简便实用的转抗除草剂基因后代植株鉴定方法——种子萌发测定法  

Seed Germination Test-A Simple but Effective Way of Analysing Transgenic Progenies Transformed with the bar Gene

作　　者：徐传祥[1] 蒯本科[2,1] 王文东 马忠华[2,1] 

机构地区：[1]复旦大学生物化学系,上海200433 [2]复旦大学生命科学学院

出　　处：《复旦学报（自然科学版）》2000年第3期318-322,共5页Journal of Fudan University：Natural Science

基　　金：上海市教育发展基金会上海市教育委员会曙光计划资助课题

摘　　要：种子萌发法是检测外源抗性基因在转基因植物中的表达和分析外源抗性基因在转基因植物中遗传行为的简单而直接的方法 .以转bar基因燕麦后代R2 种子为材料 ,用不同体积分数的除草剂Challenge处理种子 ,萌发的幼苗经PCR和Southerndotblot鉴定 ,发现在 0 0 3%的除草剂上萌发的幼苗 ,部分不含bar基因 ,而在0 0 7%和 0 0 9%的除草剂上萌发的幼苗 ,均含bar基因 .用 0 0 7%的除草剂测定 3个群体的转bar基因燕麦R2 代种子的萌发情况 ,结果表明 ,bar基因在R3 代的遗传符合孟德尔的单基因遗传分离规律 ,这与已报道的用PCR和Southerndotblot分析R2 代的结果相一致 .进一步的研究发现 ,用 0 0 5 %的除草剂分析上述 3个群体 ,结果与 0 0 7%的差异不显著 ;而用 0 0 9%的除草剂分析的结果与 0 0 7%的差异显著 ,后代遗传分离比偏离 3∶1.这可能是由于后代群体中bar基因的表达水平存在着个体间差异 .研究结果表明 ,0 0 5 %和 0 0 7%是种子萌发法分析燕麦中外源bar基因遗传分离规律适宜的除草剂的体积分数 .It was showed that Seed Germination Test (SGT) was an effective and efficient way of analysing the expression and inheritance of foreign gene in the transgenic oat R 3 progeny. The diluted solution of 0.05% to 0.07% commerical herbicide Challenge (150 g/L Glufosinate ammonium, PPT) was found to be optimum in screening the progenies of transgenic oats containing bar gene, produced via microprojectile bombardment. 0.03% of Challenge was too low whereas 0.09% was too high in these experiments. These results were confirmed by the PCR and the Southern dot blot as well as by the segregation ratio analysis. Screening transgenic progenies by the SGT is not only simple and cost effective, more importantly, it can also obtain transformants with stable highly expressed foreign genes simultaneously.

关 键 词：BAR基因 种子萌发法 转抗除草剂基因 转基因植物 

分 类 号：S332[农业科学—作物遗传育种] Q789[农业科学—农艺学]