利用农杆菌介导法将抗草甘膦基因转入玉米自交系H99的研究

Genetic transformation system of maize inbred line H99 with agrobacterium-mediated glyphosate resistance gene

【目的】利用转基因手段培育抗除草剂玉米品种以解决草害问题。【方法】采用农杆菌介导法对玉米自交系H99的幼胚进行遗传转化研究,进行单因素试验,分别设置不同胚的大小、农杆菌的菌液浓度和共培养时间处理,比较分析转基因幼胚GUS的瞬时表达率。【结果】幼胚大小1.5~2.0 mm、农杆菌的侵染浓度OD_(600)为0.4、侵染5 min、共培养3 d,GUS的瞬时表达率最高。PCR检测证明成功获得7株阳性转基因苗,且抗性鉴定实验证明转基因苗具有草铵膦抗性。【结论】为玉米转基因技术体系优化和培育抗除草剂新品种提供了理论参考。

【Objective】Breeding herbicide-resistant maize cultivars by transgenic method to solve the problem of grass damage.【Methods】Genetic transformation was conducted on immature embryos of maize inbred line of H99 by agrobacteriummediated method,which was carry out single factor test,the different size of immature embryo,concentration of agrobacterium and co-culture time were set respectively,which was compared and analyzed based on GUS transient expression rate in transgenic embryos.【Results】The results showed that the GUS transient expression rate was the highest when the size of the immature embryo was 1.5-2.0 mm,the concentration of agrobacterium OD600 was 0.4,the co-culture time was 3 d and the infection time was 5 min.PCR detection proved that 7 positive transgenic seedlings were successfully obtained,and phenotypic identification also demonstrated that transgenic seedlings were capable of resistance to glufosinate.【Conclusion】This study provides a reference for the optimization of improving maize transgenic technology system and the breeding of new herbicide-resistant varieties.