[1]张原宇,董英山,于盛竹,等.RNA干扰高油GmPEPc基因转入大豆研究[J].大豆科学,2017,36(04):514-518.[doi:10.11861/j.issn.1000-9841.2017.04.0514]
　ZHANG Yuan-yu,DONG Ying-shan,YU Sheng-zhu,et al.Introduction of RNAi-PEPcGene into Soybean[J].Soybean Science,2017,36(04):514-518.[doi:10.11861/j.issn.1000-9841.2017.04.0514]
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RNA干扰高油GmPEPc基因转入大豆研究

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第36卷 期数: 2017年04期 页码: 514-518 栏目: 出版日期: 2017-07-20
Title:
Introduction of RNAi-PEPcGene into Soybean
作者:
张原宇1; 董英山1; 于盛竹2; 邢国杰1; 李桐3; 郑惠景1; 张玲1
（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）
Author(s):
ZHANG Yuan-yu1;  DONG Ying-shan1;  YU Sheng-zhu2;  XING Guo-jie1;  LI Tong3;  ZHENG Hui-jing1;  ZHANG Ling1
(1.Jilin Academy of Agricultural Sciences, Jilin Provincial Key laboratory of Agricultural Biotechnology, Changchun 130033, China; 2. College of Resources and Environment, Northeast Agricultural University, Harbin 150030, China; 3.Life Science College of Northeast Normal University, Changchun 130024, China)
关键词:
大豆; 转基因; pepc; RNA干扰; 子叶节
Keywords:
Soybean;  Transgenic;  pepc;  RNAi;  Cotyledonary node
DOI:
10.11861/j.issn.1000-9841.2017.04.0514
文献标志码:
A
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T1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T1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:
Abstract:Soybean is one of the most important oil seed crop planted as a major source of oil and protein for human. The cultivation 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.

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备注/Memo

基金项目：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）。

第一作者简介：张原宇（1983-），男，硕士，助理研究员，主要从事转基因生物技术研究。E-mail：zyy_1983@foxmail.com。
通讯作者：张玲(1981-)，女，博士，副研究员，主要从事植物分子生物学研究。E-mail：zly_jaas@126.com。

更新日期/Last Update: 2017-08-14