Phylogenetic analysis
The ITS dataset contained 128 sequences (22 generated in this study) and comprised 604 sites, of which 284 were variable sites, and 237 of these variable sites were informative. ML and BI analyses produced identical tree topologies, and only the tree derived from the ML analyses is shown (Fig. 1).
The sequences of Melanogaster were grouped into a clade together with 4 sequences labeled as “Alpova trappei” with support values of BS = 77 and PP = 1.00, then further clustered with the other sequences of Alpova with a strong support value (BS = 100). The sequences of Alpova employed in this analysis, except the samples of “Alpova trappei”, formed two independent clades with strong support. In Melanogaster clade, a total 21 phylotypes (marked A–U) can be recognized based on a species delimitation of 98% ITS similarity and strong supported clades of the ITS-based ML tree (Fig. 1). Nine of the 21 phylotypes matched known species, while 12 phylotypes represented uncertain or undescribed species, of which 5 species were described and illustrated as new taxa from China in this study. Phylotype A was designated as M. intermedius as it contained 2 reliably identified sequences EU784372 and EU784371 of M. intermedius published by Brock et al. (2009), and had a strong support (BS = 97/PP = 1.00); Phylotype B contained 2 ectomycorrhizal (ECM) sequences of USA and Canada, one environmental sequence of Canada and 3 sequences of inaccurately identified specimens from USA, showed more than 98.18% ITS similarity among them and less than 97.23% ITS similarity with other phylotypes. It also had a strong support (BS = 99/PP = 1.00) and could represent a new taxon from North America, marked M. sp.1 here (Fig. 1); Phylotype C contained 4 sequences, 2 from ECM and environmental sample respectively, one labeled as Melanogaster ambiguus from a unpublished study and one sequence was newly obtained in the present work from a specimen of M. ambiguus (collected by Richard in Wiltshire, UK, 1987). These sequences showed more than 99.26% ITS similarity and less than 97.91% ITS similarity with other phylotypes, with a moderate support value (BS = 75). Combined the morphologic features of the specimen of M. ambiguus we therefore recognized phylotype D as M. ambiguus. Phylotype D contained 4 sequences labeled as Melanogaster ambiguus and M. broomeanus from an unpublished study about Hungary specimens. These sequences showed more than 99.02% ITS similarity and less than 97.9% ITS similarity with other phylotypes, with a strong support (BS = 94/PP = 1.00). It could represent a new taxon from Hungary, marked M. sp.2 here; Phylotype E contained one ECM sequences from Poland and 2 sequences of inaccurately identified specimens of Hungary. The 3 sequences showed more than 99.29% ITS similarity among them and less than 97.74% with other phylotypes. It could represent a new taxon from Central Europe, marked M. sp.3 here; Phylotype F was designated as M. euryspermus. For it contained one sequence of M. euryspermus pubilished by Frank et al. (2006), and the sequence showed less than 94.53% ITS sequence similarity to other Melanogaster species; Phylotype G and N contained one sequence respectively, a sequence (JN022515) of inaccurately identified specimen of USA, and an unpublished ECM sequence (JN198097). The 2 sequences showed 94.53% ITS similarity to each other even with other species of Melanogaster. The 2 phylotypes could be new taxa of Melanogaster, marked M. sp.4 and M. sp.5 respectively; Phylotype H contained 8 sequences obtained from several new collections of Shanxi Province, China, which were identified as M. spinisporus morphylogically in this study. The sequences shared more than 98.78% ITS similarity, and less than 92.03% ITS similarity to other phylotypes, convincing us that it was a distinct species. Therefore, we recognized phylotype H as M. spinisporus; Phylotype I contained 3 sequences obtained from 3 specimens newly collected from Shanxi Province, which was different from all of known species of Melanogaster morphylogically. The sequences showed more than 99.35% ITS similarity among them and less than 97.9% ITS similarity to other phylotypes. For this, the new taxon M. obovatus was proposed in the present study; Phylotype J contained 2 sequences obtained from 2 specimens newly collected from Shanxi Province, China, which could be distinguished from other species of Melanogaster morphylogically. The sequences showed more than 99.75% ITS similarity among them and less than 97.9% ITS similarity to other phylotypes. Therefore, the new taxon M. quercus was proposed for it; Phylotypes K and M contained one sequences newly extracted from holotypes of Chinese species M. shanxiensis and M. subglobisporus respectively. The two sequences showed less than 97.91% and 92.24 ITS similarity to other phylotypes respectively, clarifing their phylogenetic positions. Therefore, phylotypes K and M were identified M. shanxiensis and M. subglobisporus respectively; Phylotype L contained one sequence extracted from the specimen (BJTC FAN539) newly collected in Yunnan province, China, which was distinguished from other species of Melanogaster obviously in morphology. The sequence showed less than 95.33% ITS sequence similarity to other phylotypes, indicating it is a distinct species in Melanogaster. And the new species M. tomentellus was thus proposed for it; Phylotype O contained 15 sequences, including the holotype sequence (NR_132848) of M. rivularis, showed more than 99.02% ITS similarity among each other and less than 96.76 ITS similarity to other species of genus Melanogaster. Therefore, phylotype O was designated as M. rivularis with no doubt; Phylotype P contained 11 sequences, including two sequences (HQ714780, HQ714794) from accurately identified specimens of M. luteus published by Moreau et al. (2011), showed more than 99.41% ITS similarity among each other and less than 97.07% ITS similarity to other species of genus Melanogaster. For the above reasons, phylotype P was recognized as M. luteus; Phylotype Q was designated as M. broomeanus, containing11 sequences of which 3 sequences were newly obtained from the specimens of China in this work and a reliably identified sequence of M. broomeanus (EU784370) published by Brock et al. (2009). The 11 sequences showed more than 98.09% ITS similarity and less than 95.73% ITS similarity to other species of Melanogaster. Consequently, we recognized phylotype Q as M. broomeanus based on morphology and phylogenetic analysis; Phylotype R contained only a sequence from ectomycorrhizae (ECM) of Quercus liaotungensis Koidz in China, shared less than 94.79% ITS similarity to other species of Melanogaster. It should represent a potential new taxon from China, marked M. sp.6 here; Phylotype S contained 7 sequences of USA, one sequence from ECM of Lithocarpus densiflorus and 4 sequences extracted from the specimens labeled as “Alpova trappei”. Evidently, these sequences of “Alpova trappei” did not cluster with Alpova sequences in this analysis, implying they represented the Melanogaster species rather than Alpova species. Thus Phylotype S could represent a potential new taxon of Melanogaster from USA, marked M. sp.7 here; Phylotype T contained 2 sequences, one extracted from the specimen (BJTC FAN911) newly collected in Shanxi province, China and another sequences (JQ318611) obtained from ectomycorrhizae of Quercus liaotungensis in Loess Plateau, China. They showed 99% ITS sequence similarity and less than 91.72% ITS similarity to other species of Melanogaster, for which a new species M. minobovatus was proposed; Phylotype U was represented by one sequence newly obtained from the specimen of Sichuan province, China, which was significantly different from other species of Melanogaster morphologically. In addition, the sequence showed less than 89.89% to other species of genus Melanogaster, indicating it was a distinct species. Thus, M. panzhihuaensis was proposed for it.
