Thumbnail Image

Theses

Browse

2020 - 20214
Reset filters
Subject: Agricultural Biotechnology × End date: 2024 × Start date: 2020 × Type: Thesis × Thesis Type: M.Sc ×
Reset filters

Search Results

Now showing 1 - 4 of 4
  • Thumbnail Image
    ThesisItemOpen Access
    Evaluating the effect of zinc-based nanoparticles for drought tolerance in wheat (Triticum aestivum L.) through foliar application
    (G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2021-10) Das, Debjyoti; Lohani, Pushpa
    Wheat (Triticum aestivum L.) is an important cereal crop worldwide and drought becomes a limiting factor for its growth as well as yield. A field experiment was conducted during Rabi season of 2020-21 using randomized block design at Pantnagar Centre for Plant Genetic Resources, G.B. Pant University of Agriculture and Technology, Pantnagar, Uttarakhand to study if there is any effect of zinc-chitosansalicylic acid (ZCS) nanoparticles for providing drought tolerance in wheat without compromising the yield. The morpho-physiological and biochemical responses of wheat plants to different concentrations of ZCS nanoparticles (100, 200 and 400 ppm) under water stress condition were studied. The plants treated with 100 ppm ZCS nanoparticles showed significant increase in plant height, root length, number of leaves, chlorophyll and carotenoid content, free proline and ascorbate content under water stress condition. There was decrease in the malondialdehyde content with the application of ZCS nanoparticles at 100 ppm concentration. Specific activity of antioxidant enzymes like ascorbate peroxidase, guaiacol peroxidase, glutathione reductase and superoxide dismutase was found to increase gradually with the increase in ZCS nanoparticles concentrations. Grain yield/ plant significantly increased (≈ 50%) with foliar application of ZCS nanoparticles at 100 ppm concentration under water stress condition. Application of nanoparticles at 200 and 400 ppm concentrations did not show any promising result possibly be due to toxicity effect. From this study, it was concluded that zinc-chitosansalicylic acid nanoparticles played an important role in mitigating drought stress of wheat and it also boosted grain yield of wheat under water stress condition.
  • Thumbnail Image
    ThesisItemOpen Access
    Molecular cloning and Agrobacterium mediated transformation of Brassica spp. with full length calreticulin gene along with its n, p and c domains
    (G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2021-09) Lokshman, Milan Kumar; Pandey, Dinesh
    Biotic and abiotic stresses in crop plants can significantly affect yield under resource poor conditions. The ability of plants to grow under stress conditions is an adaptive trait that can help to survive the crop with sufficient yield and quality. Recently, Calreticulin protein is identified as one of the differentially expressed genes in plants to minimize stress induced damages. Anti-oxidative activity of Calreticulin could be responsible for decreasing disease severity. The anti-oxidative enzymes scavenge these ROS produced during infection and environmental stress in plants. Calreticulin help in faster scavenging of ROS by activating antioxidant genes and enzymes inside the cell. Therefore it could be an attractive candidate for improving defense responses in plants. Calreticulin is mainly an endoplasmic reticulum resident chaperone protein. This protein has three distinct domains namely, N, P and C domains with structural and functional specificity. However, the actual domain responsible for providing stress resistance has not been analysed and there is no systematic study on actual mechanistic pathway of functioning of these genes. Hence, the present studies were conducted to understand role of calreticulin-3 by isolating and cloning of N, P and C domains and full length Calreticulin-3 gene followed by their transformation in to two varieties of Brassica spp. (Brassica juncea cv. Varuna and Brassica rapa cv. Bhawani ) by floral-dip method with main aim of investigating antioxidative role of this gene and its domains and subsequent antioxidative and stress tolerant activities of transformed Brassica plants. In order to achieve this target, total RNA was isolated from B. juncea var. PAB9511 followed by synthesis of cDNA. PCR was performed using the synthesized cDNA as template for obtaining full length calreticulin-3, N, P and C domains. Then these constructs were cloned into pGEMT-Easy vector and digested using restriction endonuclease enzymes SpeI and BstEII. The restricted fragments of each insert were then successfully cloned into plant expression vector, pCAMBIA1302. Recombinant pCAMBIA1302 with the N, P, C domains and entire Calreticulin gene was transformed to Agrobacterium GV3101. The transformation were carried out by giving infection of Agrobacterium harbouring these four constructs, in floral tissues of Brassica genotypes Varuna and Bhawani at early inflorescence stage. Seeds were collected from each plants and they were screened to identify transformed ones by growing on selection media containing antibiotic Hygromycin B. Transformation was confirmed by performing PCR for selection marker Hygromycin B after genomic DNA extraction from leaves of seedlings growing on selection media. Transformation efficiency was calculated as approximately 1% for B. juncea and approximately 0.55% for B. rapa. These transformed Brassica plants could further be tested for its role in conferring resistance against Alternaria blight and other different pathogenic infections or environmental stresses and downstream signalling pathway involved in executing this role could further be identified and studied.
  • Thumbnail Image
    ThesisItemOpen Access
    In-silico characterization and comparative study of iron and zinc regulatory gene families NAS, YSL and ZIP in cereal crops
    (G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2021-02) Bhattacharya, Tithi; Sundip Kumar
