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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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  • ThesisItemOpen Access
    Functional validation of the potential EcCAM, EcCAX1 and EcCAX 3 genes in grain calcium accumulation through over-expression studies in Arabidopsis thaliana: Development of an efficient plant regeneration protocol towards calcium biofortification in finger millet
    (G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2019-02) Jamra, Gautam; Anil Kumar
    Mineral malnutrition is one of the major problems crippling by one-half world population mostly women’s and pre-schooled children. Calcium deficiency is also considered as major problem which may lead to defects in bones, teeth, and osteoporosis. Finger millet (Eleusine coracana) is an orphan crop, rich in calcium with high nutritional significance and antioxidant properties to grow under harsh drought conditions. Multi-pronged molecular approaches were used in our Lab to identify the candidate genes of calcium sensor and transporter families associated with differential grain calcium in different genotypes due to induction of differential calcium signaling and transport machinery. The identified genes were functionally validated in the present investigation through over-expression studies in a model plant of Arabidopsis thaliana. Complete CDS of EcCAM, EcCAX1 and EcCAX3 genes were retrieved from transcriptomic data of pooled spikes of high calcium containing genotype; GP-45. In-silico molecular characterization of these genes based on protein sequences showed the presence of conserved domains as characteristic features. The phylogenetic analysis showed the genes are closely related to Siteria italic and Oryza sativa while distinctly related with Arabidopsis. To decipher the function of EcCaM, EcCAX1 and EcCAX3 genes, the gain-offunction approach was used to generate the transformed lines in Arabidopsis thaliana and expression analysis by semi-quantitative RT-PCR of such genes carried out in T3 homozygous transgenic lines showed higher expression as compared to no expression in wild type. Phenotypic assays were performed under various abiotic conditions such as EcCAM under PEG induced drought stress, IAA induced homeostasis, NaCl induced salt stress and calcium induced ionic stress; EcCAX1 and EcCAX3 under calcium & magnesium ionic stress, and IAA induced homeostasis. The observations recorded in 7 days old seedlings suggested that over-expressed transgenic lines were more tolerant as compared to wild type as evident from root elongation with lateral growth and better physiology. The anti-oxidative potential of over-expressed transgenic lines was higher in transgenic lines showing less ROS accumulation while more ROS accumulation in wild type plants as indicated by NBT and DAB staining method. Thus, more oxidative damage was observed in wild type as compared to over-expressed transgenic lines. Finger millet is highly adaptive in harsh conditions and nutritionally superior necessitate its further improvement using transgenic technology. In order to develop superior genetically modified plants, an efficient plant tissue culture protocol is essentially required taking innate nutritional, biochemical and hormonal attributes of finger millet genotypes as indices of plant tissue culture responsiveness. In order to determine the influence of stress tolerant behavior and inherent composition on plant regeneration, four genotypes of finger millet (GP-45, GP-1 GE-1437 and GE-3385) were taken in the present study. The results indicate that GP-45 was found to show maximum stress tolerance whereas GP-1 was the least tolerant. Further estimation of endogenous total calcium, carbohydrates, protein, total phenols, total flavonoids and phytohormones (ABA and GA3) showed genotype dependent variations and high calcium is related with stress tolerance and in turn plant tissue culture responsiveness. The results of the present study clearly elucidate the importance of selection of genotypes based on biochemical indices such as innate phytonutrients, phytochemicals and phytohormones for the development of an efficient regeneration protocol in finger millet to introgres the potential genes whose functions validated through genetic transformation studies in Arabidopsis could further be harnessed for crop improvement especially for improving calcium nutrition and stress tolerance.
