摘要
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 maize inbred lines such as He344, Nongda178 and Zheng58 were used in introduction of BcBCP1 gene and some optimization of genetic transformation system mediated by three different pollen tube pathway, in cluding dropping DNA on stigma, the system of pollen-mediated introduction and dropping DNA after opening the bractand. The results indicated that the suitable DNA concentration was 100g/mL, and the highest transformation rate were obtained 18h after pollination. When using the system of drop DNA after opening the bract, the highest transformation rate were obtained 18h after pollination, while DNA concentration was 100g,/mL. Three hundreds and thirty eight maize plants resisted to herbicide, among which 87 plants showed positive PCR reaction. The positive rate of PCR was 1.99% ,0.32% and 0.23% respectively in the three different method,and the highest one was obtained when dropping DNA on stigma.
出处
《作物杂志》
CAS
CSCD
北大核心
2012年第2期51-54,共4页
Crops
基金
国家转基因生物新品种培育重大专项(2009ZX08003-015B)
转基因生物新品种培育重大专项(2011ZX08003-003)
黑龙江省青年基金项目(QC2009C54)
黑龙江省博士后科研启动基金(LBH-Q08137)
关键词
玉米
花粉管通道法
转化
BcBCP1
基因
Maize （ Zea mays L. ）
Pollen tube pathway
Transformation
BcBCP1 gene
