山葡萄抗霜霉病R基因与霜霉菌RXLR效应因子的互作机制

Downy mildew (DM), caused by oomycete pathogen called Plasmopara viticola，is one of the most damaging diseases for grapevines. The pathogen attacked young leaf tissues via the stomata at the bottom of young leaves. It can also infect other young tissues such young shoots and small berries. The disease could significantly decrease yield and berry quality if it is not controlled in a timely fashion. Fungicide is commonly used to control downy mildew disease of grapevines. However, fungicide application will not only increase the production cost, but also pollute the environment. Therefore, the alternative approach to confer the disease is to develop resistant grape cultivars. To achieve this goal, it is essential to understand the mechanism of host resistance. In this connection, we propose to use the functional genomics, transcriptomics and bioinformatics, in combination with QTL mapping and map-based cloning strategies, to identify, isolate downy mildew disease resistant genes (R genes) from the DM resistant Vitis amurensis cv. Shuanghong. In parallel with these are to indentify effector genes which are specific to ' ZJ-1-1' isolate (compatible interaction) and 'JL-7-2' isolate (incompatible interaction). Furthermore, the function validation of candidate genes and their target proteins will be analyzed using different molecular technologies, such as argo-infiltration, yeast two hybrid and grapevine transgenic system. The proposed project is designed to understand the mechanisms of disease resistance and susceptibility and their correlations against the infection and invasion of downy mildew pathogens. The overall goal is to fully understand the mechanism of host resistance. Success of this project will pave the way for developing new strategies for downy mildew disease control, and accelerate the development of downy mildew - resistant new grape cultivars.

霜霉病是危害葡萄生产最主要的病害之一。它是由葡萄霜霉菌通过叶背面的气孔入侵而导致叶片发病的。除了幼叶外，霜霉菌还可以侵染嫩稍、幼果等其它幼嫩组织。如果不及时有效的控制，将会严重影响葡萄产量和果实品质，造成巨大的经济损失。本研究选用高抗霜霉病的山葡萄'双红'及易感霜霉病的欧亚种'汤姆森无核'为寄主材料、对'双红'表现出亲和互作的菌株'ZJ-1-1'和非亲和互作的菌株'JL-7-2'为病原菌，接种后分别在不同时间点取样，通过转录组测序、生物信息学分析、基因定位和图位克隆等技术方法，挖掘山葡萄'双红'抗霜霉病基因以及"亲和"与"非亲和"菌株的特异效应因子，并通过葡萄瞬时表达及稳定转化体系对候选R基因抗葡萄霜霉病功能进行验证；同时，通过酵母双杂技术，筛选亲和互作和非亲和互作中效应因子的目标蛋白，揭示葡萄抗病和感病机理，为生产上霜霉病防治及抗病品种的培育提供新的策略和靶标。

霜霉病是危害葡萄生产最主要的病害之一。它是由葡萄霜霉菌通过叶背面的气孔入侵而导致叶片发病的。除了幼叶外，霜霉菌还可以侵染嫩稍、幼果等其它幼嫩组织。如果不及时有效的控制，将会严重影响葡萄产量和果实品质，造成巨大的经济损失。本研究选用高抗霜霉病的山葡萄‘双红’及易感霜霉病的欧亚种‘汤姆森无核’为寄主材料、用对‘双红’表现出亲和互作的菌株‘ZJ-1-1’和非亲和互作的菌株‘JL-7-2’为病原菌，接种后分别在不同时间点取样，通过转录组测序、生物信息学分析、基因定位和图位克隆等技术方法， 挖掘山葡萄‘双红’抗霜霉病基因以及“亲和”与“非亲和”菌株的效应因子，并通过葡萄瞬时表达及稳定转化体系对候选 R 基因抗葡萄霜霉病功能进行验证；同时，通过酵母双杂技术，筛选亲和互作和非亲和互作中效应因子的目标蛋白，揭示葡萄抗病和感病机理，为生产上霜霉病防治及抗病品种的培育提供新的策略和靶标。