Turkish Journal of Botany Turk J Bot (2013) 37: 1188-1194 http://journals.tubitak.gov.tr/botany/ © TÜBİTAK Research Article doi:10.3906/bot-1211-50

Three new species of basidiomycetous fungi from India

1, 1 2 Babita KUMARI *, Narender Singh ATRI , Ramesh Chandra UPADHYAY 1 Department of Botany, Punjabi University, Patiala, India 2 Directorate of Research, Chambaghat, Solan, India

Received: 24.11.2012 Accepted: 13.08.2013 Published Online: 30.10.2013 Printed: 25.11.2013

Abstract: In this paper, 2 lepiotaceous [ indica sp. nov. and Lepiota attenuispora sp. nov. (Agaricaeae)] and 1 Limacella magna sp. nov. in the are described. Lepiota indica has scurfy brownish carpophores, minute spores, and chains of golden brown-walled pileal elements. Lepiota attenuispora has a shaggy appearance, an appendiculate veil of the pileus margin, and a sheathed with soft cottony scales. Limacella magna is a species characterized by prominent brown scales on the pileus and stipe, presence of a glutinous layer, and a trichodermal pileus cutis.

Key words: Basidiomycetes, , macrofungi, India

1. Introduction in the macroscopic descriptions are from Kornerup and The Indian subcontinent is rich in natural resources Wanscher (1978). The specimens were hot air-dried and vegetation, which support the luxuriant growth of and packed in cellophane paper bags containing 1–4 different types of naturally occurring fungi. Among these dichlorobenzene pellets as an insect repellent. Macroscopic are lepiotoid mushrooms in the family with examination was carried out on fresh specimens in the field a fibrillose or floccose fibrillose surface; white or light- itself. Microscopic characters were studied from free hand colored free lamellae; a single, simple annulus; largely sections mounted in 5% KOH, stained with 1% Congo white, pink, or pale spore deposit; and sterile red. Microscopic line drawings were made with the aid of edges. During the studies on diversity of agaricoid a camera lucida at 1000×. Basidium length excludes the mushrooms in North India, 2 new species, Lepiota indica length of sterigmata. The spore shape quotient (Q = L/W) and Lepiota attenuispora belonging to family Agaricaeae, was calculated considering the mean value of length and were encountered, while 1 new species, Limacella magna, width of 20 basidiospores per collection. The specimens belonging to Limacella, an agaricoid member of were deposited in the herbarium of the Department family Amanitaceae, was found growing in old dung in a of Botany of Punjabi University, Patiala (Punjab), Pinus forest in Solan in Himachal Pradesh. This latter new India, under PUN, and the Directorate of Mushroom species is characterized by medium-sized carpophores with Research, Chambaghat, Solan (HP). The photographs and a pluteoid habit with concentrically arranged brown scales, microscopic details are given in Figures 1–7. whitish pink lamellae, globose spores, and a glutinous covering. These species are fully illustrated and described 3. Results in this paper. Identifications were conducted according Taxonomic observations to Pegler (1972, 1977, 1983, 1986), Dennis (1952), Smith 3.1. Lepiota indica B.Kumari & N.S.Atri sp. nov. (Figures (1954, 1966), Sundberg (1989), Arora (1986), Vellinga 1–3) (2001a, 2001b), Işıloğlu et al. (2010), Asef and Muradov MycoBank no.: MB 801870 (2012), and Seyidova and Hüseyin (2012). Type: India, Uttrakhand, Chakrata Road, Korba (2200 m). 21.07.2010, M.Kaur & B.Kumari (Holotype: PUN 2. Materials and methods 4329). Standard methods for collection, preservation, Diagnosis: The examined collection is close to and description of agarics were followed, using the Cystolepiota ompnera, from which it differs in having larger terminology given by Atri et al. (2005). Color notations spore size and size of squamules on the pileus surface. * Correspondence: [email protected] 1188 KUMARI et al. / Turk J Bot

A B Figure 1. A and B- Field photographs of Lepiota indica.

A B

C D

Figure 2. Lepiota indica. A- basidiospores, B- pileus hymeniform elements, C- stipe cutis elements, D- cheilocystidia. Scale bars = 10 µm.

