1. KAUTIR (Kerala Agricultural University Theses Information and Retrieval)
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Item Strain improvement of oyster mushrooms- pleurotus cystidiosus O.K. Mill and pleurotus opuntiae (Durieu and LEV.) SACC.(Department of Plant Pathology, College of Agriculture, Vellayani, 2018) Krishnapriya, P J; Geetha, DThe present study entitled “Strain improvement of oyster mushrooms: Pleurotus cystidiosus O.K.Mill and Pleurotus opuntiae (Durieu and Lev.) Sacc.” was carried out in College of Agriculture, Vellayani during 2015-2018, with the objective to standardize the techniques for production of oyster mushrooms: P. cystidiosus and P. opuntiae; and to study their morphological, physiological and cultural characteristics as well as nutritional and organoleptic qualities; and to undertake genetic improvement by protoplast fusion. The mushrooms were collected from two locations of Thiruvananthapuram and three fast growing isolates of Pleurotus spp. viz., PC2 (Vellayani), PNC1 (Chirayinkeezhu) and PO1 (Vellayani) were selected for the study. These isolates were identified as P. cystidiosus subsp. abalonus, P. cystidiosus and P. opuntiae using internal transcribed spacer (ITS) primers and subsequent sequencing; and registered at Genbank database with accession numbers KY214254, KY887023 and KY214255 respectively. The fast growing isolates of P. cystidiosus (coremial), P. cystidiosus (non-coremial) and P. opuntiae recorded maximum growth on PDPA amended with one per cent yeast under dark condition. The optimum temperatures for the growth were 30 0C, 25 to 30 0C and 25 0C respectively whereas, the optimum pH were 8, 8 and 7 to 8 respectively. Studies with different substrates and amendments for spawn production revealed that sorghum with one per cent yeast was the best for P. cystidiosus (coremial) and P. opuntiae whereas, paddy grains with one per cent yeast for P. cystidiosus (non-coremial). Experiments with different substrates and amendments for mushroom production revealed that rubber wood sawdust sprayed with 2.5 per cent of 1 M potassium dihydrogen phosphate recorded the maximum BE for P. cystidiosus (non-coremial) (192.76 per cent). P. opuntiae recorded the maximum BE in rubber wood sawdust amended either with 4 per cent neem cake (91.38 per cent) or wheat bran (91.37 per cent). Major insect pests observed in the beds of Pleurotus spp. were phorid flies, spring tails, black ants and staphylinid beetles. The competitor moulds observed were different species of Coprinus, Aspergillus, Penicillium and Trichoderma. Sporocarps soaked in one per cent CA for 15 minutes followed by mechanical drying and powdering was the best post harvest treatment for both P. cystidiosus (non-coremial) and P. opuntiae. Mycelium of P. cystidiosus (coremial) showed black coremial structures, representing its asexual stage (Antromycopsis broussonetiae Pat. & Trab.). The coremia comprised of elliptical (16.31 µm x 7.48 µm) and round conidia (8.06 to 8.49 µm). The black colour of coremia was due to melanin which was extracted (255.56 mg l-1) and characterized. The performance of long duration P. cystidiosus (non-coremial) and short duration P. opuntiae was compared with two ruling mushrooms of Kerala viz., long duration P. florida (Mont.) Singer and short duration P. eous (Berk.) Sacc. The study revealed that P. cystidiosus (non-coremial) and P. opuntiae showed higher BE compared to P. florida and P. eous, respectively. P. cystidiosus (non-coremial) recorded maximum moisture (94.05 per cent), starch (200.55 mg g-1), protein (30.2 mg g-1), fat (4.25 per cent), antioxidants (485.45 μg equivalent gram of ascorbic acid-1), beta-carotene (25.69 µg 100 mg-1), polyphenols (7.55 mg g-1) and energy (359.45 Kcal) compared to other Pleurotus spp. Sensory evaluation of mushroom products made from the species of Pleurotus was done and masala curry prepared from P. cystidiosus (non-coremial) scored the maximum value for overall acceptability. Shelf life of P. cystidiosus (non-coremial) was higher (5 days) compared to P. opuntiae, P. florida and P. eous (3 days each) in perforated poly propylene covers stored under refrigeration. Vanillin (0.05 per cent) and carbendazim (1 mM) were selected as dual biochemical markers for the PEG mediated protoplast fusion. Three days old P. cystidiosus (non-coremial) and four days old P. opuntiae recorded the maximum protoplast yield at five and four hours after incubation respectively with 0.6 M KCl and 30 mg ml-1 of enzyme consortium. Eight fusant lines with varied mycelial characters were obtained. Among fusants, F6 and F8 did not segregate in the second generation whereas, F4 segregated. F6 and F8 recorded higher BE of 168.05 and 99.95 per cent respectively compared to the parental lines and other fusants. Sporocarp of F6 and F8 was morphologically similar to P. cystidiosus (non-coremial) and P. opuntiae respectively; and F8 also exhibited low temperature adaptability. The present investigation indicated the exploitability of two promising isolates viz. P. opuntiae for tropical areas and P. cystidiosus (non-coremial) for cooler regions of Kerala using locally available materials and the standardized cultivation practices. The present study also standardized the protoplast fusion technique between P. cystidiosus (non-coremial) and P. opuntiae; and two fusant lines (F6 and F8) recorded higher BE which can be used for future breeding programmes.Item Strain evaluation and production technology of