    Micronutrient deficiency, especially iron and zinc deficiency in diet is a major reason of malnutrition in developing countries. Most of the population in developing countries is dependent on their staple crops as a source for their nutrition. Hence, developing biofortified crop varieties can be a promising way to overcome malnutrition of these minerals. Lack of genetic variation in major staples like rice and wheat is a major drawback in biofortification by conventional breeding method. Bioinformatics tools can be used as an alternative in biofortification of food grains. Iron homeostasis is a process by which the availability of iron is properly maintained in the plant body. In order to regulate this process a number of genes play a key role. Among these nicotinamine synthase (NAS), yellow stripe like (YSL) protein and zinc transporter protein (ZIP) are the most important and largely reported genes in cereal crops for iron and zinc uptake, transport and its storage. The comparative study of these iron homeostasis genes in related species helps to describe the evolutionary relationship, their nucleotide and amino acid profiling along with motif regions. During the present investigation in silico identification, characterization and comparative study of three iron homeostasis gene families viz. NAS, YSL and ZIP is conducted in five major cereal crops namely rice, wheat, maize, barley and barn yard millet. Various physico-chemical parameters such as molecular weight, isoelectric point, and aliphatic index are computed. Nicotinamine synthase gene (NAS), yellow- stripe like (YSL) protein and ZIP (ZRT-IRT) like proteins are involved in metal transport in plants. The respective sequences are retrieved from NCBI database in fasta format and different analysis are carried out. Phylogenetic tree is constructed using neighbour joining method for all the three groups together (NAS, YSL and ZIP genes) and individually for the three groups and the results are analyzed. Multiple sequence alignment study showed that amino acids namely glycine and phenylalanine are evolutionary conserved among these gene families. Phylogenetic analysis revealed that NAS and YSL gene family divided into two clusters A and B. On the other hand, the genes belonging to ZIP family divided into three clusters namely A, B and C. Conserved domain search explained that AdoMet_MTases superfamily with seven domain hits found in NAS gene family, YSL gene family contain OPT superfamily having four domain hits and ZIP gene family having five domain hits of ZIP superfamily. Physicochemical analysis is done using PROTPARAM tool. Target P-1.1 is used to determine the sub – cellular location of proteins. Some of the proteins are signal peptide, few are mitochondrial transfer peptide, while the analysis of most of the proteins show that they belong to some other location other than signal peptide, mitochondrial transfer peptide, chloroplast transfer peptide and thylakoid luminal transfer peptide. Secondary structure is predicted using Chou- Fasman secondary structure prediction software. Comparing the average percentage of secondary structure it was found that, alpha helix is the most predominant conformation revealing the higher level of conservation and stability of protein structure. Highest percentage of alpha helix was detected in ZIP family and beta strand found in YSL whereas highest percentage of turn was found in NAS gene family. The information generated out of this comparative study of iron homeostasis regulatory genes in cereal crops may be helpful for better understanding of iron homeostasis in cereals which may subsequently be utilized for iron and zinc biofortification in cereal crops.
  • Thumbnail Image
    ThesisItemOpen Access
    Evaluating the effect of zinc-chitosan-salicylic acid nanocomposite particles on germination, yield and antioxidant enzyme activity in Wheat (Triticum aestivum L.) and Indian mustard (Brassica juncea L.)
    (G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2020-12) Sahu, Shivraj; Gautam, Sneh
    In this work, zinc-chitosan-salicylic acid nanocomposite particles (ZCS NPs) have been reported as a potent promoter of seed germination, antioxidant enzyme activity and yield in wheat and Indian mustard. The experiment was conducted in rabi season of the year 2019 using Completely Randomized Design at the poly-house of Department of Molecular Biology and Genetic Engineering, College of Basic Sciences and Humanities, G.B. Pant University of Agriculture and Technology, Pantnagar, Uttarakhand. Synthesis of ZCS NPs was carried out using ionotropic gelation method. The synthesized ZCS NPs were characterized using UVvisible spectroscopy, Dynamic Light-scattering Spectroscopy, FTIR, XRD, EDX and TGA analyses. The average size of synthesized ZCS NPs was found to be 13.54 nm. Germination experiment conducted on wheat and mustard seeds soaked in different concentrations of ZC and ZCS NPs (0, 50, 100, 150, 200 ppm) showed significant improvement in seedling length, seedling dry weight, seed vigor indices (I and II) at 50 ppm concentration. The pot experiment on wheat and Indian mustard was carried out using foliar application of ZCS NPs separately at 0, 50, 100, 150 and 200 ppm concentrations. The results of DPPH radical scavenging activity, SOD, POD, CAT and APX activity along with proline content revealed that increasing concentrations of ZCS NPs significantly increased these parameters in wheat and mustard, with highest activity found at 200 ppm concentration. ZCS NPs had the highest impact on yield of wheat and mustard at 50 ppm concentration, with lower yield values on further increase in concentration. From this study, it was concluded that zinc-chitosan-salicylic acid nanocomposite particles played an important role in improving the germination, antioxidant enzyme activity and yield in wheat and mustard.