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20204
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Subject: Biochemistry × End date: 2020 × Start date: 2020 × Type: Thesis × Thesis Type: M.Sc ×
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    ThesisItemOpen Access
    In vitro and in silico evaluation for potential anti-malarial gene(s) and micropropagation standardization of Cichorium intybus
    (G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2020-12) Chanotiya, Jiya; Dubey, Ashutosh
    Cichorium intybus, perennial, native to Europe, herb with diverse subspecies which is distributed widely in different states of India, comprise tremendous medicinal and nutritional culiminary features. It has various pharmacological activities including antimicrobial, antidiabetic, anti-inflammatory, hepatoprotective character including many other ethno-medicinal activities, which are mostly due to the presence of emmense phytocompound like phenolics, alkaloids, lactone. Inulin is a compound that provide most of the nutritional activity to the plant. Present investigation was carried out to explore the different form of forage Chicory by survey in foothills of Himalaya and examine its application in fat and SNF content of cow milk by supplementing the Chicory in daily diet fodder of bovine. The direct correlation of the Chicory supplement with fat and SNF content was observed. The micropropagation protocol was standardized for in vitro establishment of Cichorium intybus, with different hormonal combination critically increased concentration of growth hormone, which declares the most appropriate hormonal combination at particular concentation for callus induction, shoot proliferation and root establishment. Five different hormonal combinations with different concentration were tested for callus induction keeping the ADS concentration constant for every hormonal combination. (IAA (13 μM)+BAP(39 μM) +ADS(14.7 μM)) combination was most appropriate for callus induction and the same combination of (IAA (26 μM)+BAP (78 μM) +ADS (14.7μM)) proved most suitable for shoot proliferation. For root establishment combination of IAA (11.4 μM)+NAA (14.8μM) demonstrated most appropriate results. To check the antimalarial potential of Cichorium intybus, Germacrene A Synthase enzyme encoding gene expression was analyzed as this enzyme is critical for synthesis of sequiterpene lactone i.e lactucin and lactopicrin. The expression of Germacrene A Synthase was done by cDNA expression analysis and presence of 1953 bp size gene was established. As the enzyme Germacrene A Synthase is known to be part of lactucin biosynthesis, the various physiochemical properties of enzyme was studied through in silico approaches where physiochemical properties of enzyme were studied along with the phylogenetic analysis. The molecular docking between Germacrene A Synthase with lactucin and another docking of lactucin and Hsp 90 protein of the malarial parasite proved the active participation of lactucin in providing antimalarial activity to the plant which mainly formed by the Germacrene A synthase enzyme encoded by Germacrene A Synthase gene. 3D model of enzyme was predicted which was not present in the Protein Data Bank database and validated by the Ramachandran plot. Thereafter the predicted 3D model was submitted in the PMDB database with PMDB id: PM0082485. The cellular localization of enzyme, active pockets on the surface along with domain of the enzyme were determined. Hence, comestible medicinal and nutritional features of Chicory were proved in the investigation, yet further studies are desicred to fully understand the significance of Chicory.
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    ThesisItemOpen Access
    Biochemical and in silico analysis for evaluation of resistance in Brassica genotypes during Alternaria Blight infection
    (G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2020-12) Najam, Syed Saif; Punetha, Himanshu
    Present investigation was carried out to evaluate the differential biochemical defence mechanisms in three genotypes of oilseed Brassicas viz. Varuna, NPJ-112 of B. juncea and Sinapis alba followed by in silico analysis for interactions between fungal proteins and major secondary metabolites of Brassicaceae family. Significant results of Biochemical analysis at 72 hours after inoculation showed maximum protein content in S. alba (66.769) and minimum in Varuna (58.145). Maximum polyphenol accumulation with respect to controls was observed in Varuna (3.322) and minimum in NPJ-112 (3.387). Maximum flavanoid content with respect to controls was observed in NPJ-112 (0.365) and minimum in Varuna (0.299). Enzymatic analysis of infected plants with respect to controls showed highest Ascorbate peroxidase (1.942), Peroxidase (11.982) and lowest Catalase activity (0.008) in S. alba. Whereas minimum APX and POD activity and maximum catalase activity was found in Varuna. Highest Superoxide dismutase activity was observed in NPJ-112 (0.019) and lowest in Varuna (0.009). Maximum docking energy (-10.3499 kcal/mol) and 10 H-Bonds resulted from docking between fungal NRPS siderophore forming domain and sinalbin followed by MD simulation of docking results showing decrease in RMSD value of protein backbone after 1.6 ns in the 2 ns simulation. Taking in to account the biochemical results, the genotypes showing satisfactory biochemical basis of resistance can be used in further breeding programmes. Structural analogs based on in silico analysis results can be used as safer fungicides.
