. 2007, Hibbett et al. 2014). As the type genus of Hydnaceae, Hydnum is the sister clade of Sistotrema confluens-subconfluens lingeage and closely related to Cantharellus as well as Craterellus in the tree (Fig. 1) which is similar to a previous study (Hibbett et al. 2014). Four subgenera, four sections and several subsections have been recognised within the genus (Niskanen et al. 2018), the four subgenera, section Hydnum, section Olympica, and five subsections get strong support in the tree (Fig. 4) which is similar to Niskanen et al. (2018).

 There are up to 900 taxa bearing the name Hydnum (http://www.indexfungorum.org), however, only ca. 40 species from Europe, North America and Asia have been described using modern molecular phylogenetic analyses and morphological features (Grebenc et al. 2009, Olariaga et al. 2012, Vizzini et al. 2013, Yanaga et al. 2015, Buyck et al. 2017, Niskanen et al. 2018, Swenie et al. 2018, Wang et al. 2018). Feng et al. (2016) estimated the global survey of diversity in Hydnum and recognised at least 31 phylogenetic species from Asia, Central America (Honduras), Europe, North America, Oceania, and South America (Venezuela) according to molecular evidence, but the samples in Africa are poorly investigated. Around less than half of the global diversity of Hydnum has been discovered and the diversity and distribution of Hydnum should be further explored (Niskanen et al. 2018).

 Until 2016, the few species of Hydnum in China that been recorded were H. repandum, H. repandum var. album and H. rufescens (Le et al. 1993, Bi et al. 1994, Zang et al. 1996). Feng et al. (2016) recognised at least 19 taxa from China based on molecular phylogeny evidence, among which around 6 taxa fell in the named-clade and 13 lineages/taxa are non-named and seem to be new taxa. Several samples were described as H. berkeleyanum, H. jussii and H. cremeoalbum (Niskanen et al. 2018, Wang et al. 2018). We added 16 samples from Feng et al. (2016) in our study and the phylogenetic tree of the genus Hydnum shows the current status of Hydnum species in China.



Hydnum subg. Brevispina T. Cao & H. S. Yuan, subgen. nov. MycoBank MB 839402; Fig. 4



Etymology: Brevispina (Lat.), following the name of the type species.



Type species: Hydnum brevispinum T. Cao & H. S. Yuan. MycoBank MB 839417.



Notes: Two species, Hydnum brevispinum and H. tenuistipitum, comprise the subgenus and they share the following features: basidiocarps small to medium, solitary or concrescent, fleshy when fresh; pilei rounded, pileal surface velutinate, smooth, azonate to subzonate, pure whitish to yellowish white or orange-white, pileal margin often entire and slightly decurved; spines non-decurrent to subdecurrent; basidia with 2–6 sterigmata; basidiospores often subglobose to broadly ellipsoid, cystidia absent, terminal cells of pileipellis hyphae thick-walled, stipitipellis hyphae slightly thick-walled and clamps present. Hydnum subgenus Alba resembles subg. Brevispinum in having whitish basidiocarps, non-decurrent to subdecurrent spines but differs by having broader basidiospores (Q avg. = 1.00–1.10). Besides, subg. Alba contains some species with large basidiocarps (e.g., H. albomagnum) while subg. Brevispinum only has (very) small to medium ones. Three other subgenera, subg. Hydnum, subg. Pallida and subg. Rufescentia can be differentiated from subg. Brevispinum by the mostly yellow or orange-coloured pileus (except for a few whitish ones).



Hydnum brevispinum T. Cao & H. S. Yuan, sp. nov. MycoBank MB 839417; Fig. 4, Fig. 5, Fig. 6, Fig. 14



Etymology: Brevispinum (Lat.), refers to the short spines.



Typus: China, Hunan Province, Sangzhi County, Badagong Nature Reserve, on soil in angiosperm forest, 23 Sep. 2020, T. Cao, holotype, Wei 10214 (IFP 019464).



Diagnosis: Differs from Hydnum minum in having shorter spines (0.2–0.8 mm 0.2–0.8 mm vs. up to 1.7 mm long), longer stipes (25 vs. 15 mm long), broader pileipellis hyphae (5–12 vs. 4–6 μm), larger basidia (20–40 × 3–10 μm 20–30 × 4–7 μm) with 4–6 sterigmata.



Description: Basidiocarps solitary or concrescent, fleshy when fresh, becoming brittle and light in weight upon drying. Pilei 10–15 mm wide, round, convex to plano-convex, shallowly depressed in the center. Pileal surface dry, velutinate, smooth, azonate to subzonate, pure white to yellowish white (4A2), drying yellowish white to greyish orange (4A2/5B4). Pileal margin thin, entire and slightly decurved, concolorous with the pileal surface. Pileal context 1–3 mm thick, pure white. Hymenophore hydnoid, spines non-decurrent to subdecurrent, crowded, evenly distributed; surface pure white when fresh, yellowish white (4A2) when dry; fibrous, subulate, acute, straight to somewhat flexuous, solitary, 0.2–0.8 mm long, shortest near the pileus margin, 5–9 per mm, brittle when dry. Stipe central or eccentric, up to 25 mm long, 2–4 mm wide, subcylindrical, solid; surface glabrous, white; stipe base enlarged and covered with a small amount of white basal mycelium. Odour mild.

 Basidiospores broadly ellipsoid, (4.5–)5–5.8(–6) × (3.5–)3.8–4.8(–5) μm, L_m = 5.28 μm, W_m = 4.16 μm, Q = 1.27–1.31 (n = 60/2), smooth, thin-walled, IKI–, hyaline, some with granular contents; hilar appendix 1 μm long. Basidia fusiform to subcylindric, ventricose, 20–40 × 3–10 μm, sometimes with large guttules or finely granulose contents; sterigmata 4–6, up to 7 μm long, 1.5 μm wide at base, somewhat curving. Basidioles numerous, ventricose, subcylindrical or subclavate, smaller than basidia, 10–35 × 3–7.5 μm. Cystidia absent. Subhymenium trama filamentous, hyphae 2–4 μm wide, thin- to slightly thick-walled, olive in KOH. Hyphae of spines 2–3 μm, thin-walled, apex cylindrical. Pileipellis composed of cylindrical hyphae, thick-walled, subparallel, occasionally branched; terminal elements rounded at apex, cells 80–145 × 5–12 μm. Stipitipellis composed of subcylindrical hyphae, slightly thick-walled, interwoven, 5–16.5 μm wide, terminal elements rounded at apex. Clamp connections present.



