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Transcriptional expression analysis of genes involved in regulation of calcium translocation and storage in finger millet (Eleusine coracana L. Gartn.)
Authors:Neelofar Mirza  Gohar TajSandeep Arora  Anil Kumar
Affiliation:Department of Molecular Biology & Genetic Engineering, College of Basic Sciences & Humanities, G B Pant University of Agriculture & Technology, Pantnagar 263145, Uttarakhand, India
Abstract:Finger millet (Eleusine coracana) variably accumulates calcium in different tissues, due to differential expression of genes involved in uptake, translocation and accumulation of calcium. Ca2 +/H+ antiporter (CAX1), two pore channel (TPC1), CaM-stimulated type IIB Ca2 + ATPase and two CaM dependent protein kinase (CaMK1 and 2) homologs were studied in finger millet. Two genotypes GP-45 and GP-1 (high and low calcium accumulating, respectively) were used to understand the role of these genes in differential calcium accumulation. For most of the genes higher expression was found in the high calcium accumulating genotype. CAX1 was strongly expressed in the late stages of spike development and could be responsible for accumulating high concentrations of calcium in seeds. TPC1 and Ca2 + ATPase homologs recorded strong expression in the root, stem and developing spike and signify their role in calcium uptake and translocation, respectively. Calmodulin showed strong expression and a similar expression pattern to the type IIB ATPase in the developing spike only and indicating developing spike or even seed specific isoform of CaM affecting the activity of downstream target of calcium transportation. Interestingly, CaMK1 and CaMK2 had expression patterns similar to ATPase and TPC1 in various tissues raising a possibility of their respective regulation via CaM kinase. Expression pattern of 14-3-3 gene was observed to be similar to CAX1 gene in leaf and developing spike inferring a surprising possibility of CAX1 regulation through 14-3-3 protein. Our results provide a molecular insight for explaining the mechanism of calcium accumulation in finger millet.
Keywords:CaMK, calmodulin dependent kinase   SAM, shoot apical meristem   qPCR, quantitative PCR   CRD, completely randomized design   CAX1, Ca  +/H+ antiporter or exchanger   TPC1, two-pore channel   CDPK, Ca  +-dependent protein kinase
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