农杆菌介导法将Bt(cryⅠA)基因导入大豆的研究  

Transformation of-Bt(cryⅠA)-Gene into Soybean via Agrobacterium-mediated Method

作　　者：曲姗姗[1,2] 刘丽君[1] 寇坤[1,2] 唐晓飞[1] 杨喆[1] 

机构地区：[1]黑龙江省农业科学院大豆研究所,黑龙江哈尔滨150086 [2]东北农业大学研究生院,黑龙江哈尔滨150030

出　　处：《大豆科学》2009年第2期186-190,194,共6页Soybean Science

基　　金：国家高技术研究发展计划(863计划)资助项目(2006AA1021F9);国家转基因重大专项资助项目(2008ZX08004-002)

摘　　要：构建含有Bt(cryⅠA)基因的植物表达载体pCAMBIA3300-Bt,以大豆子叶节为受体,通过农杆菌介导法将Bt基因导入大豆品种黑农37中,获得转基因植株。并进行大豆的再生和遗传转化系统优化的研究,以获得较高的转化率。结果表明:在6-BA浓度为1.7mg·L-1时,丛生芽分化率最高。确定该品种大豆在丛生芽分化阶段的草铵膦筛选浓度为3.5mg·L-1。获得转化质粒pCAMBIA3300-Bt的转基因植株,其中T1代PCR阳性植株19株。采用real-timePCR的方法对T1代抗性植株进行Bt基因的转录水平的分析,初步证明Bt基因已整合到受体大豆的基因组内。It is very important for diseases and insect pests controlling to transfer some resistant genes to obtain transgenic resistant varieties. In this Study,plant expression vector pCAMBIA3300-Bt containing -Bt（cry I A）- gene was constructed, and transgenie plants were obtained by Agrobacterium- mediated transformation of soybean （ Heinong 37 ） cotyledonary nodes. We regenerated plants and optimized the conditions of transformation to increase the efficiency of transformation. It had the optimal rate of shooting when the concentration of 6- BA was 1.7 mg · L^-1. The optimal selection concentration of Glufosinate-ammonium for shoot initiation was 3.5 mg · L^-1. Transformed plants containing pCAMBIA3300-Bt was obtained and 19 positive plants from Ti generation were tested by PCR. Real-time PCR was used to investigate the transcription level of Bt gene in T1 transgenie plants,it confirmed the integration of Bt genes into the genome of soybean.

关 键 词：大豆 BT基因 遗传转化 

分 类 号：S565.1[农业科学—作物学]