转Bt cry1Ah/cry1Ie双价基因抗虫玉米的研究

构建了含有人工改造的抗虫基因Bt cry1Ah、cry1Ie和耐除草剂基因2mG2-epsps的植物表达载体pMUHUESGM,利用基因枪法将表达盒片段转化玉米愈伤组织,以2mG2-epsps基因为筛选标记基因,经草甘膦异丙胺盐筛选获得24株T0代再生植株,其中PCR检测阳性植株有20株。T0和T1代植株的分子检测结果证明了外源基因已经整合到玉米基因组中并能够稳定遗传和表达,转基因株系在田间生物活性检测中表现出较好的抗虫性,这为培育抗虫玉米新品种提供了参考,同时本研究中采用了基因枪片段转化法,提高了转基因的生物安全性。

Studies on Insect-resistant Transgenic Maize(Zea mays L.) Harboring Bt cry1Ahand cry1IeGenes

A plant expression vector pMUHUESGM harboring modified Bt cry1Ah gene,Bt cry1Ie gene and glyphosate-tolerant 2mG2-epsps gene was constructed.The foreign genes cassette of the vector pMUHUESGM was transferred into maize(Zea mays L.) immature embryonic calli by microprojectile bombardment.Twenty-four T0 regenerated plants were obtained with 2mG2-epsps gene as a selectable marker gene and glyphosate isopropylamine salt as a screening agent.PCR analysis showed that 20 plants had the integration of cry1Ah,cry1Ie and 2mG2-epsps genes.T0 and T1 generation plants’ molecular detection showed that foreign genes had been not only integrated into the maize genome,but also inherited and expressed stably over generations in maize.Bioassay revealed that transgenic events had high resistance to Asia corn borer.These results provided reference for breeding of new insect-resistant maize,moreover fragment transformation improved biosafety of genetically modified organisms.