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Govind Ballabh Pant University of Agriculture and Technology, Pantnagar
After independence, development of the rural sector was considered the primary concern of the Government of India. In 1949, with the appointment of the Radhakrishnan University Education Commission, imparting of agricultural education through the setting up of rural universities became the focal point. Later, in 1954 an Indo-American team led by Dr. K.R. Damle, the Vice-President of ICAR, was constituted that arrived at the idea of establishing a Rural University on the land-grant pattern of USA. As a consequence a contract between the Government of India, the Technical Cooperation Mission and some land-grant universities of USA, was signed to promote agricultural education in the country. The US universities included the universities of Tennessee, the Ohio State University, the Kansas State University, The University of Illinois, the Pennsylvania State University and the University of Missouri. The task of assisting Uttar Pradesh in establishing an agricultural university was assigned to the University of Illinois which signed a contract in 1959 to establish an agricultural University in the State. Dean, H.W. Hannah, of the University of Illinois prepared a blueprint for a Rural University to be set up at the Tarai State Farm in the district Nainital, UP. In the initial stage the University of Illinois also offered the services of its scientists and teachers. Thus, in 1960, the first agricultural university of India, UP Agricultural University, came into being by an Act of legislation, UP Act XI-V of 1958. The Act was later amended under UP Universities Re-enactment and Amendment Act 1972 and the University was rechristened as Govind Ballabh Pant University of Agriculture and Technology keeping in view the contributions of Pt. Govind Ballabh Pant, the then Chief Minister of UP. The University was dedicated to the Nation by the first Prime Minister of India Pt Jawaharlal Nehru on 17 November 1960.
The G.B. Pant University is a symbol of successful partnership between India and the United States. The establishment of this university brought about a revolution in agricultural education, research and extension. It paved the way for setting up of 31 other agricultural universities in the country.
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ThesisItem Open 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, PushpaWheat (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.ThesisItem Open 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, DineshBiotic 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.ThesisItem Open 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 KumarMicronutrient 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.ThesisItem Open 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, SnehIn 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.ThesisItem Open Access Plasma nanoparticles treated seed induced changes in molecular and biochemical profiling of wheat grown under heat and water stress condition(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2019-08) Gandhi, Vikas; Pandey, PriyankaWheat is one of the main staple foods in India as well as in the world. The yield and grain quality of the crop are severely affected by stress conditions like heat and water stress. In the present study experiments were conducted to delineate the effect of plasma and microbial consortia under heat stress condition and effect of nano ZnO, nano chitosan and microbial consortia under water stress condition. Field experiments were done at Norman E. Borlaug Crop Research Centre of G. B. Pant University of Agriculture and Technology, Pantnagar in 2018-19, using Randomised Block Design. The “HD 2967” variety of wheat was used for seed treatment, sampling for molecular and biochemical profiling was done at three different growth stages i. e., booting, heading and grain filling stage. In both heat and water stress conditions, the contents of photosynthetic pigment, relative water content, protein, proline, sugar, and antioxidants were found to be highest in the combination plasma + MC I and nano ZnO + chitosan NPs + MC I treatment respectively, which helped wheat combat water stress condition. The grain protein, quality of grain and the yield were also best in this treatment. Maximum increase in Zn and Fe concentration was also recorded using this treatment. In case of heat stress condition, similar results were found for all the above parameters in case of plasma + MC 1 treatment. As the immunity of plant decreases during abiotic stress condition they become more susceptible to attack by biotic factors reducing plant growth, development and yield. So antimicrobial property of nano ZnO and chitosan nanoparticle was assessed in antifungal and antibacterial experiments. Both nano ZnO and nano chitosan were found to have inhibitory effect, but the combination of these two nanoparticles was observed to have maximum antimicrobial properties both in case of bacteria and fungi. Our research finding reveals that nano ZnO, nano chitosan and combination of both nanoparticles can be used to reduce the losses caused by abiotic stress in wheat.ThesisItem Open Access Evaluation of anti-fungal, growth promoting anti-oxidative and defense related activity of copper nanoparticles in Brassica juncea against Alternaria blight disease(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2019-07) Mann, Avni; Pandey, DineshAlternaria