摘要
利用子叶下胚轴为外植体，通过农杆菌Ti质粒介导途径，建立了白三叶草高效遗传转化及转基因植株高频率再生体系。PCR检测及Southern印迹鉴定结果表明，外源T—DNA片段已整合到转基因植株的基因组中，且多以1～2个少数拷贝存在。Northem印迹结果和对报告基因GusA编码蛋白的活性检测结果表明，目的基因在部分转基因植株中高水平表达。适宜条件下外植体遗传转化后的植株再生比例达58．23％～62．12％。与对照相比，转基因植株在外部形态上没有发生改变。
Cotyledon explants of T. repens were used to establish a high efficiency genetic transformation using Agrobacterium tumefaciens and transgenic plant regeneration techniques. Based on PCR and Southern blot analysis, the T-DNA fragment from the A. tumefaciens plasmid was integrated into the genome of the transgenic plants with one to two copies. Northern blot analysis and enzyme activity of the GusA protein indicated that the reporter gene in some transgenic lines was expressed at a high level. The regeneration rates of transgenic explants under suitable condition were 58.23 %--62. 12%. There were no obvious changes at plant phenotype in transgenic plants compared to the non-transformed control plants （CK）.
出处
《草业学报》
CSCD
2007年第2期84-89,共6页
Acta Prataculturae Sinica
基金
河北省自然科学基金(C2006000434)项目资助
关键词
三叶草
农杆菌
遗传转化
植株再生
Trifolium repens
Agrobacterium tumefaciens
genetic transformation
plant regeneration
