Research Article |
Corresponding author: ChengLin Hou ( chenglin-hou@cnu.edu.cn ) Academic editor: María P. Martín
© 2022 Hao Zhou, GuiQiang Cheng, XiMei Sun, RuiYi Cheng, HongLiang Zhang, YanMin Dong, ChengLin Hou.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation:
Zhou H, Cheng GQ, Sun XM, Cheng RY, Zhang HL, Dong YM, Hou CL (2022) Three new species of Candolleomyces (Agaricomycetes, Agaricales, Psathyrellaceae) from the Yanshan Mountains in China. MycoKeys 88: 109–121. https://doi.org/10.3897/mycokeys.88.81437
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Three new species, Candolleomyces incanus, C. subcandolleanus and C. yanshanensis, were found and described from Yanshan Mountains in China. The identification is based on morphological observation combined with phylogenetic analysis of ITS-LSU-Tef1α-TUB2. This study enriched the species diversity of Candolleomyces in Yanshan Mountains and provided important data support for the systematic study of Candolleomyces in the future.
molecular systematics, new taxon, Psathyrellaceae, taxonomy
Candolleomyces Wächter & A. Melzer was established in 2020, belonging to Basidiomycota, Agaricomycetes, Agaricales, Psathyrellaceae (
In previous studies of Psathyrella, there are approximately 100 taxa lacking pleurocystidia, but this feature has not been used as a key distinguishing feature (
Currently, there are 25 recognised species in Candolleomyces in the Index Fungorum website (http://www.indexfungorum.org, until Jan. 2022) and 10 species were reported in China (
Yanshan Mountains are located in North China and have a warm temperate continental monsoon climate, with higher plant diversity. The dominant plants include Quercus spp., Betula spp., Abies spp. and Pinus tabuliformis Carr. et al. (
Collections were obtained and photographed in the field from Yanshan Mountains in China from 2017 to 2020. The collected specimens were dehydrated with a dryer (Dorrex) at 50 °C and the specimens were deposited in the Herbarium of the College of Life Science, Capital Normal University, Beijing, China (
DNA extraction was achieved by the M5 Plant Genomic DNA Kit [Mei5 Biotechnology, Co., Ltd, China]. The purified DNA was dissolved in 1 × TE buffer and stored at – 20 °C for later use. The PCR amplifications were performed in Bio-Rad S1000 TM Thermal Cycler [Bio-Rad Laboratories, Inc, USA]. The primer sets ITS1/ITS4 (
The generated raw reads of the DNA sequences were used to obtain consensus sequences using SeqMan v.7.1.0 in the DNASTAR Lasergene Core Suite software (DNASTAR Inc., Madison, WI, USA). All sequences were aligned using MAFFT v.6 (
Taxa | Voucher | Locality | ITS | LSU | β-Tub | tef-1α |
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Candolleomyces aberdarensis | GLM-F116094 | Kenya | MH880928 | – | – | – |
C. albipes | DED8340 | Sao Tome | KX017209 | – | – | – |
C. badhyzensis | 79478 (TAA) Type | Turkmenistan | KC992883 | KC992883 | – | – |
C. badiophyllus | SZMC-NL-2347 | – | FN430699 | FM876268 | FN396261 | FM897252 |
C. cacao | SFSU DED 8339 | Sao Tome | NR148106 | – | – | – |
C. cacao | FP1R4 | USA | KU847452 | – | – | – |
C. cacao | MP2R2 | USA | KU847436 | – | – | – |
C. candolleanus | LAS73030 Neotype | Sweden | KM030175 | KM030175 | – | – |
C. cladii-marisci | CLUF302 Type | Italy | MK080112 | |||
C. efflorescens | Pegler2133 (K) | Sri Lanka | KC992941 | – | – | – |
C. eurysporus | GLM-F126263 Type | Germany | MT651560 | MT651560 | – | – |
C. incanus |
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China: Beijing | ON042759 | ON042766 | ON098513 | ON098508 |
C. incanus |
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China: Beijing | ON042760 | ON042767 | ON098514 | ON098509 |
C. leucotephrus | LÖ138-01 (UPS) | Sweden | KC992885 | KC992885 | KJ664865 | KJ732775 |
C. luteopallidus | Sharp20863 (MICH) Type | USA | KC992884 | KC992884 | – | – |
C. luteopallidus | HMJAU5148 | China: Jilin | MG734736 | MW301084 | MW314056 | MW314073 |
C. secotioides | UES2918 Type | Mexico | KR003281 | KR003282 | – | KR003283 |
C. singeri | HMJUA37867 | China: Jilin | MG734718 | MW301088 | MW314059 | MW314077 |
C. singeri | HMJAU37877 | China: Chongqing | MW301073 | MW301091 | MW314062 | MW314080 |
