水稻育性调控相关基因RNAi载体的构建及其表型鉴定

RNAi Vector Construction and Genotypic Identification of Genes Regulating the Fertilities in Rice(Oryza sativa L.)

利用RNAi干扰技术研究不同基因对花粉发育、卵细胞发育和合子胚发育的影响是一种重要的手段。本研究通过筛选水稻在生殖发育过程中的9个重要调控基因,构建基因的RNAi表达载体,分析转基因植株育性及相关性状表型,以期探明RNAi表达载体对靶标性状的干扰效应。其中,AT61~AT63、AT64~AT66、AT67~AT69表达载体分别靶标花粉育性、卵细胞发育以及合子胚发育的调控。结果表明,除RNAi表达载体AT64没有获得转基因植株外,其余8个RNAi表达载体均获得了转基因植株;对T0代转基因植株的花粉育性、结实率以及潮霉素筛选(40 mg/L)发芽率检测的统计结果显示,RNAi表达载体AT62(花粉发育调控)、AT65(卵细胞发育调控)和AT67(合子胚发育调控)的干扰效应较强。本研究结果将为创制新型水稻基因工程不育系提供策略和选择。

RNA interference(RNAi)technology provides an efficient technology for gene expression and gene function studies in many different genes including those involved in pollen development,egg cell development and embryo development.In this study,nine important regulatory genes involved in the reproductive development of rice(Oryza sativa L.)were screened and specific RNAi expression vectors corresponding to these genes(AT61~AT69)were constructed to generate transgenic plants.The effects of RNAi expression vectors on targeted traits related to reproductive development were examined.Results indicated that the vectors AT61,AT62 and AT63 targeted the pollen fertility,AT64,AT65 and AT66 targeted the egg cell development,and the vectors AT67,AT68 and AT69 targeted the embryo development in the transgenic rice seedlings.Moreover,except for the RNAi expression vector AT64,transgenic plants were obtained for the other 8 RNAi expression vectors.Statistical analysis of pollen fertility,seed setting rate and germination rate in hygromycin(40 mg/L)of T0-generation of transgenic plants showed that the vectors AT62(regulating pollen development),AT65(regulating egg development)and AT67(regulating embryo development)had strong interference effects.These results suggest that RNAi technology could serve as a powerful research tool for designing new strategies for the development of sterile lines for rice breeding.