Corresponding author: Zai-Wei Ge (
Academic editor: S. Redhead
Taxonomic and phylogenetic studies of
Ge Z-W, Jacobs A, Vellinga EC, Sysouphanthong P, van der Walt R, Lavorato C, An Y-F, Yang ZL (2018) A multi-gene phylogeny of
The genus
Recently, three species,
Phylogenetic studies have shown that
Based on investigations of lepiotoid fungi in China, Dominican Republic, Germany, Italy, South Africa, Thailand, United Kingdom and the United States of America, detailed morphological and molecular studies were carried out in this study. The aims were to:
1. elucidate species diversity within
2. use a combined multi-gene dataset (
3. examine diagnostic characters for recognised clades and establish an infrageneric classification that best reflects the evolutionary history of the genus.
Fifty-nine collections were newly sampled from China, Dominican Republic, Germany, Italy, South Africa, Thailand, the United Kingdom and United States of America and deposited in HMAS, PREM, HKAS (Herbarium of Cryptogams, Kunming Institute of Botany, Chinese Academy of Sciences) and MFLU. Twelve out of the 16 recognised species, plus two recently described species, as well as two putative new species and a new combination were represented in this study. Morphological characters were studied from field notes, colour images of the material and complemented with literature data. Colour names and codes are from
A small piece of dried basidiocarp was excised from a specimen and ground in an Eppendorf tube. Genomic DNA was isolated using the CTAB method (
In this study, 144 sequences were produced using standard methods and deposited in GenBank (
The
Sequences were aligned using MAFFT 6.8 (
Taxa, vouchers, geographic origin, and GenBank accession numbers of DNA sequences of
Taxon | Collection | Origin | nrITS |
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PREM 62140 | South Africa |
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PREM 62141 | South Africa |
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HKAS 101312 | Russia |
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HMAS 71678 | China: Neimenggu |
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AH31724 | Mexico |
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N/A | N/A |
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Trappe 11481 (AZ80) | USA |
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N/A | N/A |
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HKAS 101315 | Italy |
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Z. W. Ge 3112 | China: Yunnan |
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Z. W. Ge 3574 | China: Yunnan |
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Z. W. Ge 2006-1 | China: Yunnan |
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N/A | N/A |
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PREM 62147 | South Africa |
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HKAS 101317 | China: Hainan |
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Z. W. Ge 3115 | China: Yunnan |
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HKAS 90470 | China: Yunnan |
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AH45540 | Spain |
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N/A | N/A |
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AH43927 | Spain |
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N/A | N/A |
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HKAS 45051 | China: Hunan |
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Z. W. Ge 3381 | USA: Florida |
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Z. W. Ge 3146 | China: Yunnan |
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HKAS 101322 | Italy |
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Z. W. Ge 3377 | USA: Florida |
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HKAS 83208 | China: Zhejiang |
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HKAS 31587 | Germany: Marburg |
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HKAS 53466 | Germany: Marburg |
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PREM 62142 | South Africa |
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PREM 62145 | South Africa |
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HKAS 93747 | Benin: Okpara |
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AM155 | India: West Bengal |
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N/A | N/A |
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N/A |
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HMAS 66153 | China: Neimenggu |
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HMAS 71683 | China: Neimenggu |
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Z. W. Ge 3411 | USA: Florida |
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Z. W. Ge 3232 | USA: Florida |
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Z. W. Ge 3385 | USA: Florida |
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Z. W. Ge 3242 | USA: Florida |
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N/A | ||
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png012 | Thailand |
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N/A | ||
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Z. W. Ge 2175 | China: Yunnan |
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The
The
The combined data set included subsamples of the 17 species recovered in the
Emphasising both molecular data and morphological characters, a new infrageneric classification for
The genus
≡
Basidiocarps medium to large sized, stout, agaricoid, with obvious plate-like squamules. Basidiospores olive to greenish-white, thick-walled, ellipsoid to amygdaliform with a truncate apex, except for
This section contains the type species of this genus,
Differs from other sections by the slender basidiocarps with furfuraceous squamules on the pileus, the non-pored, ellipsoid basidiospores and subcylindric to slightly fusiform cheilocystidia.
≡
Basidiocarps agaricoid, small to medium sized, with furfuraceous squamules. Basidiospores ellipsoid to ovoid without germ pore. Cheilocystidia narrowly clavate to subcylindrical. Pileipellis a loose hymeniderm made up of clavate to subfusiform hyphae.
This section is represented by
≡
Basidiocarps secotioid, with inconspicuous squamules. Basidiospores thick-walled, without germ pore.
This section currently contains only one species (
Differs from other sections by the relatively smaller, porous basidiospores (less than 10 μm long) and a pileipellis composed of a palisade of hyphae with cylindrical terminal elements.
