Two new species of Agaricales, Deconica flavum and Entocybe roseoalbus, are described from Mount Tianmu, Zhejiang Province, Eastern China. Two new species are distinct and monophyletic based on morphology and phylogenetic analyses. Deconica flavum differs from other Deconica species in that the pileus is brow shallow in the center and stipe with yellowish white fibrils, scatted on litter under coniferous and broad-leaved mixed forest at 1162 m. Entocybe roseoalbus is distinguished from other species of Entocybe by nearly blue pileus and pinkish-white stipe, scatted on humus in similar forest conditions at 1025 m. The differences are discussed between the two new taxa and their similar species morphologically, and related species phylogenetically.

Entolomataceae, Strophariaceae, taxonomy

Agaricales is the largest order in the kingdom Fungi with nearly 20,000 species (Roskov et al. 2019). The members of the order play essential roles in the ecosystem as saprotrophs, ectomycorrhizae, lichens, and crops cultivated by termites and ants (Kalichman and Matheny 2020). Due to its great diversity, the order has been intensively studied worldwide (Kalichman and Matheny 2020).

The genus Deconica (W.G. Sm.) P. Karst. has been placed in the family Strophariaceae of Agaricales (Matheny et al. 2006; Ramírez-Cruz et al. 2013, 2020a) and was initially described as a subgenus of Agaricus (Smith 1870). It was subsequently raised to the genus level by Karsten (1879). But for the last several years species of Deconica were placed in the genus Psilocybe (Fr.) P. Kumm. because of the similarity in their morphology (Moncalvo et al. 2002; Noordeloos 2009). However, molecular studies have shown that the genus Deconica is phylogenetically distant from Psilocybe (Ramírez-Cruz et al. 2013). The genus Deconica is distinguished based on the absence of hallucinogenic compounds (Ramírez-Cruz et al. 2020b). The members of the genus Deconica have mycenoid, collybioid, crepidotoid, or omphaloid basidiomata occurring in/on the soil, grasses, mosses, rotten wood, trunks, and dung (Noordeloos 2009; Ramírez-Cruz et al. 2020b). Noordeloos (2011) mentioned that the genus Deconica has been divided into three sections namely, Deconica, Melanotus, and Merdariae based on characteristics of basidiomata, basidiospores, pileipellis, and cystidia. However, chrysocystidia have not been mentioned in most sections, except Deconica section (Singer 1986). Guzmán (2005) estimated around 133 species in the genus Deconica exist worldwide. However, He et al. (2019) and Kalichman and Matheny (2020) reported 44 and 45 taxa worldwide, respectively. Furthermore, Deconica was one of the largest genera with an unsequenced generic type (Kalichman and Matheny 2020).

Entocybe T. J. Baroni, Hofstetter & Largent is the genus placed in the family Entolomataceae within Agaricales (Baroni et al. 2011). Species of Entocybe were previously placed in section Turfosa, subgenus Entoloma of the genus Entoloma (Noordeloos 1992). The genus Entocybe was erected based on the morphological and molecular phylogeny-based data (Baroni et al. 2011). The basidiomata of Entocybe show slender tricholomatoid or mycenoid to collybioid habit and possess a relatively fragile, appressed fibrillose stipe (Baroni et al. 2011). The basidiospores of Entocybe are 6–10 angled in polar view with undulate-pustulate or rounded pustulate surface ornamentation, or ornamentation being composed of broken interconnected ridges with isolated pustules interspersed, which are similar to Rhodocybe (Baroni et al. 2011). Additionally, clamp connections are found on the hyphae in all tissues (Baroni and Lamoureux 2013). According to Index Fungorum (http://www.indexfungorum.org), Entocybe currently comprises around ten species.

