RNA干扰高油GmPEPc基因转入大豆研究

发布时间：2018-08-21 14:54

【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。
[Abstract]:By constructing the plant RNAi double expression vector of GmPEPc gene, and by Agrobacterium tumefaciens mediated genetic transformation of soybean cotyledon node, the related gene GmPEPc, which controls the pathway of oil and protein synthesis, was transferred into the recipient variety Shennong 9. By inhibiting the expression of endogenous GmPEPc gene in soybean and increasing the accumulation of oil, a new transgenic soybean variety with high oil content was obtained. In the process of soybean tissue culture, 407 explants of soybean were cut and 35 seedlings of T 0 generation were obtained. The transformation rate was 8.9%. The integration and expression of target genes in transgenic progenies were identified by molecular identification. Twenty-three transgenic progenies of T _ 1 generation were obtained, of which 14 were highly resistant to herbicide (300 mg mL ~ (-1). The results of PCR and Southern hybridization showed that the GmPEPc gene had been successfully inserted into the genomic DNA of transgenic soybean plants. The average oil content of transgenic soybean seeds was 9.51% higher than that of the control, and the average protein content was 5.44% lower than that of the control. These results provide a basis for screening transgenic soybean lines with high oil content and provide a germplasm basis for the selection of new high-oil soybean varieties in the next step.
【作者单位】：吉林省农业科学院吉林省农业生物技术重点实验室;东北农业大学资源与环境学院;东北师范大学生命科学学院;
【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)
【分类号】：S565.1

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