Corresponding author: Xiao-Lan He (
Academic editor: O. Raspé
In the present paper, three additional species of
He X-L, Horak E, Wang D, Peng W-H, Gan B-C (2018) Three new species of
Basidiomes of members in
In previous studies, seven species of
Fresh basidiomes were photographed in the field and described macroscopically. Colour notations follow
Genomic DNA was extracted with Biospin Fungus Genomic DNA Extraction Kit following the manufacturer’s instructions. PCR amplification was performed using DreamTaq™ Green PCR Master qMix (2×), Fermentas. The primers for RPB2 amplification were rpb2-6F and rpb2-7R, rpb2-i6f and rpb2-i7r (
Sequences used in phylogenetic analyses are listed in Table
A list of taxa, specimens and GenBank accession numbers of sequences used in this study.
Taxa | Collection No. | Origin | GenBank accessions | Remarks | |||
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ITS | LSU | RPB2 | mtSSU | ||||
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DL Largent 9641 | Australia: Queensland | – |
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GenBank ID: |
DL Largent 9663 | Australia: Queensland | – |
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GenBank ID: |
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ME Noordeloos 200314 | China: Jilin |
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DL Largent 9901 (Holotype) | Australia: Queensland | – |
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GenBank ID: |
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HMJAU 3886 (Holotype) | China: Jilin | – |
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– |
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GDGM 28821 | China: Guangdong |
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GDGM 27357 (Holotype) | China: Guangdong |
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DLL 9784 | Australia: Queensland | – |
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GenBank ID: |
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DL Largent 9623 | Australia: Queensland | – |
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GenBank ID: |
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China: Sichuan |
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– | ||
China: Sichuan |
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– | |||
China: Jilin |
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– |
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– | |||
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DL Largent 9934 (Holotype) | Australia: Queensland | – |
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GenBank ID: |
DL Largent 9900 | Australia: Queensland | – |
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GenBank ID: |
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GDGM 28818 (Holotype) | China: Jinlin |
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China: Jilin | – |
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– | – | |||
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China: Tibet |
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China: Tibet |
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China: Sichuan |
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DL Largent 9895 | Australia: Queensland | – |
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GenBank ID: |
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DL Largent 9662 | Australia: Queensland | – |
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GenBank ID: |
DL Largent 9670 | Australia: Queensland | – |
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GenBank ID: |
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DL Largent 9807 | Australia: Queensland | – |
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GenBank ID: |
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DL Largent 9811 | Australia: Queensland | – |
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GenBank ID: |
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DL Largent 9729 | Australia: Queensland | – |
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GenBank ID: |
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DL Largent 9778 | Australia: Queensland | – |
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GenBank ID: |
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TENN:068582 | USA: Tennessee |
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– | – | GenBank ID: |
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DL Largent 9794 | Australia: Queensland | – |
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GenBank ID: |
DL Largent 9834 | Australia: Queensland | – |
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GenBank ID: |
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DL Largent 9808 | Australia: Queensland | – |
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GenBank ID: |
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MEN 2004071 (Holotype) | Australia: Tasmania |
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DL Largent 9848 | Australia: Queensland | – |
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GenBank ID: |
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DL Largent 9932 (Holotype) | Australia: Queensland | – |
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GenBank ID: |
DL Largent 9949 | Australia: Queensland | – |
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GenBank ID: |
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MEN 200340 | Slovakia |
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China: Sichuan |
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China: Tibet |
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DL Largent 9809 (Holotype) | Australia: Queensland | – |
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GenBank ID: |
DL Largent 9810 | Australia: Queensland | – |
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GenBank ID: |
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K(M) 191739 (Holotype) | India: Kerala |
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– | – | |
China: Sichuan |
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China: Jilin |
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– |
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China: Jilin |
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– |
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China: Sichuan |
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– | – | |||
China: Jilin |
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– |
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China: Jilin |
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– |
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China: Jilin |
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– |
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China: Sichuan |
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China: Sichuan |
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– | – |
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080301 | China: Sichuan |
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– | – | ||
China: Sichuan |
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– |
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China: Shaanxi |
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China: Sichuan |
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China: Sichuan |
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China: Shaanxi |
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152 | Italy |
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– | – | – | |
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GDGM 28823 (Holotype) | China: Jilin |
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– | – | |
KA 12-1534 | South Korea |
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– | – | – | ||
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GDGM 28813 | China: Jilin |
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GDGM 28814 (Holotype) | China: Heilongjiang |
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P. Manomohan 645 (Holotype) | India: Kerala | – |
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GDGM 28815 | China: Yunnan |
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– |
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Sequences in bold and marked with “KU” and “MH” are newly generated in this study. Sequences marked with “GQ” were from
CHINA. SICHUAN PROV.: Yajiang County, Gexigou National Nature Reserve,
Basidiomata of the new species.
Pileus 5–15 mm broad, bluntly conic, convex or campanulate, dry, slightly hygrophanous, greyish-brown to brown (5C2–5D2), densely covered with suberect fibrils or minutely fibrillose squamules; fibrils silvery greyish, striate from entire margin to near centre. Lamellae sinuate, ventricose, distant, up to 3.5 mm wide, moderately thick, with two tiers of lamellulae, dark grey to brownish-grey, with entire and concolorous edge. Stipe 40–60 × 1–2.5 mm, central, cylindrical, equal, dry, concolorous with pileus, densely covered with grey to greyish fibrils, hollow, surface dry, with a pale yellow brownish to pale brownish strigose base. Context thin, concolorous with pileus. Odour and taste not distinctive.
