A reliable protocol for the stable transformation of non-embryogenic cells cultures of grapevine (Vitis vinifera L.) and Taxus x media

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Title: A reliable protocol for the stable transformation of non-embryogenic cells cultures of grapevine (Vitis vinifera L.) and Taxus x media
Authors: Martínez Márquez, Ascensión | Morante Carriel, Jaime | Ramírez-Estrada, Karla | Cusidó, Rosa M. | Sellés Marchart, Susana | Palazon, Javier | Pedreño, María A. | Bru-Martinez, Roque
Research Group/s: Proteómica y Genómica Funcional de Plantas
Center, Department or Service: Universidad de Alicante. Departamento de Agroquímica y Bioquímica
Keywords: Agrobacterium system | GFP | Grapevine cell culture | Resveratrol | Stable transformation | Taxus cell cultures
Knowledge Area: Bioquímica y Biología Molecular
Issue Date: 24-Jun-2015
Publisher: POL Scientific
Citation: Journal of Biological Methods. 2015, 2(2): e21. doi:10.14440/jbm.2015.51
Abstract: One of the major intent of metabolic engineering in cell culture systems is to increase yields of secondary metabolites. Efficient transformation methods are a priority to successfully apply metabolic engineering to cell cultures of plants that produce bioactive or therapeutic compounds, such as Vitis vinifera and Taxus x media. The aim of this study was to establish a reliable method to transform non-embryogenic cell cultures of these species. The V. vinifera cv. Gamay/cv. Monastrell cell lines and Taxus x media were used for Agrobacterium-mediated transformation using the Gateway-compatible Agrobacterium sp. binary vector system for fast reliable DNA cloning. The Taxus x media and Vitis cell lines were maintained in culture for more than 4 and 15 months, respectively, with no loss of reporter gene expression or antibiotic resistance. The introduced genes had no discernible effect on cell growth, or led to extracellular accumulation of phytoalexin trans-Resveratrol (t-R) in response to elicitation with methylated cyclodextrins (MBCD) and methyl jasmonate (MeJA) in the grapevine transgenic cell lines compared to the parental control. The method described herein provides an excellent tool to exploit exponentially growing genomic resources to enhance, optimize or diversify the production of bioactive compounds generated by grapevine and yew cell cultures, and offers a better understanding of many grapevine and yew biology areas.
Sponsor: This work has been supported by grants from the Spanish Ministry of Science and Innovation (BIO2011-29856-C02-01, BIO2011-29856-C02-02 and BIO2014-51861-R), European Funds for Regional Development (FEDER) and Conselleria d’Educacio, Cultura i Sport de la Generalitat Valenciana (FPA/2013/A/074). J.M.C. holds a postdoctoral grant from SENESCYT-GOVERNMENT OF ECUADOR (006-IECESMG5-GPLR-2012).
URI: http://hdl.handle.net/10045/48364
ISSN: 2326-9901
DOI: 10.14440/jbm.2015.51
Language: eng
Type: info:eu-repo/semantics/article
Rights: Creative Commons Attribution 3.0 License
Peer Review: si
Publisher version: http://dx.doi.org/10.14440/jbm.2015.51
Appears in Collections:INV - Proteómica y Genómica Funcional de Plantas - Artículos de Revistas

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