期刊
JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY
卷 24, 期 9, 页码 1162-1169出版社
KOREAN SOC MICROBIOLOGY & BIOTECHNOLOGY
DOI: 10.4014/jmb.1312.12076
关键词
5-Enolpyruvylshikimate-3-phosphate synthase; Rhizobium leguminosarum; glyphosate tolerant; transgenic Arabidopsis
资金
- Shanghai National Science Foundation [11ZR14324000]
- Youth Fund of Shanghai Academy of Agricultural Sciences [2013-11]
- Key Project Fund of the Shanghai Municipal Committee of Agriculture [2009-6-4, 2011-1-8]
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) 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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