  • ThesisItemOpen Access
    Assessment of transgenic Brassica juncea cv. Varuna harbouring MAPK3 gene against Alternaria brassicae after preconditioning with phytohormones and MAPK inhibitor
    (G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2019-09) Srivastava, Snigdha; Gohar Taj
    Alternaria blight is one of the most devastating fungal disease which hampers the productivity of Brassica juncea in India. The disease is caused by a semi-biotrophic pathogen, Alternaria brassicae. The absence of resistance source against Alternaria blight has made the plant researchers shift their interest towards de-novo sources of resistance. The MAPK 3 gene is an important gene expressed in plants during biotic stress for strengthening the defense system of plant. The MAPK 3 gene was overexpressed in Brassica. juncea cv.Varuna for the evaluation of resistance provided by transgenic lines against Alternaria blight. The two important plant hormones salicylic acid and jasmonic acid plays a major role in transducing the activation of plant defense systems against pathogen attack. These phytohormones regulate the expression of MAPK cascade genes that are the key role player of defense signaling in plants. The codon usage bias is an evolutionarily conserved phenomenon that affects the expression level of genes. The knowledge regarding the codon usage biasness pattern of genes helps us to know whether the proteins will be expressed in the heterologous system or not that guides in the development of transgenic plants. In the present investigation the copy number of MAPK 3 gene in transgenic B.juncea cv. Varuna plants were calculated using real-time PCR and Southern blotting. The effect of salicylic acid, jasmonic acid, and MAP kinase inhibitor was studied on the transgenic and non-transgenic B.juncea cv.Varuna plants at different stages of disease progression. The antioxidant activities of APX, GPX, CAT, proline was higher in phytohormones treated transgenic plants. The MAPK cascade genes viz. MAPKKK 1, MAPKK 4, MAPKK 9, WRKY 33, WRKY 22, OASTL-B, ACD-2, CSD-2 that are involved in defense pathway were observed to be expressed higher in transgenic plants as compared to the non-transgenic plants. The codon usage bias analysis of MAPKs and WRKY genes of A. thaliana and B. rapa was carried out by using the bioinformatics approaches. The studies revealed that the pattern of CUB in these genes are affected by natural selection and mutational pressure. The clustered heat map grouped the MAPKs and WRKY genes having similar RSCU values and similar functions under the same cluster. The codon AGA coding for arginine was found to be preferred in most of the MAPKs and WRKY genes. The codon usage bias in A.thaliana and B.rapa MAPKs and WRKY genes was observed to be low based on ENc values. The expression level of MAPKs and WRKY genes was observed to be low based on the RSCU values. These studies will give direction for the engineering of defense pathway so that the Alternaria blight resistant B. juncea plants could be developed.
  • ThesisItemOpen Access
    Evaluation of in vitro cytotoxicity and anti-diabetic property of Aloe spp. based on over-expression of glucose transporter (Glut-4)
    (G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2006-07) Rajiv Kumar; Anil Kumar
    The world wide epidemic of diabetes has stimulated the search of new concepts, formulations and the targets for the treatment of this disease. As existing conventional treatments for diabetes have several side effects and also they are not able to nullify diabetic complications completely. Aloe spp. claims as diabetes therapeutics has been investigated in vivo by several workers. Their results concluded that Aloe extracts possess anti-diabetic property. Their mode of action and molecular targets are yet to be determined. The objective of the present study was to analyze the influence of Aloe extracts (aqueous and ethanolic) on the cytotoxicity and to evaluate its effect on GLUT-4 gene expression on NIH/3T3 cell line as in vitro model for screening the anti-diabetic properties of plants. Under present investigation, attempts were made to investigate possible compositional differences among Aloe spp, dose-dependent cytotoxicity of Aloe extracts for determining the optimum dosage for in vitro assay and its effect on GLUT-4 gene expression. The in vitro cell culture system developed can be suitably employed for screening of anti-diabetic properties of several herbal formulations and plants. Spectral analysis of Aloe extracts indicated the presence of diverse composition and compositional variations between two Aloe species (Aloe vera & Aloe perryi). Aloe extract showed cytotoxic effects on cell in a dosedependent manner, influenced the doubling time and saturation density of NIH/3T3 cells as shown by growth kinetics in presence and absence of Aloe extracts. The cell‟s viability was also affected in a dose-dependent manner. After determination of optimal dose for in vitro study, GLUT-4 gene expression study was performed at transcript level by using semiquantitative RT-PCR with mouse GLUT-4 specific primers. Under defined culture-conditions and standardization in our Lab, Aloe extracts appear to up regulate the GLUT-4 mRNA expression as compared with untreated cells. Insulin and Metformin was used as standard references. Water soluble Aloe extracts (1 mg/ml) stimulated the GLUT-4 gene expression with their optimal effect at 36 hours of treatment. GLUT-4 expression appears to increase gradually and attained a steady-state after 36 hours of treatment and thereafter a decline in GLUT-4 expression was observed indicating the temporal down regulation perhaps due to gradual increase in apoptotic signal at later stages. Our data suggest that Aloe extract possesses the anti-diabetic activity which might be utilized for drug formulations if suitable molecules get identified. Results also explain why Aloe plants have been used in some traditional medicines to ameliorate the diabetic symptoms.
