WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES

SJIF Impact Factor 7.421 Volume 8, Issue 6, 284-293 Review Article ISSN 2278 – 4357

THREE NEW UNREPORTED MACROFUNGI

FROM JAMMU AND KASHMIR, INDIA

Munruchi Kaur*, Hilal Ahmad Rather and Naseema Aqbar Wani

Department of Botany, Punjabi University, Patiala (Punjab), India, 147002.

Article Received on ABSTRACT 24 March 2019, The present paper discusses the study of systematic survey for the Revised on 13 April 2019, Accepted on 03 May 2019, exploration of agarics from North Kashmir. The area exhibits varied

DOI: 10.20959/wjpps20196-13871 climatic and topographic conditions and provide an environment for

the lavish growth of agarics. As an outcome of fungal forays a number

*Corresponding Author of collections of different genera were made out of which three new Munruchi Kaur varieties of macrofungi that were explored and taxonomically Department of Botany, identified are Agaricus placomyces var. macrospores var. nov., Punjabi University, Patiala multisquamosa var. brevispora var. nov. and Russula (Punjab), India, 147002. cyanoxantha var. variata. All these taxa are supplemented with the field photographs, microphotographs and Camera lucida drawings.

KEYWORDS: Agaricus, Amanita, Russula, Pleurocystidia, Cheilocystidia, Etymology, Jammu and Kashmir; India.

INTRODUCTION The macrofungal diversity is decreasing rapidly due to urbanization, climate change,

deforestation and unsystematic exploitation through collection of wild mushrooms. So there

is an urgent need to collect, document and conserve this group. The work on collection and taxonomic studies of mushrooms in Jammu and Kashmir in the past is meager with only 262 larger fungi known, among which 226 are agaric taxa (Abraham, 1991). Present study was undertaken to survey the different parts of District Baramulla in different seasons for

collection of wild mushrooms, identification and documentation. Genus Agaricus L.: Fr. is

characterized in possesing a white to brown more or less yellowish or rufescent pileus; lamellae free, eventually deep colored because of the attached spores, hymenophoral trama regular when young later becoming irregular, spore print purplish brown to sepia and partial

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Kaur et al. World Journal of Pharmacy and Pharmaceutical Sciences

veil which forms a ring on the stipe. Anatomically basidiospores are smooth with a compound wall and not visibly pseudoamyloid. Genus Amanita Pers. is characterized by the presence of both annulus and saccate or cupulate volva on the stipe, hymenophoral trama bilateral divergent, basidiospores are smooth thin walled amyloid or inamyloid. The members of the genus Russula Pers. are most diverse characterised by resupinate to pleurotoid carpophores with varied colour range including yellow, red, green, brown, purple to black and verigated mixture of various colours, lamellae free to broadly adnate to adnexed, sometimes subdecurrent to decurrent and basidiospores with thick ridges, different types of cystidial apex remarkably appendiculate cystidia with long appendages.

MATERIALS AND METHODS The macrofungi specimens for the present investigation was collected from district Baramulla of Jammu and Kashmir. For this concern, extensive fungal forays were collected from various localties and sub-localties of district Baramulla. The morphological details were recorded from fresh carpophores. The field characters pertaining to gross morphology, shape, color and size of pileus, stipe and lamellae, presence or absence of annulus, etc were noted down on the ‘Field Key’, provided by Atri et al. (2005) and the color terminology used is that of Kornerup and Wanscher (1978). The specimens were hot air dried and packed in cellophane paper bags containing 1- 4, para dicholorobenzene. The microscopic details were studied by cutting free hand sections of revived part of the dried specimen and staining them in 2% Congo Red. The spores were studied from the spore print as well as from the crush mounts of the lamellae and their reaction with Melzer’s reagent was checked. The dried specimens were deposited in the Herbarium, Department of Botany, Punjabi University, Patiala, (Punjab), India under PUN for further reference.

