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以东农50大豆子叶节为材料,以大豆子叶节丛生芽黄化率和丛生芽高度为考察指标,确定氯化锂作为转基因大豆筛选剂的最佳浓度和筛选临界观察时间。在此基础上,对转基因大豆丛生芽进行GUS染色,并对转化植株进行PCR检测。通过对受体植株进行氯化锂叶片涂抹,确定适宜的表型筛选浓度。结果表明,延迟7 d进行氯化锂筛选,筛选浓度为60 mmol.L-1为宜,最佳筛选天数为10 d。抗氯化锂丛生芽的GUS染色阳性率为80%,确定叶片涂抹法的适宜筛选浓度为700 mmol.L-1,并且经氯化锂筛选得到的转基因植株后代可以正常生长发育。
Taking Dongnong50 soybean cotyledon as material, taking the cotyledons of soybean cotyledons as index, the optimum concentration of lithium chloride as screening agent for soybean was determined and the critical time of screening was determined. On this basis, GUS staining of transgenic soybean cluster shoots was carried out, and PCR analysis of the transformed plants. Appropriate phenotypic screening concentrations were determined by plating lithium chloride leaves on recipient plants. The results showed that the screening of lithium chloride was delayed for 7 days, the optimum concentration was 60 mmol.L-1, the optimal screening days was 10 days. The positive rate of GUS staining against cluster shoots of lithium chloride was 80%, and the suitable screening concentration of leaf smear method was 700 mmol.L-1. The progeny of transgenic plants screened by lithium chloride could grow and develop normally.