Functional Characterization of aroA from Rhizobium leguminosarum with Significant Glyphosate Tolerance in Transgenic Arabidopsis
文献类型: 外文期刊
作者: Han, Jing 1 ; Tian, Yong-Sheng 1 ; Xu, Jing 1 ; Wang, Li-Juan 1 ; Wang, Bo 1 ; Peng, Ri-He 1 ; Yao, Quan-Hong 1 ;
作者机构: 1.Shanghai Acad Agr Sci, Biotechnol Res Inst, Dept Shanghai Key Lab Agr Genet & Breeding, Shanghai 201106, Peoples R China
关键词: 5-Enolpyruvylshikimate-3-phosphate synthase;Rhizobium leguminosarum;glyphosate tolerant;transgenic Arabidopsis
期刊名称:JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY ( 影响因子:2.351; 五年影响因子:2.65 )
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收录情况: SCI
摘要: Glyphosate is the active component of the top-selling herbicide, the phytotoxicity of which is due to its inhibition of the shikimic acid pathway. 5-Enolpyruvylshikimate-3-phosphate synthase (EPSPS) is a key enzyme in the shikimic acid pathway. Glyphosate tolerance in plants can be achieved by the expression of a glyphosate-insensitive aroA gene (EPSPS). In this study, we used a PCR-based two-step DNA synthesis method to synthesize a new aroA gene (aroA_(R. leguminosarum)) from Rhizobium leguminosarum. In vitro glyphosate sensitivity assays showed that aroA_(R. leguminosarum) is glyphosate tolerant. The new gene was then expressed in E. coli and key kinetic values of the purified enzyme were determined. Furthermore, we transformed the aroA gene into Arabidopsis thaliana by the floral dip method. Transgenic Arabidopsis with the aroA_(R. leguminosarum) gene was obtained to prove its potential use in developing glyphosate-resistant crops.
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