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Regulation of Resveratrol Production in Vitis amurensis Cell Cultures by Calcium-Dependent Protein Kinases

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Abstract

Resveratrol is a naturally occurring plant stilbene that exhibits a wide range of valuable biological and pharmacological properties. Although the beneficial effects of trans-resveratrol to human health and plant protection against fungal pathogens are well-established, little is known about the molecular mechanisms regulating stilbene biosynthesis in plant cells. It has been recently shown that overexpression of the calcium-dependent protein kinase VaCPK20 gene considerably increased resveratrol accumulation in cell cultures of Vitis amurensis. It is possible that calcium-dependent protein kinases (CDPKs) play an important role in the regulation of resveratrol biosynthesis. In the present work, we investigated the effects of overexpression of other members of the CDPK multigene family (VaCPK9, VaCPK13, VaCPK21, and VaCPK29) on resveratrol accumulation and growth parameters of grape cell cultures. The obtained data show that overexpression of VaCPK29 increased resveratrol content 1.6–2.4-fold and fresh biomass accumulation 1.1–1.4-fold in the four independently transformed cell lines of V. amurensis compared with that in the empty vector-transformed calli. However, overexpression of the VaCPK9, VaCPK13, and VaCPK21 genes did not considerably affect resveratrol content and fresh/dry biomass accumulation in the independently transformed cell lines of V. amurensis. VaCPK29-transformed calli were capable of producing between 1.02 and 1.39 mg/l of resveratrol, while the control calli produced 0.48 to 0.79 mg/l of resveratrol. The data indicate that the VaCPK9, VaCPK13, and VaCPK21 genes are not involved in the regulation of stilbene biosynthesis in grape cells, while the VaCPK29 and VaCPK20 genes are implicated in resveratrol biosynthesis as positive regulators.

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Acknowledgments

This work was supported by a grant from the Russian Scientific Foundation (14-14-00366). We are grateful to Professor Alexander Krichevsky (State University of New York, Stony Brook, USA) for providing pSAT1 and pZP-RCS2-nptII plasmid samples and A. tumefaciens GV3101::pMP90 strain.

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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

    O. A. Aleynova, A. S. Dubrovina, K. V. Kiselev & K. V. Kiselev

  2. Mountain-Taiga Station, Far East Branch of Russian Academy of Sciences, Posyolok Gornotaezhnoe, Primorsky krai, Ussuriisky Region, 692533, Russia

    A. Y. Manyakhin

  3. Laboratory of Embryology, A.V. Zhirmunsky Institute of Marine Biology, Far Eastern Branch of the Russian Academy of Sciences, Palchevsky St. 17, 690059, Vladivostok, Russia

    Y. A. Karetin

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  1. O. A. Aleynova
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Correspondence to K. V. Kiselev.

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Aleynova, O.A., Dubrovina, A.S., Manyakhin, A.Y. et al. Regulation of Resveratrol Production in Vitis amurensis Cell Cultures by Calcium-Dependent Protein Kinases. Appl Biochem Biotechnol 175, 1460–1476 (2015). https://doi.org/10.1007/s12010-014-1384-2

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