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目的:构建水稻osvdac7基因RNA干涉表达载体,获得osvdac7表达下调的转基因水稻植株。方法:采用pMCG161双元载体,以传统方法构建水稻osvdac7基因RNA干涉表达载体;愈伤组织转化法获得osvdac7干涉转基因阳性植株,并应用实时荧光定量PCR(RT-PCR)技术检测干涉阳性植株中osvdac7的表达水平。结果:成功构建了2个osvdac7基因RNA干涉表达载体,获得了21株转osvdac7干涉载体的阳性植株,其中9株转化植株的osvdac7表达明显下调。结论:成功获得了干涉osvdac7基因表达下调的转化植株,为研究水稻osvdac基因家族的生物学功能打下基础。
OBJECTIVE: To construct RNA interference expression vector of osvdac7 in rice and obtain transgenic rice plants with down-regulated expression of osvdac7. Methods: Oscasac7 gene RNA interference expression vector was constructed by the traditional method using pMCG161 binary vector. Oscdac7 gene transfection-positive plants were obtained by transformation of callus. Oscdac7 was detected by real-time fluorescence quantitative PCR (RT-PCR) The level of expression. RESULTS: Two RNA interference expression vectors of osvdac7 gene were successfully constructed, and 21 positive transgenic plants which transduced with osvdac7 vector were obtained. The expression of osvdac7 in 9 transgenic plants was significantly down-regulated. CONCLUSION: Transformed plants that interfere with the down-regulation of osvdac7 gene expression were successfully obtained, which laid the foundation for studying the biological function of osvdac gene family in rice.