Phylogeny based nrLSU sequences were analyzed because there were the samples of Alpova trappei grouped into Melanogaster clade rather than Alpova clade in ITS-based phylogeny although there were a small number of available sequences. The nrLSU dataset contained 35 sequences (14 generated in this study) and comprise772 sites, of which 103 variable sites, and 64 of these variable sites were informative. ML and BI analyses produced identical tree topologies, and only the tree derived from the ML analyses is shown (Fig. 2). Two clades were recognized in this tree, respectively representing Melanogaster and Alpova, and the sequences labeled as “Alpova trappei” (marked with pentagram (☆) symbol) were placed in Melanogaster clade rather than Alpova clade, consistent with that of the ITS-based phylogenetic analysis (Fig. 1). The other sequences of Alpova cluster an independent clade with a strong support clade (BS = 97, PP = 1.00). These further confirmed that the samples of Alpova trappei represented the species of the genus Melanogaster. Moreover, the Chinese Melanogaster sequences were grouped in Melanogaster clade as those observed on the ITS tree (Fig. 1).
Taxonomy
Melanogaster broomeanus Berk., in Montagne, Annls Sci. Nat., Bot., sér. 2 20: 377 (1843) Fig. 3, a–f
≡Melanogaster ovoidisporus var. angustatisporus K. Tao, Ming C. Chang & B. Liu, Journal of Shanxi University, Natural Science 18(4): 449 (1995). syn. nov. Type:—CHINA. SHANXI PROV.: Jincheng City, Yangcheng County, Mang River National Nature Reserve, in soil under Quercus sp., 29 May 1988, M.-c. Chang 222 (HMAS83258 ex MHSU2022).
Description
Basidiomata 1.5–2.5 × 2–2.5 cm, hypogeous, subglobose to irregularly globose, yellow-brown, becoming blackish where handled, without distinct basal zone; rhizomorphs few, brown; odor no record. Gleba yellow brown when fresh, black when dry, with sterile white veins scattered throughout. Peridium single-layered, 150–400 µm thick, composed of hyphae, hyphae generally parallel to the surface of peridium but loosely interwoven, pale yellow to whitish, 5–12.5 µm in diam, clamp connections abundant; Trama plates of hyaline interwoven hyphae, 2.5–5 µm in diam, thin-walled, tending to be gelatinized. Basidia 16.5–22.2 × 5.0–6.7 µm, light yellow clavate-capitate when young, 4–spored, sterigmata short, collapsed and disappeared at maturity. Basidiospores ellipsoid or oblong to cylindrical, 4.4–7.8 × 3.3–4.7 µm (avX = 6.0 ± 1.0 × 3.9 ± 0.36, n = 50), Q = 1.3–1.9 (avQ = 1.5 ± 0.2, n = 50), deep brown at maturity, walls 0.5 µm thick, smooth, apex obtuse, the base broadly truncate-cupped, with very short remnants of sterigmata.
Habit, habitat and distribution
July to September, in soil under Quercus liaotungensis, Pinus bungeana Zucc. ex Endl., Lonicera japonica Thunb. Warm temperate zone and cold temperate zone of North China.
Notes: Melanogaster broomeanus is diagnosed by its yellow-brown basidiomata, single-layered peridium composed of interwoven hyphae, light brown ellipsoid or oblong to cylindrical basidiospores. The specimens (BJTC FAN909, BJTC FAN1230), newly collected from Shanxi Province, China, morphologically fit well with the characteristics of M. broomeanus, and phylogenetically their ITS sequences were clustered in the M. broomeanus clade with strong support (BS = 99/PP = 1.00) (Fig. 1), suggesting the two Chinese collections represented the Europe species M. broomeanus. M. ovoidisporus var. angustatisporus is described by Wang et al. (1995) based on the specimens from Chinese Shanxi Province, but their morphological traits described by authors is very similar to M. broomeanus. In this study, we re-examined the morphology of holotype and paratypes cited by Wang et al. (1995), and compared them with our new collections (BJTC FAN909, BJTC FAN1230) from Shanxi Province, and as a result, there are not any obvious differences to be revealed among these specimens in morphology. Unfortunately, the DNA sequences of those old specimens are not sequenced successfully, but combining the features of morphology and geography, we conclude that M. ovoidisporus var. angustatisporus should be considered synonym of M. broomeanus.
