Institutional Publications
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Item MicroRNA164e suppresses NAC100 transcription factor-mediated synthesis of seed storage proteins in chickpea(John Wiley & Sons, 2024) Chakraborty, Anirban; Singh, Baljinder; Pandey, Vimal; Parida, Swarup K.; Bhatia, SabhyataDevelopment of protein-enriched chickpea varieties necessitates an understanding of specific genes and key regulatory circuits that govern the synthesis of seed storage proteins (SSPs). Here, we demonstrated the novel involvement of Ca-miR164e-CaNAC100 in regulating SSP synthesis in chickpea. Ca-miRNA164e was significantly decreased during seed maturation, especially in high-protein accessions. The miRNA was found to directly target the transactivation conferring C-terminal region of a nuclear-localized transcription factor, CaNAC100 as revealed using RNA ligase-mediated-rapid amplification of cDNA ends and target mimic assays. The functional role of CaNAC100 was demonstrated through seed-specific overexpression (NACOE) resulting in significantly augmented seed protein content (SPC) consequential to increased SSP transcription. Further, NACOE lines displayed conspicuously enhanced seed weight but reduced numbers and yield. Conversely, a downregulation of CaNAC100 and SSP transcripts was evident in seed-specific overexpression lines of Ca-miR164e that culminated in significantly lowered SPC. CaNAC100 was additionally demonstrated to transactivate the SSP-encoding genes by directly binding to their promoters as demonstrated using electrophoretic mobility shift and dual-luciferase reporter assays. Taken together, our study for the first time established a distinct role of CaNAC100 in positively influencing SSP synthesis and its critical regulation by CamiR164e, thereby serving as an understanding that can be utilized for developing SPC-rich chickpea varieties.Item Comparative transcriptomic and metabolite profiling reveals genotype-specific responses to Fe starvation in chickpea(John Wiley & Sons, 2023) Singh, Gourav; Ambreen, Heena; Jain, Priyanka; Chakraborty, Anirban; Singh, Baljinder; Manivannan, Abinaya; Bhatia, SabhyataIron deficiency is a major nutritional stress that severely impacts crop productivity worldwide. However, molecular intricacies and subsequent physiological and metabolic changes in response to Fe starvation, especially in leguminous crops like chickpea, remain elusive. In the present study, we investigated physiological, transcriptional, and metabolic reprogramming in two chickpea genotypes (H6013 and L4958) with contrasting seed iron concentrations upon Fe deficiency. Our findings revealed that iron starvation affected growth and physiological parameters of both chickpea genotypes. Comparative transcriptome analysis led to the identification of differentially expressed genes (DEGs) between the genotypes related to strategy I uptake, metal ions transporters, reactive oxygen species (ROS) associated genes, transcription factors, and protein kinases that could mitigate Fe deficiency. Our gene correlation network discovered several putative candidate genes like CIPK25, CKX3, WRKY50, NAC29, MYB4 and PAP18, which could facilitate the investigation of the molecular rationale underlying Fe tolerance in chickpea. Furthermore, the metabolite analysis also illustrated the differential accumulation of organic acids, amino acids and other metabolites associated with Fe mobilization in chickpea genotypes. Overall, our study demonstrated the comparative transcriptional dynamics upon Fe starvation. The outcomes of the current endeavour will enable the development of Fe deficiency tolerant chickpea cultivars.Item Integrated genomic approaches delineate the novel role of ROP1 ENHANCER1 in regulating seed protein content of chickpea(Oxford University Press, 2023) Chakraborty, Anirban; Junaid, Alim; Parida, Swarup K.; Bhatia, SabhyataUtilizing a combinatorial approach of QTL-Seq and candidate gene-based association mapping, the QTLs and genes responsible for seed protein content (SPC), a major quality trait in chickpea were identified. Whole Genome Re-sequencing based QTL-Seq analysis of bulked RILs from a mapping population contrasting for SPC led to identification of two QTLs (0.94 Mb on Linkage Group (LG)5 and 1.16 Mb on LG6) encompassing three SNPs displaying the highest ΔSNP-index. These highly significant SNPs and their associated genes were validated in 211 chickpea mini-core accessions varying in SPC that revealed a tightly associated marker affecting CaREN1 (ROP1 ENHANCER1) with phenotypic variation explained of 23%. This SNP was subsequently converted into a cost effective allele specific PCR based marker that could be utilized for rapid screening of SPC during marker assisted breeding. Further, in planta functional validation via knockdown of CaREN1 led to significant reduction in SPC of chickpea. This decrease in seed protein is likely due to disruption in the formation of CaREN1 protein complexes comprising of chaperones, phosphopeptide-binding proteins and GTPases that mediate folding, transport and accumulation of seed storage proteins as indicated through AP-MS. Taken together, the information generated would expedite tailoring of chickpea cultivars with augmented SPC.
