中国野生毛葡萄转录因子VqWRKY3抗黑痘病的功能分析与调控机制研究

项目名称： 中国野生毛葡萄转录因子VqWRKY3抗黑痘病的功能分析与调控机制研究

项目编号： No.31501740

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 农业科学

项目作者： 李智

作者单位： 西北农林科技大学

项目金额： 21万元

中文摘要： 葡萄黑痘病在我国南北多雨地区都有大量发生，严重影响葡萄的产量和品质。利用野生葡萄自身的抗病基因资源，创制抗病优质葡萄新种质是最经济有效的措施之一。课题组前期通过抑制差减杂交和转录组分析从抗病材料中国野生毛葡萄‘商-24’中筛选到一个受黑痘病菌诱导的转录因子VqWRKY3。本项目拟通过超表达对葡萄感病品种‘红地球’进行遗传转化和VIGS技术对抗病材料‘商-24’进行诱导沉默，结合组织病理学、防御相关基因表达以及生理生化指标测定探讨VqWRKY3在抗葡萄黑痘病中的生物学功能；其次，通过酵母单杂和染色质免疫共沉淀筛选其上游调控基因和下游靶基因，利用凝胶阻滞实验进行体外验证，明确VqWRKY3基因的调控机制。再者，通过启动子活性分析，探讨VqWRKY3基因启动子在葡萄黑痘病菌、茉莉酸和水杨酸处理下的响应情况。本项目的开展，旨在为葡萄抗病分子育种提供基因资源，为葡萄品种改良提供理论基础。

中文关键词： 葡萄；生物逆境；基因功能；调控网络

英文摘要： Anthracnose is one of the most important grape diseases with high incidence in a wide range and very serious effect on the quality and yield of grape fruits in the south and north grape production area of China. An economic and effective method was used to create resistance germplasm resources by utilizing resistance gene resources of its own. We obtained a transcription factor induced by Elsinoe ampelina from resistance grape ‘Shang-24’ based on previous data of suppression subtractive hybridization (SSH) and transcriptome sequencing. We will obtain ‘Red globe’ (Susceptible grape) over-expression lines through agroinfiltration and ‘Shang-24’ (Resistant grape) slience lines by virus induced gene silencing (VIGS)，respectively. The functional analysis of VqWRKY3 will be preformed by disease assays, histopathology, expression of defense-related genes, and biochemical measurement. In addition, Yeast one-hybrid and ChIP-Seq assays were used to find the upstream regulation genes and downstream target gene of VqWRKY3, respectively. The candidate genes were confirmed by EMSA assay in vitro. Thirdly, the promoter activity of VqWRKY3 will be preformed by transient expression assays in grapevine to investigate the response pattern after treatment with Elsinoe ampelina, SA, and JA. The aim of this project is to provide gene resources for grape molecular breeding and to provide solid foundation for genetic improvement of grape.

英文关键词： Grape;Biotic stress;Gene function;Regulatory network