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    ThesisItemOpen Access
    Experimental evaluation of pyrolysed bio-oils for the development of anti-bacterial agent
    (G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2020-12) Badoni, Vaibhav; Verma, A.K.
    Pyrolysis bio-oils from pine needle, sugarcane leaf residue, wheat straw and rice husk was used to assess the anti-bacterial activity of various plant (Xanthomonas axonopodis, xanthomonas campestris, Xanthomonas oryzae, Erwinia tracheiphila and Ralstonia solanacearum where Pseudomonas fluorescens) and animal pathogenic bacteria (Escherichia coli, Staphylococcus aureus, Klebsiella pneumonia and Salmonella enterica). The pine needle bio-oil has about 50% of pehnolic content that may be responsible for the antibacterial activity against plant and animal pathogenic bacteria. The pine needle bio-oil showed the best antibacterial activity results against plant pathogenic bacteria with the maximum zone of inhibition of 23±0.33 mm and animal pathogenic bacteria with zone of inhibition 15.67±0.33 mm at concentration of 1000 μg/ml. Similarly the aqueous phase of pine-needle bio-oil showed the highest antibacterial activity with maximum zone of inhibition 14.33±1.09mm against plant pathogenic bacteria and 13.67±0.17 in animal pathogenic bacteria. Antibacterial activity of bio-oil was better with respect to aqueous phase in both plant and animal pathogenic bacteria. Pine-needle bio-oil was utilized for preparation of antibacterial agent. The antibacterial agent was prepared by using pine-needle bio-oil at 15%w/v, surfactant sodium-lauryl sulphate at 1%w/v, sequestering agent EDTA at 0.1%w/v and solvent ethanol at 30%v/v. The prepared antibacterial agent had a good efficacy against animal pathogenic bacteria (Escherichia coli, Staphylococcus aureus, Klebsiella pneumonia and Salmonella enterica) with ME values 5.08, 5, 4.55 and 4.33 respectively. An inhibition of bacteria was also observed in carrier based test. However, phenol coefficient was 1 for Staphylococcus aureus indicating that the antibacterial agent was as effective as phenol at the same dilutions.
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    ThesisItemOpen Access
    Biochemical analysis of amaranthus (Amaranthus caudatus L.) genotypes from different locations of Uttarakhand for selection of elite germplasm
    (G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2020-10) Joshi, Nidhi; Verma, K.C.
    From the past 20 years, little progress has been achieved in reducing child malnutrition food insecurity, and hunger. To address these problems, increased consumption of leafy vegetables is promoted as sources of both bio-active compounds and micronutrients. Amaranthus is one of the leafy vegetable which has been rediscovered as a promising food crop mainly due to its resistance to drought, heat, diseases and pests, and the high nutritional value of both seeds and leaves. They tolerate a variety of unfavorable abiotic conditions, including high salinity, acidity, or alkalinity, making them uniquely suited for subsistence agriculture. The current study was undertaken with the objective of biochemical analysis for screening of cultivated amaranthus seeds collected from different regions of Uttarakhand. The grain coat colour of amaranthus genotypes varied from dark brown to blackish brown. The colour varied from yellow, red, brown to black. Variation observed in weight of 1000 grains and it ranged from 0.81 g to 1.32 g. Seed shape was either oval or circular and seed coat was either hard or soft.Moisture content in grains of amaranthus ranged from (6.9 % - 11.1%).The results of different amaranthus genotypes for protein content ranged from ( 13.33% - 18.21% ).Amaranthus seed extracts were made in two types of solvents that are Butanol and Petroleum ether. In butanol it ranged from ( 4.32 % - 7.89 % ) while in petroleum ether it ranged from (1.04 % - 3.28 % ). Total Phenol Content results obtained by providing a direct comparison of extract content with standard sample, i.e., gallic acid. The TPC in butanol extracts of amaranthus ranged from ( 4.5 - 11.1 mg GAE/g) while in petroleum ether extract it ranged from ( 0.7 - 2.5 mg GAE/g). While the total Flavonoid Content is calculated by providing a direct comparison of the extract with standard (catechin equivalent (CE).The total flavonoid content (TFC) in butanol extracts ranged from ( 161.43- 385.53 μg CE/g ), while in petroleum ether extract it ranged from ( 13.65 - 27.47 μg CE/g ) .Among the solvents, butanol showed comparatively higher antioxidant activity than petroleum ether. The samples which have bright colours like black and red usually showed higher antioxidant activites.The test weigh to the samples was inversely proportional to the protein content.Thus, Amaranthus with higher nutritional value, antioxidant potential and a number of other health benefits is comparable to and even sometimes better than other popular crops can be a potent natural crop for blending in staple food sources. It can be exploited in nutraceutical and pharmaceutical industries.