过表达VvCYP707A1基因降低葡萄果肉中的ABA含量
Overexpression of ABA 8'-Hydroxylase(VvCYP707A1)Gene Reduces ABA Content in Grape Pulp
董昳伶 1张敏 2肖旭腾 1贾永红 1吴月燕1
作者信息
- 1. 浙江万里学院生物与环境学院,宁波,315100
- 2. 浙江万里学院生物与环境学院,宁波,315100;上海海洋大学水产与生命学院,上海,201306
- 折叠
摘要
植物中ABA(脱落酸)的含量取决于它的合成和分解代谢速度的快慢.CYP707A家族编码的ABA8'-羟化酶是ABA代谢途径的关键酶.本研究以'鄞红'葡萄果肉为材料,采用一步克隆技术构建过表达载体pCambia1302-VvCYP707A 1,并对花蕾期葡萄花序进行转化,分析葡萄果肉中VvCYP707A1基因的过表达对内源ABA含量的影响.实时荧光定量PCR(RT-qPCR)结果显示,过熟期转基因葡萄果肉中VvCYP707A1基因的表达量显著高于野生型.进一步使用酶联免疫法(ELISA)测定相同条件下的ABA含量后发现,ABA含量显著低于野生型.这些结果表明,过表达VvCYP707A1基因降低了葡萄果肉中ABA含量,推测VvCYP707A1的表达是葡萄果肉中ABA代谢的主要调控机制.研究结果为ABA分解代谢机制提供一定理论依据.
Abstract
In plants,the level of abscisic acid(ABA)depends on the rates of its synthesis and catabolic metabolism.ABA 8'-hydroxylase,a key enzyme in the ABA metabolic pathway,is encoded by the CYP707A gene family.In this study,the overexpression vector,pCambia1302-VvCYP707A1,was constructed with one-step cloning technique based on grape pulp of'Yinhong',and the inflorescence was transformed to analyze the influence of VvCYP707A1 gene overexpression on endogenous ABA content in grape pulp.Real-time quantitative PCR(RT-qPCR)results showed that the expression of VvCYP707A1 in the transgenic grape pulp at the overripe stage was significantly high-er than in the wild-type.Further analysis using enzyme-linked immunosorbent assay(ELISA)revealed that the ABA content was significantly lower than that in the wild-type under the same conditions.These results indicate that over-expression of the VvCYP707A1 gene reduces the ABA content in grape pulp,suggesting that VvCYP707A 1 plays a major role in regulating ABA metabolism in grape pulp.This study provides a theoretical basis for the mechanism of ABA catabolic metabolism.
关键词
葡萄果肉/VvCYP707A1/过表达Key words
Grape pulp/VvCYP707A1/Overexpression引用本文复制引用
出版年
2024
NETL
NSTL
万方数据