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Molecular characterization of EcCIPK24 gene of finger millet (Eleusine coracana) for investigating its regulatory role in calcium transport

Chinchole, Mahadev, Pathak, RajeshKumar, Singh, UmaM., Kumar, Anil
3 Biotech 2017 v.7 no.4 pp. 267
Eleusine coracana, amino acid composition, binding capacity, calcium, gene expression, genes, genotype, leaves, millets, molecular models, physiological transport, quantitative polymerase chain reaction, reverse transcriptase polymerase chain reaction, roots, seeds, shoots, single nucleotide polymorphism, transcriptome, transporters, vacuoles
Finger millet grains contain exceptionally high levels of calcium which is much higher compared to other cereals and millets. Since calcium is an important macronutrient in human diet, it is necessary to explore the molecular basis of calcium accumulation in the seeds of finger millet. CIPK is a calcium sensor gene, having role in activating Ca²⁺ exchanger protein by interaction with CBL proteins. To know the role of EcCIPK24 gene in seed Ca²⁺ accumulation, sequence is retrieved from the transcriptome data of two finger millet genotypes GP1 (low Ca²⁺) and GP45 (high Ca²⁺), and the expression was determined through qRT-PCR. The higher expression was found in root, shoot, leaf and developing spike tissue of GP45 compared to GP1; structural analysis showed difference of nine SNPs and one extra beta sheet domain as well as differences in vacuolar localization was predicted; besides, the variation in amino acid composition among both the genotypes was also investigated. Molecular modeling and docking studies revealed that both EcCBL4 and EcCBL10 showed strong binding affinity with EcCIPK24 (GP1) compared to EcCIPK24 (GP45). It indicates a genotypic structural variation, which not only affects the affinity but also calcium transport efficiency after interaction of CIPK-CBL with calcium exchanger (EcCAX1b) to pull calcium in the vacuole. Based on the expression and in silico study, it can be suggested that by activating EcCAX1b protein, EcCIPK24 plays an important role in high seed Ca²⁺ accumulation.