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http://dx.doi.org/10.4014/jmb.1312.12076

Functional Characterization of aroA from Rhizobium leguminosarum with Significant Glyphosate Tolerance in Transgenic Arabidopsis  

Han, Jing (Department of Shanghai Key Laboratory of Agricultural Genetics and Breeding, Biotechnology Research Institute of Shanghai Academy of Agricultural Sciences)
Tian, Yong-Sheng (Department of Shanghai Key Laboratory of Agricultural Genetics and Breeding, Biotechnology Research Institute of Shanghai Academy of Agricultural Sciences)
Xu, Jing (Department of Shanghai Key Laboratory of Agricultural Genetics and Breeding, Biotechnology Research Institute of Shanghai Academy of Agricultural Sciences)
Wang, Li-Juan (Department of Shanghai Key Laboratory of Agricultural Genetics and Breeding, Biotechnology Research Institute of Shanghai Academy of Agricultural Sciences)
Wang, Bo (Department of Shanghai Key Laboratory of Agricultural Genetics and Breeding, Biotechnology Research Institute of Shanghai Academy of Agricultural Sciences)
Peng, Ri-He (Department of Shanghai Key Laboratory of Agricultural Genetics and Breeding, Biotechnology Research Institute of Shanghai Academy of Agricultural Sciences)
Yao, Quan-Hong (Department of Shanghai Key Laboratory of Agricultural Genetics and Breeding, Biotechnology Research Institute of Shanghai Academy of Agricultural Sciences)
Publication Information
Journal of Microbiology and Biotechnology / v.24, no.9, 2014 , pp. 1162-1169 More about this Journal
Abstract
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.
Keywords
5-Enolpyruvylshikimate-3-phosphate synthase; Rhizobium leguminosarum; glyphosate tolerant; transgenic Arabidopsis;
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