转TaDREB3a大豆品系KD1遗传稳定性分析及抗旱性鉴定  

Analysis on genetic stability and drought resistance identification of transgenic TaDREB3a soybean line KD1

作　　者：孟凡立[1] 李明月[1] 洪志鹏 赵洋 王子叶 冯广慧 王旋 孔丹珣 王莞迪 张玲[2] MENG Fanli;LI Mingyue;HONG Zhipeng;ZHAO Yang;WANG Ziye;FENG Guanghui;WANG Xuan;KONG Danxun;WANG Wandi;ZHANG Ling(School of Agriculture,Northeast Agricultural University,Harbin 150030,China;Jilin Academy of Agricultural Sciences,Jilin Provincial Key Laboratory of Agricultural Biotechnology,Changchun 130033,China)

机构地区：[1]东北农业大学农学院,哈尔滨150030 [2]吉林省农业科学院/吉林省农业生物技术重点实验室,长春130033

出　　处：《东北农业大学学报》2021年第8期1-11,共11页Journal of Northeast Agricultural University

基　　金：国家转基因生物新品种培育科技重大专项(2016zx08004-002)。

摘　　要：利用基因工程技术将抗旱基因转入大豆,培育高抗旱大豆品种,可提高大豆总产量。抗旱转基因大豆KD1是通过农杆菌介导法将来源于小麦TaDREB3a基因导入大豆受体品种垦农18中获得的转基因大豆材料。利用PCR方法,对转基因大豆KD1连续3代(T_(2)~T_(4)代)转基因大豆进行特异性PCR检测验证。结果表明,TaDREB3a和bar基因在KD1后代中稳定遗传。进一步对外源基因作表达分析,连续3代(T_(2)~T_(4))qRT-PCR检测结果表明,外源基因TaDREB3a和标记基因bar在转基因大豆叶、茎、根和籽粒中均表达且稳定遗传,且TaDREB3a基因在叶片和根中表达水平较高。Western blot结果表明,TaDREB3a和bar基因翻译的外源蛋白在转基因材料后代中稳定表达。对连续2代转基因材料抗旱性分析表明,KD1在芽期具有较强抗旱性,且其抗旱性状显著高于对照且稳定遗传。对转基因大豆KD1作靶标及非靶标除草剂抗性及耐性分析得知,外源基因bar在转基因大豆中稳定表达且在受体垦农18中表达TaDREB3a基因未改变非靶标除草剂抗性。The use of genetic engineering technology to transfer drought-resistant genes into soybeans to breed high drought-resistant soybean varieties can increase the total soybean yield.The new drought-resistant transgenic soybean material KD1 is a transgenic soybean material obtained by introducing the TaDREB3a gene from wheat into the soybean recipient variety Kennong 18 through the Agrobacterium-mediated method.The PCR method was used to verify the specific PCR of the transgenic soybean KD1 for three consecutive generations(T_(2)to T_(4)generations).The results showed that the TaDREB3a and bar genes could be stably inherited in the KD1 offspring.Further analyzed the expression of exogenous genes,the results of qRT-PCR detection for three consecutive generations(T_(2)-T_(4))showed that the exogenous gene TaDREB3a and the marker gene bar were expressed in transgenic soybean leaves,stems,roots and grains and could be inherited stably,and the TaDREB3a gene expression levels were higher in leaves and roots.Western blot detection results showed that the exogenous protein translated by the TaDREB3a and bar genes could be stably expressed in the offspring of transgenic materials.The drought resistance analysis of the two consecutive generations of transgenic materials showed that the KD1 had strong drought resistance at the bud stage,and its drought resistance traits were significantly higher than those of the control and could be inherited stably.Analysis of target and non-target herbicide resistance and tolerance of genetically modified soybean KD1 showed that the exogenous gene bar could not be stably expressed in transgenic soybeans and the expression of the TaDREB3a gene in the recipient Kennong 18 did not change the non-target herbicide resistance.

关 键 词：转基因大豆 TaDREB3a 抗旱性 

分 类 号：S565.1[农业科学—作物学]