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Authors: Laya, P K
Advisor: Yamini Varma, C K
Title: Characterization of medicinal mushroom, cordyceps sp. from Kasargod district
Publisher: Department of Plant Pathology, College of Horticulture, Vellanikkara
Language: en
Type: Thesis
Pages: 94
Agrotags: null
Keywords: Mushroom, Cordyceps sp., Mycelium, Ophiocordyceps sinensis, Hirsutella, Ophiocordyceps neovolkiana, Leucopholis coneophora Burm., Coconut root grub
Abstract: Cordyceps which is an entomo-pathogenic fungus got the name from two Latin words, cord and ceps, meaning ‘club’and ‘head’. This mysterious fungus has a long reputation of being the single-most expensive raw material used in traditional Chinese medicine due to its dearth in availability. As a result of its high efficacy and potency in curing various diseases, this fungus has been recommended by medicinal practioners as a ‘panacea of all ills’. Distribution of Cordyceps sp. is cosmopolitan, but mostly confined to high Himalayan Mountains in China, Tibet, Nepal and India, at an altitude of 3000 to 5000 metres or in Asian high altitude grass land ecosystems. Cordyceps sp. which is renamed as Ophiocordyceps sp. attacks on the coconut root grub (Leucopholis coneophora Burm.) in coastal sandy areas of Kasargod district of Kerala. In this context, in the present study, ‘Characterization of medicinal mushroom, Cordyceps sp. from Kasargod district’ an attempt was made to untangle many of the mysteries of Cordyceps sp. by isolation, identification, characterization, cultivation and assessment of the proximate composition with special attention to Cordyceps sp. Purposive sampling surveys were conducted in three locations of coastal sandy tracts of Kasargod district during June to September months of 2017 and 2018. The fruiting bodies of Cordyceps sp. were collected from three locations viz., Instructional farm of College of Agriculture, Padannakkad, Valiyaparamba area of Nileshwar and Regional Agricultural Research Station, Pilicode. Maximum number of fruiting bodies were obtained from the soil under cashew trees in the Instructional farm of College of Agriculture, Padannakkad (Average of 13.86/m2) due to the richness in organic carbon (1.39%), and shaded condition in the soil under cashew orchard compared to Valiaparamba and Pilicode. Macro minerals viz., potassium, calcium and micro mineral like, manganese were in higher quantity under soil of cashew trees compared to other two regions, which may be helping in the survival and germination of spores. The fungus was isolated on PDA medium from the stroma portion of the fruiting body and the isolate obtained after pure culturing was numbered as CD-1. The isolate completed the full growth in a 9 cm Petri dish by 55 days after inoculation in PDA medium. The anamorph produced similar type of conidia as produced in the field. Characterization of Cordyceps sp. based on cultural, morphological and physiological characters were carried out. The cultural characters like colour, shape, texture and the growth rate of the mycelium were studied in different media. Among the different media tested yeast potato dextrose agar medium (YPDA) was found to be the best medium followed by potato dextrose agar media (PDA), which were significantly superior from other medium. Various macroscopic characters like shape, size and colour of sclerotia, stipe and stroma and branching pattern of the stroma in three locations were observed. Microscopic studies of both anamorph (hyphae, synnemata, conidiophores and conidia) and teleomorph (perithetia, asci and ascospores) were conducted. The molecular characterization of the fungus was carried out at Rajiv Gandhi Centre for Biotechnology (RGCB), Thiruvananthapuram and showed homology with Ophiocordyceps neovolkiana (Kobayasi) strain KC1 having 98 per cent identity. The sequence of the fungus was deposited at the Gen Bank of NCBI and the accession number obtained was MH 668282. The culture was deposited in National Fungal Culture Collection of India at MACS Agharkar Institute, Pune with culture no. NFCCI 4331. Physiological characterization of O. neovolkiana was carried out. Among eight different temperatures tested, 30oC was found to be the optimum temperature in which the growth was completed within 38 days after inoculation. The neutral pH seven was found to be optimum for mycelial growth. Among the different light conditions, 24 hrs of darkness under incubator (30oC) was preferred by this fungus. Fructose was the best source of carbon in which maximum mycelial growth was obtained within 36 days after inoculation. The best source of nitrogen was yeast extract in which maximum mycelial growth was obtained within 35 days after inoculation. KH2PO4 was the best source of macro mineral for the growth, in which the fungus completed the growth within 38 days after inoculation. Among micro minerals, ZnCl2 was the best for the growth in which the fungus completed the growth within 35 days after inoculation. Folic acid was the best source of vitamin for the growth in which the fungus completed the growth within 37 days after inoculation. One optimum media for the growth of O. neovolkiana was standardized as Yeast Potato Fructose medium supplemented with one gram of KH2PO4, 500 mg of ZnCl2, 10 mg of folic acid in one litre of distilled water which was named as Yeast Extract Potato Fructose Agar (YEPFA). The growth of O. neovolkiana in the YEPFA medium was significantly higher than the best media YPDA and PDA, so that the time required for the mycelial growth could be considerably reduced from 55.15 days to 30.85 days. The mycelial dry weight was also significantly higher in YEPFB medium than YPDB and PDB and growth completed within 32.57 days after inoculation. Different cereal grains viz., rice, wheat and sorghum were tried for artificial culturing of O. neovolkiana under in vitro conditions. The number of days taken for coverage of the mycelium in 50g of grains was 96.4 days for sorghum which was significantly different from rice (115.2 days) and wheat (142.2 days). Hence sorghum was selected as the best substrate for artificial culturing. Analysis of proximate constituents was estimated using the collected sporocarps from the field and mycelial growth obtained on the broth of YEPF medium. The carbohydrate was more in artificially cultivated mycelia than fruiting bodies (5 and 9.7% respectively). But protein, (2.9 and 1.75%) and vitamin C (3.27 and 2.5) were more in fruiting bodies than mycelia. The crude fibre was more in mycelia than the fruiting bodies (16.8 and 25%). The total minerals content was almost similar in fruiting bodies and mycelia. Present work is the first attempt in India, which resulted in a detailed systematic study on O. neovolkiana parasitizing on coconut root grub. Future line of work should be concentrated on mass production of this fungus, the estimation of its anti- cancerous components and the antifungal and antibacterial effect of this fungus on pathogens in plant disease management and its potential as a biocontrol agent in pest management.
Description: PG
Subject: Plant Pathology
Theme: Characterization of medicinal mushroom
These Type: M.Sc
Issue Date: 2018
Appears in Collections:Theses

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