葡萄果实CYP707A基因的克隆与表达分析  

Cloning and Expression Analysis of CYP707A Gene in Grape Fruit

作　　者：杨洋[1] 贾永红[2] 王锦阳 张巧 董昳伶 汪庆昊 姚芳会 吴月燕[2] Yang Yang;Jia Yonghong;Wang Jinyang;Zhang Qiao;Dong Yiling;Wang Qinghao;Yao Fanghui;Wu Yueyan(College of Fisheries and Life Science,Shanghai Ocean University,Shanghai,201306;Collge of Biological and Environmental Sciences,Zhejiang Wanli University,Ningbo,315100)

机构地区：[1]上海海洋大学水产与生命学院,上海201306 [2]浙江万里学院生物与环境学院,宁波315100

出　　处：《分子植物育种》2020年第22期7356-7363,共8页Molecular Plant Breeding

基　　金：浙江省科技项目(2016C34G2211502);宁波市科技重大专项(2019B10015);浙江省一流学科“生物工程”学生创新项目(CX2019016)共同资助

摘　　要：8’-羟化酶(8’-hydroxylase)是脱落酸代谢途径中的关键酶,由CYP707A基因家族编码。该酶降解脱落酸生成红花菜豆酸(PA),对果实生长发育及成熟有重要作用。本研究分析了‘鄞红’葡萄中CYP707A基因的结构,并对果实发育过程中幼果期、膨大期、转色期、成熟期和过熟期CYP707A基因表达量和ABA含量二者间相关性进行了探讨。结果发现,‘鄞红’葡萄中含有一个新的CYP707A基因,命名为VvCYP707A1,该基因编码区长1 995 bp,有一个1 413 bp的完整开放阅读框(ORF),编码653个氨基酸;VvCYP707A1编码的蛋白属于疏水性跨膜蛋白,且亚细胞定位于内质网,预测蛋白与该蛋白家族不同种属来源序列相似度约为54.21%,具有保守的P450半胱氨酸血红素铁配位体,属8’-羟化酶家族。在果实成熟过程中,ABA含量和VvCYP707A1基因表达量变化总趋势为上升,并呈正相关;VvCYP707A1基因表达量呈上升趋势,过熟期达到最大。本研究完善了‘鄞红’葡萄CYP707A基因家族序列,为今后探究并验证其功能和果实成熟间的联系提供了数据参考。8’-hydroxy lase is a key enzyme in the abscisic acid metabolism pathway and is encoded by the CYP707 A gene family. The enzyme decomposes abscisic acid to produce phaseic acid(PA), which plays an important role in fruit growth, development and maturity. This study analyzed the structure of the CYP707 A gene in the’Yinhong’ grape, and correlated the expression of CYP707 A gene and ABA content in the mature, young, swollen,translucent, mature, and overripe stages of the fruit. It was found that the grape vine contains a new CYP707 A gene named VvCYP707 A1. The gene coding region is 1 995 bp and has a complete open reading frame(ORF) of 1 413 bp,encoding 653 amino acids. The protein encoded by VvCYP707 A1 belongs to a hydrophobic transmembrane protein with subcellular localization in the endoplasmic reticulum. The predicted sequence simi-lt arity between the protein and the different species of the protein family is about 54.21%. It has a conserved P450 cysteine heme iron ligand, g enus 8’-hydroxylase family. Along with the process of fruit maturation, the ABA content and the expression of VvCYP707 A1 gene showed an upward trend as a whole, and there was a positive correlation between them. The expression of VvCYP707 A1 gene was the highest during the ripening stage of fruit development. This study completes the CYP707 A gene family sequence of ’Yinhong’ grapes, and provides a data reference for exploring and verifying the relationship between its function and fruit maturity in the future.

关 键 词：VvCYP707A 基因 基因克隆 葡萄 基因表达分析 

分 类 号：S663.1[农业科学—果树学]