Material examined: China, Hunan Province, Sangzhi County, Badagong Nature Reserve, on soil in angiosperm forest, 23 Sep. 2020, T. Cao, paratype, Wei 10258 (IFP 019465).



Notes: There are several almost pure white species in genus Hydnum such as H. minum, H. treui, H. zongolicense and the species in subgenus Alba (e.g., H. cremeoalbum, H. subcremeoalbum and H. albomagnum). Hydnum brevispinum is also a white species with small basidiocarps which are reminiscent of H. minum, however, H. minum differs from H. brevispinum in having longer spines (up to 1.7 mm long), shorter stipes (less than 1.5 cm long), slenderer pileipellis hyphae (4–6 μm wide), and smaller basidia (20–30 × 4–7 μm) with 5–8 sterigmata (Yanaga et al. 2015); Hydnum albomagnum can be differentiated from H. brevispinum by larger basidiocarps (pilei 60–110 mm wide), longer spines (1–6 mm long), ellipsoid basidiospores (Q up to 2.17) and slenderer pileipellis hyphae (2.5–5 μm wide) (Swenie et al. 2018); Hydnum brevispinum differs from H. zongolicense by a smaller pileus (10–15 vs. 17–35 mm wide), shorter spines (0.2–0.8 vs. 0.5–2 mm long), broadly ellipsoid and smaller basidiospores (subglobose and 5–5.8 × 3.8–4.8 μm in H. zongolicense) and basidia with 4–6 sterigmata (2–5 in H. zongolicense) (Niskanen et al. 2018). The small whitish basidiocarps and small basidiospores of H. albidum are similar to H. brevispinum, but the former species differs from the latter by having the bumpy or mottled pileal surface, longer spines (1–6 mm long) and presence of 7-spored basidia (Niskanen et al. 2018), besides, the similarity of the ITS sequences between the type materials (NR_164025 of H. albidum and MW980578 of H. brevispinum) of two species is only 92.79 %.



Hydnum flabellatum T. Cao & H. S. Yuan, sp. nov. MycoBank MB 839414; Fig. 4, Fig. 5, Fig. 6, Fig. 15



Etymology: Flabellatum (Lat.), refers to the flabelliform to semicircular pileus.



Typus: China, Liaoning Province, Xinbin County, Qingsongling Mt., on soil in angiosperm and Pinus sp. mixed forest, 5 Sep. 2020, H. S. Yuan, holotype, Yuan 14708 (IFP 019459).



Diagnosis: Differs from Hydnum subtilior in the smaller pilei (30–45 mm vs. up to 90 mm wide), shorter spines (0.6–2 mm vs. up to 8 mm long), longer basidia (35–60 μm vs. less than 44 μm) and broader pileipellis hyphae (5–16 vs. 3–7 μm).



Description: Basidiocarps solitary, fleshy and leathery when fresh, becoming soft corky and light in weight upon drying. Pilei 30–45 mm wide, flabelliform to semicircular, convex. Pileal surface dry, glabrous, irregularly bumpy or mottled, subzonate towards margin, yellowish white, pale yellow to greyish yellow (4A2/4A3/4B3), drying pale orange (5A3); scabrous, often with some brownish orange (6C7/6C8) scales. Pileal margin thin, entire and straight, concolorous with the pileal surface. Pileal context 1–5 mm thick, yellowish white (4A2). Hymenophore hydnoid, spines non-decurrent or subdecurrent, crowded, evenly distributed; surface orange-white (5A2) when fresh, greyish orange (5B5) when dry; subulate, acute, straight to somewhat flexuous, solitary, 0.6–2 mm long, shortest near the pileus margin, 3–5 per mm, brittle when dry. Stipes eccentric, up to 50 mm long, 8–13 mm wide, subcylindrical, hollow; surface glabrous, white, staining orange-white (5A2) when handled; stipe base enlarged and covered with a small amount of white basal mycelium. Odour mild.

 Basidiospores broadly ellipsoid, (7.8–)8.5–9.5(–10) × (6–)6.5–7.8(–8) μm, L_m = 9.07 μm, W_m = 7.04 μm, Q = 1.26–1.29 (n = 30/1), smooth, thin-walled, IKI–, hyaline, some with granular contents; hilar appendix 1 μm long. Basidia subcylindric, subclavate to clavate, 35–60 × 6–11 μm, sterigmata 2–5, up to 10 μm long, 1–1.5 μm wide at base, slightly curving. Basidioles numerous, subcylindrical or subclavate, smaller than basidia, 12–45 × 3.5–9 μm. Cystidia absent. Subhymenium trama filamentous, hyphae 2–5 μm wide, thin-walled, greenish yellow in KOH. Hyphae of spines 1.5–3 μm, thin-walled, apex cylindrical. Pileipellis composed of cylindrical hyphae, thin- to slightly thick-walled, subparallel, rarely branched; terminal elements rounded at apex, cells 50–185 × 5–16 μm. Stipitipellis composed of subcylindrical hyphae, thick-walled, interwoven, 5–12 μm wide, terminal elements rounded at apex. Clamp connections present.



Notes: Hydnum flabellatum was collected from a temperate forest in northeast China. It is embedded in the subgenus Pallida and shares the small to medium basidiocarps, cream-coloured to ochraceous with very pale orange pileus, stipes bruising brownish when handled, non-decurrent or subdecurrent spines and broadly ellipsoid basidiospores with other species of this subgenus (Niskanen et al. 2018). However, Hydnum flabellatum can be differentiated from H. iberidum in having larger basidiospores (avg. = 9.07 × 7.04 vs. 8.2 × 6.4 μm) and basidia with 2–5 sterigmata (Niskanen et al. 2018). Hydnum subtilior differs from H. flabellatum by having a larger pileus (up to 90 mm wide), longer spines (up to 8 mm long), shorter basidia (less than 44 μm) and slenderer pileipellis hyphae (3–7 μm wide) (Swenie et al. 2018). The subgenus type species, Europe Hydnum vesterholtii differs from H. flabellatum in having one-spored basidia (sometimes), thin-walled pileipellis hyphae with yellowish content and thin-walled and slenderer stipitipellis hyphae (4–8 μm wide) (Olariaga et al. 2012).



Hydnum flavidocanum T. Cao & H. S. Yuan, sp. nov. MycoBank MB 839415; Fig. 1, Fig. 4, Fig. 5, Fig. 6, Fig. 16



Etymology: Flavidocanum (Lat.), refers to the yellowish grey tint at the centre of the pileal surface.



Typus: China, Yunnan Province, Shizong County, Junzishan Mt., on soil in angiosperm and Pinus sp. mixed forest, 8 Aug. 2019, J. R. Yu & T. Cao, holotype, Yuan 13903a (IFP 019460).