Material examined: CHINA. SHANXI PROV.: Jincheng City, Yangcheng County, Sanglin Village, Shupigou, in soil under Lonicera japonica Thunb., Alt. 860 m, 27 August 1987, M.-c. Chang 242 (HMAS81917 ex MHSU2023); Linfen City, Xi County, Shenjiagou, in soil under Quercus liaotungensis, Alt. 1321 m, 10 September 2017, Y.-y. Xu 033 (BJTC FAN909); ibid., in soil under Pinus bungeana, X.-y. Yan 155 (BJTC FAN1230); Yuncheng City, Yuanqu County, Lishan Village, 25 July 1987, M.-c. Chang 505 (HMAS83323 ex MHSU2002); ibid., Alt. 1280 m, 8 August 1990, Y. Ma, B. Qiao, W.-k. Bai 502 (HMAS96692 ex MHSU2003). SICHUAN PROV.: Songpan County, Alt. 3100 m, 30 August 1991, M.-s. Yuan 1729 (HKAS24811).
Melanogaster fusisporus Y. Wang, in Wang, Chang, Tao & Liu, Journal of Shanxi University, Natural Science 18(4): 450 (1995) Fig. 3, g–j
Description
Basidiomata 1.1–1.6 × 1.2–3 cm, hypogeous, subglobose to irregularly globose, yellow brown to brown, without distinct basal zone; rhizomorphs few, brown; sweet odor, such as the banana flavor or the apple flavor. Gleba deep brown when fresh, black after dried, with sterile white veins scattered throughout. Peridium two-layered, epicutis 10–20 µm thick, composed of interwoven hyphae, light orange yellow, 4–5 µm in diam, smooth; subcutis 230–400 µm thick, composed of interwoven hyphae, with abundant inflated cells, irregular or puzzle-like, up to 5–40 µm wide, yellowish to pale orange yellowto ward outside surface; Trama plates of interwoven hyphae, hyaline to light yellow, thin-walled, 3–4 µm in diam, tending to be gelatinized. Basidia 9–12 × 10–23 µm, hyaline clavate-capitate when young, 2–3-spored, sterigmata short, collapsed and disappeared at maturity. Basidiospores fusiform, apex bluntly apiculate, 10.0–15.9 × 4.8–6.3 µm (avX = 12.3 ± 1.4 × 5.2 ± 0.3, n = 50), Q = 2.0–2.9 (avX = 2.3 ± 0.2, n = 50), orange brown or deep brown at maturity, 1 µm thick, smooth, the base broadly truncate-cupped, with very short remnants of sterigmata.
Habit, habitat and distribution: August to September, in soil under Quercus moglica Fish., Q. acutissima Carr. Northeast and South China.
Notes: Melanogaster fusisporus is re-described in details based on the re-examinations for paratypes cited by Wang et al. (1995) and newly added specimen HMAS83274 from type location. In this study, the beautiful illustrations were provided, intuitively presented the shape of the fusiform spore to people. The newly added specimen, HMAS83274 ex MHSU2008 used to be cited as Melanogaster fusisporus var. obovatus (this variety has been revised to Melanogaster obovatus based on present study.) in Flora Fungorum Sinicorum (Vol.7) (Liu et al. 1998). Although we sequenced unsuccessfully, we noticed this specimen was mature and fitted well with M. fusisporus in morphyology, besides it was collected from type location in Chinese Yunnan Provence. For the above reasons, we diagnosed this specimen as M. fusisporus. Melanogaster fusisporus is diagnosed by its fusiform basidiospores with a bluntly apiculate apex, orange brown or deep brown basidiospores at maturity, easy to be recognized. Melanogaster quercus differs from M. fusisporus by its brick red basidiomata and light yellowish basidiospores of 10.8–15.3 × 5.6–7.7 µm.
Material examined: CHINA. HEILONGJIANG PROV.: Ningan City, Jingpo Lake, in soil under Quercus monglica, 12 Sept 1987, K. Tao, L. Fan 193 (HMAS83095 ex MHSU2004, paratype); YUNNAN PROV.: Kunming City, Heilongtan, Kunming Botanical Garden, in soil under Q. acutissima, 10 August 1990, M.-c. Chang, L. Wang, D.-s. Wang 239 (HMAS96755 ex MHSU2005, paratype); ibid., M.-c. Chang, L. Wang 224 (HMAS83274 ex MHSU2008).
Melanogaster minobovatus L. Fan & X.Y. Yan sp.nov. MycoBank MB Fig. 3, k–n
Etymology: minobovatus, min, minor, obovatus, obovoid, referring to the size and shape of basidiospores.
Type: CHINA. SHANXI PROV.: Linfen City, Xi County, shenjiagou, in soil under Pinus bungeana, Alt. 1321 m, 10 Sept 2017, Y.-y. Xu 035 (BJTC FAN911).
