葡萄是世界果树生产中栽培面积广、产量高的最重要果树之一。全球葡萄生产主要栽培是欧洲葡萄品种，这些品种具有酿酒、鲜食、制汁及制干等广泛的用途与创造经济效益高的特性，近年的研究指出葡萄及其制品中含有的白藜芦醇对人体的健康有益。但是，欧洲葡萄多数品种不抗葡萄白粉病，仅美国每年用于防治葡萄白粉病的杀菌剂费用超过了30亿美元。从根本解决欧洲葡萄品种不抗病是利用抗葡萄白粉菌的葡萄种质资源进行抗病育种。本项目就是利用抗白粉病且白藜芦醇含量高的中国野生葡萄为材料，前期通过转录组学测序分析发现中国野生毛葡萄丹凤-2不仅抗病强且芪合成酶基因累积与表达强，又比欧洲葡萄品种多拥有6条特有的芪合成酶基因。本项目拟通过对这些特有芪合成酶基因的序列及其保守结构分析，通过农杆菌介导的方法将这些新基因转入到无此类基因的模式植物拟南芥和不抗病的欧洲葡萄品种中，在白粉菌接种诱导下，利用实时荧光定量技术分析基因的时空表达特性;利用免疫共沉淀结合质谱测序技术筛选与芪合成酶基因互作蛋白，并进行基因表达与抗病功能研究；利用酵母单杂交和体外凝胶阻滞实验EMSA筛选与特有芪合成酶基因启动子互作的转录因子，研究这些转录因子基因对特有的芪合成酶基因的调控功能；通过转芪合成酶基基因欧洲葡萄植株功能研究，分析这些基因的表达及其激活的抗病基因的特性，揭示这些基因抗白粉病的调控机理与作用。在项目实施中拟培养研究生5-8人，其中博士研究生3-5人，硕士研究生2-3人；项目研究结果拟发表7-8篇论文，其中SCI论文4-5篇；拟申请发明专利1-2件。本项目对中国野生毛葡萄6条特有的芪合成酶基因功能的研究，评价这些抗病的育种资源，通过对欧洲葡萄品种杂交育种改良抗病性与增加果实中白藜芦醇的含量有直接利作用，研究结果对利用中国野生葡萄种质，提高欧洲葡萄品种抗病性有重要的应用价值和科学意义。

Grapevine ranked the top three of the important fruit trees in the world, largely due to its economic benefits from wines, table grapes, grape juice and raisin. Vitis vinifera are main cultivars in grapevine production which have high quality and various utilizations. Resveratrol in grapevine plays important role in human health. However, most of Vitis vinifera species are susceptible to Uncinula necator, and the fungicide cost of preventing powdery mildew was over 3 billion dollars every year in America. Therefore, the key to improve the resistance of these cultivars is to make use of grapevine germplasm resources with high resistance to the disease. Resveratrol, produced by stilbene synthase in grapevines, not only enhances resistance to plant pathogens but also plays important roles in human health, including antioxidant, anti-aging and anti-cancer. In our previous studies, the result of transcriptome sequencing showed that Chinese wild Vitis quinquangularis Danfeng-2 possesses stronger pathogen resistance, and higher accumulation and expression level of stilbene synthase genes, especially, six novel stilbene synthase genes were identified in Vitis quinquangularis. In the present study, based on the analysis of the sequences and conservative structures from the six genes, they will be transfered into the plants without these specific genes, including the model plant Arabidopsis thaliana and the susceptible European grapevines by Agrobacterium mediated transformation system. After inoculation with Uncinula necator, the interacting proteins with the six stilbene synthase genes will be screened by qPCR method, and co-immunoprecipitation combined with mass spectrometry, so as to analyze gene expressions and pathogen resistance. The regulation of the stilbene synthase genes by the transcription factors will be studied by screening the transcription factors interacting with the promoters of these stilbene synthase genes using the methods, yeast one-hybrid and EMSA. Therefore, the expression net of the stilbene synthase genes and activated resistant genes will be explored in transgenic European grapevines, so as to reveal the function and regulation model of stilbene synthase genes against powdery mildew, and provide useful scientific basis for breeding novel grapevine germplasm with pathogen resistance and high resveratrol content. Nine graduate students will participate in the studies, and five of them are doctoral students and four of them are master students. Moreover, 7-8 articles will be organized and published and 1-2 invention patents will be applied. Therefore, the obtained results will be useful for improving the resistance of cultivars and increasing resveratrol content based on Chinese wild grapevines.