Diagnosis: Differs from Hydnum minum in having a subzonate pileal surface, larger basidia (33.5–55 × 5–10 vs. 20–30 × 4–7 μm) with 2–5 sterigmata, larger basidiospores (7.2–8.8 × 5.5–6.5 vs. 4.5–5.5 × 3–4.5 μm) and broader pileipellis cells (5–10 vs. 4–6 μm).



Description: Basidiocarps solitary to concrescent, fleshy and leathery when fresh, becoming soft corky and light in weight upon drying. Pilei 20–30 mm wide, round, convex to plano-convex, shallowly depressed in the center. Pileal surface dry, glabrous, smooth, subzonate, yellowish white or yellowish grey (4A2–4B2) at center and whitish towards margin when moist, azonate upon drying. Pileal margin thin, entire and slightly incurved, concolorous with the pileal surface. Pileal context 1–3 mm thick, yellowish white to pale yellow (4A2–4A3). Hymenophore hydnoid, spines non-decurrent or subdecurrent, crowded, evenly distributed; surface orange-white (5A2) when fresh, greyish orange (5B4–5B6) when dry; fibrous, subulate, acute, straight to somewhat flexuous, solitary, 0.5–2 mm long, shortest near the pileus margin, 3–5 per mm, brittle when dry. Stipes central, confluent with pilei, 25–40 mm long, 8–10 mm wide, subcylindrical, solid; surface glabrous, concolorous with the spine surface; stipe base enlarged and covered with a small amount of white basal mycelium. Odour mild and fruity.

 Basidiospores broadly ellipsoid, (7.0–)7.2–8.8(–8.9) × (5.2–)5.5–6.5(–6.8) μm, L_m = 7.75 μm, W_m = 6.01 μm, Q = 1.29–1.31 (n = 60/2), smooth, thin-walled, IKI–, hyaline, some with granular contents; hilar appendix 0.3–1 μm long. Basidia subcylindric, subclavate to clavate, 33.5–55 × 5–10 μm, sometimes with large guttules or finely granulose contents; sterigmata 2–5, up to 5 μm long, 1–3 μm wide at base, somewhat curving. Basidioles numerous, subcylindrical or subclavate, smaller than basidia, 12.5–48 × 3–10 μm. Cystidia absent. Subhymenium trama filamentous, hyphae 3–5 μm wide, thin-walled, olive yellow in KOH. Hyphae of spines 1.5–4 μm, thin-walled, apex cylindrical. Pileipellis composed of cylindrical hyphae, 5–10 μm wide, thin-walled, densely interwoven to subparallel, rarely branched; terminal elements rounded at apex, cells 20–73 × 5–10 μm. Stipitipellis composed of subcylindrical hyphae, thin- to slightly thick-walled, interwoven, 7–13.5 μm wide, terminal elements rounded at apex. Clamp connections present.



Material examined: China, Yunnan Province, Shizong County, Junzishan Mt., on soil in angiosperm and Pinus sp. mixed forest, 8 Aug. 2019, J. R. Yu & T. Cao, paratype, Yuan 13900a (IFP 019461).



Notes: Hydnum flavidocanum forms a group with H. minum. Morphologically, Hydnum minum resembles H. flavidocanum by a non-decurrent hymenophore, absence of cystidia and presence of clamps, but differs from it by the azonate pileal surface, smaller basidia (20–30 × 4–7 μm) with 5–8 sterigmata, smaller basidiospores (4.5–5.5 × 3–4.5 μm) and slenderer pileipellis cells (4–6 μm wide) (Yanaga et al. 2015).



Hydnum longibasidium T. Cao & H. S. Yuan, sp. nov. MycoBank MB 839416; Fig. 1, Fig. 4, Fig. 5, Fig. 6, Fig. 17



Etymology: Longibasidium (Lat.), refers to the long basidia.



Typus: China, Hunan Province, Sangzhi County, Badagong Mt. Nature Reserve, on soil in angiosperm forest, 24 Sep. 2020, T. Cao, Wei10383 (holotype IFP 019462).



Diagnosis: Differs from Hydnum ferruginescens in having a subzonate to zonate pileal surface, smaller basidiocarps, larger basidiospores (8.5–11 × 7.8–9.8 vs. 6–8 × 5–7.5 μm), larger basidia (45–82 × 10–14 vs. 39–56 × 7.5–9 μm) as well as thick-walled and broader (8–18 vs. 5–7 μm) pileipellis hyphae.



Description: Basidiocarps solitary to concrescent, leathery when fresh, becoming brittle and light in weight upon drying. Pilei 10–15 mm wide, irregularly round, plano-convex, shallowly depressed in the center. Pileal surface dry, subglabrous to velutinate, subzonate or zonate towards margin, orange-white to greyish orange (5A2–5B5) when fresh, greyish orange to brownish yellow (5C7–5C8) upon drying. Pileal margin entire and incurved when young, slightly decurved, straight and somewhat lobed in age, orange-white to orange-grey (6A2–6B2). Pileal context 0.5–1 mm thick, white to yellowish white (4A1–4A2). Hymenophore hydnoid, spines non-decurrent or subdecurrent, crowded, evenly distributed; surface orange-white to pale orange (5A2/5A3) when fresh, concolorous with the pileal surface when dry; fibrous, subulate, acute, straight to somewhat flexuous, solitary, 1–4 mm long, shortest near the pileus margin, 2–3 per mm, brittle when dry. Stipes central, 15–25 mm long and 3–8 mm wide, subcylindrical, solid; surface glabrous, white, staining pale yellow when handled; stipe base enlarged and white basal mycelium absent. Odour mild and fruity.

 Basidiospores broadly ellipsoid to subglobose, (8.0–)8.5–11.0(–11.5) × (7.5–)7.8–9.8(–10.0) μm, L_m = 9.81 μm, W_m = 9.03 μm, Q = 1.09–1.13 (n = 60/2), smooth, thin-walled, IKI–, hyaline, some with granular contents; hilar appendix 0.5–1 μm long. Basidia subcylindric or subclavate, 45–82 × 10–14 μm, sometimes with large guttules or finely granulose contents; sterigmata 2–4, up to 8 μm long, 1.5–3.5 μm wide at base, slightly curving. Basidioles numerous, subclavate to clavate, smaller than basidia, 15–56 × 5–14 μm. Cystidia absent. Subhymenium trama filamentous, hyphae 2–5.5 μm wide, thin- to slightly thick-walled, olive yellow in KOH. Hyphae of spines 4.5–6 μm, thin-walled, apex cylindrical. Pileipellis composed of cylindrical hyphae, 8–18 μm wide, thick-walled, interwoven to subparallel, occasionally branched; terminal elements rounded at apex, cells 40–138 × 8–18 μm. Stipitipellis composed of subcylindrical hyphae, thick-walled, subparallel, 7–25 μm wide, terminal elements rounded at apex. Clamp connections present.