Description
Basidioma 3.5 × 4.5 cm, hypogeous, subglobose, yellow-brown, without distinct basal zone; rhizomorphs invisible, Strong wine aroma or fruit aroma. Gleba black, liquefying to produce a very dark spore mass. Peridium two-layered, 20–50 µm thick, composed of interwoven hyphae, 4–7 µm in diam, smooth, clamp connections abundant; subcutis 300–450 µm thick, composed of interwoven hyphae, with a large number of inflated cells, ellipsoidal, irregular or puzzle-like, up to 6.5–17 µm wide, light yellowish to yellow toward outside surface. Trama plates of hyaline gelatinized hyphae. Basidia not observed. Basidiospores obovoid, 5.1–6.9 × 4.2–5.1 µm (avX = 6.1 ± 0.4 × 4.7 ± 0.2, n = 50), Q = 1.1–1.5 (avX = 1.3 ± 0.1, n = 50), deep brown at maturity, 0.5–1 µm thick, smooth, the base truncate-cupped, with very short remnants of sterigmata.
Habit, habitat and distribution: September, in soil under Pinus bungeana. Warm temperate zone of North China.
Notes
This species is distinguished from other Melanogaster species by its basidioma with strong wine aroma or fruit aroma, and small, obovoid and deep brown basidiospores. Melanogaster minisporus Cázares, Guevara, described from Mexico by Cázares et al. (2008), is similar to M. minobovatus in basidiospore shape and size. However, M. minisporus differs from M. minobovatus in its bright yellow basidiomata and dark brown gleba with white or yellow at maturity. Phylogenetically, an environmental sequence (Genbank accession JQ318611) associated with Quercus liaotungensis from Loess Platea, China was grouped a strong support clade (BS = 100, PP = 1.00) together with M. minobovatus (Fig. 1), and both of them shared 99% ITS similarity from each other, which supported that M. minobovatus is a distinct species, and is associated to Quercus sp. and Pinus sp.
Melanogaster obovatus (K. Tao, Ming C. Chang & B. Liu) L. Fan & X.Y. Yan comb. & stat. nov. MycoBank MB Fig. 3, o–r
≡Melanogaster fusisporus var. obovatus K. Tao, Ming C. Chang & B. Liu, Journal of Shanxi University, Natural Science 18 (4): 450 (1995).Type:—CHINA. SHANXI PROV.: Yucheng City, Yuanqu County, Lishan Town, Houwentang Village, in soil under Spiraea pubescens Turcz., 2 August 1990, M.-c. Chang 193 (HMAS83257 ex MHSU1470).
Diagnosis: Melanogaster obovatus is diagnosed by its light brick red or orange brown basidiomata, yellow green to yellow brown gleba, elongate-fusiform with rounded top to obovate and light yellow brown basidiospores. Melanogaster fusisporus differs from M. obovatus by its black gleba at maturity, fusiform with acute apex and dark brown basidiospores of (10–)11.5–13(–15)×(5–)5.5–6(–6.5) µm. Melanogaster intermedius (Berkeley) Zeller & Dodge, described from England and Amercia, is somewhat similar to M. obovatus in basidiomata appearance. However, M. intermedius is distinguished from M. obovatus by its thin peridium of 125–160 µm, slaty black gleba and wide basidiospores of 11–13 × 7.4–8 µm.
Description
Basidiomata 1.5–2.3 × 0.7–1.5 cm, hypogeous, subglobose, light brick red or orange brown, minutely tomentose, without distinct basal zone; rhizomorphs few, brown; sweet odor, like fruits flavor. Gleba gelatinous, yellow green to yellow brown, with sterile white veins scattered throughout, liquefying to become the slimy spore mass at maturity. Peridium two-layered, epicutis 50–100 µm thick, composed of interwoven hyphae, yellow brown, 4–10 µm in diam, smooth, clamp connections abundant; subcutis 300–500 µm thick, composed of interwoven hyphae, with massive inflated cells, ellipsoidal, irregular, up to 10–32 µm wide, yellowish to yellow brown toward outside surface. Trama plates of hyaline or yellowish gelatinized hyphae. Basidia 10–14.2 × 4.9–6.7 µm, light yellow clavate-capitate when young, 4–spored, sterigmata short, collapsed and disappeared at maturity. Basidiospores elongate-fusiform with rounded top to obovate, 10.3–14.8 × 4.6–7.3 µm (avX = 12.2 ± 0.9 × 6.0 ± 0.4, n = 50), Q = 1.7–2.6 (avQ = 2.1 ± 0.2, n = 50), light yellow brown, walls 0.5 µm thick, smooth, the base broadly truncate-cupped, with very short remnants of sterigmata.
Habit, habitat and distribution
June to September, in soil under Quercus sp., Pinus tabuliformis Carr. Temperate zone of North and Northeast China.
Notes
Phylogenetic analysis based on the sequences of specimens newly collected from type locality and morphologically consistent with the type specimen of M. fusisporus var. obovatus, revealed that these sequences clustered the independent clade J with a strong support (BS = 96/PP = 1.00) (Fig. 1), supporting they represented a distinct species. Therefore, we raised M. fusisporus var. obvatus to species level as M. obvatus, and an epitype (BJTC FAN1091) was designated here. Phylogenetically, M. quercus and M. shanxiensis were related to M. obovatus closely. However, they morphologically differed from M. obovatus in black gleba and elongate-fusiform spores with apiculate apex for the former and white to pale yellow basidioma and elongate-obovatus basidiospores for the latter.
