miR164c和miR168a对水稻种子活力的调控作用研究

前期的miRNA芯片分析和Real time PCR验证结果表明，人工老化处理的水稻种子中miR164c和miR168a的表达量随种子活力的下降分别呈现规律性的下降或上升变化，因此推测这两个miRNA可能参与水稻种子活力的调控。现申请本项目：（1）检测这两个miRNA的表达在水稻种子活力形成（种子发育过程）和自然老化过程中的变化规律，全面验证其与种子活力的关系；（2）构建这两个miRNA的过表达和沉默体系载体，转导水稻，获得转基因水稻种子，与野生型（非转基因受体水稻）种子相比较，探讨人工调控这两个miRNA的表达量变化对水稻种子活力的调控作用；（3）研究miRNA-靶基因-蛋白质表达-水稻种子活力之间的分子作用机制。通过本项目研究，可能从一个新的角度认识种子活力的分子调控机制，并为今后通过遗传操纵手段调控种子中miRNA的表达水平，改良水稻种子和其他植物种质资源的贮藏特性建立技术平台。

The results of miRNA microarray analysis and Real time quantitative PCR (RTqPCR) validation of the former project (NSFC, No.31071487) indicated that with artificial aging treatment, the seed vigor of rice (Oryza sativa L.) gradually declined, in the meantime the expression of miR164c and miR168a of rice seeds showed regularly falling or rising respectively, suggesting that these two miRNAs may be involved in the regulation of rice seed vigor. Hereby applying for the present project: (1) to detect the expression of these two miRNAs with vigor forming(with seed development) and in the natural aging process of rice seed, verifying the relationship between the expression of miR164c and miR168a with the seed vigor of rice; (2) to construct the over-expression and silencing system vectors of these two miRNAs, then transform them into rice and obtain transgenic seeds, by comparison with wild-type (non-genetically modified receptor rice) seeds, to explore the effect of artifially manipulating the expression of these two miRNAs on rice seed vigor; (3) to study the molecular mechanism included in the function mode of miRNAs on their target genes, and then how to affect rice seed vigor. Through this project, from a new perspective, we may reveal the molecular mechanism regulating seed vigor by miRNAs, and set up the technique platform to improve the storage quility of rice seeds and other plant germplasm resources through the genetic manipulation of miRNA expression levels of seeds in the future.