Material examined: China, Hunan Province, Sangzhi County, Badagong Mt. Nature Reserve, on soil in angiosperm forest, 24 Sep. 2020, T. Cao, paratype, Wei 10367 (IFP 019463).



Notes: Hydnum longibasidium falls in subg. Rufescentia and is closely related to Hydnum sp. (HKAS82411) (Taiwan Island), H. ferruginescens (southeastern US) and H. magnorufescens (Italian) (Feng et al. 2016, Niskanen et al. 2018). In terms of molecular and morphology characteristics, Hydnum ferruginescens and H. magnorufescens are very similar and they both resemble H. longibasidium in having an entire and incurved pileal margin when young, non-decurrent or subdecurrent spines, white basal mycelium covering stipe base, absence of cystidia and presence of clamps. However, Hydnum ferruginescens and H. magnorufescens can both be differentiated from H. longibasidium by the azonate pileal surface, larger basidiocarps (pileus up to 60 mm wide and stipes up to 40 mm long in H. ferruginescens, up to 55 mm wide and 50 mm long in H. magnorufescens), smaller spores (6–8 × 5–7.5 μm in former and 7–8.5 × 6.8–8 μm in latter), smaller basidia (39–56 × 7.5–9 μm and 38–46 × 7.5–9.5 μm respectively) as well as thin-walled and slenderer pileipellis hyphae (mostly 5–7 μm and 5–7 μm wide respectively) (Swenie et al. 2018, Niskanen et al. 2018).



Hydnum pallidocroceum T. Cao & H. S. Yuan, sp. nov. MycoBank MB 839418; Fig. 1, Fig. 4, Fig. 5, Fig. 6, Fig. 18



Etymology: Pallidocroceum (Lat.), refers to the pale orange pileal surface.



Typus: China, Xinjiang Auto. R, Kanasi National Geopark, on soil in Pinus sp. and Picea sp. forest, 6 Aug. 2019, Xu Lu & Y. H. Mu, holotype, Yuan 14023 (IFP 019466).



Diagnosis: Differs from Hydnum jussii in having a smaller pileus (25–40 vs. up to 60 mm wide), non-decurrent spines, slenderer spores (Q= 1.32–1.35 vs. 1.03–1.18) and larger basidia (42–55 × 8–11 vs. 43–48 × 7–9 μm).



Description: Basidiocarps solitary, fleshy and leathery when fresh, becoming brittle and light in weight upon drying. Pilei 25–40 mm wide, irregularly round, plano-convex, shallowly depressed in the center. Pileal surface dry, subglabrous, azonate, orange-white to pale orange (5A2–5A3). Pileal margin entire and slightly incurved or straight, concolorous with pileal surface. Pileal context 1–3 mm thick, yellowish white to yellowish grey (4A2–4B2). Hymenophore hydnoid, spines non-decurrent, crowded, evenly distributed; surface light yellow (4A4) when fresh, concolorous with pileal surface when dry, fibrous, subulate, acute, straight to somewhat flexuous, solitary, 1–5 mm long, shortest near the pileus margin, 2–6 per mm, brittle when dry. Stipes central or eccentric, 30–55 mm long, 5–10 mm wide, subcylindrical, solid; surface glabrous, concolorous with spine surface, staining brownish when handled; stipe base somewhat narrower than the apex. Odour mild and fruity.

 Basidiospores broadly ellipsoid, (7.5–)7.8–9.5(–10.0) × (5.5–)6.0–7.5(–8.0) μm, L_m = 9.09 μm, W_m = 6.72 μm, Q = 1.32–1.35 (n = 60/2), smooth, thin-walled, IKI–, hyaline, some with granular contents; hilar appendix 0.5–1 μm long. Basidia subcylindric or subclavate, 42–55 × 8–11 μm, sometimes with large guttules or finely granulose contents; sterigmata 2–5, up to 5 μm long, 1–3 μm wide at base, slightly curving. Basidioles numerous, subclavate to clavate, smaller than basidia, 15–48 × 3–10 μm. Cystidia absent. Subhymenium trama filamentous, hyphae 3–5 μm wide, thin-walled, pale yellow in KOH. Hyphae of spines 3–6 μm, thin-walled, apex cylindrical. Pileipellis composed of cylindrical hyphae, 8–23 μm wide, thin- to slightly thick-walled, interwoven to subparallel, rarely branched; terminal elements rounded at apex, cells 73–100 × 8.5–25 μm. Stipitipellis composed of subcylindrical hyphae, thick-walled, subparallel, 7–18 μm wide, terminal elements rounded at apex. Clamp connections present.



Material examined: China, Xinjiang Auto. Reg., Kanasi National Geopark, on soil in Pinus sp. and Picea sp. forest, 6 Aug. 2019, Xu Lu & Y. H. Mu, paratype, Yuan 14017 (IFP 019467).



Notes: Hydnum pallidocroceum was discovered from the Xinjiang Autonomous Region and the two samples form a strongly supported (93 % in ML and 1.00 BPP) group with the H. jussii lineage which includes two other samples from Xinjiang (Yuan 14008 and Yuan 14009). Hydnum jussii is widely distributed, being known from Finland, as well as from Xizang and Xinjiang in China. Morphologically, Hydnum jussii is closely related to the new species in having a pale orange pileal surface, thin-walled hyphae at the spine apex and stipes bruising brownish when handled but differs by the larger pileus (up to 60 mm wide), somewhat decurrent spines, broader spores (Q = 1.03–1.18) and smaller basidia (43–48 × 7–9 μm) (Niskanen et al. 2018).

 In the phylogenetic tree, Hydnum pallidocroceum, and H. jussii clustered together with H. melleopallidum as well as H. albertense. Hydnum melleopallidum is similar to H. pallidocroceum in having a convex pileus, pale orange pileal surface and thin-walled hyphae at the spine apex but can be differentiated from it by smaller basidiocarps (less than 35 mm wide and long), decurrent spines, broader spores (Q = 1.04–1.18) and smaller basidia (42–48 × 7.5–8.5 μm) (Niskanen et al. 2018). Hydnum albertense resembles H. pallidocroceum in having a convex pileus, incurved pileal margin and non-decurrent spines, however, it differs from the new species in having larger basidiocarps (pileus up to 100 mm wide and stipes up to 65 mm long), broader spores (Q = 1.06–1.22) and smaller basidia (40–47 × 6.5–8 μm) (Niskanen et al. 2018).



