DIGE analysis of proteome changes accompanying large resveratrol production by grapevine (Vitis vinifera cv. Gamay) cell cultures in response to methyl-β-cyclodextrin and methyl jasmonate elicitors ☆ M.J. Martinez-Esteso a , S. Sellés-Marchart a , J.C. Vera-Urbina a , M.A. Pedren ~ o b , R. Bru-Martinez a, ⁎ a Plant Proteomics and Functional Genomics Group, Department of Agrochemistry and Biochemistry, Faculty of Science, University of Alicante, Alicante, Spain b Plant Peroxidases Group, Department of Plant Physiology, University of Murcia, Murcia, Spain ARTICLE INFO ABSTRACT Available online 21 March 2011 We had previously shown that Vitis vinifera cv. Gamay cell suspension accumulates extracellularly large amounts of the phytoalexin trans-resveratrol (tR) in response to elicitation with methylated cyclodextrins (MBCD), which can be triplicated when the elicitor is combined with methyl jasmonate (MeJA). In parallel, new pathogenesis-related proteins accumulated in the apoplast-like extracellular space. The aim of this study was to investigate changes in the grapevine cell proteome potentially related to tR accumulation in response to the above elicitors. The DIGE technique was used to detect statistically significant changes in the cell's proteome. A total number of 1031 unique spots were detected, 67 of which were de-regulated upon elicitation. Sixty-four spots were successfully identified by nLC–MS/MS database search analysis. The tR biosynthetic pathway enzymes were up-regulated by MBCD alone or combined with MeJA, but not by treatment with MeJA alone, in agreement with tR accumulation pattern. Seven spots contained stilbene synthase encoded by four different isogenes. Likewise, four glutathione-S-transferases, potentially involved in tR trafficking within the cell and across membranes, were up-regulated in the same fashion as stilbene synthases. The relation of other de-regulated proteins with other effects caused by elicitors on grapevine cells, namely defense response and cell growth inhibition, is discussed. © 2011 Elsevier B.V. All rights reserved. Keywords: Grapevine Cell culture Proteome Resveratrol Cyclodextrin Methyl jasmonate DIGE Stilbene synthase Glutathione-S transferase class tau 1. Introduction Plant cell cultures have been shown to be adequate for the production of bioactive secondary metabolites as shown by the industrial production of shikonin, taxol or berberine which gained commercial application [1–4]. However, despite the extensive studies on the standardization of cell growth and the selection of high yielding cell lines, the number of cases where secondary metabolites are produced by cultured cells is relatively low. Elicitation of plant cell cultures with biotic or abiotic stimuli has been shown as the most efficient strategy to provoke important increases in metabolite yield [5,6]. In fact, elicitors are generally JOURNAL OF PROTEOMICS 74 (2011) 1421 – 1436 ☆ This article is part of M.J. Martinez-Esteso's PhD Thesis. ⁎ Corresponding author at: Department of Agrochemistry and Biochemistry, University of Alicante, Ctra. San Vicente del Raspeig s/n, E-03080 Alicante, Spain. Fax: +34 965903880. E-mail address: Roque.Bru@ua.es (R. Bru-Martinez). 1874-3919/$ – see front matter © 2011 Elsevier B.V. All rights reserved. doi:10.1016/j.jprot.2011.02.035 available at www.sciencedirect.com www.elsevier.com/locate/jprot