  • ThesisItemOpen 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, Priyanka
    Wheat 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.
  • ThesisItemOpen 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, Dinesh
    Alternaria 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.
  • ThesisItemOpen 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.
  • ThesisItemOpen Access
    Bioinformatics analysis of camalexin biosynthetic pathway and studies on defense responses of MAP2K4/MAP2K9/MAPK3 mutants during pathogenesis of Alternaria blight in Arabidopsis thaliana
    (G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2018-08) Gaur, Manu; Pandey, Dinesh
    Alternaria blight is one of the most destructive fungal diseases which seriously hamper productivity of Brassica oilseeds in India. Alternaria brassicae is the main pathogen of the disease which produces chlorotic, necrotic toxins and phytohormones to cause the disease. These disease determinants have been known to affect various genes/ proteins of cell cycle and cell death pathways in susceptible Brassica plants to cause the disease and enable the pathogen to follow a semibiotrophic life style. It is being speculated that Alternaria brassicae pathogen affects highly conserved MAP kinase signal transduction pathway to cause the disease. In Arabidopsis , some of the MAP kinases including MPK3, MKK4, MKK9, are responsible for synthesis of ‘Camalexin’ which is one particular phytoalexin involved in mediating defense response against the necrotrophic fungal pathogen viz. ‘Botrytis cinerea’. In the present investigation, role of MPK3, MKK4 and MKK9 in triggering Camalexin based defense response of Brassica plant towards Alternaria brassicae was studied by using Arabidopsis mutants for these kinases and Bioinformatics analysis. Following the infection of Alternaria brassicae pathogen, disease index of mkk4, mkk9 and mpk3 mutant plants was observed to be more than that of wild type plants of Arabidopsis thaliana. These observations suggested increased susceptibility of mpk3, mkk4, mkk9 mutants of Arabidopsis for Alternaria blight due to decreased camalexin biosynthesis or differential expression of proteins. In order to study differential expression of proteins, the proteins were extracted from infected leaf samples of both wild type and mutant Arabidopsis plants and protein profiles were compared at initial, middle and late stage of infection by performing one dimensional SDS PAGE analysis. In wild and mutant plants of Arabidopsis, the protein extractability decreased as the disease progressed from early to middle stage due to degradation of host proteins and increased from middle to late stage due to synthesis of new proteins. SDS PAGE based analysis of proteins extracted from leaves of wild type and mutant Arabidopsis plants indicate differential expression of proteins in form of expression of unique prroteins and downregulation or upregulation of proteins during disease progression from early to late stage of pathogenesis in Arabidopsis mutants and wild type plants. MKK4/MKK9/MPK3 signalling module involved in biosynthesis of camalexin during defense response of B. rapa towards, Alternaria brassicae pathogen was elucidated with the help of Molecular modelling, docking, and protein-protein interaction analysis of MAP kinases retrieved from Brassica rapa genome. Network Biology based in silico approach was followed to identify linkages of MKK4, MKK9 and MPK3 with 51 known defense related genes through construction of PPI network. MPK3 was observed as major hub to which large numbers of defense regulated genes/proteins are connected. This suggests that MPK3 is largely responsible for activation of large number of defense related genes besides camalexin biosynthetic genes. These studies will open up new avenues for engineering defense pathway in Brassica for preventing losses caused by Alternaria blight disease.
