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Development of Highly Efficient Genetic Transformation Protocols for Table Grape Sugraone and Crimson Seedless

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Transgenic Plants

Part of the book series: Methods in Molecular Biology ((MIMB,volume 847))

Abstract

Genetic engineering of grapevine is a powerful tool to study gene function as well as to introduce new traits into existing Vitis cultivars without altering their essential characters and identity. Agrobacterium-mediated transformation is one of the most efficient methods for gene transfer, but the efficiency of the procedure depends on several parameters such as the grapevine genotype, the selection strategy, the Agrobacterium strain, and concentration used to infect as well as the culture method among others. This chapter describes highly efficient genetic transformation protocols for seedless table grapevine cultivars Sugraone and Crimson Seedless by co-culturing embryogenic calli with Agrobacterium tumefaciens. The procedures are specific for each cultivar by adjusting the kanamycin concentration used to select transformed cells (20 mg/L and 50 mg/L kanamycin for Crimson Seedless and Sugraone, respectively) and the low Agrobacterium density used to infect the embryogenic calli (0.06 OD600 being more effective for the transformation of Crimson Seedless and 0.2 OD600 for Sugraone). Other factors that affect the transformation efficiency are the initial amount of embryogenic calli used to co-culture with Agrobacterium and the culture method of calli.

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Acknowledgements

The authors wish to thank Dr. A. J. López-Pérez and M. Pazos-Navarro for their contribution in the development of this grape genetic transformation protocol. This research was supported by grants from the Instituto Murciano de Investigación y Desarrollo Agrario y Alimentario (PR06-002) and from Fundación Séneca. Mercedes Dabauza was supported by the European Social Fund.

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Authors and Affiliations

  1. Instituto Murciano de Investigación y Desarrollo Agrario y Alimentario (IMIDA), Murcia, Spain

    Mercedes Dabauza

  2. Instituto Murciano de Investigación y Desarrollo Agrario y Alimentario (IMIDA), La Alberca, Murcia, Spain

    Leonardo Velasco

Authors
  1. Mercedes Dabauza
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  2. Leonardo Velasco
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Correspondence to Mercedes Dabauza .

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Editors and Affiliations

  1. School of Biological Sciences, University of Reading, Reading, Berkshire, RG6 6AS, United Kingdom

    Jim M. Dunwell

  2. Dept. of Agricultural Botany, School of Biological Sciences, University of Reading, Reading, United Kingdom

    Andy C. Wetten

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Dabauza, M., Velasco, L. (2012). Development of Highly Efficient Genetic Transformation Protocols for Table Grape Sugraone and Crimson Seedless. In: Dunwell, J., Wetten, A. (eds) Transgenic Plants. Methods in Molecular Biology, vol 847. Humana Press. https://doi.org/10.1007/978-1-61779-558-9_19

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  • DOI: https://doi.org/10.1007/978-1-61779-558-9_19

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-557-2

  • Online ISBN: 978-1-61779-558-9

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