In the past decade, new species, combinations, and records of Deconica and Entocybe have been reported across the world (Baroni et al. 2011; Baroni and Lamoureux 2013; da Silva et al. 2013, 2014; Park et al. 2017; Ramírez-Cruz et al. 2020a, 2020b). In China, diverse macrofungal resources have been documented (Wu et al. 2019). However, there is limited research on Deconica and Entocybe. During the present study, specimens were collected from the National Nature Reserve of Mount Tianmu located in Zhejiang Province, China from July to September 2022. Two new species, Deconica flavum and Entocybe roseoalbus within Agaricales, are described and illustrated based on morphological and phylogenetic evidence.

The analyses of Deconica generated four supported clades, labeled a–d (Fig. 1). Clade a (BS = 99%, PP = 1.00) grouped D. flavum, D. cokeriana, D. thailandensis, D. overeemii, D. magica, D. bayliasiana, D. novae-zelandiae, D. milvispora, D. micropora, D. phyllogena, and D. umbrina. The clade contained six taxa with chrysocystidia i.e. D. flavum, D. cokeriana, D. thailandensis, D. overeemii, D. milvispora, and D. umbrina. Moreover, D. flavum, D. cokeriana, D. thailandensis and D. overeemii formed a subclade with strong bootstrap supports (BS = 94%, PP = 0.98) in clade a. Deconica flavum differs from D. cokeriana (MK965913), D. cokeriana (MK965914), D. cokeriana (KC669315) and D. sp. (MK607606) by 7 bp, 7 bp, 8 bp and 9 bp respectively. In the clade, all the species have small basidiomata, ellipsoid, rhomboid, hexagonal, or rhomboid-nodulose basidiospores. Two collections formed an independent lineage in the trees with strong bootstrap supports (BS = 100%, PP = 0.99) in clade a (Fig. 1). Clade b (BS = 100%, PP = 1.00) included D. protea, D. horizontalis, D. citrispora, D. hartii, and D. coprophila. Clade c (BS = 95%, PP = 0.97) included D. chionophila, D. montana, and D. coprophila. In clade d (BS = 97%, PP = 1.00), Deconica sp. and D. pratensis got clustered into one branch.

The analyses of Entocybe resulted in two well-supported clades, clade a and clade b (Fig. 2). Clade a mainly consists of Entocybe species (BS = 99%, PP = 1.00). Clade b is composed of most Entoloma species used in this study (BS = 99%, PP = 1.00). The trees showed that the new species identified as Entocybe formed a well-supported clade (BS = 100%, PP = 1.00) in clade a (Baroni et al. 2011).

Taxonomy

 Deconica flavum Y.Y. Shen & Y.B. Song, sp. nov.

Index Fungorum: IF901538

Facesoffungi Number: FoF16635

Figs 3, 4

Etymology

The specific epithet flavum (Lat.) refers to the species having stramineous color in the center of the pileus.

Figure 3. 

Deconica flavum (holotype) a–c basidiomata d–e basidiospores f basidium g pleurocystidia type chrysocystidia h pleurocystidia type leptocystidium i cheilocystidia. Scale bars: 10 mm (a–c); 10 µm (d, g, h); 5 µm (e, f, i).

Holotype

• China, Zhejiang Province, Hangzhou, National Nature Reserve of Mount Tianmu at 1162 m a.s.l., 30°21'N, 119°26.4'E (DDM), grew on litter under coniferous and broad-leaved mixed forest, 2 July 2022, 2381 (holotype), GenBank accessions: OR906279 (ITS), OR906277 (LSU).

Figure 4. 

Micromorphological features of Deconica flavum (holotype) a basidiospores b basidia c pleurocystidia type leptocystidium d cheilocystidia e pleurocystidia type chrysocystidia. Scale bars: 10 µm.

Description

Pileus convex-campanulate, commonly papillate, light brown (6D8) or sunburn (6D5), center stramineus (5C8), (8–)10–20 mm; margin striate, invariable color when bruised, surface flocculose or with white fibrillose patches of veil on half of the pileus. Context fleshy on disk, pale yellowish (6A3). Lamellae adnexed to adnate, with decurrent tooth, distant, pompeian yellow (5C6), brown shellow (5C8), or gold brown (5D7). Stipe central, cylindrical, equal to broader at apex, hollow, cartilaginous, flocculose, 11–14 × 1.2–2.7 mm; dark blonde (5D4) to light brown (6D5), with yellowish white fibrils. Smell indistinct.

