RNAi介导的OsBTF3基因沉默及其对水稻籽粒相关性状的影响  

RNA Interference-mediated Gene Silencing of OsBTF3 Results in Functional Defects in Grain-related Traits of Rice

作　　者：李广旭[1,2] 陈华民[1] 刘维振[1] 吴静[1] 吴茂森[1] 何晨阳[1] 

机构地区：[1]中国农业科学院植物保护研究所、植物病虫害生物学国家重点实验室,北京100193 [2]辽宁省农业科学院果树研究所,熊岳115009

出　　处：《生物技术通报》2012年第5期126-131,共6页Biotechnology Bulletin

基　　金：国家转基因生物新品种培育重大专项(2009ZX08009-044B;2011ZX08001-002)

摘　　要：为了创制OsBTF3基因沉默的水稻植株、验证该基因在水稻籽粒相关性状中的功能、评价其在水稻遗传改良中潜在的应用价值,设计和合成OsBTF3基因序列的引物、扩增部分基因片段,构建RNAi基因沉默载体、通过农杆菌介导转化愈伤组织、植株再生、潮霉素抗性筛选和PCR验证、定量分析OsBTF3基因表达量,测定转基因水稻籽粒相关性状。结果表明,成功地获得了20个T1代OsBTF3-RNAi转基因株系,OsBTF3基因表达量得到显著的抑制和干扰,抑制效果平均达到85%;与野生型对照株相比,5个所测定RNAi转基因株系的穗长、穗粒数、千粒重和穗粒重等籽粒相关性状明显地减小或降低。因此,RNAi介导的基因沉默导致了OsBTF3基因表达水平抑制以及在籽粒性状中的功能缺失;OsBTF3可能是一个调控水稻籽粒相关性状重要的功能基因。In order to generate the transgenic rice lines with RNA interference （ RNAi ） -silenced OsBTF3 gene, demonstrate the functions of gene in grain-related traits and evaluate its potential use in genetic improvement of rice, the fragment amplification of OsBTF3 gene by using the specific primers, RNAi construct generation, Agriobacterium-mediated transformation of callus tissues, Hyg screening, regeneration of transgenic rice plants and its PCR validation were conducted. The transcription of OsBTF3 gene was assayed by using quantitative real-time PCR, and the grain-related traits of transgenic rice with RNAi-silenced OsBTF3 gene were analyzed. The results showed 20 of T1 generation transgenic lines were successfully generated and validated respectively, in which the significant suppression of OsBTF3 gene transcription occurred. Significant decreases in panicle length, seeds per panicle, thousand seed weight and seed weight per panicle were found in the RNAi transgenic lines compared with the controlled untransgenic plants. Therefore, RNAi-mediated gene silencing of OsBTF3 results in suppression of its transcription and function defects of grain-related traits of rice. Therefore, OsBTF3 might he a key regulatory gene that functions in expression of grain-related traits of rice.

关 键 词：水稻 OsBTF3基因 RNA干扰 基因沉默 籽粒相关性状 

分 类 号：S511[农业科学—作物学]