C. subcacao | HMJAU37807 Type | China: Henan | MW301064 | MW301092 | MW314063 | MW314081 |
C. subcacao | HMJAU37808 | China: Henan | MW301065 | MW301093 | MW314064 | MW314082 |
C. subcacao | HFJAU1014 | China: Jiangxi | MW559218 | – | – | – |
C. subcacao | HFJAU1274 | China: Jiangxi | MW559219 | – | – | – |
C. subcacao | HFJAU1488 | China:Anhui | MW559220 | – | – | – |
C. subcandolleanus |
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China: Tianjin | ON042755 | ON042762 | ON098510 | ON098505 |
C. subcandolleanus |
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China: Tianjin | ON042756 | ON042763 | – | – |
C. subminutisporus | HMJAU37801 Type | China: Hubei | MW301066 | MW301094 | MW314065 | MW314083 |
C. subminutisporus | HMJAU37916 | China: Henan | MW301067 | MW301095 | MW314066 | MW314084 |
C. subsingeri | HMJAU37811 Type | China: Jilin | MG734715 | MW301097 | MW314067 | MW314085 |
C. subsingeri | HMJAU37913 | China: Jilin | MG734725 | MW301098 | MW314068 | MW314086 |
C. sulcatotuberculosus | GB:LO55-12 | – | KJ138422 | KJ138422 | – | – |
C. sulcatotuberculosus | HFJAU1515 | China: Fujian | MW375696 | – | MW382967 | MW382965 |
C. sulcatotuberculosus | Chiarello 07-10-2013 | – | KJ138423 | – | – | – |
C. trinitatensis | TL9035 (C) | Ecuador | KC992882 | KC992882 | KJ664863 | – |
C. trinitatensis | ADK4162 (BR) | Togo | KC992886 | KC992886 | – | – |
C. yanshanensis |
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China: Beijing | ON042757 | ON042764 | ON098511 | ON098506 |
C. yanshanensis |
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China: Beijing | ON042758 | ON042765 | ON098512 | ON098507 |
Candolleomyces sp. | BAB-4773 | India | KP686450 | – | – | – |
Candolleomyces sp. | BAB-5172 | India | KR349656 | – | – | – |
Candolleomyces sp. | BAB-4748 | India | KR154977 | – | – | – |
Candolleomyces sp. | BAB-4747 | India | KR154976 | – | – | – |
Candolleomyces sp. | BAB-5202 | India | KT188611 | – | – | – |
Psathyrella multipedata | LÖ237-04 | Sweden | KC992888 | KC992888 | KJ664867 | KJ732777 |
Maximum Parsimony analysis was performed by a heuristic search option of 1000 random-addition sequences with a tree bisection and reconnection (TBR) algorithm. Maxtrees were set to 1000, branches of zero length were collapsed and all equally parsimonious trees were saved. Other calculated parsimony scores were tree length (TL), consistency index (CI), retention index (RI) and rescaled consistency (RC) (
Maximum Likelihood analysis was performed with a GTR site substitution model (
The combined alignment and phylogenetic tree were submitted on TreeBASE (www.treebase.org, study 29579).
For the ITS-LSU- Tef1α-TUB2 sequence dataset, a total of 3459 characters including gaps (694 for ITS, 1316 for LSU, 1023 for Tef1α, and 426 for TUB2) were included in the phylogenetic analysis. Using RAxML, MrBayes and PAUP to construct ML, Bayesian and MP phylogenetic trees, the results show that the topology and branching order were similar and the Bayesian tree is shown in this paper (Fig.
Multi-gene phylogenetic tree obtained from the Bayesian analysis. Numbers above branches are Bayesian posterior probability (pp) values, Maximum Likelihood bootstrap (MLB) and Maximum parsimony bootstrap (MP) values. Asterisks (*) denote branches with pp = 1.00, MLb = 100% and MPb = 100%. Numbers above branches represent strongly and moderately support (pp ≥ 0.95, MLb ≥ 50% and MPb ≥ 50%). The red font indicates the position of the new species.
Based on the results, six specimens were assigned to three branches and were described as three new species. The three new species (Candolleomyces yanshanensis, C. subcandolleanus, C. incanus) and a known species (Candolleomyces badiophyllus (Romagn.) D. Wächt. & A. Melzer etc.) clustered together in the phylogenetic tree. The three new species clustered into together (pp = 0.99, MLbs = 82%,MPbs = 74%), but three new species separately formed three subclades with high support value. Candolleomyces yanshanensis, C. subcandolleanus and C. incanus can be distinguished by the phylogenetic tree, sequence base differences and morphological characteristics.
yanshanensis referred to the locality where the type specimen was collected.