shittake mushroom ( Lentinula edodes ( Berk. ) pegler)(Department of Plant Pathology, College of Agriculture, Vellayani, 2016) Deepa Rani, C V; Lulu DasThe present investigation on "Strain evaluation and production technology of Shiitake mushroom (Lentinula edodes (Berk.) Pegler' was conducted at Department of Plant Pathology, College of Agriculture, Vellayani, Thiruvananthapuram during the period 2012-2015. The aim of the experiment was to exploit various strains of Lentinula spp. for novel production technology and their phylogeny analysis through physiological and molecular studies. Surveys were collected during pre and post monsoon periods of May to December from different parts of Thiruvananthapuram, Kollam, Wayanad, Idukki, Pathanamthitta, Kannur and Kasargode districts. Six isolates of sp. (VLYN- 1 to VLYN-13) obtained during the survey were identified and compared with procured reference strains of Lentinula edodes (LE-1 to LE-5 from GB Pant University of Agricultural and Technology, Pantnagar, Uttarakhand) and LE-6 strain (Maharana Pratap University of Agriculture and Technology, Udaipur) . Morphologically the native isolates of Lentinus spp. had concave, funnel and convex pileus with varying colors and were leathery in nature.L. edodes strains in contrast had convex pileus with chocolate brown and golden yellow sporocarps which were fleshy and edible. Phylogenetic analysis of all six strains of L. edodes using RAPD markers confirmed the variability between the strains. Maximum similarity coefficient of 74.10 per cent was observed between LE-2 and LE-6 strains while LE-2 and LE-4 strains showed a minimum similarity coefficient of 35.70 per cent. Further studies by ITS sequencing showed that all the L. edodes strains tested in the study showed 99- 100 per cent similarity with the known sequences off L. edodes available in NCBI database while that of native isolates showed 99- 100 per cent similarity to Lentinus tuber-regium and Lentinus connatus thus confirming the variability between Lentinus and Lentinula sp. All the six strains of L. edodes, showed maximum mycelial growth in malt extract peptone dextrose agar in solid and oat meal broth in liquid medium. L. edodes strains preferred temperature of 20 °C with an acidic pH of 6. Dark and ambient light conditions favored maximum mycelial growth and biomass production for L. edodes culture. Although a minimum period of 16.33 days was required for full mycelial run in maize grains but due to comparatively less contamination rate in paddy grains which took 18.33 days for completion of mycelial run were selected as best substrate for further studies. Different substrates were evaluated for the development of a cultivation package for shiitake mushroom. Results showed that LE-1 strain took minimum of 71.00 days for initiation of sporocarp in sawdust supplemented with 20 per cent wheat bran. Hard wood sawdust especially of teakwood was used in the study. The substrate based on paddy straw and banana pseudo stem were not found effective for pinhead initiation and thus failed to produce sporocarps. LE-1 produced maximum sporocarp (11.33) in sawdust + 20 per cent wheat bran which was followed by LE-3 (10.63) in sawdust + 20 per cent rice bran. Maximum yield of 290.66 g/ 500 g substrate was obtained in sawdust + 20 per cent wheat bran by LE-6 strain. Maximum biological efficiency of 58.13 per cent was also recorded in LE-6 in sawdust supplemented with 20 per cent wheat bran substrate. Substrates like paddy straw and sawdust amended with 20 per cent wheat bran substrates were evaluated for the development of native isolates of Lentinus tuberregium and Lentinus connatus . Results showed that maximum biological efficiency of 58.00 per cent was obtained by Lentinus tuber-regium whereas 36.60per cent biological efficiency by Lentinus connatus in sawdust amended with 20 per cent wheat bran substrate. Nutrient analysis of all the six strains showed that carbohydrate content ranged between 35.29 per cent to 40.23 per cent, protein 18.33 per cent to 21.66 per cent, crude fibre 22.33 per cent to 27.33 per cent, Vitamin- C 2.53 per cent to 3.50 per cent, ash 2.70 per cent to 4.40 per cent and lipid 2.46 per cent to 3.60 per cent. Mineral content of L. edodes included Ca (11.00 mg to 19.00 mg/ 100 g), Mg (0.46 to 1.10 mg/ 100 g), Fe (1.36 mg to 1.80 mg/ 100 g), Mn (1.53 mg to 2.63 mg), P (1.65 mg to 2.87 mg), K (16.33 mg to 25.20 mg), Na (13.00 mg to 23.66 mg) and Zn (19.66 mg to 28.33 mg/ 100 g). Sensory evaluation of mushroom products made from L. edodes was carried out by a panel of judges for various characters of which mushroom masala scored maximum for texture, taste, flavor and overall acceptability when compared to other recipes like mushroom cutlet, scramble, soup, baji and biscuit. As part of the study, paddy grain was found to be the most suitable substrate for spawn production of L. edodes and teakwood sawdust amended with 20 per cent wheat bran was the most efficient bed substrate. LE-6 strain was superior in terms of yield and biological efficiency. Therefore findings of the above investigation recommends the adoption of a suitable cultivation package for shiitake mushroom by using low cost substrates (hardwood sawdust) available in Kerala in plains and hilly regions.Item Effect of cluster beans (Cyamopsis tetragonoloba (L.) Taub.) on dietary management of life style disorders(Department of Community Science, College of Agriculture Vellayani, 2017) Aiswarya, L; Suma Divakar