Description. Carpophores: 1.6–7.0 cm in height. are more prominent in young specimens; margin incurved Pileus 0.2–3.5 cm in diameter, subumbonate, yellowish with pileal remnants, nonstriate, irregular, splitting at white (4A2) with brown umbo (6E4), surface covered by maturity; cuticle fully peeling; flesh whitish to creamish, dark brown (6F5) scurfy squamules to spiny scales, which unchanging, up to 0.2 cm thick; taste mild, odor heavily

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C A D

B

F

E Figure 3. Lepiota indica. A- carpophore, B- basidiospores, C- basidia, D- cheilocystidia, E- pileus trichodermal elements, F- stipe cuticle projecting hyphae. Scale bars: A = 3 cm, B–F = 20 µm. aromatic. Lamellae free, close, unequal, of 3 lengths, 0.2 Lamella edges sterile (Figure 3). Cheilocystidia 27.5–34.0 cm broad, yellowish white (4A2); edges lacerate. Stipe × 6.6–7.7 µm, septate, clavate to pyriform, scattered central, 1.2–6.2 cm long, 0.2–0.5 cm broad, yellowish (Figures 2 and 3). Pleurocystidia absent. Pileus: surface white (4A2), to orange white (5A2), covered by brown a disrupted trichoderm consisting of ascending or erect, (6E4) to dark brown (6F5) scales starting just below the inflated hyphae 4.0–12.0 µm in width and loosely arranged attachment of stipe, aggregated more prominently towards cylindrical to claviform closely septate, clamped elements the base, brownish on bruising; annulus pruinose fibrillose, with blunt to ampullaceous tips measuring 15.0–25.0 µm evanescent, attached to the upper half of the stipe (Figure in width, hyphae thin-walled with golden brown pigments 1). Basidiospores: 4.0–5.6 × 2.4–3.2 (Q = 1.7), ellipsoid, (Figures 2 and 3); context homoiomerous; hymenophoral ochre to light brown spores when accumulated, light brown trama regular, tramal hyphae measuring 4.0–20.0 µm in watery mount, dextrinoid, cyanophilous, congophilous, in width; subhymenium pseudoparenchymatous. Stipe not metachromatic in cresyl blue (Figures 2 and 3). Basidia cuticle formed of septate parallel-running hyphae 12.8–19.3 × 3.2– 6.4 µm, clavate, tetrasporic; with well- measuring 10.0–16.0 µm in width with some projecting developed sterigmata measuring 3.2–5.6 µm in length. loosely arranged cylindrical to clavate elements with

1190 KUMARI et al. / Turk J Bot yellowish brown pigment, measuring 10.0–12.0 µm in width, somewhat similar to those of pileal covering (Figures 2 and 3). Clamp connections present throughout. Habitat: Growing scattered under Cedrus deodara and Eucalyptus trees on soil among decaying leaf litter in mixed Cedrus deodara forests. A Additional collection examined (Paratype): India, Himachal Pradesh, Barot, 2500 m, 11.07.2010, B.Kumari B (PUN 4335). Etymology: The name of the species is based on the name of the Indian nation. C Remarks: The present collections belong to Lepiota section Echinatae in having pileal covering with hyphae gradually transitioning into globose or ellipsoid elements (Vellinga, 2001a). This species is easily recognized by its brownish carpophores covered by small spiny scales, minute spores, and chains of golden brown-walled pileal elements. In its outward morphology it appears quite close to Cystolepiota ompnera as described by D Pegler (1986), which mainly differs from the presently examined collection in having lamellulae of 3 lengths, larger spore size, and appearance and size of squamules on the pileus surface. It also bears a resemblance to Cystolepiota furfuracea in external morphology, except in having pruinose fibrillose annulus and lacking hyaline ellipsoid refractive guttules in oblong or subcylindrical basidiospores (Kumar and Manimohan, 2009b). 3.2. Lepiota attenuispora B.Kumari & N.S.Atri sp. nov. (Figures 4 and 5) E MycoBank no.: MB801871 Figure 5. Lepiota attenuispora. A- carpophore, B- basidia, C- basidiospores, D- cheilocystidia, E- pileus trichodermal elements. Scale bars: A = 3 cm, B–E = 20 µm.

