RNA干扰高油GmPEPc基因转入大豆研究  

Introduction of RNAi-PEPc Gene into Soybean

作　　者：张原宇[1] 董英山[1] 于盛竹 邢国杰[1] 李桐[3] 郑惠景 张玲[1] 

机构地区：[1]吉林省农业科学院吉林省农业生物技术重点实验室,吉林长春130033 [2]东北农业大学资源与环境学院,黑龙江哈尔滨150030 [3]东北师范大学生命科学学院,吉林长春130024

出　　处：《大豆科学》2017年第4期514-518,共5页Soybean Science

基　　金：国家自然科学基金青年基金(31501327);吉林省科技发展计划-青年科研基金项目(20150520123JH)

摘　　要：通过构建GmPEPc基因的植物RNAi双元表达载体,并通过农杆菌介导的大豆子叶节遗传转化方法将控制油脂和蛋白合成途径的相关基因GmPEPc转入受体品种沈农9号中,通过抑制大豆内源GmPEPc基因的表达,增加油脂积累,从而获得高油的转基因大豆新品种。在大豆组织培养过程中,共切取大豆外植体407块,获得T0代转化苗35株,转化率8.9%。通过对转基因后代中目的基因的整合及表达情况进行分子鉴定。获得23株T_1代转基因后代,其中高抗草丁膦除草剂(喷施浓度300 mg·mL^(-1))14株,通过PCR检测结果表明其中12株为PCR阳性;PCR和Southern杂交检测表明,GmPEPc基因已经成功插入到转基因大豆植株基因组DNA中。对T_1代测定结果显示,转基因大豆籽粒的平均含油量比对照高9.51%,平均蛋白质含量下降5.44%。这些研究结果为筛选高油脂含量的转基因大豆新株系提供了依据,为下一步高油新品种的选育提供了种质基础。Soybean is one of the most important oil seed crop planted as a major source of oil and protein for human. The cul- tivation of high-oil soybean varieties has been one of the important issues for soybean breeding. In this study, we expressed the pepc-RNAi into the receptor varieties Shennong 9 by soybean cotyledonary nodes method. Though inhibiting the soybean endogenous GmPEPc gene expression to gain high oil of genetically modified soybean varieties. We co-cultivated about 407 explants, and got 35 transgenic plants, conversion rate 8. 9%. Stock of glufosinate（300 mg. L- 1 ） was painted on half the upper surface of T1 soybean leaves, and we had got 23 transgenic plants. PCR analysis showed that 12 of 14 transgenic plants were positive. Southern blot analysis of PCR-positive plants indicated the integration of the pepc-RNAi gene into the soybean genome. The results of T1 generation showed that the average oil content of transgenic soybean grain was 9. 51% higher than that of the control, and the average protein content decreased by 5.44%. The results of these studies provide a basis for the selection of new varieties of high oil content, which provides the basis for the selection of new varieties of high oil.

关 键 词：大豆 转基因 PEPC RNA干扰 子叶节 

分 类 号：S565.1[农业科学—作物学]