Publications of NIPGR Scientists
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Item Iono-metabolomic guided elucidation of arsenic induced physiological and metabolic dynamics in wheat genotypes(Elsevier B.V., 2023) Saini, Himanshu; Panthri, Medha; Rout, Biswaranjan; Pandey, Ashutosh; Gupta, MeetuDespite the growing concerns about arsenic (As) toxicity, information on wheat adaptability in such an aggravating environment is limited. Thus, the present investigation based on an iono-metabolomic approach is aimed to decipher the response of wheat genotypes towards As toxicity. Wheat genotypes procured from natural conditions were characterized as high As-contaminated (Shri ram-303 and HD-2967) and low As-contaminated (Malviya-234 and DBW-17) based on ICP-MS As accumulation analysis. Reduced chlorophyll fluorescence attributes, grain yield and quality traits, and low grain nutrient status were accompanied by remarkable grain As accumulation in high As-contaminated genotypes, thus imposing a higher potential cancer risk and hazard quotient. Contrarily, in low As-contaminated genotypes, the richness of Zn, N, Fe, Mn, Na, K, Mg, and Ca could probably have supported less grain As accumulation, imparting better agronomic and grain quality traits. Additionally, from metabolomic analysis (LC-MS/MS and UHPLC), abundances of alanine, aspartate, glutamate, quercetin, isoliquiritigenin, trans-ferrulic, cinnamic, caffeic, and syringic bestow Malviya-234 as the best edible wheat genotype. Further, the multivariate statistical analysis (HCA, PCA, and PLS-DA) revealed certain other key metabolites (rutin, nobletin, myricetin, catechin, and naringenin) based genotypic discrimination that imparts strength to genotypes for better adaptation in harsh conditions. Out of the 5 metabolic pathways ascertained through topological analysis, the two main pathways vital for plant's metabolic adjustments in an As-induced environment were: 1. The alanine, aspartate and glutamate metabolism pathway, and 2. The flavonoid biosynthesis pathway. This is also evident from network analysis, which stipulates amino acid metabolism as a prominent As regulatory factor closely associated with flavonoids and phenolics. Therefore, the present findings are useful for wheat breeding programs to develop As adaptive genotypes that are beneficial for crop improvement and human health.Item In silico characterization of Thinopyrum elongatum-derived PsyE1 gene and validation in 7D/7E bread wheat introgression lines open avenues for carotenoid biofortification in wheat(Springer Nature Publishing AG, 2023) Padhy, Asish Kumar; Kaur, Parampreet; Singh, Baljinder; Kaur, Ravinder; Bhatia, Sabhyata; Shamshad, M.; Sharma, Himanshu; Kaur, Satinder; Srivastava, Puja; Sharma, AchlaCurrent global scenario demands agricultural productivity of food grains to be kept at abreast with burgeoning population. Cereals constitute major food stuff for millennia and biofortification of new cereal varieties provides an opportunity to tackle global-scale malnutrition deficiencies without doing major shifts in the diets. Carotenoid biofortification in wheat grains has recently caught the attention of breeders owing to a myriad of health benefits offered by this micronutrient. Thinopyrum elongatum-derived PsyE1 gene encoding for Phytoene Synthase encoding Y gene, is a jackpot to enhance the carotenoid content in wheat. The present study is the first report deciphering detailed in silico characterization of Thinopyrum elongatum-derived PsyE1 gene and its protein. Promoter analysis of chloroplast localized PsyE1 gene provides clues about its possible role in stress resistance along with enhancing the carotenoid content in both durum and bread wheat. Homology, phylogeny and protein modelling studies of PsyE1 revealed its closer evolutionary relationship with barley and wheat, as well as provided a preliminary insight into catalytic and secondary structure of the protein. PCR validation of PsyE1 in 7D/7E bread wheat introgression lines further facilitated development of functional marker that could be used to track its introgression in elite bread wheat varieties. Overall, these detailed insilico insights into structure, function and validation of PsyE1 open doors for its deployment in to produce carotene biofortified hexaploid wheat through facilitating development of functional markers and MAS, as well as to elucidate its mechanism of action and regulation in response to external stimuli.Item Functional analysis of SCD1 gene involved in pathogenicity of spot blotch disease of wheat causing fungus Bipolaris sorokiniana(Springer Nature Publishing AG, 2022) Singh, Kartar; Aggarwal, Rashmi; Verma, Praveen K.; Verma, Sandhya; Sharma, Sapna; Manjunatha, C.; Choudhary, Manoj; Kulshreshtha, Deepika; Rawat, KirtiSpot blotch of wheat is an important disease of wheat mainly of warm and humid areas. Melanin produced by various fungi has an important role to play in survival and longevity of fungal propagules against various environmental stresses through providing structural strength. Melanin production is positively correlated with virulence and conidiogenesis that varies in different isolates of plant pathogenic fungus Bipolaris sorokiniana. Scytalone dehydratase gene (SCD1) plays an important role in synthesis of fungal dihydroxynaphthalene melanin. In this study, the SCD1 gene was isolated, characterized and its DNA sequence along with flanking regions was determined. Structural analysis of gene revealed that 516 bp un-interpreted open reading frame. Sequence analysis showed that SCD1 gene encodes a 180 amino acids putative protein. A comparison of the predicted amino acid sequence and actual nucleotide sequence of the SCD1 gene was made with other fungal reductase genes involved in melanin biosynthesis which indicates 56–98% sequence identity. Targeted disruption of SCD1gene revealed that this gene is necessary for melanin biosynthesis in B. sorokiniana. SCD1 gene mutant isolate lack melanin synthesis and showed significant reduction in pathogenicity on wheat plants. This study expands our knowledge and provides comprehensive understanding of the pathogenesis of B. sorokiniana and insight of fungal phytopathogens.Item Identification of novel SNP in promoter sequence of TaGW2-6A associated with grain weight and other agronomic traits in wheat (Triticum aestivum L.)