Type: India, Punjab: Patiala, Punjabi University Campus near Botany Department, 250 m, 26.08.2010, B.Kumari (Holotype: PUN 4626). Diagnosis: The newly proposed species is quite close to Lepiota clypeolaria, from which it differs in having small carpophores and small spores with apical attenuation. Description. Carpophores: 2.2–4.2 cm in height. Pileus 1.5–5.0 cm in diameter, convex; surface composed of appressed fibrillose brownish gray (6C2) to orange white (6B2) scales over yellowish white (4A2) background; margin irregular, shaggy, splitting at maturity, feebly striate at maturity; cuticle fully peeling; flesh 4.0–7.0 mm thick, white, unchanging; taste mild; odor sour; pileal veil appendiculate. Lamellae free, unequal, with lamellulae of 3 lengths, crowded, up to 0.6 cm broad, white to yellowish white (4A2), changing to yellowish pink to brownish on bruising; lamella edges lacerate to wavy; spore deposit yellowish white (2A2). Stipe central, 1.8–3.9 cm long, Figure 4. Carpophores of Lepiota attenuispora in natural habitat. 3.0–5.0 mm broad, yellowish white (4A2), concolorous

1191 KUMARI et al. / Turk J Bot with pileus, unchanging, cartilaginous, slightly tapering Description. Carpophores: up to 25.0 cm in height. downwards, hollow, pruinose fibrillose sheathed with soft Pileus 20.0 cm in diameter, convex, and finally applanate; cottony scales below the annulus, annulate; annulus single, surface moist, grayish yellow (4B3) to light orange (5A4) persistent, attached on the lower half of the stipe (Figure covered with small appressed dark brown (6F6) to brown 4). Basidiospores: 4.0–6.4 × 2.4–3.8 µm (Q = 1.67), (6E5) scales, margin irregular, splitting at maturity; subamygdaliform with a slight apical attenuation; aporous, reflexed and rolled. Lamellae adnexed, crowded, pinkish brown spores when accumulated, brown in watery white, lanceolate to normal, 1.6–2.0 cm broad, unequal of mount, dextrinoid, cyanophilous, congophilous (Figure 5–6 lengths; edges serrate to dentate. Context off white, 5). Basidia 11.3–19.3 × 3.2–4.8 µm, clavate, thin-walled, 1.2 cm thick, unchanging; taste mild; odor strong, very inamyloid, 4-spored; sterigmata up to 1.6 µm long (Figure unpleasant. Spore deposit white. Stipe central, 22.0 cm 5). Lamella edges sterile. Cheilocystidia clavate, septate, long, 2.5–2.7 cm broad, near upper most region, 1.8 cm hyaline 17.7–27.4 × 4.0–6.4 µm (Figure 5). Pleurocystidia in the middle portion, 3.0 cm near the base, off-white, absent. Pileus: surface a disrupted trichoderm consisting becomes pale orange (5A3) and light orange (5A4) near of upright, thin-walled elements measuring 20.0–55.8 µm the base, obclavate, having distinctly bulbous base, surface long, 2.4–4.2 µm in diameter, hyphae thin-walled with covered with brownish scales, slimy, very sticky, pruinose, light brown parietal pigments (Figure 5). Pileus context annulate; annulus superior, single, movable, woolly, homoiomerous; lamella trama parallel. Stipe cuticle thick, hanging yellowish fragments, creamish (Figure hyphae parallel, with light yellow pigment 3.2–11.3 µm 6). Basidiospores: 7.0–12.5 (14.0) × 5.0–7.0 µm (Q = broad. Clamp connections present. 1.2), globose to subglobose, thin-walled, amyloid, light Habitat: Scattered in small groups on sandy soil among yellowish spores when accumulated, yellowish in watery grasses around Pinus roxburghii and Eucalyptus trees. mount, weakly cyanophilous and congophilous, apiculate, Additional collection examined (Paratype): India, aporous with oil content (Figure 7). Basidia 30.0–50.0 × Punjab, Rajpura road, Shambhu, 250 m, 9.03.2009, 10.0–14.0 µm, subglobose to cylindrical with basal clamp B.Kumari (PUN 4627), 26.08.2010, B.Kumari (PUN 4628). connections, granular; sterigmata 1.5–4.0 µm long (Figure Etymology: The species has been named on the basis 7). Lamella edges fertile. Cystidia absent. Pileus: surface of attenuation in basidiospores. a trichoderm consisting of clavate, cylindrical, tubular, Remarks: It is an interesting mushroom with a shaggy obtuse, and slightly lageniform elements measuring 10.3– appearance of the pileus margin and stipe sheathed 32.0 µm in width with septation and clamp connections; with soft cottony scales below the veil, and absence of hyphae thin-walled with light brown pigments (Figure 7); distinct annulus. In these features, it closely resembles Le. clypeolaria. However, the basidiospores are much smaller, only 4.0–6.4 × 2.4–3.8 µm instead of 11.0–18.5 × 4.5–6.0 µm as documented by Vellinga (2001a), and with slight apical attenuation, which makes the presently examined collections interesting and unique. It resembles in external morphology Le. magnispora and Le. spheniscispora, differs mainly in having yellow-brown color in Le. spheniscispora, more orange or rusty brown in Le. magnispora (Vellinga, 2001b). This species is also quite distinct from Le. shveta as documented by Kumar and Manimohan (2009a) in lacking discoloration of stipe to pale brownish orange on bruising and large 6.0–9.5 × 3.5–5.0 µm oblong to subcylindrical spores instead of slight apical attenuation, which makes the here-examined collections interesting and unique. 3.3. Limacella magna B.Kumari & R.C.Upadhyay sp. nov. (Figures 6 and 7) MycoBank no.: MB 801283 Type: India, Himachal Pradesh: Solan, 1650 m, 15.07.2010, B.Kumari (Holotype: PUN 5072). Diagnosis. Pileus 20.0 cm broad; surface capucine buff, covered with tawny to brown fibrillose scales. Lamellae adnexed, crowded, pinkish white. Basidiospores globose to subglobose, amyloid, aporous. Cheilocystidia none. Carpophore surface with characteristic glutinous covering. Figure 6. Field photograph of Limacella magna.