Material examined: CHINA. JILIN PROV.: Dunhua city, Dashitou county, under Quercus mongolica Fisch. ex Ledeb., 15 September 1987, L. Fan, K. Tao 226 (HMAS83269 ex MHSU2010). SHANXI PROV.: Jinzhong city, Lingshi county, Houhuigou, 27 June 1987, under Q. liaotungensis Koidz., T.-l. Shangguan 234 (HMAS81918 ex MHSU2015); Yuncheng City, Yuanqu county, Lishan Town, Houwentang Village, Alt. 1280 m, 2 August 1990, Y. Ma, B. Qiao, X.-k. Bai 198 (HMAS83094 ex MHSU2006); ibid., X.-k. Bai, B. Qiao, Y. Ma 215 (HMAS83267 ex MHSU2007); ibid., Y. Ma, B. Qiao, X.-k. Bai 225 (HMAS83265 ex MHSU2009); ibid., Y. Ma, X.-k. Bai, B. Qiao 227 (HMAS83266 ex MHSU2011); ibid., B. Qiao, Y. Ma, X.-k. Bai 501 (HMAS83268 ex MHSU2018); ibid., 1 August 1990, Y. Ma, X.-k. Bai, B. Qiao 231 (HMAS83270 ex MHSU2014); ibid., Y. Ma, B. Qiao, X.-k. Bai 503 (HMAS83272 ex MHSU2019); ibid., under Q. liaotungensis, 27 July 1987, M.-c. Chang 228 (HMAS83271 ex MHSU2012); ibid., Alt. 1900 m, 24 July 1987, M.-c. Chang 230 (HMAS83264 ex MHSU2013); ibid., 20 July 1987, M.-c. Chang, Y.-h. Chang 240 (HMAS83273 ex MHSU2017); ibid., 25 July 1987, M.-c. Chang 236 (HMAS83275 ex MHSU2016); Yuncheng City, Xia County, Sijiao Town, Yujialing Village, Alt. 970 m, 27 Octomber 2017, in soil under Q. variabilis, T. Li 055 (BJTC FAN1081); ibid., X.-y. Yan 123 (BJTC FAN1091, epitype, designated here); in soil under Q. variabilis Bl., Y.-y. Xu 072 (BJTC FAN1102-A); ibid., Y.-y. Xu 072 (BJTC FAN1102-B); ibid., in soil under Pinus tabuliformis, K.-b. Huang 156 (BJTC FAN1118); ibid., 19 August 2019, in soil under Quercus sp., Y.-y. Xu 150 (BJTC FAN1208).
Melanogaster panzhihuaensis L. Fan & X.Y. Yan sp.nov. MycoBank MB Fig. 3, s–v
Etymology: panzhihuaensis, referring to the type locality, where the type was collected.
Type: CHINA. SICHUAN PROV.: Panzhihua City, in soil under Pinus yunnanensis Franch., 1995, Z.-j. Gu 6 (HMAS 81915 ex MHSU2266).
Diagnosis: Melanogaster panzhihuaensis is diagnosed by its oval basidiospores with blunt and narrow apex and reddish brown basidiospores at maturity.
Description: Basidioma1.5 × 1.7 cm, hypogeous, subglobose, brown to dark brown when dry; no rhizomorphs visible; odor no record. Gleba pale yellow-brown when fresh, firm, black after dried, with sterile white veins scattered throughout. Peridium single-layered, 150–220 µm thick, compactly interwoven hyphae, 2.5–5 µm, with a large number of inflated cells, puzzle-like or irregular, up to 7.5–13 µm wide, light yellowish to yellow walls toward surface. Trama plates of interwoven hyphae, pale orange–yellow, thin-walled, 2–5 µm in diam. Basidia not observed. Basidiospores oval, apex blunt and narrow, 8.7–12.4 × 5.2–7.6 µm (avX = 10.4 ± 0.8 × 6.1 ± 0.6, n = 45), Q = 1.4–2.0 (avX = 1.7 ± 0.2, n = 45), reddish brown, the base truncate-cupped, walls 1 µm thick, smooth, the base broadly truncate-cupped, with very short remnants of sterigmata.
Habit, habitat and distribution
in soil under Pinus yunnanensis, Southwest China.
Notes
In present study, the sequence of M. panzhihuaensis showed less than 89.89% ITS similarity to other species and formed an independent clade U (Fig. 1), indicating it was a distinct species. In addition, the sequences of M. minobvatus formed sister clade with M. panzhihuaensis. However, M. minobvatus is differentiated from M. panzhihuaensis by its yellow-brown basidiomata, obovoid and small basidiosporesof 5.1–6.9 × 4.2–5.1 µm. Morphyologically, Melanogaster natsii is similar to M. panzhihuaensis in the color of gleba, but M. natsii differs from the latter in its broadly citriform or obovatus and pale yellow brown basidiospores.
Melanogaster quercus L. Fan & X.Y. Yan sp.nov. MycoBank MB Fig. 4, a–d
Etymology: quercus, referring to the host of this species.
Type: CHINA. SHANXI PROV.: Lvliang City, Jiaocheng County, Pangquangou, in soil under Quercus sp., Alt. 1879 m, 6 September 2017, Y.-y. Xu 011 (BJTC FAN809).
Diagnosis: Melanogaster quercus is diagnosed by brick red appearance, black gleba and light yellow elongate-fusiform basidiospores with apiculate apex. M. obovatus is closely related to M. quercus. However, M. obovatus differs from M. quercus in its yellow green to yellow brown gleba and elongate-fusiform with rounded top to obovate spores.
Description: Basidiomata 1.8–2 × 2–2.5 cm, hypogeous, subglobose, brick red, without distinct basal zone; no rhizomorphs visible; slightly sweet odor. Gleba gelatinous-firm, black, irregular glebal chambers separated by sterile white veins. Peridium two-layered, epicutis 50–100 µm thick, composed of interwoven hyphae, yellow, 4.5–10 µm in diam, clamp connections visible; subcutis 400–500 µm thick, composed of interwoven hyphae, with massive inflated cells, ellipsoidal, puzzle-like or irregular, 10–50 µm, thin-walled, yellowish to yellow toward ouside surface. Trama plates of hyaline or yellowish gelatinized hyphae. Basidia not observed. Basidiospores elongate-fusiform, apex apiculate, 10.8–15.3 × 5.6–7.7 µm (avX = 13.1 ± 1.1 × 6.5 ± 0.5, n = 50), Q = 1.6–2.4 (avX = 2.0 ± 0.2, n = 50), light yellowish, 0.5 µm thick, smooth, the base truncate-cupped, with very short remnants of sterigmata.
