The role of glutathione-S-transferases in resveratrol transportout of grapevine cells
Martínez-Márquez Aa, Martinez-EstesoMJa, VilellaMa, Sellés-MarchartSa,b, Morante-Carriel Jac,Hurtado Ea, Almagro Ld, Palazon Je, Pedreño MAd, Bru-Martínez Ra
aPlant Proteomics and Functional Genomics Group, Department of Agrochemistry and Biochemistry, Faculty of Science, University of Alicante, Alicante, Spain.bResearch Technical Facility, Proteomics and Genomics Division, University of Alicante, Alicante, Spain. cBiotecnology and Molecular Biology Group, Quevedo State Technical University, Quevedo, Ecuador. dPlantPeroxidases Group, Department of Plant Physiology, University of Murcia, Murcia, Spain. eLaboratory of Plant Physiology, Faculty of Pharmacy, University of Barcelona, Joan XXIII sn, E-08028 Barcelona, Spain.
Vitisvinifera cell culturesresponse to elicitors synthesizingand extracellularly accumulatingstilbenoidphytoalexins, particularly, large amounts of trans-resveratrol (tR) are produced when cells are challenged with methylatedcyclodextrins (MBCD) alone or combined with methyl jasmonate (MeJA).The pathways for transport of tR to the extracellular medium in grapevine cells, being of utmost biotechnological relevance, are mostly unknown.Glutathione-S-transferases (GSTs) are a large protein family with different classes that carry out an array of functions including the trafficking and accumulation of plant secondary metabolites. Upon MBCD/MeJA treatments, both microarray transcriptome analysis(Almagro et al. 2014) and,especially DIGEproteome analysis(Martinez-Esteso et al. 2011; unpublished results), display strong abundance profile correlation of several subclass GSTs with several stilbenesynthase (StSy), and phenylalanine ammonia lyase (PAL) isoforms and paralogs and the tRmetabolite itself. Further, qRT-PCR gene expression analysis provides evidence of co-expression of 5 StSyparalogs and 2 specific -GSTs subclass paralog genes but not with 2 GSTs from other clades. This accumulated evidence strongly points to a role for GST in tR accumulation, potentially be related to the transport of the metabolite.To analyze the functionality of these specific GSTs, we have cloned one of the elicitor-induced grapevine GST genes and used for the generation of stably transformed grapevine cv Gamay cell lines using Agrobacterium-mediated transformation. In contrast to wild lines, only transformed cell lines were shown to accumulate extracellular of tRfor more than 12 days when an absorbent compound such as PVP or beta-CD was added in the culture medium at low concentration. As a conclusion, the co-expression GSTs in relation to StSy and accumulation extracellular of tRprovide strong evidence of itsrole in tR trafficking within the cell in its way to the extracellular medium.
Acknowledgements:Work supported by Spanish Ministry of Science and Innovation (BIO2011-29856-C02-01, BIO2011-29856-C02-02 and BIO2014-51861-R), FEDER and Conselleria d’Educacio, Cultura I Sport de la GeneralitatValenciana (FPA/2013/A/074). J.M.C. holds a postdoctoral grant from SENESCYT-GOVERNMENT OF ECUADOR (006-IECESMG5-GPLR-2012)
References cited
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