大豆再生植株抗草铵膦的筛选方法研究

Comparison on Methods Screening of Transgenic Soybean Resistant to Glufosinate

为解决农杆菌介导的大豆子叶节转化试验中草铵膦筛选所得转基因后代假阳性高的问题,采用浸种法、根吸法、叶片涂抹法、叶片喷洒法4种筛选方法和多种筛选浓度对大豆再生植株及其后代抗性进行筛选。结果采用1∶200草铵膦浓度的叶片喷洒法效果最佳。叶片喷洒法筛选6802株转化植株,获得了9株抗性植株,其后代的分子检测表明均为阳性植株。因此,采用叶片喷洒法对大豆抗草铵膦植株进行筛选的方法简便可行。

Four screening methods and several screening concentrations were used to screen soybean regeneration plant and the resistance of its progeny as well as to resolve the problem of high false-positive regeneration plants in agrobacterium-mediated transformation of soybean cotyledons after glufosinate screening.After screening the 6802 transgenic plants by spraying the leaf,9 resistant plants were obtained,and the molecular detection of their progeny showed that they were all positive transgenic plants.Therefore,the results indicated that the method that screening the transgenic plant by the leaf spraying was feasible.