Habit, habitat and distribution
September, in soil under Quercus sp. Warm temperate zone of North China.
Notes
Phylogenetically, the sequences of Melanogaster quercus clustered together and branched as the sister group to the sequence of M. shanxiensisis with no support value, implying that M. quercus is a distinct species and related to M. shanxiensisis. However, M. shanxiensis differs from M. quercus in its white to pale yellow basidiomata and elongate-obovate basidiospores. Morphyologically and phylogenetically, M. obovatus is closely related to M. quercus. However, M. obovatus differs from M. quercus in its yellow green to yellow brown gleba and elongate-fusiform with rounded top to obovate spores. In addition, the sequences of the two species were grouped in two different branches (I and J) with a high support value (BS = 95/PP = 0.95) and (BS = 99/PP = 1.00) respectively.
Material examined: CHINA. SHANXI PROV.: Lvliang City, Jiaocheng County, Pangquangou, in soil under Quercus sp., Alt. 1879 m, 6 September 2017, Y.-y. Xu 010 (BJTC FAN808); ibid., in soil under Q. liaotungensis, T. Li 015 (BJTC FAN832).
Melanogaster shanxiensisi B. Liu, K. Tao & Ming C. Chang, Acta Mycol. Sin. 8(3): 210 (1989) Fig. 4, e–h
Description: Basidiomata 2–2.9 × 1.5–1.8 cm, hypogeous, subglobose, white to pale yellow when fresh, without distinct basal zone, smooth to minutely tomentose when dry; rhizomorphs visible; odor no record. Gleba firm, pale brown when young to brown to brownish black when mature, irregular glebal chambers separated by sterile yellowish brown veins. Peridium two-layered, epicutis 30–70 µm thick, composed of interwoven hyphae, deeply orange-yelow, 5.0–5.5 µm in diam, clamp connections visible; subcutis 150–350 µm thick, composed of interwoven hyphae, with massive inflated cells, ellipsoidal to irregular, 7–25 µm, thin-walled, yellowish to yellow toward ouside surface. Trama plates of hyaline or yellowish gelatinized hyphae. Basidia not observed. Basidiospores elongate-obovate to narrowly oblong, apex obtuse, 10.5–14.5 × 5.0–6.5 µm (avX = 12.2 ± 1.0 × 6.1 ± 0.4, n = 50), Q = 1.6–2.3 (avX = 2.0 ± 0.2, n = 50), light yellow brown, 1 µm thick, smooth, the base truncate-cupped, with very short remnants of sterigmata.
Habit, habitat and distribution
September, in soil under Lespedeza bicolor Turcz. Warm temperate zone of North China.
Notes
Melanogaster shanxiensisi can be easily distinguished from other Melanogaster species by its white to pale yellow basidiomata and yellow brown elongate-obovate to narrowly oblong basidiospores 10.5–14.5 × 5.0–6.5 µm. In present study, the sequence newly obtained from holotype (HMAS81910) of Melanogaster shanxiensisis showed less than 97.91% ITS similarity to other species, indicating it is a distinct species. Phylogenetic analysis reveals this sequence is sister clustered with M. quercus with no support value, implying that M. shanxiensisis is a distinct species and related to M. quercus. Morphyologically, M. quercus are differentiated from M. shanxiensisi by brick red basidiomata and elongate-fusiform basidiospores.
Material examined. CHINA. SHANXI PROV.: Yuncheng City, Yuanqu County, Lishan township, in soil under Lespedeza bicolor Turcz., 10 September 1987, M.-c. Chang 197 (HMAS81910 ex MHSU909, Holotype).
Melanogaster spinisporus Y. Wang, in Wang, Chang, Tao & Liu, Journal of Shanxi University, Natural Science 18(4): 450 (1995), emend. L. FAN & X.Y. Yan Fig. 4, i–n
Description
Basidiomata 3–3.5 × 4–5 cm, hypogeous, sometimes exposed, subglobose, lignt brown with some red brown or brick red when fresh, light brown to brown when dry, without distinct basal zone; rhizomorphs few, brown; strong wine aroma. Gleba dark red-brown to black when fresh, firm, gelatinous, with sterile white veins scattered throughout, trend toward liquefaction, black and hard when dry. Peridium single-layered, 360–750 µm thick; compactly interwoven hyphae, 5–7.5 µm in diam, thin-walled, light yellowish or yellow, clamp connections abindant. Trama plates of gelatinized hyphae. Basidia 23.0–36.5 × 8.0–14.5 µm, light brown clavate-capitate when young, 4-spored, sterigmata short, collapsed and disappeared at maturity. Basidiospores broadly ellipsoid to obovoid, subglobose, bilaterally symmetric, appearing smooth or decorated with sparse spine under light microscope (LM), 0.5–1 µm high, short strip, even connected into small pieces, irregularly verrucose under scanning electron micro-scope (SEM); 7.4–12.0 × 5.2–9.2 µm (avX = 9.6 ± 1.0 × 7.3 ± 0.7, n = 50), Q = 1.1–1.7 (avX = 1.3 ± 0.1, n = 50), deep brown at maturity, walls 0.5–1 µm thick the base truncate-cupped, with very short remnants of sterigmata.
