中文摘要：棉铃象鼻虫是南美洲最具破坏性的棉花害虫，本文首次对Cry10Aa转基因棉花棉铃象鼻虫的高抗性进行了研究，通过苏云金芽孢杆菌基因查杀虫蛋白编码，成功开发出了对棉铃象鼻虫具有高抗性的Cry10Aa转基因棉花。转基因植物转化载体中含有Cry10Aa基因被泛素化修饰uceA1.7引子启动，并导入到通过基因枪转化的巴西棉品种中。定量PCR检测结果显示，Cry10Aa在T-0转基因棉花叶和花芽组织中转录水平很高，通过Northern杂交和基于Delta Delta Ct计算方法的定量PCR检测分析显示：T-0转基因植物有一个或两个转基因拷贝；Cry10Aa蛋白表达的定量和定性分析显示，T-0转基因植物的花蕾和叶组织中的可变蛋白表达水平约是3.0-14.0 μg/g新鲜组织；棉铃象鼻虫生物敏感性测定显示其具有显著的昆虫毒性和高水平的棉铃虫的死亡率；分子分析表明，对T-1代棉铃象鼻虫转基因的稳定性和昆虫毒性很高，保持在4.05-19.57μg/g新鲜组织范围内，且棉铃象鼻虫死亡率在100%。总之，Cry10Aa转基因抗虫棉是目前棉铃象鼻虫防治的一大进步，可对棉花产业造成巨大的影响。
外文摘要：Genetically modified (GM) cotton plants that effectively control cotton boll weevil (CBW), which is the most destructive cotton insect pest inSouth America, are reported here for the first time. This work presents the successful development of a new GM cotton with high resistance to CBW conferred by Cry10Aa toxin, a protein encoded by entomopathogenic Bacillus thuringiensis (Bt) gene. The plant transformation vector harbouring cry10Aa gene driven by the cotton ubiquitination-related promoter uceA1.7 was introduced into a Brazilian cotton cultivar by biolistic transformation. Quantitative PCR (qPCR) assays revealed high transcription levels of cry10Aa in both T-0 GM cotton leaf and flower bud tissues. Southern blot and qPCR-based 2(-Delta Delta Ct) analyses revealed that T-0 GM plants had either one or two transgene copies. Quantitative and qualitative analyses of Cry10Aa protein expression showed variable protein expression levels in both flower buds and leaves tissues of T-0 GM cotton plants, ranging from approximately 3.0 to 14.0 mu g g(-1) fresh tissue. CBW susceptibility bioassays, performed by feeding adults and larvae with T-0 GM cotton leaves and flower buds, respectively, demonstrated a significant entomotoxic effect and a high level of CBW mortality (up to 100%). Molecular analysis revealed that transgene stability and entomotoxic effect to CBW were maintained in T-1 generation as the Cry10Aa toxin expression levels remained high in both tissues, ranging from 4.05 to 19.57 mu g g(-1) fresh tissue, and the CBW mortality rate remained around 100%. In conclusion, these Cry10Aa GM cotton plants represent a great advance in the control of the devastating CBW insect pest and can substantially impact cotton agribusiness.
外文关键词：Gossypium hirsutum, genetic transformation, Anthonomus grandis, insect pest, plant resistance, Bt toxin
作者：Ribeiro, Thuanne Pires; Monteiro Arraes, Fabricio Barbosa; Lourenco-Tessutti, Isabela Tristan; Silva, Marilia Santos; Lisei-de-Sa,
作者单位：Embrapa Genet Resources & Biotechnol, Brasilia, DF, Brazil
期刊名称：PLANT BIOTECHNOLOGY JOURNAL
期刊影响因子：7.443
出版年份：2017
出版刊次：8
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