白叶枯病菌未知主效TALE的鉴定与广谱抗病水稻新种质的创制

白叶枯病菌（Xoo）毒性变异常导致水稻抗病性丧失。前期发现，Xoo菌株LN4在3个OsSWEET感病基因EBE(effector-binding element)被修饰的水稻上仍具有致病性，推测该菌株存在不结合3个EBE位点的未知主效TALE(transcription-activator like effector)。拟完成LN4菌株的基因组，分离该菌株所有tal基因和构建单一tal菌系，通过致病性测定，鉴定该菌株的未知主效TALE；利用RNA-seq、EBE分析、qRT-PCR、GUS测定和EMSA等途径，鉴定其在水稻中的感病基因，明确感病基因的性质和功能；在3个OsSWEET感病基因EBE修饰的基础上，利用基因编辑技术对该感病基因的EBE进行精准编辑，创制广谱抗白叶枯病的水稻新种质。预期研究结果不仅揭示Xoo毒性变异和水稻抗病性丧失的的机理，而且还可为水稻白叶枯病抗病育种提供新种质。

The pathogenicity variation of Xanthomonas oryzae pv. oryzae (Xoo) often lead to the loss of rice resistance to bacterial leaf blight (BLB). Previously we found that Xoo strain LN4 was compatible in rice varieties with EBEs (effector-binding elements) of three OsSWEET genes modified, suggesting that there are unknown major TALEs (transcription-activator like effectors) that don’t bind these EBEs in this strain. In this proposal, we are going to clone and identify the unknown major TALEs of LN4 strain by genome sequencing, isolating tal genes, constructing strains with individual tal genes and pathogenicity assays, to identify the susceptible genes and determine the characteristics and functions of them in rice through RNA-seq, EBE analysis, qRT-PCR, GUS assays and EMSA, to generating new rice varieties with broad-spectrum resistant to BLB by precisely modifying the EBEs of the newly identified susceptible genes in the rice varieties with EBEs of three OsSWEET genes mutated using gene editing technology. The expected results from the mentioned above will not only reveal the mechanism of pathogenicity variation of Xoo and loss of rice resistance to BLB, but also provide new germplasms for resistance breeding of BLB.