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Co-transformation of Grapevine Somatic Embryos to Produce Transgenic Plants Free of Marker Genes

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Book cover Transgenic Plants

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

Abstract

A cotransformation system using somatic embryos was developed to produce grapevines free of selectable marker genes. This was achieved by transforming Vitis vinifera L. “Thompson Seedless” somatic embryos with a mixture of two Agrobacterium strains. The first strain contained a binary plasmid with an egfp gene of interest between the T-DNA borders. The second strain harbored the neomycin phosphotransferase (nptII) gene for positive selection and the cytosine deaminase (codA) gene for negative selection, linked together by a bidirectional dual promoter complex. Our technique included a short positive selection phase of cotransformed somatic embryos on liquid medium containing 100 mg/L kanamycin before subjecting cultures to prolonged negative selection on medium containing 250 mg/L 5-fluorocytosine.

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Author information

Authors and Affiliations

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

    Manjul Dutt

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

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

Authors
  1. Manjul Dutt
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  2. Zhijian T. Li
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  3. Sadanand A. Dhekney
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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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Dutt, M., Li, Z.T., Dhekney, S.A., Gray, D.J. (2012). Co-transformation of Grapevine Somatic Embryos to Produce Transgenic Plants Free of Marker Genes. 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_17

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  • DOI: https://doi.org/10.1007/978-1-61779-558-9_17

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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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