抗亚洲玉米螟、抗草甘膦转基因玉米的培育  

Breeding of Transgenic Maize with Resistance to the Asian Corn Borer(Ostrinia furnacalis) and Tolerance to Glyphosate

作　　者：孙越[1] 刘秀霞[1] 李丽莉[2] 官赟赟[1] 张举仁[1] 

机构地区：[1]山东大学生命科学学院,济南250100 [2]山东省农业科学院植物保护研究所,济南250100

出　　处：《农业生物技术学报》2015年第1期52-60,共9页Journal of Agricultural Biotechnology

基　　金：国家转基因生物新品种培育重大专项课题(No.2011ZX08003)

摘　　要：5-烯醇式丙酮酸莽草酸-3-磷酸合酶基因(5-enolpyruvyl-shikimate-3-phosphate synthase,epsps)是一个高抗草甘膦的基因,苏云金芽胞杆菌(Bacillus thuringiensis)的杀虫蛋白基因(Bt基因)cry1Ac M是一个能在单子叶植物中高效表达的抗亚洲玉米螟基因。为了获得具有抗虫、抗除草剂优良复合性状的转基因玉米,本研究构建出质粒p CAMBIA1302-P35S::epsps-Tnos-Pubi::cry1Ac M-Tnos,通过农杆菌(Agrobacterium tumefacien)介导的玉米(Zea mays)茎尖遗传转化法将目标基因转入玉米。通过分子检测、草甘膦抗性筛选和田间接种亚洲玉米螟(Ostrinia furnacalis(Guenée))试验筛选出转基因株系,在此基础上进行了转基因植株室内抗亚洲玉米螟试验和田间草甘膦抗性分析,并采用RT-PCR检测和Western blot方法确定了抗虫蛋白在转基因植株中稳定表达。从大量转基因株系中优选出6个遗传稳定且抗亚洲玉米螟、抗除草剂草甘膦的转基因玉米株系,这些株系的抗亚洲玉米螟、抗草甘膦特性完全满足大田应用的需求。综上所述,将改造后的Bt抗虫基因cry1Ac M和抗草甘膦基因epsps一同转入玉米,赋予两种玉米骨干自交系植株抗玉米螟、抗草甘膦特性,为我国抗亚洲玉米螟抗草甘膦转基因玉米的大面积推广培育出了优良自交系。5-enolpyruvyl-shikimate-3-phosphate synthase （epsps） is a new gene of high resistance to glyphosate and Bt crylAcM as an insect-resistent gene can expresse at high level in monocotyledon. In order to breed transgenic maize with stacking traits of insect-resistance and glyphosate-tolerance, the plant expression vector pCAMBIA1302-P35S：：epsps-Tnos-Pubi：：crylAcM-Tnos was constructed and then introduced into maize （Zea mays） inbred lines by Agrobacterium mediated shoot apical meristem transformation. After herbicide screening, survived plants were confirmed by PCR, and the expression of crylAcM gene in plants was detected by RT-PCR and Western blot, respectively. The resistance of transgenic plants to Asian corn borer（Ostrinia furnacalis （Guen6e））, was examined by indoor and field experiments. And the tolerance of transgenic plants to glyphosate was evaluated by the field trials. The analysis of transgenic maize resistances demonstrated that the plants were highly resistant to herbicide and the Asian com borer. We obtained 6 transgenic maize lines with genetic stability, highly Asiatic corn borer-resistance and glyphosate-tolerance, which achieved the criterion ofactual application. Among them lines Z1-Z3 came from elite inbred line Zheng58, and lines C1-C3 came from elite inbred line Chang7-2. In summary, the introduction of crylAcM and epsps genes into maize elite inbred lines endued the Asiatic corn borer-resistance and glyphosate-tolerance of transgenic plants, which created new corn transgenic lines with excellent complex traits. It is very useful for large-scale planting of the insect-resistance and glyphosate-tolerance transgenic maize in China.

关 键 词：5-烯醇丙酮酸莽草酸-3-磷酸合酶基因(epsps) Bt cry1AcM基因 转基因玉米 抗亚洲玉米螟 抗草甘膦 稳定遗传 5-enolpyruvyl-shikimate-3-phosphate synthase gene (epsps) 

分 类 号：S435.132[农业科学—农业昆虫与害虫防治]