RBSDV P5-2与水稻转录因子ZFP207互作在病毒侵染中的作用机理

Rice black-streaked dwarf disease (RBSDV) is a serious virus disease of rice in China in recent years. In previous studies, pathogenic factor P5-2 protein interacted with host factor ZFP207, playing an important role during the rice anti-virus progress. In this project, overexpression and RNAi silencing transgenic rice of P5-2 and ZFP207 will be gotten to investigate the role of P5-2 and ZFP207 in the pathogenicity of. The rice proteins interacting with rice protein- ZFP207 will be screened by using yeast two hybrid assay. RNA-seq and CHIP will be taken to identify the target genes controlled by ZFP207, and mutants will be used to verify the function of target genes in the RBSDV infection process. The results are of great importance for the research of explain the mechanisms of P5-2 in RBSDV infection process, and would give fresh ideas for controlling rice black-streaked dwarf disease at the molecular level.

水稻黑条矮缩病（RBSDV）是近年来我国广大稻区重要的病毒病害。前期研究证实RBSDV 致病因子P5-2蛋白与寄主转录因子ZFP207互作，初步证实ZFP207在水稻的抗病毒过程中起着重要的作用。本项目拟通过构建P5-2过量表达和基因沉默水稻，研究P5-2在病毒侵染致病过程中的作用；构建ZFP207过量表达和基因沉默水稻，研究ZFP207在水稻抗病毒侵染过程中的作用；酵母双杂交筛选验证ZFP207水稻体内互作因子；利用RNA-seq、CHIP等手段获得ZFP207调控的目的基因，并通过突变体验证目的基因在RBSDV侵染中的作用；最终阐明RBSDV致病因子P5-2与ZFP207互作在病毒侵染致病过程中的作用机理。研究结果对阐明致病因子P5-2在RBSDV侵染水稻中的作用机理意义重大，为从分子水平上来制定水稻黑条矮缩病防控策略提供新的思路。

水稻黑条矮缩病（RBSDV）是近年来我国广大稻区重要的病毒病害。前期研究证实RBSDV 致病因子P5-2蛋白与寄主转录因子ZFP207在细胞核互作，但ZFP207在RBSDV侵染过程中的作用还尚不清楚。本项目用ZFP207的过表达植株（ZFP207 OE）和crispr-cas9突变体植株（zfp207-KO）研究了ZFP207在RBSDV侵染过程中发挥的作用。验证转基因植株后，接种RBSDV，发现过表达ZFP207植株的发病率显著低于野生型，RT-qPCR和Western blot分析表明ZFP207 OE植株体内的RBSDV S6 RNA水平和P6蛋白积累都显著低于野生型植株；而zfp207-KO突变体植株接种RBSDV之后，病害症状比野生型严重，RT-qPCR和Western blot分析表明zfp207-KO植株体内的RBSDV S6 RNA水平和P6蛋白积累水平显著高于野生型植株。表明水稻锌指蛋白ZFP207正调控水稻对RBSDV的抗性。此外，水稻ZFP207抑制赤霉素合成基因OsGA20ox2的表达，RBSDV侵染后水稻中的OsGA20ox2被诱导表达，另外在zfp207-KO突变体植株中OsGA20ox2表达显著高于野生型，这暗示ZFP207可能通过负调控OsGA20ox2，从而降低植株对RBSDV的抗性。研究结果为深入解析水稻锌指蛋白ZFP207在在水稻防御RBSDV过程中的作用机制提供了坚实的基础，为从分子水平上来制定水稻黑条矮缩病防控策略提供新的思路。