China, Beijing, Changping District, Beitaizi Village, 40.272906°N, 116.4203°E, alt. 149 m, 14 Aug 2019, coll. X.Y. Shen, H Zhou and R.T. Zhang,
Candolleomyces yanshanensis, pileus 20–60 mm, flabellate, flattening with age, hygrophanous. Basidiospores 5.8–8.2 × 3.3–5.4 μm, often with germ pore. Subglobose cell, irregular oval, (18) 20–27μm broad.
Pileus 20–60 mm, flabellate, flattening with age, hygrophanous, slightly dirty white (#e3dac9) to pale brown (#deb887). Veil white (#ffffff), fibrils in young, evanescent. Context 1.0–2.0 mm broad at centre, same colour as pileus. Lamellae sparsely to moderately, adnate, slightly dirty white (#e3dac9) to champagne (#fad6a5), edge white (#ffffff) as spores mature. Stipes 50–130 × 3–6 mm, smooth, fibrils on the base, cornsilk (#f0ead6) to white (#ffffff).
Basidiospores 5.8–8.2 × 3.3–5.4 μm, Q = 1.4–2.0, ellipsoid to long ellipsoid, ovoid to ellipsoid, partly triangular at base, dark brown (#b8860b) to brown (#b06500) in water, smooth, abundant, multi-guttules, often with germ pore. Basidia 17–31 × 5.8–7.5 μm, short clavate, hyaline, 4-spored. Cheilocystidia 22–35 (40) × 8–11 (15) μm, irregular utriform or claviform, apex obtuse or broadly obtuse or often subcapitate, rarely with deposits. Pileipellis consists of 2–3 cells deep layer of irregular subglobose cell, irregular oval, (18) 20–27μm broad.
. Clumped on the ground with rich humus in broad-leaved forests or broad-leaved shrubs.
China, Beijing, Changping District, Tailing, 40.327397°N, 116.21916°E, alt. 172 m, 17 Aug 2020, coll. X.Y. Shen, H Zhou and X.B. Huang,
subcandolleanus referred to its morphological similarity to Candolleomyces candolleanus (Fr.) D. Wächt. & A. Melzer.
China, Tianjin, Jizhou District, Sanjiebei, 40.227984°N, 117.43354°E, alt. 235 m, 17 Aug 2019, coll. X.Y. Shen, H. Zhou and R.T. Zhang,
Candolleomyces subcandolleanus, pileus 5–20 mm. Basidiospores 5.5–6.7 × 3.2–4.5 μm, germ pore absent. Cheilocystidia 21–28 (30) × 8–12 (15) μm. Subglobose cell, irregular oval or long oval, (13) 16–25 μm broad.
Pileus 5–20 mm, campanulate to conical, smooth, fibrils in young, evanescent, brown (#b06500) to golden brown (#996515). Veil white (#ffffff), fibrils in young, evanescent. Context 0.2–0.5 mm broad at centre, same colour as pileus. Lamellae moderately to normally, adnate, slightly dirty white (#e3dac9) to white (#ffffff), edge white (#ffffff) as spores mature. Stipes 20–60 × 1–3 mm, smooth, fibrils on the base, cornsilk (#f0ead6) to white (#ffffff).
Basidiospores 5.5–6.7 × 3.2–4.5 μm, Q = 1.4–2.0, ellipsoid to ovoid, pale cream (#fffff0) to pale lemon (#fffacd) in water, smooth, multi-guttules, germ pore absent. Basidia 18–27 × 5–10 μm, short clavate, hyaline, 4-spored. Cheilocystidia 21–28 (30) × 8–12 (15) μm, utriform or claviform, apex obtuse or broadly obtuse or often subcapitate, rarely with deposits. Trama of gills irregular. Pileipellis consists of irregular subglobose cell, irregular oval or long oval, (13) 16–25 μm broad.
Clumped on the ground with rich humus in broad-leaved forests or broad-leaved shrubs.
China, Tianjin, Jizhou District, Huangyaguan Great Wall, 40.245615°N, 117.44047°E, alt. 235 m, 17 Aug 2019, coll. X.Y. Shen, H. Zhou and R.T. Zhang,
incanus referred to the basidiomata appears incanus.
China, Beijing, Changping District, Sidaohe Village, 40.246374°N, 116.4406°E, alt. 114 m, 16 Aug 2020, coll. X.Y. Shen, H Zhou and X.B. Huang,
Candolleomyces incanus, pileus 5–25 mm, hemispherical to conical. Basidiospores 6.0–7.0 × 3.2–4.5 μm. Stipe 40–70×4–6 mm, smooth, germ pore absent. Subglobose cell, irregular oval or long oval, (22) 25–32 μm broad.
