摘要
9-顺式-环氧类胡萝卜素双加氧酶(NCED)为脱落酸(ABA)合成途径的关键基因,参与果实生长发育。为明确NCED基因家族对葡萄果实成熟的调控功能,以鄞红葡萄为材料,在开花后50 d进行环割处理,以不环割为对照,分析其果实成熟期、内源脱落酸含量和NCED家族中6个成员表达的变化;通过克隆这6条基因编码区目的片段,构建了pCambia1302过表达植物载体,在花蕾期用农杆菌侵染液浸泡花序,以空载和野生型为对照,分析农杆菌侵染后的果肉中ABA含量和基因表达变化。结果表明,环割后鄞红葡萄果实较对照提前成熟13 d左右;实时荧光定量PCR分析结果显示,环割处理7 d后的果肉中VvNCED1、VvNCED3、VvNCED4和VvNCED7表达量显著上调,与葡萄果肉中内源ABA变化相关;农杆菌侵染花穗后阳性转基因果肉中VvNCED1、VvNCED3、VvNCED4和VvNCED7基因表达量和ABA含量显著高于野生型。综上可知,VvNCED1、VvNCED3、VvNCED4和VvNCED7的表达具有调控葡萄果实成熟的作用。本研究结果为葡萄成熟期的分子调控提供了理论基础。
9-cis-epoxycarotenoid dioxygenase(NCED) is a key gene in the ABA(abscisic acid) synthesis pathway that regulates fruit growth and development, but how these genes function in regulation of the fruit ripening is yet unclear. In this study, Yinhong grape was used as materials. Girdling was carried out at 50 days after flowering, and no-girdling was used as the control to analyze the changes in fruit maturity, endogenous ABA content and the expressions of 6 genes of the VvNCED family. The pCambia1302 overexpression plant vector was constructed by cloning the target fragments of the coding regions of the six genes. The inflorescence was soaked with Agrobacterium infection at the bud stage, the changes of ABA content and gene expression in the pulp after Agrobacterium infection were analyzed with the empty load and wild type as controls. The results showed that the Yinhong grape berries had higher ABA content and matured about 13 days earlier in girdling veins than no-girdling grapes. Real-time quantitative PCR analysis also showed that the expression of VvNCED1, VvNCED3, VvNCED4 and VvNCED7 in the grape pulp after 7 days of girdling were significantly enhanced, which was related to the change of endogenous ABA in grape pulp. The gene expression and ABA content of VvNCED1, VvNCED3, VvNCED4 and VvNCED7 in the positive transgenic pulp after Agrobacterium-infected panicle were significantly higher than those of the wild type. These results indicated that the expression of VvNCED1, VvNCED3, VvNCED4 and VvNCED7 have roles in regulating the fruit ripening of grapes, which could provide a theoretical basis for researching on the molecular regulation on fruit ripening of grapes.
作者
董昳伶
肖旭腾
张敏
沈乐意
陈天池
贾永红
吴月燕
DONG Yiling;XIAO Xuteng;ZHANG Min;SHEN Leyi;CHEN Tianchi;JIA Yonghong;WU Yueyan(College of Biological&Enviromental Sciences,Zhejiang Wanli University,Ningbo,Zhejiang 315000)
出处
《核农学报》
CAS
CSCD
北大核心
2022年第7期1339-1349,共11页
Journal of Nuclear Agricultural Sciences
基金
浙江省重点研发项目(2021C02053)
宁波市科技创新2025重大专项(2019B10015)
省一流学科学生创新项目(CX2020006)。
作者简介
董昳伶,女,主要从事葡萄新种质培养研究。E-mail:1095186301@qq.com;通讯作者:吴月燕,女,教授,主要从事植物新种质培养与育种研究。E-mail:wyy2000@zwu.edu.cn。
