| 多启动子驱动苘麻epsps优化基因植物表达载体的构建及其转化 |
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| 引用本文： | 张莎莎,张锐,周焘,郭三堆.多启动子驱动苘麻epsps优化基因植物表达载体的构建及其转化[J].中国农业科技导报,2013,15(1):48-54. |
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| 作者姓名： | 张莎莎 张锐 周焘 郭三堆 |
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| 作者单位： | (中国农业科学院生物技术研究所, 国家农作物基因资源与基因改良重大科学工程, 北京 100081) |
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| 基金项目： | 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 |
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| 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。
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| 关 键 词： | 棉花转化 草甘膦epsps 启动子 载体构建 |
Construction of a Vector for an Optimized epsps Gene from Abutilon theophrasti Medic Promoted by Multi-types of Promoters and its Transformation |
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| ZHANG Sha-sha,ZHANG Rui,ZHOU Tao,GUO San-dui.Construction of a Vector for an Optimized epsps Gene from Abutilon theophrasti Medic Promoted by Multi-types of Promoters and its Transformation[J].Journal of Agricultural Science and Technology,2013,15(1):48-54. |
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| Authors: | ZHANG Sha-sha ZHANG Rui ZHOU Tao GUO San-dui |
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| Institution: | (Biotechnology Research Institute, National Key Facility of Crop Gene Resources and Genetic Improvement, ;
Chinese Academy of Agricultural Sciences, Beijing 100081, China) |
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| Abstract: | 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 coexpression, including constitutive promoter CaMV35S, cotton reproductive organs referential expression promoter arf1 and glyphosate induced promoter ag2. The construct was introduced into cotton via Agrobacterium based flower dipping approach. Eight T0 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. |
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| Keywords: | cotton genetic transformation glyphosate epsps promoter vector construction |
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