Isolation and Functional Analysis of Convolvulus arvensis EPSPS Promoter
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A 1142-bp upstream sequence (named CaEPSPS-P, GenBank accession number: KC107822) of the EPSPS gene from Convolvulus arvensis L was obtained by genome walking. The full-length sequence of the CaEPSPS-P and four deletion mutants were fused to the β-glucuronidase (GUS) gene and introduced into Arabidopsis via Agrobacterium-mediated transformation. Histochemical GUS staining of the transgenic plants showed that the CaEPSPS-P could drive GUS expression in the roots, stems, and leaves, but no visible GUS staining was detected in seeds. Further deletion analysis revealed two positive regulatory regions (−900 to −632 and −632 to −418) responsible for the basal activity of the EPSPS promoter. GUS activity assays indicated that GUS expression can be stimulated by light and glyphosate. In addition, a region between −632 and −400 was necessary for light-induced expression, while a region from −900 to −632 was necessary for glyphosate-induced GUS expression. These results suggested that the CaEPSPS-P was modulated by multiple cis-regulatory elements in distinct and complex patterns to regulate transgene expression.
KeywordsGlyphosate Arabidopsis thaliana Promoter Transgenic plant
5-enolpyruvylshikimate-3 phosphate synthase
Cetyl trimethyl ammonium bromide
Quantitative reverse transcription polymerase chain reaction
Murashige and Skoog
We thank Li Cui from the Chinese Academy of Agriculture Sciences for helpful discussions and critical reading of this manuscript. This work is funded by Special Fund for Agro-scientific Research in the Public Interest (201303022).
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