参与TuMV复制的不结球白菜寄主因子的筛选及抗病基因功能验证

项目名称： 参与TuMV复制的不结球白菜寄主因子的筛选及抗病基因功能验证

项目编号： No.31272172

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 农业科学

项目作者： 侯喜林

作者单位： 南京农业大学

项目金额： 80万元

中文摘要： 芜菁花叶病毒病(TuMV)是不结球白菜的重要病害，TuMV病毒基因组的复制需要大量的寄主因子参与，分离和克隆这些寄主因子，通过基因工程手段使这些寄主因子基因沉默，可以使寄主产生对病毒的抗性，虽然近年来，对TuMV复制机理以及寄主因子的研究取得了一定的进展，但TuMV与寄主互作的复制机理以及复制复合体模型还远没有研究清楚，因此本研究利用蛋白免疫共沉淀结合质谱分析技术筛选参与TuMV复制复合体的不结球白菜寄主基因，并通过酵母双杂交技术筛选与TuMV复制相关基因直接互作的不结球白菜寄主基因，然后通过基因共定位、BIFC、酵母双杂交等技术验证后，得到参与TuMV复制的不结球白菜寄主基因，总结和完善TuMV复制复合体模型，最后研究参与TuMV复制的寄主基因沉默后对TuMV侵染的影响，探讨TuMV复制与寄主互作机理，发现抗病毒的新策略，并有望获得具有独立知识产权的抗TuMV的不结球白菜新基因。

中文关键词： 不结球白菜；芜菁花叶病毒；寄主因子；抗病基因；基因功能验证

英文摘要： Turnip mosaic virus (TuMV) is the most prevalent viral pathogen infecting non-heading Chinese cabbage(Brassica campestris ssp. chinensis Makino). TuMV has a relatively small genome and must depend on host factors for replication. Isolation of host factors and subsequently silencing of a host factor gene could lead to the generation of genetic resistance to TuMV. In spite of many recent advances, our current understanding of TuMV replication and virus-host interactions is far from complete. In this research, we isolate the host factors of the TuMV replication complex (VRC) by co-immunoprecipitation with LC-MS/MS and screen the host factors interacted directly with TuMV replication genes by yeast two hybrids in non-heading Chinese cabbage. After these host factors are verified by colocalization, BIFC and yeast two hybrids, we will complete the model of TuMV replication complex. Finally we study the affection on TuMV replication and multiplication after host factors gene silencing. These researches should lead to a better understanding of viral replication and the interactions between host factors and TuMV and might also help develop novel antiviral strategies and potential resistant genes of non-heading Chinese cabbage.

英文关键词： non-heading Chinese cabbage；Turnip mosaic virus；host factor；disease resistant gene；gene function verification