Liu Zhi, Zhu Zhen, Zhang Tianzhen
National Key Laboratory of Crop Genetics and Germplasm Enhancement, Cotton Research Institute, Nanjing Agricultural University, Nanjing, PR China.
Methods Mol Biol. 2013;958:199-210. doi: 10.1007/978-1-62703-212-4_17.
Elite cotton cultivar Sumian16 was transformed with p7RPSBK-mGNA-NPTII containing Bt (CryIA(c)), Galanthus nivalis agglutinin (GNA) resistance genes and selectable marker NptII gene via the pollen tube pathway method and two fertile transgenic Bt + GNA plants were obtained in the present study. The integration and expression of the Bt and GNA genes were confirmed by molecular biology techniques and insect bioassays. Insect bioassays showed that the transformed plants were highly toxic to bollworm larvae as well as obviously retarding development of aphid populations. PCR analyses and identification of resistance to Kanamycin and bollworm showed that the resistance to bollworm for the two transgenic plants was dominantly inherited in a Mendelian manner and the two resistance genes and selectable marker co-segregated from primary transformed parents to the first self-fertilized progeny plants.
通过花粉管通道法,用含有Bt(CryIA(c))、雪花莲凝集素(GNA)抗性基因和选择标记NptII基因的p7RPSBK-mGNA-NPTII转化优良棉花品种苏棉16,本研究获得了两株可育的转基因Bt + GNA植株。通过分子生物学技术和昆虫生物测定证实了Bt和GNA基因的整合与表达。昆虫生物测定表明,转化植株对棉铃虫幼虫具有高毒性,对蚜虫种群的发育也有明显的抑制作用。PCR分析以及对卡那霉素和棉铃虫抗性的鉴定表明,这两株转基因植株对棉铃虫的抗性以孟德尔方式显性遗传,两个抗性基因和选择标记从最初的转化亲本到第一代自交后代植株共分离。