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来自细菌Nocardia sp.AB2253的蛋氨酸砜N-乙酰基转移酶基因(methionine sulfone Nacetyltransferase gene,Mat)编码产物具有N-乙酰基转移酶活性,能解除灭生性除草剂草铵膦的毒性,但在植物中表达效率较低。本研究用水稻(Oryza sativa)偏爱密码子优化的Mat#基因转化籼稻品系9K(Oryza sativa ssp.indica),经过PCR和Southern杂交验证,证明该基因已经整合至水稻基因组中。除草剂抗性检测结果显示,该转化体的芽期草铵膦耐受浓度至少为600 mg/L、秧苗期草铵膦耐受浓度至少为1 000 mg/L,对草铵膦的抗性水平不低于转双丙氨酰膦抗性基因(bialaphos resistance gene,Bar)水稻9KA2。酶活性测定结果显示,该转化体叶片中的N-乙酰基转移酶活性约为非转基因对照中的6.6倍。说明优化后的Mat#基因增强了草铵膦抗性,可作为转化筛选标记和抗除草剂目的基因应用于转基因作物育种。
Methionine sulfone Nacetyltransferase gene (Mat) encoded by the bacterium Nocardia sp. AB2253 has N-acetyltransferase activity, which can release the toxicity of the herbicide herbicide glufosinate. However, In the lower efficiency of expression. In this study, the Oryza sativa preferred codon-optimized Mat # gene was transformed into Oryza sativa ssp. Indica and verified by PCR and Southern blotting that the gene was integrated into rice genome. Herbicide resistance test results showed that the tolerant glufosinate-resistant transformants at the bud stage of at least 600 mg / L, the seedlings of glufosinate-resistant concentration of at least 1 000 mg / L, resistance to glufosinate Not lower than the level of bialaphos resistance gene (Bar) rice 9KA2. The result of enzyme activity assay showed that the N-acetyltransferase activity in the transformant leaves was about 6.6 times of that of the non-transgenic control. The results showed that the optimized Mat # gene enhances glufosinate resistance, and can be used as a marker for transformation screening and a target gene for herbicide resistance in transgenic crop breeding.