葡萄抗根瘤蚜侵染转录因子基因WRKY表达与功能研究

项目名称： 葡萄抗根瘤蚜侵染转录因子基因WRKY表达与功能研究

项目编号： No.31201609

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 植物保护学、园艺学与植物营养学

项目作者： 杜远鹏

作者单位： 山东农业大学

项目金额： 21万元

中文摘要： 葡萄根瘤蚜为专性寄生害虫，05年至今在我国南北已均有分布，一旦传播扩散将导致我国自根栽培葡萄产业的毁灭。WRKY转录因子是一类重要的参与植物防御反应的蛋白。在前期基因芯片研究基础上，我们以砧木140Ru受侵染后表达显著上调的2个WRKY转录因子为目标，利用RACE技术获得cDNA全序列；通过组织定位、亚细胞定位及qRT-PCR和Western杂交研究其组织细胞表达特性；对不同抗性葡萄资源接种根瘤蚜，研究WRKY在转录和翻译水平上的表达变化与抗根瘤蚜能力的相关性；对病毒诱导瞬时转化抗性葡萄根系接种根瘤蚜，研究对抗根瘤蚜能力的影响：包括根瘤蚜生长发育、根结数量、防御途径相关酶活性及关键酶基因表达量、次生代谢和侵染位点组织结构响应；最后通过检测基因沉默植株中另一个WRKY基因的表达响应，分析两个成员间的功能关系。为了解WRKY在防御根瘤蚜侵染中的功能和抗性改良奠定基础，支持葡萄产业可持续发展。

中文关键词： 葡萄；根瘤蚜；转录因子；WRKY；

英文摘要： Grape phylloxera (Daktulosphaira vitifolia Fitch) is an obligate parasites insect, distributed in the southern and northern part of China since 2005 till now. Once spread, it will destroy our self-root viticulture industry. Transcription factor gene WRKY is an important defence protein. Based on the DNA microarray analysis of rotstock 140Ru during phylloxera infestation, two obviously up-regulated WRKY genes are used as research target, the full length cDNA is cloned by RACE (rapid amplification of cDNA ends); the expression characteristics are analyzed by tissue localization, subcellular localization, qRT-PCR and Western blot; Different phylloxera-resistace grape varieties are inoculated with phylloxera to study the correlation of changes in WRKY expression and phylloxera resistance; Virus induce gene silence of WRKY in phylloxera-resistance varieties are inoculated with phlloxera to study the WRKY function on phylloxera resistace, include phylloxera growth and development, nodosities numbers and size, defence pathway enzymes actuvuty and key enzyme genes expression, secondary metabolites and feeding site tissue structure responses; Finally, we analyze the other WRKY gene expression of the virus induce one WRKY silence plant to study the functions between the two WRKY genes. We aim to understand the function of

英文关键词： Grape；Phylloxera；Transcription factor；WRKY；