Basidiospores fusiform, ellipsoid to ovoid, yellowish brown under light microscopy with germ pore, the middle part concave under scanning electron microscopy, (3.0) 3.3–4.7 (5.1) × (2.2) 2.6–3.3 (4.3) µm, Q = 0.8–1.8, Q_av = 1.4 (concave side), (3.6) 4.2–4.9 (5.1) × (2.9) 3.1–3.9 (4.0) µm, Q = 1.0–1.5, Q_av = 1.3 (convex side) in frontal view. Basidia cylindrical or claviform with median constriction, 4-spored, hyaline, thin-walled, 11.7–17.1 × 3.8–5.7 µm. Pleurocystidia type chrysocystidia clavate to broadly clavate, apex mucronate or rostrate, thin-walled, hyaline, with hyaline content, 15.2–26.8 × 5.6–13.0 µm. Pleurocystidia type leptocystidium narrowly utriform, hyaline, thin-walled, abundant, 7.7–17.9 × 3.3–5.7 µm. Cheilocystidia widely utriform, cylindrical, hyaline, 12.3–20.5 × 3.6–5.2 µm. Pileipellis a gelatinous cutis 2.0–4.5 µm diam, hyaline, and thin-walled, with clamp connections. Stipitipellis a cutis 4.7–10.6 µm diam, hyaline, thin-walled, with clamp connections.

Habitat and distribution

Scattered on litter under coniferous and broad-leaved mixed forests at 1162 m a.s.l., currently only known from Zhejiang Province, China.

Additional material examined

(paratype). • China, Zhejiang Province, Hangzhou, National Nature Reserve of Mount Tianmu at 1162 m a.s.l., 30°21'N, 119°26.4'E (DDM), grew on litter under coniferous and broad-leaved mixed forest, 2 July 2022, 2382, GenBank accessions: OR9066280 (ITS), OR906278 (LSU).

 Entocybe roseoalbus Y.Y. Shen & Y.B. Song, sp. nov.

Index Fungorum: IF902333

Facesoffungi Number: FoF16636

Figs 5, 6

Etymology

The specific epithet roseoalbus (Lat.) refers to the pinkish-white stipe.

Holotype

• China, Zhejiang Province, Hangzhou, National Nature Reserve of Mount Tianmu at 1025 m a.s.l., 30°20.4'N, 119°26.4'E (DDM), grew on humus under coniferous and broad-leaved mixed forest, 2 September 2022, 3461 (holotype), GenBank accessions: PP974446 (ITS) and PP974447 (LSU).

Figure 5. 

Basidiomata of Entocybe roseoalbus (holotype) a habitat of Entocybe roseoalbus b whole basidiomata c lamellae of Entocybe roseoalbus d stipe of Entocybe roseoalbus. Scale bars: 10 mm.

Description

Pileus umbonate, undulating, occasional dehiscence in the middle, and slight dehiscence at the edge when mature, not hygrophanous, not translucent-striate, surface finely felted with densely appressed-fibrillose or matted-fibrillose, rivulose, blackish blue (20F7 or 20F8) in the middle, gradually lighter, becoming dark blue (20E6), 47–68 mm diam. Context white, 1.8–1.9 mm thick above the stipe. Lamellae unequal, adnate, margin slightly serrate, 23–26 × 9.3–11.2 mm (length × breadth), at first pinkish white (7A2) then pastel red (7A4) to pale red (7A3) with basidiospore maturity. Stipe central, 69–72 mm long, 7.2–7.7 mm (apex)–7.1–7.5 mm (middle)–8.3–8.8 mm (base) diam, equal but slightly thinner in the upper middle, hollow and splits longitudinally with ease, pinkish white (10A2), white at the base, fragile. Odor not distinctive. Taste not recorded.

