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以根癌土壤杆菌 (Agrobacteriumtumefaciens)介导法分别将植物表达载体pBinMoBc和pBinoBc导入陆地棉(GossypiumhirsutumL .)栽培品种“新陆早 1号”、“晋棉 7号”、“晋棉 12号”和“冀合 32 1”。pBinMoBc携带有高效启动子复合OM启动子控制下的cry1Ac3基因 ,pBinoBc携带有 35S启动子控制下的cry1Ac3基因。经过共培养、卡那霉素筛选抗性愈伤组织及体细胞胚的诱导 ,得到了再生植株。对T2 代的PCR、Southernblotting、ELISA检测及Westernblotting证明cry1Ac3基因已整合入受体棉花基因组并得到表达。抗虫性检测表明转基因后代对棉铃虫 (Heliothisarmigera )具有良好的抗性 ,转pBinMoBcT2 代与转pBinoBcT2 代相比 ,对棉铃虫具有更快的致死速度。本研究建立了一套高效的陆地棉栽培品种转化体系 ;进一步的检测结果表明 ,复合OM启动子可以提高外源基因的表达量从而增强转基因棉的抗虫性。
Plant expression vectors pBinMoBc and pBinoBc were introduced into Gossypium hirsutum L. cultivars “Xinluzao 1”, “Jinmian 7” and “Jinmian 12” respectively by Agrobacterium tumefaciens-mediated method “Jihe 32 1”. pBinMoBc carries the cry1Ac3 gene under the control of a highly efficient promoter complex OM promoter, and pBinoBc carries the cry1Ac3 gene under the control of the 35S promoter. After co-culture, kanamycin screening of resistant callus and somatic embryo induction, obtained regenerated plants. PCR analysis of T2 generation, Southern blotting, ELISA and Western blotting confirmed that the cry1Ac3 gene has been integrated into the recipient cotton genome and expressed. Insect resistance test showed that transgenic progeny had good resistance to Heliothis armigera. The transgenic plants pBinMoBcT2 had faster lethality than the pBinoBcT2 transgenic plants. In this study, a set of efficient transformation system of upland cotton cultivars was established. Further test results showed that the composite OM promoter can increase the expression of foreign genes and enhance the insect resistance of transgenic cotton.