摘要
转基因抗草甘膦作物是商业化最广泛的转基因作物之一,但是棉花的雄蕊对草甘膦敏感,在四叶期后喷施草甘膦会造成抗草甘膦棉花雄性败育,不能正常授粉,最终导致产量降低。为降低草甘膦对棉花育性的不良影响,有针对性的构建了棉花生殖器官优势表达的arf1启动子、棉花草甘膦高效诱导表达的ag2启动子和组成型启动子CaMV35S驱动苘麻epsps优化基因的串联三表达盒植物表达载体,采用农杆菌喷花法将其转入棉花,对T0代喷施草甘膦筛选获得8株对草甘膦有较强抗性并且生殖发育正常的植株;PCR和RT-PCR分析表明外源基因已整合至棉花基因组并正确表达。为探索用不同类型启动子驱动相同基因以扩大外源基因在植物中的表达范围和培育具有自主知识产权的抗草甘膦转基因棉花打下基础。
Genetically modified glyphosate-resistant （GR） crops are widely commercialized GM crops in the world. However, cotton stamens are sensitive to glyphosate treatment, resulting in poor pollination and boll abortion and less cotton yield. To further improve the glyphosate-resistance trait of cotton and reproductive abnormalities in GR cotton, we employed a strategy in which the optimized epsps gene from Abutilon theophrasti Medic were driven by 3 promoters of tandem eoexpression, including constitutive promoter CaMV35S, cotton reproductive organs referential expression promoter arf/and glyphosate-indueed promoter ag2. The construct was introduced into cotton via Agrobacterium based flower dipping approach. Eight TO generation glyphosate-resistant plants were obtained after glyphosate screening, and their reproductive development were normal. PCR and RT-PCR results suggested that the epsps gene had successfully integrated into the cotton genome and had expressed correctly. These results indicated the possibilities of broadening the expression scope about a gene promoted by different types of promoters, so as to obtaining our own genetically modified GR cotton.
出处
《中国农业科技导报》
CAS
CSCD
北大核心
2013年第1期48-54,共7页
Journal of Agricultural Science and Technology
基金
国家自然科学基金项目(31201252)
国家转基因生物新品种培育重大专项(2008ZX08005-004
2009ZX08005-004B
2008ZX08001-006)资助
关键词
棉花转化
草甘膦epsps
启动子
载体构建
cotton genetic transformation
glyphosate epsps
promoter
vector construction
