水稻反温敏雄性不育基因RTMS3的功能及其作用机理研究

反温敏雄性不育是培育两系杂交水稻的重要遗传资源，其育性转换模式与温敏雄性不育相反，表现为高温可育、低温不育。我们获得了2种表型相似的水稻反温敏雄性不育突变体。遗传分析与定位结果表明，这两种突变体是核基因RTMS3的不同等位突变。利用混合分离测序分析和遗传互补实验验证，确定RTMS3蛋白可能是一种与分支氨基酸代谢有关的酶。初步推测，RTMS3蛋白可能与温度敏感因子互作，调节水稻小穗发育过程中的氨基酸含量及其代谢产物，从而影响花药和花粉的发育。在此基础上，本项目拟综合利用遗传学、蛋白组学、基因组学等相关的技术、方法，检测与水稻育性相关的基因，筛选与RTMS3互作的蛋白质，分析不同温度条件引起rtms3突变体育性改变的原因，阐明RTMS3基因调节水稻育性的功能与作用机制，为揭示水稻雄性不育的分子调控机制提供新的见解，也为利用该基因拓宽两系杂交水稻的应用途径奠定基础。

Reverse thermo-sensitive male sterility is an important genetic resource for breeding two-line hybrid rice. The mode of its fertility conversion is opposite to that of thermo-sensitive male sterility, behaving as fertile under high temperature and sterile under low temperature. We obtained two reverse thermo-sensitive male sterile mutants with similar phenotypes. Genetic analysis and gene mapping showed that the two mutants were two different allelic mutations of the nuclear gene RTMS3. The results of bulked segregant sequencing analysis and genetic complementary experiments confirmed that RTMS3 protein is probably a key enzyme involved in the metabolism of branched amino acids. It is preliminarily speculated that RTMS3 protein probably interacts with thermo-sensitive factors to regulate the amino acid content and its metabolites during spikelet development, so as to regulate anther and pollen development. Based on these results, the present project aims to detect the genes related to rice fertility, screen the proteins interacting with RTMS3, and analyze the causes of fertility changes in rtms3 mutant due to different temperature conditions, by comprehensively using the related techniques and methods of genetics, proteomics and genomics, so as to elucidate the function and mechanism of RTMS3 gene in regulating the fertility of rice. The results of this study will provide new insights into the molecular regulation mechanism of male sterility in rice, and lay the foundation for widening the application of two-line hybrid rice in the future.