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Open Access
Response of iron and zinc nano oxides in Picroside I biogenesis during in vitro growth and development of Picrorhiza kurroa Royle ex Benth.
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2022-01) Rawat, Geeta; Gaur, A.K.
Picrorhiza kurroa Royle ex Benth is one among human and animal health associated valuable herbs. Several iridoid glycosides, as well as their derivatives with diverse pharmacophore, have been documented in this plant which have been undertaken for detail studies for their potential in a variety of modern and traditional healthcare systems toward managing a variety ailment since ancient times. Commercially available formulations for cure of disorders are Livocare, Livomap, Livplus, Katuki and Arogya. Due to business potential, an increasing global demand of this plant, this important plant resource has received much attentions of various researchers. Focus in this study have been provided with special reference to optimize protocols of in vitro regeneration of Picrorhiza kurroa. The influence of nanoparticles (ZnO-NPs and Fe2O3- NPs) during callus cultures, upon secondary metabolites specially on picroside I and levels of antioxidative enzymes namely superoxide dismutase (SOD), Catalase (CAT), Guaiacol peroxidase (GPOX), Ascorbate peroxidase (APOX). Biological assays against potent harmful microbes were also undertaken. Laboratory scale parameters were established for developing optimum suspension cultures for picroside I in order to develop large-scale production parameters. Elicitation response for picrosides production and related genes expression analysis of picroside metabolic pathway in suspended microcalli were observed. Zinc oxide nanoparticles (ZnO NPs) and Iron oxide nanoparticles (Fe2O3 NPs) responses were noticed to recognise specific identification of pathway genes with special reference to Hydroxy-methyl glutaryl CoA reductase (HMGR), Mevalonate diphosphate synthase (MVDD), Isopentyl diphosphate isomerase (IPPI), 3-Deoxy- D-arabino heptulosonate 7-phosphate synthase (DAHPS), 5-enolpyruvylshikimic acid-3-phosphate synthase (EPSPS), Dehydroquinate dehydratase (DQS), Geraniol diphosphate synthase (GDPS), 1-deoxy- D-xylulose-5-P-synthase (DXPS), Hydroxy methyl butenyl-4-diphosphate reductase (ISPH), Cytochrome P-450 monooxygenase (CPM), 2-hydroxy isoflavanone dehydratse (2HFD), Chorosmate mutase (CM) and Phenylalanine ammonia lyase (PAL) for picroside I only as an attention for its biogenesis. These observations imply that such an induced callus cultures under response of nanoparticles are suitable for picroside I production from Picrorhiza kurroa. Exploration of mass manufacturing particles at least of picroside I is possible by establishing suitable parameters in a simulated bioreactor system based on laboratory scale parameters identified from this particular study.
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, 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.
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, 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.
Open Access
Characterization of novel mutants of hexaploid wheat (Triticum aestivum L.) for isolation of mutants with higher content of micronutrients and lodging tolerance
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2021-03) Bisht, Darshana; Sundip Kumar
Producing nutritious and safe foods sustainably and sufficiently is the ultimate goal of modern agriculture. Mutagenesis is one of the powerful genetic strategies for crop improvement programmes. A chemically induced mutated genetic resource for detecting novel variations has been developed in bread wheat (Triticum aestivum) cultivar DPW-621-50. 1536 EMS-induced wheat mutant lines along with 4 checks grown during the Rabi season of 2019/2020 were evaluated for variability parameters, correlations and path coefficients for thirteen quantitative traits i.e., plant height, number of tillers per plant, flag leaf length, flag leaf width, flag leaf area, ear length, peduncle length, number of spikelets per spike, number of grains per spike, grain length, grain width, 1000 grain weight and grain yield per plant. Significant genotypic differences were observed for all the traits studied indicating considerable amount of variation among mutant lines for each character. The cluster analysis grouped these mutant lines into 40 different clusters. Grain yield per plant showed highly significant positive correlation with number of tillers per plant, number of grains per spike, thousand grain weight, number of spikelets per spike, plant height, grain width, grain length, ear length peduncle length. Path coefficient analysis revealed that number of tillers