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.
出处
《大豆科学》
CAS
CSCD
北大核心
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)
关键词
大豆
BT基因
遗传转化
Soybean
Bt gene
Genetic transformation
