Journal of Biotechnology 128 (2007) 681–692 The rolB gene-induced overproduction of resveratrol in Vitis amurensis transformed cells K.V. Kiselev a,c,∗ , A.S. Dubrovina a,c , M.V. Veselova b , V.P. Bulgakov a , S.A. Fedoreyev b , Y.N. Zhuravlev a a Institute of Biology and Soil Science, Far East Branch of Russian Academy of Sciences, 159 Stoletija Str., Vladivostok 690022, Russia b Pacific Institute of Bioorganic Chemistry, Far East Branch of Russian Academy of Sciences, Vladivostok 690022, Russia c Far Eastern State University, 690090 Vladivostok, Russia Received 3 June 2006; received in revised form 28 September 2006; accepted 10 November 2006 Abstract Resveratrol is a stilbene, which prevents carcinogenesis at stages of tumor initiation, promotion and progression. In the present investigation, we developed cell cultures of wild-growing grape (Vitis amurensis Rupr.). The cultures produced low levels of resveratrol, up to 0.026% dry wt., i.e., comparable to levels reported for other plant cell cultures previously established. Different methods commonly used to increase secondary metabolite production (cell selection, elicitor treatments and addition of a biosynthetic precursor) only slightly enhanced cell productivity. Transformation of V. amurensis V2 callus culture by the rolB gene of Agrobacterium rhizogenes resulted in more than a 100-fold increase in resveratrol production in transformed calli. The rolB-transformed calli are capable of producing up to 3.15% dry wt. of resveratrol. We show that the capability to resveratrol biosynthesis is tightly correlated with the abundance of rolB mRNA transcripts. Tyrosine phosphatase inhibitors abolished the rolB-gene-mediated stimulatory effect, thus documenting for the first time the involvement of tyrosine phosphorylation in plant secondary metabolism. © 2006 Elsevier B.V. All rights reserved. Keywords: Vitis amurensis; Resveratrol; rolB gene; Callus culture Abbreviations: MeJA, methyl jasmonate; PAO, phenylarsine oxide; Phe, phenylalanine; PFP, p-fluoro-dl-phenylalanine; SA, sal- icylic acid; SNP, sodium nitroprusside; BAP, 6-benzylaminopurine; NAA, -naphthaleneacetic acid; IAA, indoleacetic acid; 2,4-D, 2,4- dichlorophenoxyacetic acid ∗ Corresponding author at: Institute of Biology and Soil Science, Far East Branch of Russian Academy of Sciences, 159 Stoletija Str., Vladivostok 690022, Russia. Tel.: +7 4232 312129; fax: +7 4232 310193. E-mail address: kkv5@mail.ru (K.V. Kiselev). 1. Introduction Stilbenes play an important role in protecting plants against fungal infections (Dixon and Harrison, 1990) and constitute the main group of phytoalexins within the Vitaceae (Jeandet et al., 2002). In recent years, much attention has been emphasized to the effects of stilbenes, especially resveratrol, on human health. Although resveratrol synthesis has been reported in 0168-1656/$ – see front matter © 2006 Elsevier B.V. All rights reserved. doi:10.1016/j.jbiotec.2006.11.008