OsDCL3b基因调控稻穗生长发育的遗传机理研究

项目名称： OsDCL3b基因调控稻穗生长发育的遗传机理研究

项目编号： No.31260313

项目类型： 地区科学基金项目

立项/批准年度： 2013

项目学科： 农业科学

项目作者： 李绍波

作者单位： 南昌大学

项目金额： 42万元

中文摘要： 在前期研究中，我们获得了OsDCL3b下调表达影响稻穗生长发育的转基因突变株、突变株与对照幼穗中差异表达的ncRNA（no-coding RNA）和OsDCL3b过量表达转基因T0代植株。在此基础上，本项目利用测序、双向电泳和质谱技术分析OsDCL3b基因沉默和过量表达转基因突变株与对照幼穗中的mRNA表达谱及蛋白质组差异，结合网上资源和已有的ncRNA结果，利用生物信息学分析获得影响稻穗生长发育的候选ncRNA、差异表达的mRNA和蛋白质，经Northern blot、荧光定量PCR和Western blot验证其表达后，进行转基因分析获得1-2对重要的ncRNA和差异表达蛋白基因的功能，初步确定OsDCL3b基因、ncRNA、蛋白基因、稻穗生长发育这4者间的相互关系，进而揭示OsDCL3b基因调控稻穗生长发育的可能遗传途径，为深入分析OsDCL3b基因的功能奠定重要基础。

中文关键词： OsDCL3b基因功能；水稻结实率；小分子非编码RNA；mRNA；可变剪接

英文摘要： Transgenic mutant rice plants with down-regulated expression of OsDCL3b and abnormal development and growth of panicles, ncRNAs(non-coding RNAs) expressing differentially in young panicles of the transgenic mutant and control, and transgenic T0 plants with over-expressing of OsDCL3b have been obtained in our previous research. In this program, the differentially expressed mRNAs and proteins in the young panicles of transgenic mutant and control will be screened by sequencing, two-dimensional gel electrophoresis and mass spectrometry. Besides, some candidate ncRNAs and protein genes, which may affect the development and growth of rice panicles, will also be discerned by bioinformatics analyses of the results herein and those on the internet. Expressions of the candidate ncRNAs and protein genes/proteins will further be detected by Northern blot, real-time fluorescent quantitative PCR and Western blot. Functions of one to two pairs of ncRNAs and protein genes are to be confirmed by transgenic analysis respectively, which will facilitate the understanding of the relationships among OsDCL3b, ncRNA, protein genes and the development and growth of rice panicles, revealing the possible genetic pathways that OsDCL3b controls the development and growth of rice panicles. The results obtained herein will provide crucial ba

英文关键词： Functions of OsDCL3b；Seed-setting rate of rice；Small non-coding RNA；mRNA；Alternative splicing