per plant had the highest direct effect on grain yield per plant. Prior efforts have focused on raising crop yields, but enhancing the concentrations of mineral micronutrients has become an urgent task because about half of the world population suffers from the malnutrition of iron and zinc. The overall impact of this high concentration of micronutrients is directly based on their bioavailability which is influenced by antinutritional (tannins and phytic acid) and nutritional (ascorbic acid) factors. This mutant population was used to isolate the mutants containing higher contents of minerals (Fe and Zn) in grains. Mutant lines namely, BSM 410, BSM 531, BSM 659, BSM 580, BSM 531, BSM 5, BSM 804, BSM 485, BSM 518, BSM 644, BSM 611, BSM 1004, BSM 390, BSM 990, BSM 609, BSM 97, BSM 567 and BSM 380 were found to possess high Fe and Zn and low phytic acid content. BSM 379, BSM 11, BSM 462, BSM 558, BSM 647, BSM 344, BSM 49, BSM 54, BSM 44, BSM 269, BSM 348, BSM 560, BSM 386, BSM 354, BSM 556, BSM 373, BSM 3, BSM 583, BSM 419 and BSM 580 were found to contain high Fe and Zn and low tannic acid content. BSM 549, PMW-2016-1, BSM 348, BSM 483, BSM 455, BSM 558, BSM 591, BSM 523, BSM 74, BSM 31, BSM 863, BSM 347, BSM 360, BSM 531, BSM 532, BSM 24, BSM 373 and BSM 408 were found to contain high Fe and Zn and high ascorbic acid content in comparison to parent. From the newly developed mutant population, we have also identified a putative lodging tolerant plant (PMW-2016-1). Comparative anatomical studies of the culm of lodging tolerant plant (PMW-2016-1) and parent (DPW-621-50) were carried out by means of various microscopic techniques coupled with chemical studies. It was found that the culm of PMW-2016-1 possessed greater stem diameter, thicker width, higher thickness-diameter ratio and higher percentage of mechanical tissue, comparing with its parent. Furthermore, the lignin content was determined by Klason’s method. The result showed that Klason lignin content in the novel mutant (PMW- 2016-1) was higher than that in its parent genotype (DPW-621-50). Estimation of mechanical stem strength with Universal Tensile Machine exhibited that the force required to break the straw of lodging tolerant mutant wheat PMW-2016-1 is greater as compared to the straw of control (DPW-621-50). Bending of stems of mutant PMW-2016-1 and control (DPW-61-50) was analyzed by using the prostate tester. Stem strength of mutant PMW-2016-1 was found to be significantly higher than the control (DPW-61-50). Therefore, it is suggested that newly identified mutant PMW-2016-1 can be used as variety or as a donor parent for the development of lodging tolerance in wheat cultivars. This mutant population may serve as new genetic resource for functional genomics studies and novel variants for different traits in elite germplasm can be made available to the plant breeders for wheat improvement.
Open Access
Effect of differential nitrogen fertilization and nano zinc seed priming on yield and seed protein quality in finger millet genotypes
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2020-02) Gururani, Kavita; Pandey, Dinesh
The “green revolution” based modern agriculture helped in increasing crop production and averting the hunger with the use of Nitrogenous fertilizers mainly. However, excessive use of fertilizers has threatened agriculture and sustainability of environment; and reduced the diversity of food, which is a major cause of malnutrition. Agronomic biofortification through direct application of fertilizer is an effective method to overcome the problem of malnutrition; however, fertilizers should be used with caution for both commercial and environmental reasons. Therefore there is need to optimize the dose of fertilizer and to use naotechnological interventions so as to produce nutritionaly rich food without compromising yield and damaging the environment. Such approaches will not only biofortify the crops but also ensure environment-friendly agriculture by promoting low-input sustainable crop cultivation. Finger millet (Eleusine coracana), holds immense agricultural and economic importance due to its high nitrogen use efficiency and richness in proteins being good source of essential amino acids and minerals. Two genotypes of finger millet viz. GE-1437 and GE-3885 were taken for the study to observe the effect of differential nitrogen fertilization without and with nano zinc seed priming (5ppm) on agro-morphological parameters, seed protein content, seed zinc content, expression analysis of seed storage proteins, regulatory genes of essential amino acid metabolism and accumulation of seed storage proteins. Results of agro-morphological data analysis showed the maximum biomass, grain yield and zinc