Habit, habitat and distribution
Summer and Autumn (July, September to October), in soil under Quercus sp, Pinus bungeana, sometimes exposed. Warm temperate zone of North China.
Notes. Melanogaster spinisporus is diagnosed by its light brown with some red brown or brick red tints basidiomata, broadly ellipsoid to obovoid or subglobose basidiospores, decorated with short strip, even connected into small pieces, irregularly verrucose under SEM. Melanogaster spinisporus was original described obovoid spore, spinose and spine 0.5 µm high by Wang et al. (1995). However, we checked the holotype of Melanogaster spinisporus (HMAS27263 collected from Beijing of China in 1958 by Shuqun Deng), confirmed the spore ornamented by verrucose under SEM, but failed to obtain sequence. Phylogenetic analysis based on the sequences of specimens (BJTC FAN936, BJTC FAN938, BJTC FAN941, BJTC FAN1085, BJTC FAN1092) newly collected from Shanxi Province and morphologically consistent with the type specimen of M. spinisporus revealed that these sequences clustered in independent clade H with a strong support (BS = 99/PP = 1.00), supporting they represented a distinct species (Fig. 1). Therefore we recognized it as M. spinisporus, emended the description and an epitype (BJTC FAN941-A) was designated here. Phylogenetically, M. obovatus, M. quercus, M. shanxiensis, M. tomentellus were related to M. spinisporus. However, the formers could be distinguished obviously from the latter by their smooth elongate–fusiform or elongate-obovate spores. Morphologically, Melanogaster utriculatus Y. Wang, Castellano & Trappe have obovatus basidiospores with decoration described by Wang et al. (2005) from Japan is similar to M. spinisporus. However, M. utriculatus differs from M. spinisporus by its bigger spores (9–)11–15(–17) × (7–)8–11 µm and appressed or inflated utricle on the spore surface .
Material examined: CHINA. BEIJING: Beijing City, Mentougou District, Tantuo Temple, on the ground of Quercus sp. forest, July 1958, S.-q. Teng 4809 (HMAS27263, Holotype). SHANXI PROV.: Linfen City, Xi County, shenjiagou, in soil under Pinus bungeana, Alt. 1321 m, 10 September 2017, K.-b. Huang 110 (BJTC FAN936-A); ibid., K.-b. Huang 110 (BJTC FAN936-B); ibid., K.-b. Huang 110 (BJTC FAN936-C); ibid., Alt. 1326 m, in soil under Pinus bungeana, K.-b. Huang 112 (BJTC FAN938); ibid., K.-b. Huang 115 (BJTC FAN941-A, epitype, designated here); ibid., K.-b. Huang 115 (BJTC FAN941-B); Yuncheng City, Xia County, Sijiao Town, Yujialing Village, in soil under Quercus variabilis, Alt. 970 m, 27 October 2017, T. Li 059 (BJTC FAN1085); ibid., X.-y. Yan 124 (BJTC FAN1092).
Melanogaster subglobisporus K. Tao, Ming C. Chang & B. Liu, in Wang, Chang, Tao & Liu, Journal of Shanxi University, Natural Science 18(4): 451 (1995), emend. L. Fan & X.Y. Yan
Description
Basidiomata 1.5–2 × 0.5–1.5 cm, hypogeous, subglobose, yellow brown to brown when fresh, rust brown to deep brown when dry; rhizomorphs few, brown; sweet odor, like fruits flavor. Gleba firm, black, with sterile white veins scattered throughout. Peridium two-layered, epicutis 60–100 µm thick, composed of interwoven hyphae, yellow brown, 5–7.5 µm in diam, clamp connections visible; subcutis 100–200 µm thick, compactly interwoven hyphae, with some inflated cells, ellipsoidal, irregular, up to 7.5–15 µm wide, light yellowish to yellowish brown walls toward surface. Trama plates of interwoven hyphae, hyaline to pale yellow, 2.5–4 µm in diam. Basidia not observed. Basidiospores obovate, appearing smooth under LM, irregularly tiny verrucose under SEM, 8.7–10.6 × 6.6–8.0 µm (avX = 9.8 ± 0.5 × 7.0 ± 0.4, n = 50), Q = 1.1–1.5 (avX = 1.4 ± 01, n = 50), yellowish brown to brown, walls 1 µm thick.
Habit, habitat and distribution
Summer (July), in soil under Quercus sp. Warm temperate zone of North China.
Notes: Melanogaster subglobisporus is diagnosed by its yellow brown to brown basidiomata and obovate basiodiospores decorated with irregularly tiny verrucose under SEM. Melanogaster subglobisporus has obovoid or subglobose and smooth basidiospores in original descriptions (Wang et al. 1995). However, afterchecking the holotype and two specimens of M. subglobisporus (collected from type location) cited in Flora Fungorum Sinicorum (Vol. 7) (Liu et al., 1998), we confirmed its spores were ornamented with irregularly tiny verrucose under SEM. Phylogenetically, the sequence of holotype (HMAS83329 ex MHSU2001) clustered in the independent clade M and showed less than 91.49% ITS similarity with other species. Therefore we recognized the phylogenetic position of M. subglobisporus and emended the description with illustration. Phylogenetically and Morphyologically, M. spinisporus is related to M. subglobisporus with similar spore shape and size. However, M. spinisporus differs from M. subglobisporus by its light brown with some red brown or brick red basidiomata and basidiospore ornamented by short strip, even connected into small pieces, irregularly verrucose under SEM.