Hydnum pallidomarginatum T. Cao & H. S. Yuan, sp. nov. MycoBank MB 839419; Fig. 1, Fig. 4, Fig. 5, Fig. 6, Fig. 19



Etymology: Pallidomarginatum (Lat.), refers to the light-coloured pileal margin.



Typus: China, Yunnan Province, Yimen County, on soil in angiosperm forest, 10 Aug. 2019, H. S. Yuan, holotype, Yuan 13928a (IFP 019468).



Diagnosis: Differs from Hydnum ibericum in having broader stipes (10–15 vs. 5–8 mm wide), basidia with 2–4 sterigmata and larger basidiospores (8.2–9.8 × 6.5–7.8 vs. 7.5–8.5 × 6–7 μm).



Description: Basidiocarps solitary to concrescent, sometimes multipileate, fleshy and leathery when fresh, becoming hard and light in weight upon drying. Pilei 20–35 mm wide, irregularly round or semicircular, infundibuliform, depressed in the center. Pileal surface dry, glabrous, smooth, orange-white to pale orange (6A2/6A3), with a light colour zone towards center, drying azonate. Pileal margin entire to incised, straight or slightly decurved, whitish, obviously lighter than the pileal surface. Pileal context 3–5 mm thick, yellowish white to pale yellow (4A2–4A3). Hymenophore hydnoid, spines decurrent, crowded, evenly distributed, surface orange-white to pale orange (5A2–5A3) when fresh, brownish orange (5C6) when dry; subulate, terete or flattened, straight to somewhat flexuous, solitary, 0.5–2 mm long, shortest near the pileus margin, 2–3 per mm, brittle when dry. Stipes central or eccentric, confluent with pilei, 25–30 mm long, 10–15 mm wide, subcylindrical, solid; surface glabrous, concolorous with the spine surface, bruising brownish when handled; stipe base slightly enlarged and covered with white basal mycelium. Odour mild and fruity.

 Basidiospores broadly ellipsoid, (8.0–)8.2–9.8(–10.0) × (6.0–)6.5–7.8(–8.2) μm, L_m = 8.75 μm, W_m = 6.99 μm, Q = 1.25–1.28 (n = 60/2), smooth, thin-walled, IKI–, hyaline, some with granular contents; hilar appendix 0.3–0.5 μm long. Basidia subcylindric, subclavate to clavate, 32–65 × 6.5–11 μm, sometimes with large guttules or finely granulose contents; sterigmata 2–4, up to 10 μm long, 2–5 μm wide at base, slightly curving. Basidioles numerous, subcylindrical or subclavate, smaller than basidia, 23–60 × 3–10 μm. Cystidia absent. Subhymenium trama filamentous, hyphae 2.5–6 μm wide, thin-walled, pale yellow in KOH. Hyphae of spines 2–4 μm, thin-walled, apex cylindrical. Pileipellis composed of cylindrical hyphae, 6–10 μm wide, slightly thick-walled, interwoven, rarely branched; terminal elements rounded at apex, cells 35–110 × 6–11.5 μm. Stipitipellis composed of subcylindrical hyphae, thick-walled, densely interwoven to subparallel, 4.5–18 μm wide, terminal elements rounded at apex. Clamp connections present.



Material examined: China, Yunnan Province, Yimen County, on soil in angiosperm forest, 10 Aug. 2019, T. Cao, paratype, Yuan 13940a (IFP 019469).



Notes: The two samples of Hydnum pallidomarginatum form a fully supported lineage in the phylogenetic tree. They are embeded in subgenus Pallida and grouped with H. ibericum, H. vesterholtii, H. subtilior as well as another new Chinese species, H. flabellatum. Morphologically, the Spanish species. Hydnum ibericum, is closely related to H. pallidomarginatum in having a pileal surface with pale orange tints, decurrent spines, stipes bruising brownish and presence of clamps. However, it can be differentiated from the new species by the slenderer stipes (5–8 mm wide), basidia with 3–5 sterigmata and smaller basidiospores (7.5–8.5 × 6–7 μm). Additionally, H. ibericum grows on the soil of an angiosperm forest whereas H. pallidomarginatum grows in coniferous mixed forests (Niskanen et al. 2018). Hydnum vesterholtii is described from France and resembles H. pallidomarginatum in having zonate pileus with whitish margin, broadly ellipsoid basidiospores and absence of cystidia, but differs from it by the non-decurrent to slightly decurrent spines, basidia with 3–5 sterigmata, thin-walled and broader (up to 14 μm wide) terminal cells of pileipellis and slenderer stipitipellis hyphae (4–8 μm wide) (Olariaga et al. 2012). Hydnum subtilior is similar to H. pallidomarginatum in having a zonate pileal surface, broadly ellipsoid basidiospores, absence of cystidia and presence of clamps, but differs from it by a larger pileus (up to 90 mm wide), longer spines (up to 8 mm long), larger stipes (up to 60 mm long and 21 mm wide), smaller basidia (less than 44 μm long and 9 μm wide) and thin-walled terminal cells of pileipellis (Swenie et al. 2018).

 Hydnum flabellatum and H. pallidomarginatum are found in the temperate and subtropical forests of China, respectively, and they share the pale orange-tinted pileal surface. However, the former species differs from H. pallidomarginatum in having a scabrous pileal surface, longer stipes (up to 50 mm long), basidia with 2–5 sterigmata, broader pileipellis hyphae (up to 16 μm wide) and slenderer stipitipellis hyphae (less than 12 μm wide). The two species from subtropical China, H. pallidomarginatum and H. flavidocanum, share the zonate pileal surface, enlarged stipe base, absence of cystidia and presence of clamps. However, H. pallidomarginatum can be differentiated from H. flavidocanum by the shorter but wider stipes (25–30 × 10–15 mm), longer basidia (up to 65 μm long) with 2–4 sterigmata, larger basidiospores (8.2–9.8 × 6.5–7.8 μm) and slightly thick-walled and larger terminal cells (35–110 × 6–11.5 μm) of pileipellis.



Hydnum sphaericum T. Cao & H. S. Yuan, sp. nov. MycoBank MB 839420; Fig. 1, Fig. 4, Fig. 5, Fig. 6, Fig. 20



Etymology: Sphaericum (Lat.), refers to the subglobose pileus.



Typus: China, Hunan Province, Sangzhi County, Badagong Mt. Nature Reserve, on soil in angiosperm forest, 23 Sep. 2020, T. Cao, holotype, Wei10243 (IFP 019470).