content was obtained at 60kg/h urea without nano zinc seed priming; and 40kg/h urea with nano zinc seed priming. Nine regulatory genes involved in metabolism of essential amino acids have been identified from the transcriptome of finger millet spike. Promoter analysis revealed the presence of various cis-regulatory elements in the regulatory genes i.e. GCN4 motif, O2-site, G-box and GATA-motif, P-box etc. which are found to play an important role in N metabolism, SSP synthesis, light response, hormone response or stress response and directly or indirectly may regulate the biosynthesis of essential amino acids. Among the treatments without nano zinc seed priming the maximum expression of all the seed storage proteins was obtained with 40kg/h urea at later stages of spike development. On the other hand, for the treatments with nano zinc seed priming, the maximum expression of all the SSPs was obtained with control and 80kg/h urea at later stages of spike development. Both the genotypes showed higher expression of most of the regulatory genes with farm yard manure without and with nano zinc seed priming. The comparative analysis showed the higher expression of globulin and prolamin with the treatments without nano zinc seed priming and albumin with the treatments with nano zinc seed priming at later stages of spike development. SDS-PAGE further confirmed the increase in globulin and prolamin with the treatments without nano zinc seed priming and albumin with the treatments with nano zinc seed priming in mature seeds. Results of present investigation have suggested that high productivity/yield along with high Zn content of Finger millet can be obtained upon application of 40kg/h urea with nano zinc seed priming. Though this treatment can be used to solve Zn malnutrition problem among the masses yet it compromises with quality of protein and protein content in grains. In order to solve protein malnutrition problem, however, one can apply 40kg/h urea without nano zinc seed priming to enhance the seed protein content and quality in terms of essential amino acids.
Open Access
Isolation, cloning and in silico analysis of drought responsive EcMyb1 transcription factor gene from Eleusine coracana
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2021-01) Bhatt, Megha; Lohani, Pushpa
Myb genes are members of a family of transcription factors that play an important role in cell division, differentiation, response to abiotic stress etc. It has been reported that MYB transcription factor play an important role as a master regulator and modulates the expression of various downstream genes involved in drought stress response. The partial sequence of Ecmyb1 gene was isolated and submitted to NCBI with accession number JN107890.1. In present study, full length Ecmyb1 gene was isolated and cloned. The cloned fragment was sequenced and submitted to NCBI with accession number MT312253. For expression analysis of Ecmyb1 gene, finger millet drought tolerant variety PRM 6107 was given three different abiotic stress treatment i.e. drought, salt and heat stress. The expression of the gene increased from 12 to 28 fold under drought stress as compared to control. Similarly, in salt and heat treated plants, expression increased from 7 to 29 fold and 19 to 46 fold respectively. The cloned gene was introgressed into Arabidopsis thaliana plant by floral dip method. The transformed plants were selected by hygromysin screening and confirmed by PCR using hygromycin primers. Insilco analysis of Ecmyb1 gene was carried out using various softwares. Promoter analysis of Ecmyb1 gene revealed the presence of various regulatory elements like ABA responsive elements (CACT GG), stress responsive elements (AAGG GG), Myb recognition sites (CAACAG), Myb binding sites (CAAC TG), CpG island and tandem repeats. Homology and phylogenetic analysis of Ecmyb1 gene and amino acid sequences showed similarity with other MYB proteins such as Sateria italica, Hordeum vulgare, Saccharum barberi and Oryza sativa etc. Motif analysis predicted the presence of HTH domain in EcMYB1 protein. Subsequently, it was computed that EcMYB1 protein is water soluble, thermostable and slightly acidic in nature. Additionally, it was observed that EcMYB1 protein contains 28% alpha helix, 5.43% extended strand, 2.57% beta turn and 64% random coil. 3-D structure of EcMYB1 using swiss model predicted that 90% aminoacid residues were in Ramachandran favored region with no bad angles. It indicated that EcMYB1 protein may exist in nature.
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 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.
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, 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.
