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Isolation and Functional Analysis of Convolvulus arvensis EPSPS Promoter

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Abstract

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.

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Abbreviations

EPSPS:

5-enolpyruvylshikimate-3 phosphate synthase

CTAB:

Cetyl trimethyl ammonium bromide

cDNAs:

Complementary DNAs

qRT-PCR:

Quantitative reverse transcription polymerase chain reaction

GUS:

β-Glucuronidase

MS:

Murashige and Skoog

WT:

Wild type

4-MU:

4-Methylumbelliferone

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Acknowledgments

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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Authors and Affiliations

  1. Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, 100193, China

    Zhaofeng Huang, Guirong Wang, Hongjuan Huang, Shouhui Wei, Xinxin Zhou, Jinyi Chen, Jingchao Chen & Chaoxian Zhang

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  1. Zhaofeng Huang
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  2. Guirong Wang
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  3. Hongjuan Huang
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  4. Shouhui Wei
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  8. Chaoxian Zhang
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Correspondence to Chaoxian Zhang.

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Huang, Z., Wang, G., Huang, H. et al. Isolation and Functional Analysis of Convolvulus arvensis EPSPS Promoter. Plant Mol Biol Rep 33, 1650–1658 (2015). https://doi.org/10.1007/s11105-015-0861-2

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  • DOI: https://doi.org/10.1007/s11105-015-0861-2

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