水稻种子特异表达基因OsEnS97的功能与作用机理研究

稻米是人类最重要的粮食之一，它来自于水稻籽粒中的胚乳。水稻胚乳的发育主要包括细胞分裂与分化、储藏物质的合成与积累以及淀粉胚乳细胞的PCD等过程。生长素在种子发育过程中发挥着关键的作用，生长素受体基因（AUX/IAA）可能参与到生长素的信号转导调控。前期本课题组创建了一个种子特异表达AUX/IAA类基因OsEnS97的遗传材料，RNAi T2代家系的种子大小和千粒重都显著低于野生型，并产生一定比例的皱缩种子。本项目采用表达分析和原位杂交已证实OsEnS97为种子特异表达基因；将通过生物信息学分析、RNAi、CRISPR和超表达研究OsEnS97的生物学功能；从蛋白定位、IAA处理和流式细胞检测等分析其作用方式；最后对水稻种子酵母文库进行筛选，获得调控蛋白，并通过遗传学实验验证。研究OsEnS97在水稻种子中的功能，将有助于进一步解析生长素信号转导途径在胚乳发育中的作用和机制。

Rice is one of the most important foods for human. It is mainly from the endosperm of rice seeds. The development processes of endosperm were composed of different phases including mitosis, endoreduplication, accumulation of many nutrients, as well as the programmed cell death (PCD) of starch. Earlier studies have shown that auxin plays a key role in rice seed development and AUX/IAA protein involved in the signal transduction of auxin regulation, but the mechanism remained poorly understood. Our group identified a AUX/IAA protein OsEnS97, and the transgenic materials were obtained by RNAi, CRISPR and over-expression techniques. In T2 OsEnS97-RNAi lines, we found that the seed size and 1000-grain weight were significantly reduced compared with the WT, and a certain percentage of shrunken seeds were produced. Transcriptional profiling and in situ hybridization analysis confirmed that OsEnS97 was exclusively expressed in developing seed. Then study its biological function through bio-information analysis, RNAi, CRISPR and over-expression strategies. Further, the auxin treatment and flow cytometry were performed to analyze its function mechanism. Finally we propose to screen the interacting protein of OsEnS97 through the Yeast two-hybrid technology and Transcriptome Sequencing, and then proved by genetic experiments. Completion of this project will determine the molecular mechanism of the signal transduction of auxin in rice seed, and will provide resources for genetic improvement of yield and quality of rice.