blight is one of the most catastrophic fungal diseases which hamper the productivity of Brassica oilseeds in India. Alternaria brassicae is the main pathogen of the disease which produces chlorotic, necrotic toxins and phyto hormones to cause the disease. At present the disease is controlled mainly by using toxic chemicals such as fungicides which have hazardous effect on environment and human health. So, nano technological interventions are necessary to find out some novel kind of substances that can act as anti –microbials because nanoparticles have high surface area with respect to volume. The surface area of nanoparticles is directly proportional to their interaction with the microbes to carry out antimicrobial activities. Copper based compounds (Bordeaux mixture) have been used as fungicides since ancient times. Therefore, copper nanoparticles (CuNPs) can act as effective antimicrobial agents. In present study it was observed that disease index during Alternaria blight disease was reduced to 22% on application of 75 ppm as compared to 86% control untreated pants. CuNPs induces the expression of COI1, receptor of Jasmonic acid but does not modulate signal transduction pathway because of absence of JA signal so Defensin gene expression is not triggered. Under effect of CuNPs Cu/Zn SOD, anti-oxidant enzyme gene was significantly over expressed. Along with anti – oxidative roles, Copper nanoparticles have anti-fungal properties too. It was observed that 75 ppm CuNPs cause a reduction of 23% during Alternaria growth and 71% inhibition during spore germination due to higher penetration power than salts, CuNPs shows high inhibitory effect on growth and development of Alternaria by damaging their protein structure and cell membrane 75ppm of CuNPs showed substantial growth promotory effect on seed germination, plant height and fresh weight by increasing anti-oxidant enzyme activities. The reason behind decrease in disease deverity could be anti-oxidantive activity of CuNPs. ROS produces by hypersensitive reactions on infection leads to oxidative stress in plants. The anti – oxidative enzymes scavenges these ROS. Hence CuNPs help in faster scavenging of ROS by activating anti – oxidant genes and enzymes inside the cell. CuNPs shows growth promoting activities also and thus help plants to escape the pathogen infecting stage at higher rate. CuNPs have significant effect on reducing disease severity of Alternaria blight in Brassica by trigerring anti-oxidative machinery of plant cell. Therefore CuNPs based formulations can effectively be utilized to counter Alternaria blight of Brassica. This would help in increasing the yield of oilseeds by reducing the losses due to disease.ThesisItem Open Access Studies on cytopathogenicity of egg drop syndrome-76 virus(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2005-08) Srivastava, Gaurangi; Rao, V.D.P.In the present investigation, an attempt was made to study the cytopathogenicity of Egg drop Syndrome-76 (EDS-76) virus. In this study, VN1 isolate of EDS-76 virus was cultivated in 9-10 days old duck embryo. When infected allantoic fluid was given to laying birds as an antigen, affected birds appeared dull and depressed with diarrhea. The important characteristic was drop in egg production coinciding with production of soft and thin shelled eggs. The virus was isolated from infected allantoic fluid in primary chicken embryo liver (CEL) cell culture. The cytopathic effects characterized by rounding and degeneration of cells were evident from the first passage itself. May Grunwald and Giemsa stained preparations of infected CEL cells culture revealed eosinophilic intranuclear inclusion bodies with distorted nuclei. AGPT performed using infected duck allantoic fluid and standard and experimentally raised hyperimmune sera against EDS-76 virus, revealed precipitin lines. But CIE was unable to detect the presence of virus in infected duck allantoic fluid and infected cell culture supernatants. FAT performed to detect the presence of EDS-76 virus in infected CEL cell culture exhibit intense intranuclear yellow fluorescence. The haemagglutination property of the virus was studied in infected duck allantoic fluid and infected CEL cell culture. The HI test was performed using hyperimmune serum to confirm the specificity of HA test and a 100% correlation was obtained.ThesisItem Open Access Molecular Cloning, Sequencing and In-silico Analysis of N, P, C domains and full length Calreticulin-3 gene of Brassica juncea for understanding its role in downstream regulation(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2019-02) Meenu; Pandey, DineshRapeseed and mustard (Brassica juncea) holds immense agricultural and economic importance. The productivity of rapeseed and mustard is severely affected by Alternaria blight disease which is caused by the fungal pathogen, Alternaria brassicae. So far there is no source of resistance against this disease. However, biotechnological approaches could be used to develop disease resistance provided that the key candidate genes/proteins involved in imparting defence towards A. Brassicae pathogen are identified. In the recent past, the role of calreticulin has been demonstrated in during defence responses of plants towards various pathogens. Therfore it could prove an attractive candidate for improving defense response of Brassica plants towards Alternaria blight disease. Hence, the present studies