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采用DNA柱头滴加法、花粉粒携带法和开苞叶导入法等3种不同的花粉管通道遗传转化方法,将抗逆基因BcBCP1转入3个玉米自交系合344、农大178和郑58中,对3种方法的部分影响因素进行了优化,旨在筛选出转化效率高的方法,为玉米转基因育种提供技术支持。结果表明:DNA柱头滴加法的DNA溶液浓度100g/mL,授粉后18h转化率最高。花粉粒携带法最佳处理时间为10min。开苞叶导入法在授粉后18h转化率最高,DNA浓度为100μg/mL。T0代共筛选出338株除草剂抗性植株,其中87株PCR呈阳性。DNA柱头滴加法的平均PCR阳性率最高,为1.99%;而花粉粒携带法的平均PCR阳性率最低,为0.23%;开苞叶导入法平均PCR阳性率居中,为0.32%。
Three different pollen tube pathway genetic transformation methods, such as DNA stigma dropping method, pollen grain carrying method and bract introduction method, were used to transform the resistance gene BcBCP1 into three maize inbred lines 344, Nongda 178 and Zheng 58 , Some influencing factors of the three methods were optimized in order to screen out the methods with high transformation efficiency and provide technical support for the transgenic breeding of maize. The results showed that the DNA solution concentration of 100g / mL and the highest conversion rate 18h after pollination. Pollen grain carrying the best treatment time is 10min. The method of introducing bracts into leaves was the highest at 18h after pollination, and the DNA concentration was 100μg / mL. T0 generations were screened a total of 338 herbicide-resistant plants, of which 87 were PCR-positive. The average PCR positive rate of DNA was 1.99%, while the average PCR positive rate of pollen carrier was the lowest (0.23%). The average PCR positive rate was 0.32%.