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采用花粉管通道途径,将合成的Bt杀虫蛋白基因导入棉花优良品种泗棉3号和中棉所12,获得了转基因植株。组织化学分析表明,GUS标记基因已在R1代植株中表达。根据对GUS阳性转基因植株的PCR分析,证明Bt杀虫蛋白基因出现在R1代植株中,并通过自交传递至R2代。经抗虫性鉴定,获得了5株对棉铃虫有高度毒杀作用的R1代转基因抗虫植株S545、S591、S636、S1001(泗棉3号+Bt/GUS)和Zh1109(中棉所12+Bt/GUS),对棉铃虫幼虫的致死率分别达到91.6%、93.8%、92.3%、85.7%和75.0%。通过自交和选育,获得了R5代转基因抗虫棉,并进行了抗虫性和转基因的跟踪鉴定,表明通过基因工程方法获得了抗棉铃虫的棉花新种质。
The pollen tube pathway was used to transfer the synthesized Bt insecticidal protein gene into cotton elite variety Simian 3 and Cummins 12, and transgenic plants were obtained. Histochemical analysis showed that the GUS marker gene was expressed in R1 plants. Based on PCR analysis of GUS-positive transgenic plants, it was demonstrated that the Bt insecticidal protein gene was present in the R1 plants and was selfed to the R2 generation. Five insect-resistant plants R1, S545, S591, S636, S1001 (Simian 3 + Bt / GUS) and Zh1109 (CIMC 12 + Bt / GUS) ), The lethal rates of H. armigera larvae reached 91.6%, 93.8%, 92.3%, 85.7% and 75.0% respectively. Through selfing and breeding, the R5 transgenic cotton was obtained, and the insect resistance and transgene tracing identification were carried out. The results showed that new cotton germplasms resistant to cotton bollworm were obtained through genetic engineering.