TAXONOMIC OBSERVATIONS Agaricus placomyces Peck Ann. Rep. NY. St. Mus. Nat. Hist. 29: 40, 1878 var. macrospores var. nov. Fig.1,2 & 7(A-G).

Etymology: The name of the variety is based on the large sized basidiospores.

Carpophore up to 7.5 cm in height, campestroid. Pileus up to 9.2 cm broad; plano-convex; umbo absent; margin iregular; splitting at maturity; non striate; surface yellowish brown (5E8) with yellowish gray (2B2) scales on the margin; scaly, scales appressed fibrillose towards margins, squamulose in centre, dry; cuticle half peeling; flesh up to 0.4 cm thick, www.wjpps.com Vol 8, Issue 6, 2019. 285

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white, unchanging; odor soury. Pileal veil appendiculate. Lamellae free, crowded, unequal, not in series, moderately broad (up to 0.5 cm), dark brown to brownish grey, unchanging; gill edges smooth; lamellulae present. Spore print brown (6E8). Stipe central, up to 5.5 cm long, up to 2.1 cm broad above, up to 2.0 cm broad at base, equal in diameter throughout with a clavate bulbous base; surface yellowish brown (5E8), yellow towards base, unchanging; solid, scaly, scales smooth above annulus, scaly below, white; annulate, annulus patchy but fleshy, hard textured.

Basidiospores 7.1 – 8.05 (8.95) × 5.37 – 6.26 (7.16) μm (excluding apiculus), Q = 1.32, ellipsoid, double walled, smooth, non guttulate; outer wall thick and darker than inner; apical pore absent; apiculate, apiculus 0.9 - 1.8 μm long, excentric. Basidia 17.9 – 39.0 × 7.0 – 11.0 μm, clavate, hyaline, thin walled, tetrasterigmate; sterigmata 3.6 - 5.0 μm long, hyaline. Pleurocystidia absent. Cheilocystidia 21.0 – 24.0 × 6.0 – 9.0 μm, clavate, thin walled, granular towards apex; gill edges sterile. Hymenophoral trama regular. Pileus cuticle hyphal, made up of 6.7 – 11.0 μm broad, septate, interwoven, gelatinized hyphae giving rise to 50.0 – 63.0 × 12.0 – 16.0 μm, encrustrated, granular pileocystidiod elements, abundant. Stipe cuticle hyphal, made up of longitudinally arranged, 6.7 – 13.5 μm broad, septate hyphae, caulocystidia not seen. Clamp connections absent throughout.

Collection Examined: Jammu and Kashmir, Baramulla, Sain, (2048 m), 340 16.840 N – 074016.349 E growing solitary on humicolous soil in mixed coniferous forest, Hilal Ahmad Rather, PUN 5180, May 29th, 2012 (Holotype).

Remarks: The presently examined collection matches well in its morphology and anatomy to the details provided for Agaricus placomyces Peck as given by Murrill (1922) and Freeman (1979). The only difference being in the size of basidiospores which is very large i.e, 7.1 – 8.05 (8.95) × 5.37 – 6.26 (7.16) μm in present collection, as compared to (4.5) 5.3 – 6.0 (7.0) × (3.0) 4.5 μm given by Murrill (1922) and Freeman (1979) for Agaricus placomyces. On the basis of much variation in basidiospore size, a new variety A. placomyces var. macrosporus var. nov. is proposed. This species is earlier reported from Lucknow (Uttar Pradesh) by Ghosh et al. (1974), Southwest India by Sathe and Rahalkar (1978), Sharma et al. (1978) from Himachal Pradesh, Punjab Plains by Saini et al. (1991) and from Maharashtra by Borkar et al. (2015). Presently this species is reported first time from Jammu and Kashmir, while a new variety of this species is proposed.

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Amanita multisquamosa Peck Annual Report on the New York State Museum of Natural history, 53: 840,1900. var. brevispora var. nov. Fig. 3,4 & 8(A-G).

Etymology: The name of the variety is based on its basidiospores.