Figure 6. 

Entocybe roseoalbus (holotype) a marginal cell b basidiospores under the light microscope in oil (1000×) c basidiospores under SEM (5200×) d–e basidia f–i cheilocystidia j pileocystidia k pileipellis l stipitipellis. Scale bars: 10 µm.

Basidiospores distinctly angular (6–8 angled) to some indistinctly and faintly rounded pustulate, ornamentation composed of broken ridges under an SEM, (4.0) 4.73–5.6 (6.5) × (4.6) 4.8–5.5 (6.1) µm, Q = 0.7–1.3, Q_av = 1.0 in side-view. Basidia clavate, 3– or 4– sterigmate, filled with refractive oil bodies, 22.3–32.8 × 7.1–9.9 µm. Hymenial cystidia absent. Hymenophoral trama subregular, made up of cylindrical to slightly inflated elements, 36.9–92.5 × 11.6–22.8 µm. Lamella edge crowded with tufts of cheilocystidia. Cheilocystidia clavate, hyaline, abundant, 12.7–25.1 × 2.3–7.1 µm. Hymenial cystidia absent. Pileipellis multi-layered cutis, cylindrical, pigments intracellular, with special long and curved hyphae, 30.2–60.8 × 8–20.9 µm. Pileocystidia (terminal cells) narrowly cylindric to clavate, pigments intracellular, subtended by inflated cells of the pileal trama, 15.7–30.8 × 2.8–4.5 µm. Stipitipellis multi-layered cutis, similar to pileipellis, cylindrical hyphae, pigments intracellular, with special long and curved hyphae, 7.7–12.4 µm diam. Caulocystidia absent. Clamp connections present in all tissues.

Habitat and distribution

Scatted on humus under coniferous and broad-leaved mixed forests at 1025 m a.s.l., currently only known from Zhejiang Province, China.

Additional material examined

(paratype). • China, Zhejiang Province, Hangzhou, National Nature Reserve of Mount Tianmu at 1025 m a.s.l., 30°20.4'N, 119°26.4'E (DDM), scatted on humus under coniferous and broad-leaved mixed forest, 2 September 2022, 3462, GenBank accessions: PP974445 (ITS) and PP974448 (LSU).

Deconica flavum is characterized by small and convex basidiomata with ellipsoid to ovoid basidiospores, two types of pleurocystidia, chrysocystidia and leptocystidia. It was growing on litter as other Deconica species. Entocybe roseoalbus is peculiar in having isodiametric basidiospores with 6–8 angles and broken ridges, pileipellis and stipitipellis with intracellular pigment, and abundant clamp connections in all tissues, as shown by the other Entocybe species. Furthermore, phylogenetically in ML and BI trees, the specimens formed two distinct lineages within Deconica and Entocybe, respectively (Figs 1, 2).