≡
Basidiocarps small to medium-sized, agaricoid, covered with large squamules contrasting in colour with the background. Basidiospores relatively small (less than 10 μm long), with germ pore, forming a truncated apex. Cheilocystidia clavate to mucronate clavate. Pileipellis a palisade of hyphae with cylindrical terminal elements. Hyphae without clamp connections.
This section contains the species from south-eastern North America (
Differs from other sections by the stout basidiocarps with plate- like squamules on the pileus and white lamellae, basidiospores with wide germ pore and pileipellis composed of a tightly packed hymeniderm of cylindrical and flexuous, or narrowly clavate or narrowly lageniform elements.
≡
Basidiocarps medium to large sized, stout, agaricoid, with plate like squamules, basidiospores with wide germ pore, forming a truncated apex. Cheilocystidia clavate to sphaeropedunculate. Pileipellis a tightly packed hymeniderm of cylindrical and flexuous, or narrowly clavate or narrowly lageniform elements.
This section contains
Differs from other sections by the nonporous, globose to subglobose basidiospores and gasteroid basidiocarps or globose to subglobose basidiospores and a hymenodermal pileipellis made up of loosely arranged clavate to broadly clavate elements when the basidiocarps are agaricoid.
Basidiocarps agaricoid or gasteroid, covered with inconspicuous squamules. Basidiospores subglobose to globose without germ pore (with rounded apex). Cheilocystidia (if present) clavate to broadly clavate. Pileipellis a hymeniderm made up of loosely arranged clavate to broadly clavate elements.
This section contains the agaricoid
This species is distinguished from other
SOUTH AFRICA. 2229 BB Beit Bridge, Farm Matolege 133 MS (
Basidiocarps of representative species of
Micro-morphological features of
Pileus 30–50 mm broad, hemispherical to convex when young, expanding to broadly convex or applanate with age, sometimes with a prominent umbo; margin sulcate striate; surface covered with thin, yellow grey (4B2–4B3), orange grey (6B2) to grey orange (5B4) furfuraceous squamules, these remaining intact on the disc but elsewhere diffracted-scaly with expansion and receding from pileus margin. Lamellae free and remote from stipe, white to off white, crowded, narrow, up to 6 mm deep, with 1–2 series of lamellulae; edges entire, white. Stipe 35–60 × 3–6 mm, subcylindric, slightly enlarged at base, glabrous, white to light brown, hollow, nearly stuffed, with a simple annulus about 10–15 mm from top of the stipe. Context white, 1–2 mm thick in pileus, discolouring brown to red where bruised or handled, with strong mushroom odour, taste mild. Spore print white to cream.
Basidiospores [100,5,2] (7.5)8.0–9.0 × (5.5) 6.0–6.5(7.0) μm (mean 8.2 ± 0.4 × 6.2 ± 0.3 μm), Q = (1.2)1.3–1.4 (1.5), Qav = 1.3 ± 0.05, ellipsoid, occasionally ovoid in side view or in frontal view, with rounded apex, smooth, hyaline, congophilous, dextrinoid, with one guttule in most cases, without germ pore, slightly thick walled, becoming purplish-red (14A6–14A7) in cresyl blue. Basidia 29–33 × 10.0–11.0 µm, clavate, hyaline, 4-spored, rarely 2-spored. Cheilocystidia 28–50 × 6.0–10.0 µm, cylindric to slightly fusiform, hyaline. Pleurocystidia not observed. Lamella trama regular to slightly interwoven, made up of subcylindrical hyaline hyphae, 7.0–12.0 μm diam. Pileipellis a hymenidermal layer made up of greyish-yellow (1B4, 2B3) to dull yellow (3B3), clavate elements of 21–50 × 9.0–14.0(16.0) µm, slightly thick walled, with greyish-yellow vacuolar pigments; wall greyish-yellow; terminal elements mostly narrowly clavate. Clamp connections not observed.
So far, only known from South Africa.
Saprotrophic, solitary to scattered, terrestrial.
(L.) in reference to Africa where it is collected.
SOUTH AFRICA. 2229 BB Beit Bridge, Farm Matolege 133MS,
This species is distinguished from other
SOUTH AFRICA. 2229 BD Kamkusi, Farm Ludwigslust 163MS (
Micro-morphological features of
Pileus 50–100 mm broad, hemispherical to convex at first, expanding to convex to broadly convex with age; surface covered with fibrillose, tufted, reddish-white (7A2) to brownish-orange (6C3) squamules at the margin and brownish-grey (6C2), orange grey (6B2), to greyish-brown (7D3) plate-like squamules at the centre. Lamellae free and remote from stipe; white to off-white when young, whitish to greenish-white (26A2) when mature, crowded, 6–11 mm deep, with 1–2 series of lamellulae. Stipe 16–90 × 3.5–8 mm, subcylindrical, with slightly enlarged base, straight or curved, white; hollow, nearly stuffed, with an annulus at the middle part of the stipe. Context white, 4–6 mm thick in pileus, white in pileus and stipe, discolouring pastel pink (7A4) when drying, with a distinct mushroom smell, taste mild. Spore print greyish-green (30B3–30B4).