Carpophore 10.4 – 14 cm in height. Pileus 6.1- 12.6 cm cm broad; convex to plano convex; umbo absent; margin regular; splitting at maturity; striate; surface silver brown to light silver

(4E2) with a brownish (6E5) centre; scaly, scales appressed to recurved fibrillose, more concentrated at center; moist; cuticle fully peeling; flesh up to 0.5 cm thick, white, unchanging; odor mild. Pileal veil appendiculate absent. Lamellae free, close, unequal, not in

series, broad (up to 0.7 cm), creamy white (2A1), when young, yellowish white (3A2) at maturity, unchanging; gill edges smooth; lamellulae present. Spore print off white. Stipe central, 8.8 – 13.3 cm long, up to 1.5 cm broad above, up to 3.7 cm broad at base, equal in

diameter throughout with a bulbous base; surface creamy white (2A1), unchanging; solid, annulate, annulus skirt like, persistant, superior; volva in the form of whitish, flocullose broken rings.

Basidiospores 7.16 – 12.53 × 6.26 – 10.74 μm (excluding apiculus), Q = 1.16, subglobose to broadly ellipsoid, thin walled, smooth, non guttulate, inamyloid; apical pore absent; apiculate, apiculus 1.8 μm long, centric. Basidia 34.0 – 62.0 × 8.0 – 14.0 μm, clavate, hyaline, thin walled, basal clamps absent, tetrasterigmate; sterigmata 5.0 - 6.0 μm long, hyaline. Lamellae edge cells, 29.0 – 32.0 × 13.0 – 14.0 µm, clavate to pyriform with rounded blunt tips, granular; gill edges heteromorphous. Hymenophoral trama bilateral divergent. Pileus cuticle hyphal, ixocutis, made up of 4.0 – 6.0 µm broad, subhorizontally tangled, septate, granular, hyphae with tubular ends; pilocystidia absent; context made up of 6.0 - 8.0 µm broad, hyaline, septate, loosely interwoven, inflated, acrophysalides rarely present. Stipe cuticle hyphal, made up of longitudinally arranged 4.0-10.0 µm broad, septate, hyphae; context hyphal, made up of loosely interwoven 10.0 – 16.0 µm broad, septate, hyphae. Volval elements made up of 2.5 - 6.64 µm broad, septate, hyphae, intermixed with 4.0 – 10.0 µm, subglobose, hyaline, inflated cells. Clamp connections absent throughout.

Collections examined: Jammu and Kashmir, Baramulla, village Soyan Saidnar (1771 m) growing solitary in the mixed coniferous forest during spring season, Hilal Ahmad Rather, PUN (5197), May 15th, 2014; Baramulla, village Soyan Saidnar (1700 m) growing solitary in

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Pinus and Cedrus forest during spring season, Hilal Ahmad Rather, PUN, (5176), May 16th, 2012.

Remarks: The presently examined collection belongs to section Amanita because of the presence of inamyloid basidiospores and the stipe with bulbous base. The morphoanatomical characters of this collection is in complete agreement with details provided for Amanita multisquamosa Peck by Jenkins (1978) and Saini and Atri (1993). This species is very close to A. pantherina (DC.: Fr.) Krombh. but differs from the present species in possessing deep brown or brown-bistre to brown to hazel-brown to pale ochraceous brown cap color which otherwise silver brown to light silver in the present collection. Further in the present collection the basidiospore size is quite larger varying from 7.16 – 12.53 × 6 – 10.74 µm as compared to the actual basidiospore size 9.0 – 11.0 × 7.0 – 9.0 µm, given for of A. multisquamosa by Jenkins (1978). Due to these variations in the basidiospore size a new variety Amanita multisquamosa var. brevispora var. nov. has been proposed.

Russula cyanoxantha var. variata (Bann.) Sing.Fedde. Rep., 33: 351, 1934.