  • ThesisItemOpen Access
    Delineating the redox signaling network under oxidative stress through over expression of Ecapx1 in Eleusine coracana
    (G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2018-12) Pande, Anjali; Arora, Sandeep
    Global warming has emerged as one of the biggest threats to agriculture in the last two decades. In arid and semi-arid regions, where temperatures are already close to the physiological maxima for crops, higher temperatures may increase the incidence of heat stress in crops; negatively affecting the growth and productivity. Under such conditions, development of climate resilient crops is a dire need. As reactive oxygen species are a common denominator in the effect of various environmental stresses, therefore it is important to understand the signaling events in plants that are implicated in activating the anti-oxidative defense in plants and providing redox homeostasis. Towards this objective, we successfully developed Exapx1 over-expressing E. coracana plants. The transgenic lines were developed by using Agrobacterium mediated transformation method. The transgenic lines with increased ascorbate peroxidase activity where used to delineate the redox sensing mechanisms that are involved in providing protection from oxidative stress. The transgenic lines were morphologically similar to the wild type plants and had similar PS-II activity. Increased expression of apx1 gene in the transgenic lines could induce higher expression of superoxide dismutase and monodehydroascorbate reductase genes, under stress; but no significant increase in dehydroascorbate reductase and glutathione reductase gene expression was recorded. Over-expression of Ecapx1 gene could selectively (and not universally) induce the expression and activity of other anti-oxidant enzymes of the AsA-GSH pathway, indicting the existence of a cross talk or signaling mechanism in plants, for coordinated expression of these enzymes. On exposure to stress, the transgenic lines accumulated lesser H2O2 as compared to the wild type plants and also had lower proline levels. An effective control over H2O2 levels (through Ecapx1 over-expression) prevented undue activation of specific antioxidative enzymes like DHAR and GR, while helping to maintain the redox balance of the cells under stress. Incidentally, the transgenic lines had higher reduced to total ascorbate ratio, than the wild type plants, that was responsible for sensing the increased influx of ROS and activate the cellular anti-oxidative defense network, through down-stream signaling.
  • ThesisItemOpen Access
    Effect of zno and fe2o3 nanoparticles as released from an alternate nutrient sources in in vitro culture media on various biomolecules Picrorhiza kurroa Royle ex Benth
    (G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2018-12) Bameta, Alka; Gaur, A.K.
    Genus Picrorhiza and their different species from varied habitat have been recognized and proven for various ailments, significant contribution from India has a potential due to the enormous biodiversity with special reference to native species collected from either from cultivated at various micro and macro environment or otherwise. Picrorhiza kurroa Royle ex Benth is one of the precious herbs of genus Picrorhiza. Several iridoid glycosides are present in this plant which includes their derivatives having various pharmacophore, are being subjected for their utilization in several modern and traditional medicines systems to cure number of ailments since ancient time. The plant is endemic and has been vastly explored. At present it is well recognized for hepatoprotection. It is essential to apply biotechnological approaches to enhance the production of various pharacophore either by extracts and their purifications or semi synthesis from in vitro as well as in vivo. Application of nanoscience related technologies from the bio mass through various culturing practices in various environments, have provided versatility with the secondary metabolites. During the present research work, various attempts have been made to optimized protocols for in vitro regeneration of Picrorhiza kurroa along with the effect of ZnO and Fe2O3 nanoparticles as an alternate nutrient source to recognize some facts about the synthesis of secondary metabolites, antioxidative enzymes (SOD, CAT, POX, APOX) and expression of rate limiting genes ((CAM, HMGR, DXS, EPSPS, GDPS, PAL)) involved in the synthesis of Picroside II. Nanoparticles study results indicated that it might be possible that IPP for biogenesis of different iridoid glycoside and their derivatives under application of their precursor the mevalonate, non mevalonate and phenyl-propanoid pathways might enhance the Picroside II. ZnO and Fe2O3 nanoparticles have been proved better method for eliciting their effect in order to enhance picrosides production.