(PLOS, 2015) Jaiswal, Vandana; Gahlaut, Vijay; Mathur, Saloni; Agarwal, Priyanka; Khandelwal, Manoj Kumar; Khurana, Jitendra Paul; Tyagi, Akhilesh K.; Balyan, Harindra Singh; Gupta, Pushpendra KumarTaGW2 is an orthologue of rice gene OsGW2, which encodes E3 RING ubiquitin ligase and controls the grain size in rice. In wheat, three copies of TaGW2 have been identified and mapped on wheat homoeologous group 6 viz. TaGW2-6A, TaGW2-6B and TaGW2-6D. In the present study, using as many as 207 Indian wheat genotypes, we identified four SNPs including two novel SNPs (SNP-988 and SNP-494) in the promoter sequence of TaGW2-6A. All the four SNPs were G/A or A/G substitutions (transitions). Out of the four SNPs, SNP-494 was causal, since it was found associated with grain weight. The mean TGW (41.1 g) of genotypes with the allele SNP-494_A was significantly higher than mean TGW (38.6 g) of genotypes with the allele SNP-494_G. SNP-494 also regulates the expression of TaGW2-6A so that the wheat genotypes with SNP-494_G have higher expression and lower TGW and the genotypes with SNP-494_A have lower expression but higher TGW. Besides, SNP-494 was also found associated with grain length-width ratio, awn length, spike length, grain protein content, peduncle length and plant height. This suggested that gene TaGW2-6A not only controls grain size, but also controls other agronomic traits. In the promoter region, SNP-494 was present in 'CGCG' motif that plays an important role in Ca2+/calmodulin mediated regulation of genes. A user-friendly CAPS marker was also developed to identify the desirable allele of causal SNP (SNP-494) for use in marker-assisted selection for improvement of grain weight in wheat. Using four SNPs, five haplotypes were identified; of these, Hap_5 (G_A_G_A) was found to be a desirable haplotype having significantly higher grain weight (41.13g) relative to other four haplotypes (36.33-39.16 g).Item Advances in wheat genomics and its potential in ensuring food security in the scenario of climate change(Indian National Science Academy, 2014) Muthamilarasan, Mehanathan; Parida, Swarup K.; Prasad, ManojPrevalence of hunger and under-nutrition among the world population at alarming rates challenges the food security. Further, the instability in global wheat production in past two years and the projection of continuing decrease in wheat inventories predominantly due to climate change add-up to food insecurity. The global climate change imposing biotic and abiotic stress made the crop improvement more challenging. However, the timely release of wheat genome sequence holds the promise of evading these tribulations, provided the researchers and breeders pertinently utilize the genome information. In view of this, this short-review highlights the intact insights for the wheat research community on how to implement the genome sequence data in breeding and engineering transgenic wheat with enhanced biotic and abiotic stress tolerance.Item Transcript profiling identifies novel transcripts with unknown functions as primary response components to osmotic stress in wheat (Triticum aestivum L.)(Springer, 2013) Garg, Bharti; Puranik, Swati; Misra, Shrilekha; Tripathi, Bhumi Nath; Prasad, ManojOsmotic stress induced by dehydration and salinity, is among the major abiotic stresses that adversely impacts crop productivity and plants often display cultivar-dependent response against osmotic imbalance. To better understand the molecular mechanisms underlying differential responses to dehydration, transcriptome changes of two contrasting wheat (Triticum aestivum L.) cultivars were evaluated in plants grown under unfavorable osmotic conditions. A total of 107 non-redundant transcripts were identified. Of these, most had unknown functions (31; ~30 %) signifying the existence of putative stress-specific genes in wheat, reported here for the first time. Upon comparing with previous transcriptomic studies, 43 (40 %) of the osmotically-responsive transcripts were found not to be documented. These new transcripts may therefore signify unexplored gene sources for specific responses towards short-term osmotic stress in wheat. Through macroarray analysis, 69 (~64 %) transcripts were found to be differentially expressed (≥3-fold) and expression of 14 transcripts (with known or unknown functions) was further confirmed by quantitative real time PCR. Expression analysis of the seven unknown transcripts also revealed their tissue- and stress-specific regulation. Comparative in silico mapping of these 107 wheat transcripts against available mapping data for rice (40; ~37 %), maize (34; ~32 %), and sorghum (33; ~31 %) revealed presence of wheat orthologous sequences in these cereal crops. This study provides an interesting