转Cry1A.401基因抗虫玉米CM8302和CM8303对亚洲玉米螟的抗性研究

Evaluation of Resistance of Transgenic Cry1A.401 Maize CM8302 and CM8303 to the Ostrinia furnacalis

采用农杆菌介导法获得转Cry1A.401基因抗虫玉米CM8302和CM8303,通过多代回交与自交获得BC_(4)F_(1)、BC_(4)F_(2)和BC_(4)F_(3)世代的转基因玉米材料。利用qRT-PCR和ELISA方法明确转基因抗虫玉米叶片、花丝和子粒均有Cry1A.401基因表达,不同世代间目的基因表达量无显著差异。田间接虫鉴定结果表明,CM8302和CM8303三个世代心叶期和吐丝期的抗性等级均为1级,抗性水平为高抗。室内生测结果表明,亚洲玉米螟幼虫取食CM8302心叶、花丝和子粒第6天存活率分别为0、4.0%和4.0%;取食CM8303心叶、花丝和子粒第6天存活率分别为0、4.0%和2.0%。分析显示转基因抗虫玉米CM8302和CM8303的心叶、花丝和子粒均具有显著杀虫效果且遗传稳定。

Through the utilization of agrobacterium mediated method and new insect-resistant gene Cry1A.401,transgenic insect resistant maize CM8302 and CM8303 were developed.Three generations of transgenic materials,BC_(4)F_(1),BC_(4)F_(2)and BC_(4)F_(3)were obtained through backcross screening.The methods of qRT-PCR and ELISA were used to determine that Cry1A.401 gene was expressed in the heartleaves,silks,and grains,and no significant difference in the expression amount of the target gene among three generations.Field test results with insect-artificial inoculation showed that the resistance level of CM8302 and CM8303 at the heart leaf and the silking stage was high.The results of laboratory bioassay showed that the survival rates of Ostrinia furnacalis larvae on the 6th day after feeding CM8302 and CM8303 heartleaves,silks and grains were 0,4.0%and 4.0%,and 0,4.0%and 2.0%respectively.In conclusion,the heartleaves,silks,and grains of transgenic insect-resistant maize CM8302 and CM8303 have significant insecticidal effects and are genetically stable.