Deconica species are distributed worldwide, with notable records in Europe, America, South Asia, and Oceania (GBIF, https://www.gbif.org/search?q=Deconica). Many species of the genus Deconica have been described recently based on the ITS phylogenetic analysis (Ramírez-Cruz et al. 2013, 2020a, 2020b). In the present research work, ITS and LSU sequences of D. flavum were generated. Due to the lack of other gene sequences (LSU), we perform only single gene (ITS) phylogenetic analysis. The phylogenetic analysis of 58 ITS sequences from Deconica including two newly generated sequences formed four clades with strong bootstrap supports (Fig. 1). In clade a (BS = 99%, PP = 1.00), six taxa possess chrysocystidia, four of which are clustered into a subclade with strong bootstrap supports (BS = 94%, PP = 0.98). The four species included D. flavum, D. cokeriana, D. thailandensis and D. overeemii. Although the phylogenetical distances of D. flavum, and D. cokeriana are close in ITS trees, they have distinct differences in morphological and microscopic characteristics. Deconica flavum is lighter in pileus color than D. cokeriana, and has no discoloration when bruised. Interestingly, the center of the pileus is brow shellow, similar to “egg yolk” in D. flavum. Moreover, D. cokeriana stipe is yellowish white to light brown, with white to brownish fibrils, which become darker when bruised. While D. flavum is dark blonde to light brown, with yellowish-white fibrils. Microscopically, the pleurocystidia type chrysocystidia and leptocystidia of D. cokeriana (17–40 × 6.5–11 µm; 14–24 × 3.5–7 µm) are longer than D. flavum (15.2–26.8 × 5.6–13.0 µm; 7.7–17.9 × 3.3–5.7 µm). D. thailandensis and D. overeemii were originally described as Psilocybe species (Horak and Desjardin 2006; Horak et al. 2009). It is easy to differentiate D. thailandensis and D. flavum by basidiospores. Basidiospores of Deconica thailandensis and D. overeemii, originally described as Psilocybe species (Horak and Desjardin 2006; Horak et al. 2009), are rhomboid. Basidiospores of D. flavum are ellipsoid to ovoid, and the middle part is concave under SEM (Horak et al. 2009).

The nearly blue species in Entolomataceae, Entocybe haastii, E. nitida, Entoloma alcedicolor, E. eugenei, E. hochstetteri, E. mengsongense, E. tadungense and E. virescens have similar color in pileus and stipe (Noordeloos and Hausknecht 2007; Alves and do Nascimento 2012; Bergemann et al. 2013; Ediriweera et al. 2017). Entocybe roseoalbus is unique in having nearly blue pileus and yellowish grey stipe. Entocybe species are distributed worldwide, focused on Eastern Europe, the East and West coasts of North America, and Oceania (GBIF, https://www.gbif.org/search?q=Entocybe). The ITS–LSU phylogenetic analysis of Entocybe resulted in clade a and clade b with strong bootstrap support (Fig. 2). The new species, Entocybe roseoalbus is close to Entocybe nitida, E. haastii, Entoloma alcedicolor, E. contrastans, E. fuligineoviolaceum and E. coeruleogracilis in the phylogenetical trees, which are clustered into one branch (BS = 88%, PP = –). They have distinct differences in morphological and microscopic characteristics. Compared with Entocybe roseoalbus, Entocybe nitida has comparatively smaller pileus (20–40 mm), darker stipe (grayish-blue), bigger spores (7–9 × 6–8 µm), and no cheilocystidia (Noordeloos 2004). On the other hand, Entocybe haastii has a robust and blackish blue stipe and bigger pileocystidia (54.1–81.1 × 2.5–7.8 µm) (Bergemann et al. 2013). Entoloma alcedicolor has steel blue pileus and stipe and garlic odor (Noordeloos 2004). Entoloma contrastans has smaller mycenoid basidiomata, white pileus (8–20 mm) with a slightly darker brown center, violaceus stipe (30–50 × 2 mm), and bigger spores (6.0–8.0 × 5.5–7.5 µm) (Noordeloos 2004). Entoloma fuligineoviolaceum has darker blue pileus and stipe, dark brown-violet to violet lamellae, and bigger spores (5.5–7.5(–8) × 5.5–6.5(–7) µm) (Noordeloos 2004). Entoloma coeruleogracilis has deeper blue basidiomata, smaller pileus (8–22 mm) and longer stipe (30–60 × 1–3 mm), and bigger spores (6.0–8.0 × 5.5–7.5 µm) (Noordeloos 2004).

In conclusion, sufficient evidence from morphological and molecular phylogenetic analyses supports the distinction of D. flavum and Entocybe roseoalbus from other recorded species of the respective genus.

We are grateful to Xing-Qiang Wang and Jia-Jie Shi for collecting samples. We are grateful to Yu Yang for her help in experimentation. We appreciate Fang-Min Fei’s help in writing.