Diagnosis: Differs from Hydnum repandum in the smaller pileus (20–35 vs. 110 mm wide), non-decurrent to subdecurrent spines and larger basidia (37–65 × 8–13.5 vs. 35–45 × 6–8 μm).



Description: Basidiocarps solitary to concrescent, fleshy and leathery when fresh, becoming brittle and light in weight upon drying. Pilei 20–35 mm wide, subglobose when young, becoming irregularly round with age. Pileal surface dry, subglabrous, azonate, orange-white (6A2) when moist, greyish orange to brownish orange (5B5–6C7) upon drying. Pileal margin entire and strongly incurved when young, slightly decurved and undulate in age, concolorous with the pileal surface. Pileal context 1–3 mm thick, white to yellowish white (4A1–4A2). Hymenophore hydnoid, spines non-decurrent to subdecurrent, crowded, evenly distributed; surface white (3A1–4A1) when fresh, brownish orange (6C4–6C6) when dry; fibrous, subulate, acute, straight to somewhat flexuous, solitary, 0.5–3 mm long, shortest near the pileus margin, 3–6 per mm, brittle when dry. Stipe central or eccentric, 18–25 mm long, 5–8 mm wide, subcylindrical, solid; surface glabrous, concolorous with the spine surface; stipe base enlarged and covered with a small amount of white basal mycelium. Odour mild and fruity.

 Basidiospores broadly ellipsoid, (7.5–)8.0–8.8(–9.0) × (6.0–)6.5–7.5(–8.0) μm, L_m = 8.36 μm, W_m = 6.94 μm, Q = 1.20–1.23 (n = 60/2), smooth, thin-walled, IKI–, hyaline, some with granular contents; hilar appendix 0.5 μm long. Basidia subcylindric or subclavate, 37–65 × 8–13.5 μm, sometimes with large guttules or finely granulose contents; sterigmata 4–5, up to 8 μm long, 2–2.5 μm wide at base, slightly curving. Basidioles numerous, subclavate, smaller than basidia, 15–53 × 4–11 μm. Cystidia absent. Subhymenium trama filamentous, hyphae 3–5 μm wide, thin- to slightly thick-walled, pale yellow in KOH. Hyphae of spines 3–5 μm, thin-walled, apex cylindrical. Pileipellis composed of cylindrical hyphae, 5–13 μm wide, thick-walled, interwoven to subparallel, occasionally branched; terminal elements rounded at apex, cells 70–105 × 5–14.5 μm. Stipitipellis composed of subcylindrical hyphae, thick-walled, subparallel, 8.5–20 μm wide, terminal elements rounded at apex. Clamp connections present.



Materials examined: China, Hunan Province, Sangzhi County, Badagong Mt. Nature Reserve, on soil in angiosperm forest, 23 Sep. 2020, T. Cao , paratype, Wei 10262 (IFP 019471); Wei 10300 (IFP 019472).



Notes: The three samples of Hydnum sphaericum cluster with HKAS51070 and all were collected from subtropical China. Although the coverage between the ITS sequences of Wei 10243 (holotype of H. sphaericum, 647 bp) and HKAS51070 (556 bp) is 85 %, they got 98.74 % similarity. The four samples form a separate lineage with strong support (97 % ML, 1.00 BPP) and we describe them as a new taxon.

 In the phylogenetic tree, Hydnum sphaericum fell in subg. Hydnum. The genus type species H. repandum is also nested in this subgenus and it is closely related to H. sphaericum in having a cream colour to orange pileal surface, crowded spines, thin-walled hyphae in the spines, and presence of clamps. However, it differs from H. sphaericum in having a larger pileus (up to 110 mm wide), larger stipes (35–60 × 7–14 mm), mostly decurrent spines and smaller basidia (35–45 × 6–8 μm) (Niskanen et al. 2018).

 Species of Hydnum subg. Hydnum usually have medium to large (pilei 40–110 mm wide) basidiomata (Niskanen et al. 2018), however, H. sphaericum has small ones (pilei 20–35 mm wide). Another relatively small species is H. olympicum (pilei 30–65 mm wide) and it resembles the new species in having whitish pileal surface, crowded spines, broadly ellipsoid basidiospores and presence of clamps, but H. olympicum differs from H. sphaericum in having larger basidiocarps (pileus up to 65 mm wide and stipes up to 60 mm long), smaller basidia (37–43 × 6.6–8.6 μm) with 4 sterigmata, and presence of pleurocystidia (Niskanen et al. 2018).



Hydnum tangerinum T. Cao & H. S. Yuan, sp. nov. MycoBank MB 839421; Fig. 4, Fig. 5, Fig. 6, Fig. 21



Etymology: Tangerinum (Lat.), refers to the orange tint of pileal surface.



Typus: China, Hunan Province, Sangzhi County, Badagong Nature Reserve, on soil in angiosperm forest, 23 Sep. 2020, T. Cao, holotype, Wei 10245 (IFP 019473).



Diagnosis: Differs from Hydnum melitosarx in having shorter stipes (60 vs. 70 mm long), broadly ellipsoid basidiospores and basidia with 3–6 sterigmata.



Description: Basidiocarps solitary or concrescent, fleshy and leathery when fresh, becoming soft corky and light in weight upon drying. Pilei 10–50 mm wide, flabelliform, or irregularly round, convex to plano-convex, shallowly depressed in the center. Pileal surface dry, velutinate, smooth, azonate, pale orange, light orange, orange to brownish orange (5A3/5A4/6B8/6C8), drying greyish orange (5B5/5B6/6B5); scabrous, often with some scales towards center. Pileal margin thin, entire and undulate, concolorous with the pileal surface. Pileal context 1–5 mm thick, yellowish white (4A2). Hymenophore hydnoid, spines non-decurrent, crowded, evenly distributed; surface orange-white (6A2) when fresh, greyish orange (5B5/5B6) when dry; subulate, acute, straight to somewhat flexuous, solitary, 2–6 mm long, shortest near the pileus margin, 2–3 per mm, brittle when dry. Stipes central, up to 60 mm long, 3–7 mm wide, subcylindrical, solid; surface glabrous, white, staining pale orange (5A3) when handled; stipe base enlarged and covered with a small amount of white basal mycelium. Odour mild and fruity.

