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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
    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
    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.
  • ThesisItemOpen 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, Dinesh
    Rapeseed 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.
  • ThesisItemOpen Access
    Genotyping-by-sequencing and functional genomics approaches for identification and validation of SNP markers/genes associated with grain calcium trait in finger millet
    (G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2018-07) Sharma, Divya; Anil Kumar
    Finger millet (Eleusine coracana (L.) Gaertn), holds immense agricultural and economic importance due to its richness in calcium and its proteins being good source of essential amino acids. Hence, research towards such indigenous, mineral nutrient rich crop will surely help in accomplishing the challenges of mineral malnutrition. Markertrait studies in finger millet could be useful in the detection of markers associated with high grain calcium and, eventually, in selection of superior genotypes. With an aim to identify markers linked to grain yeild or its component traits and calcium content, initially phenotyping was done for fourteen agro-morphological traits and grain calcium content amongst finger millet global germplasm collection at two different environments viz. Almora and Pantnagar. Analysis of variance for both the environments resulted in highly significant differences among accessions for most of the traits, which showed the existence of sufficient variability. Also the wide range of variation in the agronomic performance of the accessions suggested that these traits could be considered as good candidates for marker-trait associations. The calcium content varied from 53mg/100g seed to 454mg/100g seed with an average of 277mg/ 100 g seed at E1 and from 42mg/100g seed to 466mg/100g seed with an average of 274mg/ 100 g seed at E2, which indicates presence of sufficient variability for grain calcium trait amongst the selected finger millet genotypes. Further, to study the genetic makeup of 202 diverse finger millet accessions Genotyping by Sequencing (GBS) analysis was done, which resulted in a genome wide set of several thousand SNPs identified within every accession. Genetic diversity and population structure analysis was conducted on a natural collection of 202 finger millet genotypes. Neighbour joining cluster analysis grouped the population into 3 clusters ‘A’, ‘B’ and ‘C’ based on their geographical location. The major cluster A consisted of broadly all the Indian genotypes whereas cluster B and C comprised of exotic genotypes. A majority of accessions of Asian origin showed their tendency to cluster together with accessions of African lowland countries, which fits the hypothesis that finger millet was first domesticated in African highlands and from there it was introduced to India, thereby proving the origin of finger millet. There was good congruence between the phylogenetic tree and the population structure identified using STRUCTURE software. SNPs through GBS were used for association mapping to identify reliable marker(s) linked to grain yield or its component traits and calcium content. Five SNP markers showed homology to candidate genes of Oryza sativa (Rice) and Setaria italica, which play an important role in flowering, maturity and grain yield. Two potential SNP markers were found to be significantly associated to grain calcium content. The orthologous regions of the two identified SNP markers were found in cereal crops like Setaria italica encoding genes for Calmodulin Binding Protein (CBP) and CBL-Interacting Protein Kinase 7(CIPK7), which might play important role in grain calcium accumulation process. Promoter analysis of these genes revealed that they contain many stress responsive elements. Since, finger millet is a drought tolerant crop and also accumulate high amounts of calcium, the promoter region of these genes contain stress responsive elements, thus imparting stress tolerance to finger millet along with high grain calcium content as calcium ions play a central role in stress signaling. Differential expression analysis by qPCR showed that the two genes (EcCBP and EcCIPK7) were highly expressed in high calcium genotypes as compared to medium and low calcium genotypes. EcCIPK7 gene identified in the present study might be involved in regulating the activity of CaX exchanger and consequently increases the calcium storage in vacuoles, thus forming the basis of high grain calcium accumulation in finger millet. Since calmodulin (CaM) expresses abundantly in developing spikes and thus regulates the Ca2+ ATpase transporter located in vacuolar membrane, therefore, higher expression of EcCBP gene in later stages of spike development might be responsible for its interaction with CaM and might be responsible for higher accumulation of calcium in finger millet grains. Therefore, these genes could be considered as strong candidates for higher grain yield, Ca accumulation and its further use for Ca bio-fortification. However, exploration of its spatial distribution within seed, over expression and knockout studies will help in understanding the exact role of these genes in high seed calcium accumulation. Since, there is very limited information on genetic analysis of calcium content in finger millet grains, results from the present study would provide a fundamental basis for future research on genetic improvement of calcium content in finger millet and other crops.