IN-SILICO AND MOLECULAR CHARACTERIZATION OF YSL1 GENE IN WHEAT
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Date
2019
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Dr. Rajendra Prasad Central Agricultural University, Pusa, Samastipur
Abstract
Wheat is the staple food for the majority of people in the world and thus forms an important component of human nutrition and hence its bio-fortification concerning mineral nutrients is a major concern in the scientific world. In this study, the biofortification of wheat with respect to grain Fe and Zn content has been taken into consideration. Twenty-five wheat genotypes were sown under normal and late sown conditions (heat stress). Estimation of Fe and Zn content in wheat grains was done by AAS after wet digestion by Nitric acid: Perchloric acid:: 1:3. The data about Fe and Zn content in grains of wheat sown under normal and late conditions were analyzed statistically and it was found that there was a reduction of Fe and Zn content in the late sown than the normal sown conditions in most of the genotypes under study. Contrarily, 5 genotypes showed increased Fe accumulation and seven genotypes showed increased Zn accumulation in wheat grains under late sown condition, thus exhibiting negative HSI values. This distribution of Fe and Zn content over two dates of sowing was subjected to cluster analysis and were classified into three groups Fe content and 4 groups based on Zn content in seed.
The YSL gene family plays an important role in Fe and Zn homeostasis into the grains. In this study, all 63 members of the YSL gene family were identified in the wheat genome. Out of all the TaYSL genes, TaYSL2, in particular, was characterized in silico and was found to be expressed in the spike and involved directly in metal-NA loading into the seeds. Three homeologous copies of the gene TaYSL2 namely TaYSL2-6A, TaYSL2-6B and TaYSL2-6D, were identified on the 6th chromosome of each of the three genomes. 6 introns and 7 exons were found in each of the homeologous copies of the gene. In-silico expression of TaYSL2-6A increased many folds under prolonged drought conditions, TaYSL2-6B increased many folds under heat stress and combined stress of heat and drought increases the expression of TaYSL2-6B to many folds and TaYSL2-6D significantly. TaYSL2 gene was taken up for molecular characterization using gene-based genome-specific primers as well as gene-based SSRs primer in 12 genotypes selected from the varied Fe and Zn content data. Amplification was obtained successfully in all the designed primers. A total of 67 alleles including 37 unique alleles, were obtained from all the primers. The PIC value of primers ranged from 0.542 to 0.904. Maximum polymorphism was observed in B genome followed by A and D genomes respectively. Dice's similarity coefficient computed from the molecular data analysis ranged from 0.821 to 0.672. Based on the similarity coefficient, the genotypes were classified into 4 clusters and were successfully correlated with the Fe and Zn content data.