 Basidiospores broadly ellipsoid, (7–)7.2–8.8(–9) × (5.5–)5.8–7(–7.5) μm, L_m = 8.11 μm, W_m = 6.19 μm, Q = 1.23–1.31 (n = 60/2), smooth, thin-walled, IKI–, hyaline, some with granular contents; hilar appendix 1 μm long. Basidia subcylindric, subclavate to clavate, 29–45 × 5.5–10 μm, sometimes with large guttules or finely granulose contents; sterigmata 3–6, up to 6 μm long, 1.5 μm wide at base, somewhat curving. Basidioles numerous, subcylindrical or subclavate, smaller than basidia, 12.5–43.5 × 3–10 μm. Cystidia absent. Subhymenium trama filamentous, hyphae 3–5 μm wide, thin-walled, greenish yellow in KOH. Hyphae of spines 2–3.5 μm, thin-walled, apex cylindrical. Pileipellis composed of cylindrical hyphae, thin-walled, subparallel, occasionally branched; terminal elements rounded at apex, cells 75–210 × 4–18 μm. Stipitipellis composed of subcylindrical hyphae, slightly thick-walled, interwoven, 3.5–13 μm wide, terminal elements rounded at apex. Clamp connections present.



Material examined: China, Hunan Province, Sangzhi County, Badagong Nature Reserve, on soil in angiosperm forest, 23 Sep. 2020, T. Cao, paratype, Wei 10249 (IFP 019474); Wei 10250 (IFP 019475).



Notes: Hydnum tangerinum is nested in the subgenus Rufescentia and shares the small to medium basidiocarps, orange-tinted pileal surface, bruising stipes when handled, non-decurrent spines and broadly ellipsoid basidiospores, with most of the species in the subgenus. Hydnum melitosarx and H. tangerinum form a weakly supported lineage in the tree (Fig. 4), the former species is similar to H. tangerinum in the medium basidiocarps, orange-tinted pileal surface and non-decurrent spines but differs by having longer stipes (up to 70 mm long), subglobose basidiospores (Q avg. = 1.11) and basidia with 3 sterigmata (Niskanen et al. 2018). Hydnum mulsicolor and H. submulsicolor both share medium basidiocarps, orange-tinted pileal surface and non-decurrent spines with H. tangerinum, however, H. mulsicolor can be differentiated from H. tangerinum by having shorter basidiospores (L_m = 7.5); H. submulsicolor differs from H. tangerinum in having subglobose basidiospores (Q avg. = 1.13) and basidia with 3–4 sterigmata (Niskanen et al. 2018).



Hydnum tenuistipitum T. Cao & H. S. Yuan, sp. nov. MycoBank MB 839422; Fig. 4, Fig. 5, Fig. 6, Fig. 22



Etymology: Tenuistipitum (Lat.), refers to the slender stipes.



Typus: China, Hunan Province, Sangzhi County, Badagong Nature Reserve, on soil in angiosperm forest, 23 Sep. 2020, T. Cao, holotype, Wei 10410 (IFP 019476).



Diagnosis: Differs from Hydnum treui in having longer stipes (uo to 60 mm vs. 15–20 mm long), and larger basidia (45–63 × 3–12 μm 35–42 × 6–7 μm) with 2–6 sterigmata



Description: Basidiocarps solitary or concrescent, fleshy when fresh, becoming brittle and light in weight upon drying. Pilei 10–30 mm wide, round, convex to plano-convex, shallowly depressed in the center. Pileal surface dry, glabrous, smooth, azonate, yellow-white to orange-white (4A2/5A2), drying pale orange to greyish orange (5A3/5B4). Pileal margin thin, entire and slightly decurved, concolorous with pileal surface. Pileal context 1–5 mm thick, white to yellowish white (4A2). Hymenophore hydnoid, spines non-decurrent to subdecurrent, crowded, evenly distributed; surface orange-white (5A2) when fresh, light brown (6D7/6D8) when dry; subulate, acute, straight to somewhat flexuous, solitary, 1–3 mm long, shortest near the pileus margin, 3–4 per mm, brittle when dry. Stipes central, up to 60 mm long, 2–6 mm wide, subcylindrical, solid; surface glabrous, white, staining pale orange (5A3) when handled; stipe base enlarged and covered with a small amount of white basal mycelium. Odour mild and fruity.

 Basidiospores subglobose, (6.5–)6.8–7.2(–7.5) × (5.2–)5.5–6.5(–6.8) μm, L_m = 7.08 μm, W_m = 6.09 μm, Q = 1.07–1.16 (n = 60/2), smooth, thin-walled, IKI–, hyaline, some with granular contents; hilar appendix 1 μm long. Basidia subcylindric, subclavate to clavate, 45–63 × 3–12 μm, sometimes with large guttules or finely granulose contents; sterigmata 2–6, up to 10 μm long, 1.5 μm wide at base, slightly curving. Basidioles numerous, subcylindrical or subclavate, smaller than basidia, 12–50 × 3–10 μm. Cystidia absent. Subhymenium trama filamentous, hyphae 3–5 μm wide, thin-walled, greenish yellow in KOH. Hyphae of spines 2–4 μm, thin-walled, apex cylindrical. Pileipellis composed of cylindrical hyphae, thick-walled, subparallel, frequently branched; terminal elements rounded at apex, cells 98–260 × 4–15 μm. Stipitipellis composed of subcylindrical hyphae, slightly thick-walled, interwoven, 3.5–13 μm wide, terminal elements rounded at apex. Clamp connections present.



Material examined: China, Hunan Province, Sangzhi County, Badagong Nature Reserve, on soil in angiosperm forest, 23 Sep. 2020, T. Cao, paratype, Wei 10417 (IFP 019477).



Notes: Hydnum tenuistipitum and H. brevispinum form a well-supported lineage in the tree (Fig. 4) so, we suggest subgenus Brevispina; the five samples in the subgenus are all collected from a subtropical forest in China. Hydnum tenuistipitum resembles H. brevispinum in the whitish and smooth pileal surface, non-decurrent to subdecurrent spines, thick-walled pileipellis hyphae and slightly thick-walled stipitipellis hyphae, but the latter species differs from H. tenuistipitum by the smaller pileus (less than 15 mm wide), shorter stipe (less than 25 mm wide), smaller (5–5.8 × 3.8–4.8 μm) and broadly ellipsoid basidiospores and shorter basidia (20–40 μm long). The Australasian Hydnum treui belongs to subtropical/tropical forest and is similar to H. tenuistipitum in having a whitish pileal surface, non-decurrent to subdecurrent spines and subglobose basidiospores, however it can be distinguished from the latter species by shorter stipes (15–20 mm long), and smaller basidia (35–42 × 6–7 μm) with 4 sterigmata (Niskanen et al. 2018).



