利用RNA干涉技术构建水稻DDB1(Damaged DNA binding protein 1)基因不同启动子驱动植物表达载体,DDB1-RNAi和DDB1-glu-RNAi.通过根癌农杆菌介导转入水稻愈伤,经组织培养成功获得转基因植株.半定量RT-PCR分析显示,与野生型植株相比,两组转基因植株幼叶内DDB1的表达量均有不同程度的降低.花粉染色结果显示,转基因植株花粉染色异常,败育率高;植株表现出明显的雄性半不育;秕谷率均有不同程度的提高;并且以上表型在DDB1-RNAi转基因植株中均比在DDB1-glu-RNAi转基因植株中明显.因此,推断DDB1基因与水稻的生殖发育有着密切的关系.
In this study,two RNA interference(RNAi) vectors of rice DDB1 gene,DDB1-RNAi and DDB1-glu-RNAi,droved by different promoters were constructed.They were introduced into rice cv.Nipponbare by Agrobacterium-mediated transformation.The analysis of semi-quantitative RT-PCR revealed the variation of suppression of DDB1 in both RNAi transgenic lines compared to that of wild type.I2-KI indicated abnormally stained pollens,high level male semi-sterile in two sets of transgenic plants,and DDB1-RNAi transformed lines showed more severe sterility compared with that of DDB1-glu-RNAi ones.Consequently,DDB1 may be involved in reproductive development in rice.