Material examined: CHINA. SHANXI PROV.: Jincheng City, Yangcheng County, Yangzhuang river, Alt. 580–600 m, 16 July 1990, Y. Ma, M.-c. Chang 504 (HMAS83329 ex MHSU2001, Holotype); ibid., Y. Ma, M.-c. Chang 243 (HMAS81919 ex MHSU2107, epitype, designated here); ibid., Alt. 600–620 m, under Quercus sp., 13 July 1990, M.-c. Chang 500 (HMAS81921 ex MHSU2110).
Melanogaster tomentellus L. Fan & X.Y. Yan sp. nov. MycoBank MB Fig. 4, s–w
Etymology: tomentellus, referring to the tomentose surface of the basidiomata.
Type: CHINA. YUNNAN PROV.: Kunming City, Xishan District, Haikou Town, in soil under Pinus sp., Alt. 1900 m, 8 January 2016, J.-z. Cao 1707 (BJTC FAN539).
Diagnosis: Melanogaster tomentellus could be easily diagnosed by its minutely green tomentose surface of basidiomata, small and elongate-fusiform spores.
Description
Basidiomata 1.1–2.2 × 1.9–2.5 cm, hypogeous, globose to subglobose, brown to deep brown, surface minutely green tomentose, without distinct basal zone, few brown rhizomorphs; odor strong turpentine flavor. Gleba black, solid, gelatinous, with sterile white veins scattered throughout. Peridium single-layered, 250–350 µm thick, textura globosa to textura prismatica structure (pseudoparenchymatous), made of short, subglobose, ellipsoidal or irregular round hyphae, 5–30 µm, thin-walled, light yellowish to yellow brown walls toward outside surface. Cystidia up to 15–40 µm long and 5–7.5 µm broad at base arising from some of the outermost cells of the peridium, clamp connections visible, thick-walled at apical tips. Trama composed of gelatinized hyphae. Basidia not observed. Basidiospores elongate fusiform, 8–11.4 × 3.8–5.6 µm (avX = 9.4 ± 0.8 × 4.5 ± 0.4, n = 50), Q = 1.7–2.7 (avX = 2.1 ± 0.2, n = 50), brown, walls 0.5 µm thick, smooth, the base broadly truncate-cupped, with very short remnants of sterigmata.
Habit, habitat and distribution
Winter (January), in soil under Pinus sp., plateau subtropical evergreen forests of Southern China.
Notes
Phylogenetically, the sequence of holotype (BJTC FAN539) clustered in the independent clade L with strong support (BS = 97/PP = 1.00) and showed less than 95.33% ITS similarity with other species. Therefore, we proposed M. tomentellus as a new species from China. M. obovatus, M. quercus and M. shanxiensis are related to M. tomentellus phylogenetically. However, M. obovatus and M. quercus differ from M. tomentellus in their light brick red or orange brown surface and yellowish or yellow brown basidiospores. M. shanxiensis differs from M. tomentellus in its white to pale yellow surface, yellow brown and big basidiospores (10.5–14.5 × 5.0–6.5 µm).
Key to species of Melanogaster from China
1. Basidiospores verrucose……………………………………………………………………………….2
1. Basidiospores smooth…………………………………………………………...…………………….3
2. Basidiospores appearing smooth or decorated with sparse spine under light microscope (LM), but short strip, even connected into small pieces, irregularly verrucose under scanning electron micro-scope (SEM), 7.4–12.0 × 5.2–9.2 µm………………………………...………….M. spinisporus
2. Basidiospores appearing smooth under LM, irregularly tiny verrucose under SEM, 8.7–10.6 × 6.6–8.0 µm…………………………………………………………………...……...M. subglobisporus
3. Basidiospores light yellow to yellow brown…………………………………………………………..4
3. Basidiospores brown to deep brown or reddish brown……………………….……………………….7
4. Basidiospores ovoid to ellipsoid, small, 5–7 µm long ……………..…..……...……...M. ovoidisporus
4. Basidiospores broadly citriform or obovatus to fusiform, large, 9.5–16 µm long, ………...........……5
5. Basidiospores elongate-obovatus to narrowly oblong, 10.5–14.5 × 5.0–6.5 µm…….....M. shanxiensis
5. Basidiospores broadly citriform or obovatus, 9.6–13.3 × 4.8–9.8 µm…………..…....………M. natsii
5. Basidiospores elongate-fusiform, 10–15 × 4.6–7.7 µm………………….....………..…………………6
6. Gleba yellow green to yellow brown, basidiospores elongate-fusiform with rounded top, or obovate …………………………………………………………………………...………….M. obovatus
6. Gleba black, basidiospores elongate-fusiform with apiculate apex ……………...………...M. quercus
7. Basidiospores reddish brown……………………………………………………….M. panzhihuaensis
7. Basidiospores yellow brown to deep brown…………………………………………………………..8
8. Basidiospores ellipsoid, oblong to cylindrical, 4.4–7.8 × 3.3–4.7 µm…….……..……M. broomeanus
8. Basidiospores fusiform………………………...……………………………………………………...9
8. Basidiospores obovoid……………………………………………………………………………….10
9. Basidiospores fusiform, apex bluntly apiculate, 10.0–15.9 × 4.8–6.3 µm…………..…...M. fusisporus
9. Basidiospores elongate fusiform, apex apiculate, 8–11.4 × 3.8–5.6 µm………….…….M. tomentellus
10. Gleba yellowish to brown, basidiospores obovoid, sometimes narrowly oblong or ellipsoid, 6.5–8 × 3.8–4.7 µm……………………………………………………………...…..……..M. obovatisporus
10. Gleba black, basidiospores obovoid, 5.1–6.9 × 4.2–5.1 µm………………...……….M. minobovatus