Open Access
Genotyping-by-sequencing and functional genomics based approaches for identification and nano-minerals (Fe and Zn) mediated modulation of candidate genes involved in iron and zinc homeostasis in finger millet
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2020-08) Chandra, Ajay Kumar; Pandey, Dinesh
Micronutrient deficiencies of iron and zinc are mainly responsible for significant amount of malnutrition among children and women across the world. One of the approaches to solve this problem is through biofortification of staple food crops provided that key candidate genes and markers associated with iron and zinc accumulation are identified. Being very good accumulator of minerals such as iron and zinc, finger millet is an ideal crop for identifying the molecular mechanism of iron and zinc homeostasis and molecular markers linked to high iron or zinc content in finger millet. With an aim to identify potential markers linked to grain iron and zinc content, initially phenotyping of 202finger millet genotype was done for fourteen morphological traits along with grain iron and zinc content at two different environments viz. Pantnagar (L1) and Almora (L2). 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 grain iron content varied from 26.72 to 78.59 mg.kg-1 seed with an average of 44.34 mg.kg-1 seed at L1 whereas, the GIC at L2 varied from 29.83 to 76.40 mg.kg-1 seed with an average of 46.14 mg.kg-1 seed. Similarly, the grain zinc content varied from 16.28 to 45.17 mg.kg-1 seed with an average of 32.18 mg.kg-1 seed at L1 whereas, the GZC at L2 varied from 12.40 to 47.18 mg.kg-1 seed with an average of 32.36 mg.kg-1 seed which indicates presence of sufficient variability for grain iron and zinc trait amongst the selected finger millet genotypes. Further, 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 and grain iron and zinc content. The major cluster A consisted broadly of all the Indian genotypes whereas cluster B and C comprised of exotic genotypes. Thus, there was good congruence between the phylogenetic tree and the population structure. SNPs through GBS were used for association mapping to identify reliable marker(s) linked to grain iron and zinc content. A total of five SNP markers, three for grain iron [TP413281 associated with ferritin (Fer1), TP566120 with iron-regulated transporter-like protein (IRT2), and TP1431395 associated with yellow stripe-like 2 proteins (YSL2)] and two for grain zinc [TP1316808 associated with zinc transporter ZIP1 protein and TP784188 associated with zinc transporter ZTP29-like protein], showed homology to candidate genes of Oryza sativa and Setaria italica, which might play an important role in grain iron and zinc homeostasis process in finger millet. Further, a total of fifteen genes (EcDMAS1, EcFER1, EcIRT2, EcNAAT6, EcNAC2, EcNAS1NAS2, EcNRAMP2, EcNRAMP6, EcPDR9, EcTOM1, EcTOM2, EcYSL1, EcYSL2, EcZIP1, and EcZTP29) orthologous to CGs of Oryza sativa and Setaria italica putatively involved in iron and zinc homeostasis in finger millet were identified through Genome-Transcriptome transition approaches. Structural and functional annotation of identified genes further highlights its significance that the genes are putatively involved in iron and zinc homeostasis process in finger millet. Furthermore, it was studied whether nano-treatments have any influencing role in epigenetic behavior so that it could enhance the molecular machineries of Fe and Zn enrichment in the plants. Molecular modeling and docking potential of identified proteins with nano-minerals (Fe3O4 NPs and ZnO NPs) confirmed that these nano-minerals may significantly influence the binding potential and modulation of regulatory genes. In addition, comparative In vitro studies for nano-minerals augmentation using seed priming approach resulted that the lower concentrations of Fe3O4 NPs (100 ppm) and ZnO NPs (5 ppm) play a significant and promotory effect on fourteen quantitative traits. Besides, the mean iron content in nano seed primed harvested grains, significantly increased by 12.26% and, the zinc by 13.96% concerning controlled harvested grains. Thus to confirm the genetic and molecular basis of identified genes and their modulation in response to nano-minerals, the Transcriptome based expression profiling revealed that of these 15 genes, EcFER1, EcIRT2, EcYSL2, EcZIP1, and EcZTP29 are the major genes which might play an important and direct role while the rest genes may indirectly involved in differential iron and zinc homeostasis in finger millet. The result was further validated by quantitative real-time PCR analysis. Structural and functional validation of set of genes involved in strategy-I (EcIRT2, EcFER1, and EcZIP1), strategy-II (EcYSL2), combined i.e., strategy-I and strategy-II (EcIRT2, EcFER1, EcYSL2, and EcZIP1) and the expression of EcZTP29 (stress-responsive gene) in finger millet tissues suggested that being a stress-resilient crop finger millet utilizes a combination of strategies I and II of iron and zinc homeostasis pathway. Since, there is not yet available any information on genetic and molecular basis of Fe and Zn homeostasis in this crop, therefore the results of present investigations have unlocked new avenues for effective utilization of finger millet in future iron and zinc biofortification programs.

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