论文部分内容阅读
我国棉花抗虫基因大都为Cry1Ab/c,抗性风险日趋增加。本研究依据棉花密码子偏好,人工合成Bt-Cry5Aa抗虫基因,通过花粉管通道法转入棉花,并通过卡那霉素法及PCR方法对不同世代转化株进行鉴定,同时进行了抗虫性测试。结果表明,通过花粉管通道法成功获得转Bt-Cry5Aa基因植株,通过田间卡那霉素鉴定,阳性株率T1为7.76%,T2为73.1%,T3为95.5%;PCR检测显示,T1阳性率为2.35%,T2为55.8%,T3为94.5%;田间抗性试验分析,转Bt-Cry5Aa株系对第2、3、4代棉铃虫校正死亡率分别达到85.42%、75.35%和62.79%,其抗虫性与GK19相比差异不显著;Bt-Cry5Aa能够部分替代目前主流鳞翅目抗虫基因,是棉铃虫的新抗源。
China’s cotton insect-resistant genes are mostly Cry1Ab / c, increasing the risk of resistance. In this study, Bt-Cry5Aa gene was synthesized based on cotton codon preference and transformed into cotton by pollen tube pathway. The transformants of different generations were identified by kanamycin method and PCR method, test. The results showed that the Bt-Cry5Aa transgenic plants were successfully obtained by pollen tube pathway. The positive rate of T1 was 7.76%, T2 was 73.1%, and T3 was 95.5%. The positive rate of T1 T2 was 55.8% and T3 was 94.5%. Correlation analysis showed that the corrected mortality rates of transgenic Bt-Cry5Aa lines were 85.42%, 75.35% and 62.79%, respectively, Compared with GK19, Bt-Cry5Aa can partially replace the current dominant Lepidoptera insect-resistant gene and is a new anti-cotton bollworm resistance.