1. KAUTIR (Kerala Agricultural University Theses Information and Retrieval)

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Subject: search.filters.subject.Phylogenetic analysis

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    Phylogenetic analysis of the endemic and threatened bat Hipposideros pomona K. Andersen, 1918 (Chiropetra: Hipposideridae) in the Western Ghats
    (College of Forestry, Vellanikkara, 2023-03-02) Abhin M Sunil; Nameer, P O
    Hipposideros pomona K. Andersen, 1918 is a small to medium sized insectivorous bat species endemic to the southern Western Ghats (SWG). According to IUCN, the species is endangered due to its restricted distributional. It is found mainly in thick forests of mid to high elevations of the SWG. Biogeographic barriers can act as a barrier to species dispersal, resulting in reproductive isolation and thus aiding speciation. The effect of the barriers depends on the motility of the animal. Actively moving and robust animals, such as higher vertebrates, may disperse across more minor barriers, while less mobile animals could be restricted due to the same. The 30 km wide Palghat gap in the SWG acts as a barrier to the species movement for many forest depended species, consequently acting as a barrier for gene flow, which can result in speciation as well as increase the chances of gene pool isolation. This barrier could affect the movement between subpopulations of H. pomona in the south and north of the Palghat gap. The current study was conducted in Kerala part of the SWG, on the locations where H. pomona was previously recorded. Bat sampling was carried out using harp traps of dimension 1.5m width, 2.2m height, 7.5cm between four frames and 2.5 cm between vertical monofilament fishing lines. Sixteen standard morphological measurements were taken on the bats captured in the harp trap (to the nearest 0.1mm) using a digital vernier calliper. Wing punches of 3 mm diameter were taken with a sterile biopsy punch, preserved in molecular grade ethanol, and stored in a deep freezer. DNA extraction was carried out using the standard protocol. Polymerase chain reaction performed to amplify the mitochondrial cytochrome oxidase 1 (COI), mitochondrial cytochrome b (Cyt b). Bat acoustic recordings were made using an ultrasound detector and analysed using the software BatSound Touch. Phylogenetic analysis was performed using software Molecular Evolutionary Genetics Analysis Version 11 (MEGA 11) and Population Analysis with Reticulate Trees (POPART). Morphometric analyses, acoustic analysis and principal component analysis (PCA) of morphometric parameters were done in R software. The current known distribution of H. pomona is restricted to the SWG, and the species was elevated to endangered status in IUCN red list due to the current restricted range. However, no genetic studies have been conducted to confirm the split. Present study results confirm that H. pomona and H. gentilis are genetically distinct species. Within the SWG, the Palghat gap is a potential biogeographic barrier for H. pomona. The present study compared the forearm length (FA), length of first phalanx of third metacarpal (1PH3MT), and length of fourth metacarpal (4MT) – (Morphometry), and frequency of maximum energy FmaxE (Acoustics) of H. pomona subpopulations in the north and south of the Palghat gap and found significant variation, indicating that they are phenotypically distinct. However, the PCA result indicated that these subpopulations are cryptic. Overall, phylogenetic analysis of H. pomona, which was recently split from H. gentilis solely based on morphology, was done by comparing mitochondrial cytochrome oxidase 1 genome, which showed a clear difference between the two species. Acoustic and morphometric analysis of the H. pomona subpopulations in the north and south of the Palghat gap revealed that they are phenotypically distinct. The PCA done on the morphometric parameters revealed that the subpopulations of H. pomona across the Palghat gap are morphologically cryptic.
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    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 Das
    The 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.