Basidiospores [100,5,3] (8.0)8.5–11.0(12.0) × (6.0)7.0–9.0(10.0) μm (mean 9.8 ± 0.9 × 8.0 ± 0.8 μm), Q = 1.0–1.4, Qav = 1.2 ± 0.05, ellipsoid, oblong in side view or in frontal view, with rounded apex, smooth, hyaline when young, greenish-white (27A2), olive to brownish (in KOH) when mature, congophilous, dextrinoid, without germ pore, slightly thick-walled; mature basidiospores staining purplish-red (14A6–14A7) in cresyl blue; immature basidiospores staining bluish-violet (18B7) in cresyl blue. Basidia 29–33 × 10.0–12.0(15.0) µm, clavate, hyaline, 4-spored. Cheilocystidia (13)20–55(63) × 10.0–15.0(20.0) µm, clavate, rarely broadly clavate or narrowly clavate, brownish to fuscous brown, sometimes septate. Pleurocystidia absent. Lamella trama slightly interwoven, made up of subcylindrical hyaline hyphae, 8–14 μm diam. Pileipellis a trichoderm made up of filamentous or cylindrical hyphae, slightly interwoven, interspersed with brown to tea brown hyphae, 8–14 µm in diam., thick-walled, with brownish vacuolar pigments; wall brownish-yellow; terminal elements mostly slightly enlarged to narrowly clavate, rarely cylindrical. Clamp connections present on basal septa of young basidia and tissue of annulus, but not common.
Known from Benin and South Africa in Africa and from China in Asia.
Saprotrophic, solitary to scattered, terrestrial.
(L.) with reference to distribution of this species in the Old World tropics.
BENIN. Okpara: countryside of North-eastern Parakou, 15 km from Parakou, alt. ca 330 m, 18 June, 2015, G. Wu 1370 (HKAS 93747). CHINA. Hainan Province: Sanya, Yalongwan, on man-made lawn near seaside, 29 June 2010, Z.W. Ge 2519 (HKAS 60195). SOUTH AFRICA. 2229 BB Beit Bridge, Farm Wimpsh 139 MS, 12 February 2014, Van Der Walt, R891 (PREM 62144), growing in cleared area, near water hole, among
VIETNAM. Hanoi: Tonkin, M. Demange 236 (Herb. Patouillard, FH 4244–holotype!).
Pileus small to medium-sized, 2.5–8.5 cm in diam. (Figure
Basidiospores [45/2/1] (7.5) 8.0–10.0 (12.5) × (5.0) 5.5–7.0 (7.5) μm, 8.7 ± 0.4 × 6.3 ± 0.3 μm, Q= (1.3)1.4–1.7 (1.8), Qav =1.5 ± 0.09; ellipsoid, hyaline, strongly dextrinoid, slightly thick-walled, apex lacking germ pore but somewhat thinner than other areas. Basidia 25–30 × 7–9 μm, clavate, 4-spored. Squamules on pileus (pileus disc of the smaller slice of the holotype) composed of clavate to narrowly clavate cells, 45–66 × 11–15 μm, hyaline to very pale brownish in KOH. Clamp connections not observed.
Known from China and Vietnam in Asia.
Saprotrophic, solitary to scattered, terrestrial.
CHINA. Yunnan Province: Xishuanbangna Prefecture, Mengla County, Mengyuan, alt. ca. 770 m, July 2, 2014, Z.W. Ge 3574 (HKAS 84412); same locality, K. Zhao 494 (HKAS 89157); Honghe Prefecture, Gejiu City, Manghao town, September 25, 2011, Z.W. Ge 3112 (HKAS 70616).
The distinctive characters of
CAMEROON. Yaoundé, alt. 780 m, growing on humus in shade under tree, 1 November, 1996, D.
Basidiocarps medium to large-sized (Figure
Basidiospores [40,2,2] (10.5)11.5–12.0 (12.5) × (8.0) 8.5–9.0 μm (mean 11.8 ± 0.4 × 8.7 ± 0.3 μm), Q = 1.3–1.4 (1.5), Qav = 1.4 ± 0.05, broadly amygdaliform in side view, ovoid in frontal view, with truncate apex, smooth, greenish-white (28A2), congophilous, dextrinoid, thick-walled (Figure
Micro-morphological features of
Known from Cameroon, Nigeria and South Africa in Africa and from China, India and Thailand in Asia.
Saprotrophic, solitary to scattered, terrestrial.