Pileus 5–25 mm, hemispherical to conical, hygrophanous, incanus (#f2f3f4) to nude (#fdf5e6). Veil white (#ffffff), fibrils in young, evanescent. Context 0.5–1.0 mm broad at centre, same colour as pileus. Lamellae moderately to normally, adnate, off-white (#f2f3f4) to white (#ffffff), edge white (#ffffff) as spores mature. Stipes 40–70 × 4–6 mm, smooth, hygrophanous, cornsilk (#f0ead6) to white (#ffffff).
Basidiospores 6.0–7.0 × 3.2–4.5 μm, Q = 1.4–1.9, ellipsoid, floral white (#fffaf0) to dark yellow (#eedc82) in water, smooth, abundant, multi-guttules, germ pore absent. Basidia 15–20 × 5–8 μm, short clavate, hyaline, 4-spored. Cheilocystidia 17–27 (31) × 7–11 (13) μm, utriform, apex obtuse or broadly obtuse or often subcapitate, rarely with deposits. Trama of gills irregular. Pileipellis consisted of irregular subglobose cell, irregular oval or long oval, (22) 25–32 μm broad.
Clumped on the ground with rich humus in deciduous broad-leaved or deciduous coniferous forests.
China, Beijing, Yanqing District, Yudu Mountain, 40.54399°N, 115.893984°E, alt. 860 m, 12 Sep 2018, coll. C.L. Hou, H Zhou and J.Q. Li,
In this study, three new species were identified by morphology and phylogeny. It is very interesting that the three new species C. yanshanensis, C. subcandolleanus and C. incanus formed a stronger supported clade and they clustered with Candolleomyces badiophyllus (Romagn.) D. Wächt. & A. Melze, Candolleomyces candolleanus, Candolleomyces badhyzensis (Kalamees) D. Wächt. & A. Melzer, Candolleomyces trinitatensis (R.E.D. Baker & W.T. Dale) D. Wächt. & A. Melzer and Candolleomyces cladii-marisci (Sicoli, N.G. Passal., De Giuseppe, Palermo & Pellegrino) J.Q. Yan together in the phylogenetic tree. In addition, three new species were weakly sister to the known species C. badiophyllus in the phylogenetic tree.
Candolleomyces yanshanensis and C. subcandolleanus are different in macroscopic morphology of basidiomata. Candolleomyces yanshanensis is lighter in pileus colour and C. yanshanensis has larger spores (5.8–8.2 × 3.3–5.4 vs. 5.5–6.7×3.2–4.5 μm) and longer cheilocystidia (22–35 × 8–11 vs. 21–28 × 8–12 μm) than those of C. subcandolleanus. Moreover, C. yanshanensis spores often have a germ pore. Candolleomyces subcandolleanus is very easily confused with C. candolleanus in the field because of their similar macroscopic characteristics. In particular, two species in these sections possess the combined characteristics of small basidiomata. C. candolleanus is the type species of Candolleomyces, with early studies on this species being based on the number of pleats and other characteristics, but this also led to confusion in the identification of this species. Candolleomyces subcandolleanus can be distinguished from C. candolleanus by the smaller spores (5.5–6.7 × 3.2–4.5 vs. 7–9 × 4–5 μm) (
Candolleomyces incanum, C. badiophyllus, C. candolleanus and C. badhyzensis are close to each other in the phylogenetic tree. However, the four species show significant differences in morphology. These species can be distinguished as follows: C. incanus has smaller and narrower spores (6.0–7.0 × 3.2–4.5 μm), whereas C. candolleanus, C. badhyzensis and C. badiophyllus have larger spores (Spores of C. candolleanus were 7.0–9.0 × 4.0–5.0 μm, spores of C. badhyzensis were 10.2–11.5 × 5.5–6.5 μm, spores of C. badiophyllus were 10–14 × 5–6 μm). In addition, C. incanus has smaller cheilocystidia (17–27 × 7–11 vs. 34–51 × 10–15 μm) than those of C. badhyzensis (
Except for morphological differences, the three new species in this study can also be distinguished by sequence similarity. Candolleomyces yanshanensis (
According to the research of
It is considered that the natural growth of Candolleomyces may be related to precipitation. However, the investigation and specimen collection in this study were carried out in the rainy season in July to August, with no collection in other periods. Therefore, more species of Candolleomyces might be expected in Yanshan Mountains.
This study was supported by the Biodiversity Survey and Assessment Project of the Ministry of Ecology and Environment, China. (2019HJ2096001006) and National Natural Science Foundation of China (No. 31870629).