核糖体蛋白基因OsRPP0调控水稻育性发育研究

项目名称： 核糖体蛋白基因OsRPP0调控水稻育性发育研究

项目编号： No.31500254

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 生物科学

项目作者： 陈竹锋

作者单位： 深圳市作物分子设计育种研究院

项目金额： 20万元

中文摘要： 水稻雄性不育不仅在杂交水稻生产上具有重要的应用价值，而且是研究植物花器官发育和形态建成的模式材料。前期我们筛选获得一个水稻无粉型不育突变体h6231，并利用改进的MutMap方法克隆到控制该雄性不育性状的基因OsRPP0，该基因编码核糖体P0蛋白，目前水稻中该类基因生物学功能研究尚未见报道。本项目拟采用CRISPR技术在水稻中定点敲除失活OsRPP0基因，进一步验证该基因突变体对雄器官发育的影响并观察其他表型变化；同时通过等位基因的遗传转化进行功能互补试验验证候选基因的功能，并利用石蜡包埋切片和染色技术以及透射电镜扫描技术进行细胞学观察，明确突变体发生花粉败育的时期及细胞学变化，从而确定OsRPP0基因的组织细胞学功能；采用原位杂交技术及实时荧光定量等方法研究目的基因的表达模式、基因表达调控情况，深入揭示OsRPP0基因调控水稻育性发育的分子机理，进一步丰富和完善水稻雄性不育基因调控网络。

中文关键词： 水稻；雄性不育；核糖体蛋白；基因表达调控

英文摘要： Rice male sterile lines are critical not only for hybrid breeding, but also for the understanding of floral organ development and morphology in rice. Recently, we screened a rice male sterile mutant named H-6231S. Using a modified MutMap method, we identified a mutation in an unreported gene OsRPP0, which encodes a 60S acidic ribosomal protein P0, responsible for the male sterility phenotype. However, the biological function of this gene in rice development remains largely unexplored. In this project, we intend to further verify the function of OsRPP0 mutation in mediating the male-sterile phenotype by targeted knockout of OsRPP0 using the CRISPR/Cas9 system, and by genetic complementation of the mutant phenotype using wild-type allele. We will examine the transverse sections of the mutant anthers at different developmental stages using light microscopy and transmission electron microscopy to obtain a detailed understanding of the developmental and cellular defects in H-6231S mutant. RNA in situ hybridization and real-time quantitative PCR will be performed to analyze the expression pattern and regulation pathway of OsRPP0 gene during rice anther development. Further study on the role of OsRPP0 will contribute to a better understanding of the mechanisms underlying rice male reproductive development.

英文关键词： Rice;Male sterility;Ribosomal Protein;Gene expression and regulation