Cloning，Identification，Sequence Analysis and Spatio-temporal Expression Characteristics of Rs Gene in Kyoho Grape

doi:10.7668/hbnxb.201751391

Abstract

Abstract: To explore the potential role of Resveratrol synthase(Rs) in fruit development of Vitis vinifera cv. Kyoho, the structure and function of Rs sequence were analyzed, and the spatio-temporal expression specificity at different developmental stages of fruit was identified. The bioinformatics analysis and qRT-PCR method were used to analyze the sequence of Rs gene in Kyoho grape. The structure and subcellular localization of Rs gene were predicted, and its expression in different tissues of fruit at different developing stages was analyzed. The full-length cDNA of Rs gene was 1 539 bp, with an open reading frame(ORF) of 1 179 bp, encoding 392 amino acids. The predicted molecular weight was 42.88 ku, and the theoretical pI was 6.09. The gene contained chalcone, stilbene synthase activity sites and complete stilbene synthase family characteristic sites. The protein interaction predicted that Rs interacts with OMT2.1, which catalyzes resveratrol biosynthesis of Pterotilbene. The results of subcellular prediction showed that the Rs gene mainly expressed in the cytoplasm and chloroplast. The promoter analysis showed that the expression of Rs gene might be regulated by light, MYB, fungi and hormones, presenting a certain tissue specificity. qRT-PCR analysis showed that the Rs gene of grape expressed at different stages of the peel and flesh, but the expression level was highest in the 25th day after anthesis. Combined with the role of Rs gene accumulation in resveratrol, it could be inferred that there might be a high accumulation of resveratrol in the fruit peel during the early period of fruit development. The Rs gene of Kyoho grape had a high conservation during evolution, and its expression might be regulated by environment, fungi and hormone. It had the interaction with OMT2.1 and had a certain specificity in different tissues and development periods.

Key words: Grape, Rs gene, Clone, Sequence and expression analysis

CLC Number:

S663.1

CUI Tengfei, WANG Chen, TAN Hongyu, JIA Haifeng, BAI Yunhe, WANG Wenran, FANG Jinggui. Cloning，Identification，Sequence Analysis and Spatio-temporal Expression Characteristics of Rs Gene in Kyoho Grape[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2019, 34(3): 59-67. doi: 10.7668/hbnxb.201751391.

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