Hydnum ventricosum T. Cao & H. S. Yuan, sp. nov. MycoBank MB 839423; Fig. 1, Fig. 4, Fig. 5, Fig. 6, Fig. 23



Etymology: Ventricosum (Lat.), refers to the ventricose basidia.



Typus: China, Liaoning Province, Xinbin County, Gangshan Mt., on soil in angiosperm and Pinus sp. mixed forest, 12 Aug. 2020, H. S. Yuan, holotype, Yuan 14536 (IFP 019478).



Diagnosis: Differs from Hydnum berkeleyanum in having a smaller pileus (28–35 vs. up to 80 mm wide), shorter spines (1–5 mm vs. up to 9 mm long), shorter but broader (46 × 14 vs. 60 × 9 μm) ventricose basidia and slightly thick-walled pileipellis hyphae.



Description: Basidiocarps solitary to concrescent, fleshy and leathery when fresh, becoming soft corky and light in weight upon drying. Pilei 28–35 mm wide, irregularly round, plano-convex, shallowly depressed in the center. Pileal surface dry, subglabrous, azonate, orange (6A7) when moist, brown (6D6–6D7) upon drying. Pileal margin thin, entire and incurved, concolorous with the pileal surface. Pileal context 1–2.5 mm thick, yellowish white to orange-white (4A2–5A2). Hymenophore hydnoid, spines non-decurrent, crowded, evenly distributed; surface orange-white (5A2–6A2) when fresh, brownish orange (6C4-6C6) when dry, subulate, straight to somewhat flexuous, solitary, 1–5 mm long, shortest near the pileus margin, 2–4 per mm, brittle when dry. Stipes central or eccentric, 30–35 mm long, 10–15 mm wide, subcylindrical, solid; surface glabrous, concolorous with the spine surface, bruising brownish when handled; stipe base somewhat narrower than apex and white basal mycelium absent. Odour mild and fruity.

 Basidiospores subglobose, (7.5–)8.2–9.0(–9.5) × (7.0–)7.5–8.5(–9.0) μm, L_m = 8.64 μm, W_m = 8.17 μm, Q = 1.05–1.09 (n = 60/2), smooth, thin-walled, IKI–, hyaline, some with granular contents; hilar appendix 0.3–1 μm long. Basidia fusiform to subcylindric, ventricose, 30–46 × 7.5–14 μm, sometimes with large guttules or finely granulose contents; sterigmata 2–4, up to 8 μm long, 1–3 μm wide at base, somewhat curving. Basidioles numerous, subclavate, smaller than basidia, 13–35 × 3–11 μm. Cystidia absent. Subhymenium trama filamentous hyphae 3–7.5 μm wide, thin- to slightly thick-walled, brownish yellow in KOH. Hyphae of spines 2–5 μm, thin-walled, apex cylindrical. Pileipellis composed of cylindrical hyphae, 5–10 μm wide, slightly thick-walled, interwoven to subparallel, rarely branched; terminal elements rounded at apex, cells 38–95 × 6–10 μm. Stipitipellis composed of subcylindrical hyphae, thick-walled, subparallel, 7.5–18 μm wide, terminal elements rounded at apex. Clamp connections present.



Material examined: China, Liaoning Province, Xinbin County, Gangshan Mt., on soil in angiosperm and Pinus sp. mixed forest, 26 Aug. 2020, H. S. Yuan, paratype, Yuan 14601 (IFP 019479).



Notes: The two samples of Hydnum ventricosum cluster with HKAS61795. This cluster forms a separate branch with strong support (100 % ML and 0.99 BPP) and all samples were collected from a temperate forest in China. We describe them as a new taxon.

 In the phylogenetic tree, Hydnum ventricosum falls in subg. Rufescentia Niskanen & Liimat and formed a subgroup with H. berkeleyanum, H. rufescens and H. subrufescens. Morphologically, the Indian species H. berkeleyanum is closely related to H. ventricosum in having a light orange pileal surface, entire pileal margin, subglobose basidiospores, 2–4 sterigmata and presence of clamps, but differs from it in having a larger pileus (up to 80 mm wide), longer spines (up to 9 mm long), longer but slenderer basidia (up to 60 μm long and less than 9 μm wide) and thin-walled pileipellis hyphae (Wang et al. 2018). Hydnum rufescens is the type species of subg. Rufescentia, and is similar to H. ventricosum in having the non-decurrent spines and presence of clamps. However, H. rufescens can be distinguished from the new species by the larger pileus (up to 65 mm across), deep reddish orange pileal surface, longer stipes (up to 55 mm long), slenderer basidia (less than 9 μm wide) and smaller basidiospores (7–8.5 × 6–7.2 μm) (Niskanen et al. 2018). Hydnum subrufescens is a species described from Canada and is a synonym of H. aerostatisporum according to Swenie et al. (2018). It is related to H. ventricosum in having an orange-tinted pileus surface and non-decurrent spines. But H. subrufescens can be differentiated from the latter by the larger pileus (up to 60 mm wide), longer stipes (up to 40 mm long), slenderer basidia (less than 9 μm wide) and smaller basidiospores (7.4–8.8 × 6.4–7.8 μm) (Niskanen et al. 2018).



Hydnum albomagnum Banker, Bull. Torrey Bot. Club 28: 207. 1901. MycoBank MB 141572.



Notes: Phylogenetic analyses based on ITS and nLSU sequences and morphological characteristics confirmed the new record. The two Chinese samples and the American Hydnum albomagnum formed an isolated lineage with strong support (98 % in ML and 0.99 BPP). For a detailed description of H. albomagnum, see Banker (1901) and Yanaga et al. (2015). This species was originally described from the USA and recorded in Japan by Yanaga et al. (2015).



Materials examined: China, Hunan Province, Sangzhi County, Badagong Mt. Nature Reserve, on soil in angiosperm forest, 21 Sep. 2020, W. M. Qin, Wei 10194 (IFP 019480); 23 Sep. 2020, W. M. Qin, Wei 10247 (IFP 019481).



Hydnum minum Yanaga & N. Maek., Mycoscience 56: 435. 2015. MycoBank MB 808762.



Notes: Morphological and phylogenetic analyses based on ITS and nLSU sequences confirmed the new record, which is described in detail by Yanaga et al. (2015). This species was originally described from Japan (Yanaga et al. 2015) and so far, has only been collected in two East Asian countries, China and Japan (Yanaga et al. 2015, Niskanen et al. 2018).



Materials examined: China, Hunan Province, Sangzhi County, Badagong Mt. Nature Reserve, on soil in angiosperm forest, 23 Sep. 2020, T. Cao, Wei 10252 (IFP 019482), 10260 (IFP 019483).