基因枪介导的玉米NPR1基因的遗传转化

[目的]探索玉米基因枪转化的条件。[方法]以玉米自交系7239的幼胚胚性愈伤组织为受体材料,研究了轰击距离、气体压力、真空度和轰击次数对转化率的影响。[结果]转化时以金粉用量100μg/枪、发射点与靶细胞距离9 cm、气体压力1 350 psi、真空度25In.Hg、轰击次数2次的组合最佳。经潮霉素(Hygromycin)筛选,获得了再生植株,PCR分析及Southern杂交结果表明NPR1基因已整合到玉米基因组中,平均转化率为1.76%。[结论]该研究为玉米优良抗病品种的培育奠定了基础。

Genetic Transformation of NPR1 Gene into Maize by Microprojectile Bombardment

[Objective] The aim was to explore the conditions of transformation in maize by microprojectile bombardment.[Method] Immature embryonic callus of maize inbred line 7239 were used as explants to study the effects of shoot distance,helium pressure,vacuum and bombardment frequency on the transformation efficiency in the particle bombardment system of maize.[Result] Considering the transformation efficiency,particle bombardment with 100 μg/P of golden particles,at a shoot distance of 9 cm from the target cells,under helium pressure of 1 350 psi and vacuum 25 In·Hg,and bombarding twice could achieve relatively ideal results.After selection on media supplemented with different concentration of hygromycin,some regenerated plants were obtained.The results of PCR and Southern blotting analysis demonstrated that the NPR1 gene had been integrated into the genome of transgenic maize plants,with an average transformation efficiency of 1.76%.[Conclusion] The study had laid the foundation for the cultivation and breeding of excellent resistant varieties of maize.