Fig. 5,6 & 9(A-I) Carpophores 5.5 – 9.1 cm in height. Pileus upto 7.5 cm broad; flattened depressed; umbo absent; margin irregular, wavy, unpellucid, reflexed; splitting at maturity; non striate; surface

light gray (1C1) when young, grayish (1E1) at maturity with light brown (6D5) center and greenish tings over margins; moist; cuticle fully peeling; flesh upto 0.4 cm thick, white,

changing to yellowish white ( 4B2 ) on bruising; odor mild. Pileal veil absent. Lamellae

adnate, close, unequal, not in series, broad (up to 0.6 cm), light yellow (4A4), unchanging;

gill edges smooth; lamellulae present. Spore print yellowish white (3A2). Stipe excentric upto 7.0 cm long, up to 1.8 cm broad above, up to 2.0 cm broad at base, cylindric obclavate;

surface creamy white (3A1) changing to yellowish white (3A2) on exposure, unchanging; hollow; smooth, veined; exannulate.

Basidiospores 58.05 – 11.63 × 7.16 – 8.95 µm (excluding ornamentation), Q = 1.29, ellipsoid to broadly ellipsoid, warts upto 0.17µm high, blunt, conical; wart to wart connections few, mostly isolated, reticulum not formed, ornamentation type IV,V,VI; apiculate, apiculus 0.9 – 1.79µm long, nodulose type. Basidia 35.0 – 48.0 × 11.0 – 14. 0 µm, clavate, hyaline, tetrasterigmate; sterigmata 4.5 – 6.26 µm long, hyaline. Pleurocystidia: macrocystidia 52.0 – 61.0 × 11.0 – 12.0 µm, fusoid with blunt tubular tips, granular in the upper half, deeply www.wjpps.com Vol 8, Issue 6, 2019. 288

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seated. Cheilocystidia 45.0 – 52.0 × 11.0 – 12.0 µm, fusoid ventricose with broad and narrow apices, granular; gill edges heteromorphous.

Hymenophoral trama heteromerous. Pileus cuticle hyphal, undifferentiated into epicutis and subcutis, cuticle ixocutis, made up of 2.2 – 9.0 µm broad, broad, subhorizontally tangled closely septate hyphae, giving rise to a regular turf of filamentous, branched, tubular tipped hyphae; pilocystidia 27.0 – 39.0 × 5.0 – 7.0 µm, clavate with capitate to acute tips; context made up of rosettes of sphaerocysts intermingled with 6.7 – 13.0 µm broad septate hyphae. Stipe cuticle hyphal, made up of longitudinally tangled, septate up 2.2 – 6.7µm broad hyphae, giving rise to scattered septate to aseptate 38.0 – 45.0 × 5.0 – 7.0 µm broad caulocystidioid elements; context made up of rosettes of sphaerocysts and tangles of septate 3.5 - 5.5 µm broad hyphae. Clamp connections absent throughout.

Chemical Colour Reaction: Pileus surface turns strong brown in KOH solution while as

stipe surface turns greenish in FeSO4.

Collection examined: Jammu and Kashmir, Baramulla, village Takya Panzulla (1929m), growing in groups in the mixed coniferous forest, Hilal Ahmad Rather, PUN (5193), May 19th, 2012.

Edibility: It is an edible , costs high in Europe markets. It is hard crisp buttons difficult to beat in omelets and with scalloped potatoes (Arora, 1986).

Remarks: The above examined collection belongs to R. cyanoxantha var. variata (Bann.) Sing. The external and internal details of this collection are in accordance with the description given by Singer (1957), Romagnesi (1967) and Arora (1986). Singer (1957) reported it from Florida. This species was found in the month of September growing in shady broad leaved woods on calcarious ground (Romagnesi, 1967). Further, this species was also found under deciduous trees as reported by Rayner (1970). Pala et al. (2012) found R. cyanoxantha growing scattered under broad leaved trees from Hirpora, South Kashmir. Ao et al. (2016) reported R. cyanoxantha growing under Lithocarpus and Castanopsis in sub- tropical forests of Nagaland. Presently, this variety has been recorded for the first time from India.

