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用含合成的Cry1Ac杀虫蛋白嵌合基因 (Bt2 9K)及慈菇蛋白酶抑制剂B(API B)基因表达框的双抗虫基因植物表达载体 ,通过根癌农杆菌介导转化棉花生产品种冀合 32 1,获得了一批抗卡那霉素的转化再生植株。在对转化再生植株进行PCR、卡那霉素抗性和抗虫性测定的基础上 ,经 6代筛选培育出棉铃虫抗性 90 %以上且农艺性状优良的 9个转双抗虫基因棉纯合品系。各纯合株系子代植株的抗虫性及部分序列检测结果进一步表明上述Cry1Ac嵌合蛋白基因是能稳定遗传的。这些品系具有很好的农艺性状 ,其结铃性和纤维比强度明显高于转化受体冀合 32 1,这些纯合系可直接用于生产或作为双抗虫棉种质利用。
By using the plant expression vector of the double-insect resistance gene containing the synthetic CrylAc insecticidal protein chimeric gene (Bt2 9K) and the arrowhead glycoprotein inhibitor B (API B) gene expression vector, the Agrobacterium tumefaciens-mediated transformation of cotton varieties A total of 32 1, access to a number of anti-kanamycin transformation of regenerated plants. On the basis of PCR, kanamycin resistance and insect resistance determination of transformed and regenerated plants, six transgenic cotton plants with more than 90% of bollworm resistance and excellent agronomic traits were obtained after six generations of screening Co-product line. The resistance of the progeny plants of each homozygous strain and partial sequence test further indicated that the above Cry1Ac chimeric protein gene can stably inherit. These lines have good agronomic traits, their boll-forming ability and fiber-specific strength are significantly higher than that of the transformation receptor Jihe321, and these homozygous lines can be directly used for production or utilization as double insect resistant cotton germplasms.