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Involvement of DNA methylation in the regulation of STS10 gene expression in Vitis amurensis

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Abstract

DNA methylation is known to play an important role in various developmental processes and defense mechanisms in plants and other organisms. However, it is not known whether DNA methylation is implicated in the genetic regulation of plant secondary metabolism, including resveratrol biosynthesis. Resveratrol is a naturally occurring polyphenol that is present in grapes, peanuts, and other plant sources, and it exhibits a wide range of valuable biologically active properties. The transformation of the wild-growing grape Vitis amurensis with the oncogene rolB from Agrobacterium rhizogenes has been demonstrated to considerably increase resveratrol production. To investigate whether DNA methylation regulates resveratrol biosynthesis, we treated both rolB transgenic and empty vector control V. amurensis cell cultures with the DNA demethylation agent 5-azacytosine (azaC). The azaC treatment significantly increased stilbene synthase 10 gene (VaSTS10) expression and resveratrol content in the V. amurensis cell cultures. Using bisulfite sequencing, we examined the methylation status of VaSTS10 in cell cultures under normal conditions and after azaC treatment. Both the promoter and 3′-end of the protein coding region of the VaSTS10 gene were hypermethylated (54–67 %) in the control cell culture. The rolB transgenic cell culture had high levels of resveratrol and lower hypermethylation levels of the VaSTS10 gene (20–47 %). The azaC treatment resulted in reduction in the DNA methylation levels in the promoter and coding regions of the VaSTS10 gene in both cell cultures. These data suggest that the DNA methylation may be involved in the control of resveratrol biosynthesis via the regulation of STS genes expression.

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Abbreviations

azaC:

5-Azacytidine

DW:

Dry weight

STS:

Stilbene synthase

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Acknowledgments

The authors express their thanks to Alexandra S. Dubrovina for helpful comments on the manuscript. This work was supported by grant of the Russian Foundation for Basic Research (10-04-00189-a and 12-04-33069-mol_ved), by grants of the Far East Division of the Russian Academy of Sciences.

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Authors and Affiliations

  1. Laboratory of Biotechnology, Institute of Biology and Soil Science, Far East Branch of Russian Academy of Sciences, Vladivostok, 690022, Russia

    Konstantin V. Kiselev, Alexey P. Tyunin & Yuri N. Zhuravlev

  2. Department of Biochemistry and Biotechnology, Far Eastern Federal University, 690090, Vladivostok, Russia

    Konstantin V. Kiselev, Alexey P. Tyunin & Yuri N. Zhuravlev

Authors
  1. Konstantin V. Kiselev
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  2. Alexey P. Tyunin
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  3. Yuri N. Zhuravlev
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Correspondence to Konstantin V. Kiselev.

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Kiselev, K.V., Tyunin, A.P. & Zhuravlev, Y.N. Involvement of DNA methylation in the regulation of STS10 gene expression in Vitis amurensis . Planta 237, 933–941 (2013). https://doi.org/10.1007/s00425-012-1806-8

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  • DOI: https://doi.org/10.1007/s00425-012-1806-8

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