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B A

C

D E Figure 7. Limacella magna. A- carpophores, B- basidiospores, C- basidia, D- pileus trichodermal elements, E- stipe basal elements. Scale bars: A = 3 cm, B–E = 20 µm. context homoiomerous; lamella trama bilateral divergent; glutinous layer over the entire surface, clamp connections, tramal hyphae 2.0–12.0 µm in width, septate, hyaline; hymenophoral trama bilateral with divergent lateral strata, subhymenium pseudoparenchymatous, well-developed, and pileus covering trichodermial hyphae embedded 9.0–24.3 µm wide. Stipe surface hyphae parallel-running in a gelatinous matrix. Presently Li. magna sp. nov. is throughout with light yellow pigment measuring 17.1– described from Himachal Pradesh, India. Li. magna 27.9 µm in width. Clamp connections present throughout. clearly differs from the other known species of Limacella Habitat: Growing scattered on old dung in Pinus forest by a combination of characters, including the large-sized and growing scattered on soil among decaying Pinus carpophore with appendiculate margin, relatively large roxburghii leaves. globose to subglobose basidiospores, and a pileus covering Additional collection examined (Paratype): India, composed of a trichoderm of clavate, cylindrical obtuse, Himachal Pradesh, Solan, Kather, 1650 m, 10.07.2010, and slightly lageniform elements. Li. magna resembles B.Kumari and R.C.Upadhyay (DMR 25/10). Li. glischra (Murril, 1914) because both species bear a Etymology: The name of the species is based on the pileus with brownish scales with appendiculate margin large size of basidiospores and carpophore. with patches of universal veil. However, Li. glischra has Remarks: Genus Limacella is basidiomycetes of smaller basidiospores of 3.0–5.0 µm, instead of 7.0–12.5 family Amanitaceae, represented by 20 species worldwide (14) × 5.0–7.0 µm in Li. magna. The presently examined (Kirk et al., 2008). Morphologically, it is characterized by a species is quite different from Li. furnacea in possessing

1193 KUMARI et al. / Turk J Bot large-sized basidiospores measuring 7.0–12.5 (14.0) × Acknowledgements 5.0–7.0 µm in size and trichodermal elements 10.3–32.0 The authors wish to thank the Head of the Department µm instead of 4.0–8.0 × 6.0–2.0 µm basidiospores and of Botany at Punjabi University, Patiala, for providing narrow trichodermal hyphae 3.0–5.0 µm as documented laboratory facilities, and the University Grants Commission by Pazyman and Bers (1988/1989) in Li. furnacea, and it for financial assistance under the SAP-III program of DRS, does not resemble any collections throughout the world in which mycology and plant pathology is one of the thrust due to its unique, large spore size and big carpophores. areas.

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