Basidiospores (13–) 13.5–17.5 × 8–9.5 (–10.5) µm (
Scattered or gregarious on soil and amongst leaf litter in broadleaf forest dominated by
CHINA. SICHUAN PROVINCE: CHINA: SICHUAN PROV. Yajiang County, Gexigou National Nature Reserve,
Morphologically,
Microscopic structures of
CHINA. TIBET: Linzhi, Lulang,
Pileus 5–20 mm broad, hemispherical, convex, bluntly conic to broadly campanulate, dry, not hygrophanous, greyish-brown to brown (4D3–4E3), densely covered by suberect hispid or minutely squamulose overall, denser in centre; fibrils dark grey, pale grey brownish or concolorous with pileal surface (4D2–4D3), striate from entire margin to near centre. Lamellae sinuate with short decurrent tooth, ventricose, distant, moderately thick, up to 3 mm broad, with two tiers of lamellulae, dark grey to brownish-grey, with entire and concolorous edges. Stipe 20–50 × 0.7–1.5 mm, cylindrical, equal, dry, concolorous with pileus, densely covered with pale yellow brownish flocculose hairs, hollow, with a dirty yellowish to pale yellow brownish strigose base. Context thin, concolorous with pileus. Odour and taste not distinctive.
Basidiospores 12.5–15.5 (–17) × (6.5–) 7.5–9 (–9.5) µm (
Scattered on soil amongst decaying litter in mixed conifer-broadleaf forest dominated by
CHINA. TIBET: Linzhi, Lulang,
The greyish-brown pileus covered by suberect hispid or minutely squamulose, the brown external encrustations on pileipellis and stipitipellis and the cylindrical terminal cells of pileipellis and stipitipellis indicate
Microscopic structures of
CHINA: SICHUAN PROV.: Yajiang County, Gexigou National Nature Reserve,
Pileus 7–20 mm broad, conical-convex, truncate conical to broadly campanulate, dark reddish-brown (8D2–8D3) at first, becoming greyish-orange to pale beige brownish (5B2–5C2), dry, slightly hygrophanous, densely covered by reddish-brown erect or suberect squamules and fibrils; fibrils much denser at disc, margin not striate or very slightly striate only. Lamellae adnate to sinuate, ventricose, up to 2.5 mm wide, relatively thick, with two tiers of lamellulae, brownish-pink when mature, with concolorous and entire edges. Stipe 40–73 × 0.8–2 mm, central, cylindrical, hollow, densely covered with rust reddish hairs or fibrils, very dark brown strigose at base. Odour and taste not distinctive.
Basidiospores (12.5–) 13–17 × 7.5–9.5 µm (
Scattered on soil amongst decaying litter in broadleaf forest dominated by
CHINA. SICHUAN PROV.: Yajiang County, Gexigou National Nature Reserve,
The setiform terminal cells of pileipellis and stipitipellis place
Microscopic structures of
1 | Pileus reddish-brown or greenish-brown |
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– | Pileus greyish-brown |
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2 | Pileus covered with reddish-brown suberect fibrils |
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– | Pileus greenish-brown with reddish tinge, zonate |
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3 | Pileus covered with appressed or suberect silvery fibrils |
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– | Pileus squamulose or covered with suberect brownish fibrils |
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4 | Pileus covered with appressed silvery fibrils |
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– | Pileus covered with suberect silvery fibrils |
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5 | Pileus fibrillo-squamulose or squamulose-tomentose, growing in tropical forest |
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– | Pileus covered with erect or suberect fibrils |
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6 | Pileus pale brownish with pinkish tinge, basidiospores larger, average (16.8 ± 0.5) × (10.8 ± 0.3) µm |
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– | Pileus greyish-brown, basidiospores smaller, average (15.5 ± 0.5) × (8.8 ± 0.3) µm |
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7 | Average spore length less than 13 µm |
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– | Average spore length more than 13 µm |
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8 | Pileus greyish-brown, striate from entire margin to near centre |
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– | Pileus peach brown, not striate |
|
A total 76 sequences were generated in this study and they were deposited in GenBank. The combined dataset in the molecular analyses is composed of 62 specimens and 2925 aligned sites. MP, ML and Bayesian analyses produced almost the same topologies except for the unsupported branches and the MP tree is shown (Fig.
Phylogenetic relationships of
For the ITS sequences used in the analyses, both the size of the entire ITS1-5.8-ITS2 region as well as for ITS1 and ITS2 were separately compared. It is remarkable that the sizes of the entire ITS region and ITS1 were significantly divergent. In general, the total length of ITS sequences in subgen.
The three new species in this study were placed in different clades, showing they are quite different from each other.
In the present study, three new species of
In the phylogenetic analyses, 35 taxa in subgen.
The length of the ITS sequence of the species nested in Clade I, Clade II and Clade IV is relatively short, ranging from 591 bp to 709 bp. Available ITS sequences of
We express special thanks to Ms. Bo Zhang (Jilin Agricultural University) and Dr. Hai-Xia Ma (Chinese Academy of Tropical Agricultural Sciences) for assistance during fieldwork. This study is funded by the National Natural Science Foundation of China (Nos. 31400023, 31770020) and the Sichuan Provincial Innovation Ability Promotion Engineering (2016ZYPZ-028).