基因枪介导小麦成熟胚遗传转化的影响因素
Study on Influencing Factors of Mature Embryos of Wheat by Particle Bombardment
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摘要: 小麦成熟胚作为转化受体可克服小麦幼胚存在的受季节和幼胚发育阶段限制的缺点。以湖北省小麦品种‘鄂麦12’和模式品种‘Bobwhite’为材料,成熟胚为转化受体,优化基因枪转化法的轰击压力、轰击距离、选择剂等因素,建立以小麦成熟胚为转化受体的高效转化系统。结果表明:小麦成熟胚作为转化受体时,适宜轰击压力和轰击距离组合是900 psi、6 cm;成熟胚对选择剂G418的敏感性强于幼胚,轰击后需要延长恢复时间,选择剂G418的适合浓度为20~40 mg/L。在以上优化条件下小麦成熟胚转化频率达0.3%~0.9%,已初步建立基因枪介导的小麦成熟胚遗传转化系统。Abstract: Mature embryos of wheat as explants have many advantages for transformation.They can overcome some of the immature embryos’ disadvantages,which is difficult to obtain throughout the year and strictly limited by their suitable stage for culture.Mature embryos of wheat from cultivar EM12 in Hubei province and mode variety Bobwhite were used as target materials.The effects of bombardment pressure,bombardment distance,geneticin etc.on mature embryos of wheat by particle bombardment have been optimized in order to obtain a high frequency of plant transformation system.The results show that optimal values of bombardment parameters were found to be a bombardment pressure of 900 psi at a distance of 6 cm for mature embryos.Mature embryos were more sensitive to geneticin G418 than immature embryos,and they had to be grown for longer time before application of the selectant.The optimal selective concentration of G418 for mature embryos is 20-40 mg/L.Under these optimum conditions,the frequency of transformation for mature embryos of wheat was up to 0.3%-0.9%.The transformation system of mature wheat embryos using particle bombardment has been set up preliminarily.