CHINA. Yunnan Province: between Yuanmou and Yongren, 28 June 2006, Z.W. Ge 2006-1 (HKAS 52741). SOUTH AFRICA. 2229 BD Kamkusi, Farm Ludwigslust 163 MS,
1 | Basidiocarps sequestrate (either secotioid or gasteroid), basidiospores statismosporic |
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– | Basidiocarps agaricoid, basidiospores ballistosporic |
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2 | Basidiocarps secotioid, the margin of the pileus does not break free from the stipe, hymenophore (gleba) labyrinthiform to sub-lamellate |
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– | Basidiocarps gasteroid, stipe absent or rudimentary with a thick whitish mycelial cord, gleba crossed by a columella and capillitium |
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3 | Columella not fully developed; basidiospores 7–12 µm in diam |
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– | Columella well-developed, reaching halfway up the basidiocarp; basidiospores 10–14(–15) × 10–13(–14) µm |
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4 | Basidiocarps overall white; basidiospores without germ pore, with rounded apex |
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– | Basidiocarps with distinct dark brown patches and squamules; basidiospores with germ pore; apex truncated |
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5 | Basidiospores subglobose to globose; cheilocystidia clavate to broadly clavate |
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– | Basidiospores ellipsoid; cheilocystidia subcylindric to slightly fusiform |
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6 | Spore print greyish-green, known from the palaeotropical regions |
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– | Spore print white, known from temperate region in Northern China |
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7 | Basidia 2-spored; widely distributed in Africa, America and Asia |
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– | Basidia 4-spored; known from palaeotropics |
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8 | Pileus with obvious umbo, basidiospores measuring (7.5) 8.0–10.5 (12.5) × (5.0) 5.5–7.0 (7.5), known from Southeast Asia |
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– | Pileus without obvious umbo, basidiospores (7.5) 8.0–9.0 × (5.5) 6.0–6.5(7.0) μm), known from South Africa |
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9 | Basidiospores less than 10 µm long; terminal elements of pileipellis cylindrical, basidiocarps grown in bamboo forest in east Asia or under oaks of Florida in south-eastern U.S.A |
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– | Basidiospores longer than 10 µm; terminal elements of pileipellis clavate to narrowly clavate; basidiocarps in various habitats (meadows, pastures, lawns, greenhouse, natural forests) |
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10 | Cheilocystidia clavate, without apical excrescences; clamp connections present at base of basidia and cheilocystidia; distributed in Asia |
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– | Cheilocystidia clavate, some mucronate or with apical excrescences; clamp connections absent at base of basidia and cheilocystidia; distributed in North America |
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11 | Spore print green; lamellae completely greenish with age |
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– | Spore print white or off-white; lamellae whitish or brownish with age, never totally green; sometimes a bluish-green shade is present near the stipe |
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12 | Cheilocystidia sphaeropedunculate to broadly clavate, often catenate |
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– | Cheilocystidia narrowly clavate to clavate |
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13 | Pileus squamules of similar colour as background, olivaceous brown to greyish-brown |
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– | Pileus squamules brown (different shades) on white to cream background, which is distinctly paler than squamules |
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14 | Clamp connections absent at base of basidia and cheilocystidia, cheilocystidia clavate to fusiform; annulus relatively simple |
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– | Clamp connections present at base of basidia and cheilocystidia, cheilocystidia narrowly clavate to fusiform; annulus relatively simple or complex |
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15 | Basidiocarps with abruptly to marginately bulbous stipe base; annulus relatively simple, without a double crown, but with a tough brown patch on the underside |
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– | Basidiocarps with widened base of stipe, but not abruptly so; annulus complex, with double crown |
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16 | Spore print yellowish-white to pale yellow to greyish-green, basidiospores greenish-white, 8–11 × 5–6 (7) µm |
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– | Spore print white, basidiospores hyaline, 10–10.7 (11) × (7) 8–8.5 (9.5) µm |
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In the present study, based on the extensive dataset comprising 75 % of all known species, four gene regions were used to clarify the evolutionary relationships of
The morphological diversity within
This study constitutes the first multigene molecular phylogenetic analysis of the genus
We are very grateful to Drs Z.H. Chen and P. Zhang of Hunan Normal University and Drs G. Wu, K. Zhao and Q. Zhao of Kunming Institute of Botany (KIB), Chinese Academy of Sciences for providing collections. We would like to thank Dr T.Z. Wei of the Institute of Microbiology, Chinese Academy of Sciences for sending us specimens on loan and Ms Shu Yao for assistance with lab work. This study was supported by the National Natural Science Foundation of China (No. 31461143031 and 31670024).
Figure S1. Maximum Likelihood tree showing the monophyly of
molecular data
Bootstrap values (>50) are indicated along nodes. The clade where