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Effects of the Calmodulin Antagonist W7 on Resveratrol Biosynthesis in Vitis amurensis Rupr.

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Abstract

The calmodulin antagonist N-(6-Aminohexyl)-5-chloro-1-naphthalenesulfonamide (W7) binds to calmodulzin and inhibits Ca2+/calmodulin-regulated enzyme activities. In plant cells, W7 inhibits the activity of calcium-dependent protein kinases (CDPKs)—the major calcium sensors in plants. In the present study, we examined the effect of W7 on increased resveratrol biosynthesis and expression of CDPK and stilbene synthase (STS) genes in a cell culture of Vitis amurensis Rupr. We used coumaric acid (CA), salicylic acid (SA), and phenylalanine (Phe) to increase the content of resveratrol in V. amurensis calli, since its content is low under standard conditions. W7 significantly decreased resveratrol production and expression of STS genes in CA-, SA-, and Phe-treated grape cells. Also, treatment of the V. amurensis calli with SA, Phe, or CA considerably increased expression of VaCDPK1a (with SA, Phe), VaCDPK1L (with SA, Phe), VaCDPK2a (with Phe) genes, and decreased expression of VaCDPK3a (with CA). Addition of W7 to CA-, SA-, and Phe-treated grape cells reversed this effect, resulting in increased VaCDPK3a expression and decreased VaCDPK1a, VaCDPK1L, and VaCDPK2a expression. The results obtained suggest that CDPK activities might play an important role in resveratrol biosynthesis.

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Fig. 1
Fig. 2

Abbreviations

CA:

Coumaric acid

CDPK:

Calcium-dependent protein kinase

Phe:

Phenylalanine

SA:

Salicylic acid

DW:

Dry weight

STS:

Stilbene synthase

W7:

N-(6-Aminohexyl)-5-chloro-1-naphthalenesulfonamide

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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 (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 Eastern Branch of the Russian Academy of Sciences, Vladivostok, 690022, Russia

    K. V. Kiselev & O. A. Shumakova

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

    K. V. Kiselev

  3. Mountain-Taiga station, Far Eastern Branch of the Russian Academy of Sciences, Posyolok Gornotaezhnoe, Primorsky krai, Ussuriisky region, 692533, Russia

    A. Y. Manyakhin

Authors
  1. K. V. Kiselev
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  2. O. A. Shumakova
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  3. A. Y. Manyakhin
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Correspondence to K. V. Kiselev.

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Kiselev, K.V., Shumakova, O.A. & Manyakhin, A.Y. Effects of the Calmodulin Antagonist W7 on Resveratrol Biosynthesis in Vitis amurensis Rupr.. Plant Mol Biol Rep 31, 1569–1575 (2013). https://doi.org/10.1007/s11105-013-0620-1

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