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Fig. 1. Agaricus placomyces var. Fig. 2. Agaricus placomyces var. macrospores var. nov. macrospores var. nov.

Fig. 3. Amanita multisquamosa Fig. 4. Amanita multisquamosa var. brevispora var. nov. var. brevispora var. nov.

Fig. 5. Russula cyanoxantha var. Fig. 6. Russula cyanoxantha var. variata variata

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Fig 7: (A-G) Agaricus placomyces var. macrospores var. nov.

Fig. 8 (A-G) Amanita multisquamosa var. brevispora var. nov.

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Fig. 9 (A-I) Russula cyanoxantha var. variata.

CONCLUSION The present study thus illustrates the unexplored biodiversity of macro fungi from Jammu and Kashmir. As a result of which three new varieties viz. Agaricus placomyces var. macrospores var. nov., Amanita multisquamosa var. brevispora var. nov; and Russula cyanoxantha var. variata has been proposed. Thus, the present investigations have contributed three new varieties reported first time from Jammu and Kashmir.

ACKNOWLEGEMENTS The authors are grateful to the Head, Department of Botany, Punjabi University, Patiala, India for providing laboratory facilities during the present course of work.

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4. Atri, NS, Kaur, A. and Kour, H. Wild mushrooms- Collection and identification. In Frontiers Mushroom Biotechnology, R.D. Rai, R.C. Upadhyay and S. R. Sharma (eds.), 2005; 9-26. 5. Borkar, P. Doshi, A. and Navathe, S. Mushroom diversity of Konkan region of Maharashtra, India. Journal of Threatened Taxa. ISSN 0974-7907 (Online), 2015; 7(10): 7625-7640. 6. Freeman, AEH. Agaricus in North America: Type studies. Mycotaxon, 1979; 8(1): 1-49. 7. Ghosh, RN. Pathak, NC. and Singh, BP. Studies on Indian II Proc. Nat. Acad. Sci. India, 1974; 44: 125-128. 8. Jenkins, DT. A study of Amanita types. I. Taxa described by C.H. Peck. Mycotaxon, 1978; 7: 23-44. 9. Kornerup, A. and Wanscher, JH. Methuen Handbook of Colour, 3rd ed., Eyre Methuen, London, 1978. 10. Murrill, WA. Dark spored agarics-III Agaricus. Mycologia, 1922; 14: 200-221. 11. Pala, SA. Wani, AH. and Mir, RA. Diversity of macrofungal genus Russula and Amanita in Hirpora Wildlife Sanctuary, Southern Kashmir Himalayas. Biodiversitas, 2012; 13: 65-71. 12. Rayner, RW. Keys to the British species of Russula pt. III. Lists of species presenting specified diagnostic features and descriptions. Bull. Brit. Mycol. Soc, 1970; 4(1): 19-46. 13. Romagnesi, H. Les Russules d´ Europe et de 1´ Afrique du Nord. Bordas. Paris, 1967; 1-998. 14. Saini, SS. and Atri, NS. North Indian Agaricales-IV. Indian J. Mycol. Pl. Pathol, 1993; 23(3): 250-54. 15. Saini, SS. Atri, NS. and Gupta, AK. Additional studies on North-West Indian Agarics. In: Indian Mushrooms 1991, M.C. Nair (ed.), Kerala Agricultural University, Velenikkara, 680654, India, 1991; 7-12. 16. Sathe, AV. and Rahalkar, SR. Agaricales from South-West India-III. In: Indian Mushroom Science, C. K. Atal, B. K. Bhat and T.N. Kaul (eds.). Indo American Literature House, USA, 1978. 17. Sharma, AD. Jandaik, CL. Munjal, RL. and Seth, PK.. Some fleshy fungi from Himachal Pradesh-I. Indian J. Mush, 1978; 4: 1-4. 18. Singer, R. New and interesting species of Basidiomycetes V. (Descriptions of Russulae). Sydowia Ann. Mycol, 1957; 11(1-6): 141-272.

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