were conducted to understand the similar role of Brassica juncea calreticulin-3 with the objective of isolating and cloning of N, P and C domains and full length Calreticulin-3 gene of Brassica juncea. Total RNA was isolated from 30-day old leaves of B. juncea var. PAB9511 followed by synthesis of cDNA and its confirmation by amplification of internal control Actin. The primers were designed from the mRNA sequence of Calreticulin-3 of Brassica rapa using Primer3 and BLAST tools. The PCR conditions were optimised for obtaining 1300 bp, 645 bp, 288 bp and 339 bp amplicons of full length calreticulin-3, N, P and C domains respectively. The N, P, C domains and full length calreticulin-3 gene were cloned into TA cloning vector, pGEMT-Easy vector and confirmed by colony PCR and restriction digestion using enzymes Xba I and EcoR I . The full length calreticulin 3 gene along with N, P and C domains were sequenced. The restricted fragments of each insert were then successfully cloned into mammalian expression vector, pCDNA3.1 and again confirmed by colony PCR and restriction digestion. The BLASTn results revealed that the full length calreticulin-3 of Brassica juncea showed 98% sequence identity with the mRNA sequence of calreticulin-3 of Brassica rapa subspecies pekinensis. Also, the phylogenetic tree revealed more than 90% sequence similarity between calreticulin-3 of Brassica juncea to the calreticulin-3 of Brassica rapa, Brassica napus and Brassica oleracea. The SmartBLAST analysis revealed 86% homology between calreticulin-3 protein of Brassica juncea and Arabidopsis thaliana. The domain analysis assured the presence of calreticulin domain in this sequence and motif analysis revealed the presence of many motifs having significant involvement in responding to various biotic and abiotic stresses. The in-silico prediction of phosphorylation sites revealed the presence of 17 serine residues, 5 tyrosine residues and 9 threonine residues that could be involved in phosphorylation/ dephosphorylation indicating their involvement in signaling transduction.The prediction of interacting partners of calreticulin-3 of Brassica rapa showed that it could interact with 70 KDa heat shock protein, Brassinosteroid insensitive-1 and dolichyl-diphospho-oligosaccharide-protein glycotransferase. The role of these proteins in plant response to various stresses has been demonstrated by recent studies. These could also be hypothesized to be the interacting partners for calreticulin-3 of Brassica juncea. The experimental studies of effect of over expression of calreticulin-3 in Brassica plants could be tested for its role in conferring resistance against Alternaria blight and other different pathogenic infections or environmental stresses.ThesisItem Open Access Comparative expression analysis of defense-related genes in both wild and transgenic Brassica juncea (var.) Varuna harboring mapk3 gene in response to infection by Albugo candida(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2018-05) Modak, Annayasa; Gohar TajMustard and Rapeseed are the third leading source of vegetable oil in the world after soybean and oil palm. According to Department of Agriculture & Farmers welfare 2017-2018 annual production of Rapeseed and mustard is 75.40 Lakh tones. Production in India is still not enough to meet the domestic requirement. The potential yield of Brassica is very high but there is a wide gap between the potential and realized yield of Brassica and it is largely because of biotic & abiotic stresses. White rust is one of the most important disease of Brassica and it is caused by pathogen Albugo candida.Even 60% reduction in yield of Brassica juncea is due to its high susceptibility to white rust. Plants defend themselves against most potent pathogens through various defense mechanisms. Activation of MAPK signalling cascade is one of major defense pathways by which extracellular stimuli are transduced into intracellular responses in all the eukaryotes. MAPK cascade also play key role in regulation of various defense related genes in response to pathogens. Therefore our investigation was to analyse expression level of potential MAPK cascade and defense related genes in wild and transgenic Brassica juncea (var.) Varuna harboring overexpressed MAPK3 gene to access tolerance against Albugo candida. Lower disease index in transgenic Brassica suggests its tolerance to Albugo candida. Higher expression level of MAPK6 than MAPK3 in the transgenic Brassica suggests that MAPK6 mimics a portion of MAPK3 signalling pathway and work in a direct cascade for production of defense related proteins. Overexpression of MAPK3 gene was observed in downregulating the expression of MAPK4 in transgenic Brassica juncea . The expression of WRKY33 and WRKY29 was also analysed in the present study. Expression levels of both WRKY33 and WRKY29 were found to be higher in transgenic plant overexpressing MAPK3. Expression of 3 Defense related genes (OASTL-B, ACD2, and CSD2) were found higher in transgenic plants horboring overexpressed MAPK3 gene & expression level of CYP20-3 was found higher in wild Brassica plant. The defense related genes (OASTL-B, ACD2, CSD2, and CYP20-3) were also subjected to In-silico study. Insilico study showed the Secondary structure, tertiary structure and putative phosphorylation sites of these 4 defense related genes & also derived protein-protein interaction of potential MAPKs cascade and WRKY (29, 33) with OASTL-B, ACD2, CSD2 & CYP20-3.
