利用农杆菌介导合子胚转化法将Bar基因转入玉米自交系的研究

Study on Agrobacterium-mediated Zygote Transformation of Bar Gene into Maize Inbred Line

基于农杆菌介导的合子胚转化方法是一种具有自主知识产权的转基因新方法。利用该方法转化玉米合子胚,可直接获得转化种子。以东北春玉米区玉米自交系8902、Pa91、丹340作为转化受体,通过合子转化法进行抗草丁膦除草剂基因Bar的转基因研究。在获得的3 560份材料中,通过对T_1代种子苗叶面喷施草丁膦除草剂Basta,共获得153株抗性植株,抗性频率为4.3%。PCR鉴定阳性植株为102株,转化频率为2.87%。对PCR检测出的阳性植株进行自交获得T_2代,对材料继续进行抗性筛选,对抗除草剂表型的分离比率进行抗性遗传分析。

Agrobacterium-mediated zygote transformation is a new transgenic method with independent intellectual property rights. Transgenic seeds could be obtained by this method. Bar gene with the resistance to Basra herbicide was transferred into 3 northeast elite maize inbred lines of 8902, Pa91, Dan340 via this method. The 153 resistant plants were obtained from 3 560 seeds by the spraying of Basta herbicide and the resistant rate was 4.3%. In 153 plants, the 102 positive plants were screened by PCR analysis and the transformed frequency was 2.87%. The positive plants were self-pollinated. T2 generation was obtained and used to further screen resistant plants and evaluate the genetic dissections of resistances to herbicide based on phenotyping segregation ratio.