account on several novel genes, besides those with known functions, which may regulate stress response dynamics and thus, may be used as potential candidates to improve stress adaptability through genetic and molecular studies.Item A physiologically regulated multidomain cystatin of wheat shows stage-dependent immunity against karnal bunt (Tilletia indica)(Springer, 2012) Purwar, Shalini; Sundaram, Shanthy; Verma, Praveen K.; Srivastava, Shaili; Kumar, AnilTo identify novel components of basal resistance against the Tellitia indica of wheat, breeding for disease resistance was carried out on resistant and susceptible genotype of Karnal Bunt. The different members of wheat cystatin gene families were cloned, and their role in triggering differential resistance through co-expression was analyzed in our lab. The multidomain wheat cystatin (WCM) is a proteinase inhibitor characterized by cloning the gene from susceptible (WH542) and resistant genotype (HD 29). A WCM cDNA was isolated from both genotypes and sequenced. The WCM had a highly conserved N-terminal cystatin domain and a long C-terminal extension containing a second region, which exhibited similarity to the cystatin domain. The expression level was significantly (P > 0.001) higher in resistant compared to susceptible genotype at all the physiological stages of wheat spikes. In order to characterize the biochemical properties of WCM, the coding sequence was expressed in Escherichia coli using pET expression vector. The recombinant WCM was purified from soluble fraction of the cell extract by using affinity chromatography. WCM, with 23 KDa molecular mass, showed cysteine proteinase inhibitory activity against papain (Ki 3.039 × 10−7 M) as determined by using BAPNA as substrate. Furthermore, it was able to arrest the fungal mycelial growth of T. indica. Hyphae growth was inhibited, and morphological changes such as swelling and fragmentation of the fungus were observed. Overall, these observations suggest an endogenous high expression of cystatin, possibly associated with the resistance of wheat against Karnal bunt.Item A comprehensive study on dehydration-induced antioxidative responses during germination of Indian bread wheat (Triticum aestivum L. em Thell) cultivars collected from different agroclimatic zones(Springer Science, 2012) Garg, Bharti; Jaiswal, Jai P.; Misra, Shrilekha; Tripathi, Bhumi Nath; Prasad, ManojTo explore the adaptability of bread wheat to dehydration stress, we screened 28 cultivars collected from different agroclimatic zones, on the basis of malonaldehyde content as biochemical marker in roots of wheat seedlings during germination and classified them as highly tolerant, tolerant, sensitive and highly sensitive. From this primary screening, ten cultivars that showed differential responses to dehydration stress were selected to understand the biochemical and physiological basis of stress tolerance mechanisms. The highly tolerant cultivars showed lower levels of lipid peroxidation, less membrane damage, increased levels of antioxidants, enzymes like catalase, ascorbate peroxidase, glutathione reductase activities, and maintained higher relative water content in comparison to sensitive cultivars, indicating better protection mechanism operating in tolerant cultivars. Correspondingly, highly tolerant cultivars exhibited more accumulation of proline and less H2O2 content across different time points of polyethylene glycol treatments in comparison to sensitive ones. The above biochemical and physiological parameters were further validated through northern analysis of catalase (CAT1) gene, that showed differential expression patterns in tolerant and sensitive cultivars largely in confirmation with the biochemical and physiological analyses. Our study positively correlates the differences in the redox status and antioxidant defense system between tolerant and sensitive cultivars for the establishment of wheat seedlings in typical dehydration conditions.Item Identification and characterization of high temperature stress responsive genes in bread wheat (Triticum aestivum L.) and their regulation at various stages of development(Springer Science, 2011) Chauhan, Harsh; Khurana, Neetika; Tyagi, Akhilesh K.; Khurana, Jitendra P.; Khurana, ParamjitTo elucidate the effect of high temperature, wheat plants (Triticum aestivum cv. CPAN 1676) were given heat shock at 37 and 42°C for 2 h, and responsive genes were identified through PCR-Select Subtraction technology. Four subtractive cDNA libraries, including three forward and one reverse subtraction, were constructed from three different developmental stages. A total of 5,500 ESTs were generated and 3,516 high quality ESTs submitted to Genbank. More than one-third of the ESTs generated fall in unknown/no hit category upon homology search through BLAST analysis. Differential expression was confirmed by cDNA macroarray and by northern/RT-PCR analysis. Expression analysis of wheat plants subjected to high temperature stress, after 1 and 4 days of recovery, showed fast recovery in seedling tissue. However, even after 4 days, recovery was negligible in the developing seed tissue after 2 h of heat stress. Ten selected genes were analyzed in further detail including one unknown protein and a new heat shock factor, by quantitative real-time PCR in an array of 35 different wheat tissues representing major developmental stages as well as different abiotic stresses. Tissue specificity was examined along with cross talk with other abiotic stresses and putative signalling molecules.