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Transgenic Plants pp 215–225Cite as

Initiation and Transformation of Grapevine Embryogenic Cultures

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

Abstract

Protocols for the production and transformation of grapevine embryogenic cultures are described. Embryogenic cultures are initiated from leaves or stamens and pistils and transformed with Agrobacterium containing an enhanced green fluorescent protein/neomycin phosphotransferase II (egfp/nptII) fusion gene. Cultures are transferred to induction medium in the dark for callus formation and proliferation. Resulting cultures are transferred to somatic embryo development medium to induce secondary embryogenesis and formation of transgenic somatic embryos. Transgenic embryos identified on the basis on GFP fluorescence and kanamycin resistance are transferred to germination medium to regenerate transgenic plants. The presence of transgenes in independent plant lines is confirmed by PCR.

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Author information

Authors and Affiliations

  1. Mid-Florida Research and Education Center, University of Florida/IFAS, Apopka, FL, USA

    Sadanand A. Dhekney, Zhijian T. Li & Dennis J. Gray

  2. Citrus Research and Education Center, University of Florida/IFAS, Lake Alfred, FL, USA

    Manjul Dutt

Authors
  1. Sadanand A. Dhekney
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  2. Zhijian T. Li
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  3. Manjul Dutt
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  4. Dennis J. Gray
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Corresponding author

Correspondence to Dennis J. Gray .

Editor information

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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© 2012 Springer Science+Business Media, LLC

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Dhekney, S.A., Li, Z.T., Dutt, M., Gray, D.J. (2012). Initiation and Transformation of Grapevine Embryogenic Cultures. 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_18

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  • DOI: https://doi.org/10.1007/978-1-61779-558-9_18

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-557-2

  • Online ISBN: 978-1-61779-558-9

  • eBook Packages: Springer Protocols

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