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Transgenic plants from shoot apical meristems of Vitis vinifera L. “Thompson Seedless” via Agrobacterium-mediated transformation

  • Genetic Transformation and Hybridization
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Abstract

Shoot apical meristem explants of Vitis vinifera “Thompson Seedless” were used for Agrobacterium-mediated genetic transformation. It was determined that the meristems had to be subjected to a dark growth phase then wounded to obtain transgenic plants. Morphological and histological studies illustrated the role of wounding to expose apical meristem cells for transformation. A bifunctional egfp/nptII fusion gene was used to select kanamycin resistant plants that expressed green fluorescent protein (GFP). Kanamycin at a concentration of 16 mg L−1 in selection medium resulted in recovery of non-chimeric transgenic plants that uniformly expressed GFP, whereas 8 mg L−1 kanamycin allowed non-transgenic and/or chimeric plants to develop. Polymerase chain reaction (PCR) and Southern blot analyses confirmed the presence of transgenes and their stable integration into the genome of regenerated plants. Up to 1% of shoot tips produced stable transgenic cultures within 6 weeks of treatment, resulting in a total of 18 independent lines.

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Abbreviations

BA:

Benzyladenine

GFP:

Green fluorescent protein

NAA:

Napthyleneacetic acid

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Acknowledgments

We thank Marilyn Van Aman and Diann Achor for invaluable technical support and advice during this project. Use of the Electron Microscope Laboratory at the UF/IFAS Citrus Research & Education Center is gratefully acknowledged. This research was supported by the Florida Agricultural Experiment Station and the Florida Department of Agriculture and Consumer Services’ Viticulture Trust Fund. D.J. Gray and Z.T. Li own stock in Florida Genetics LLC, which may license resulting patents and, as such, may benefit financially as a result of the outcome of research reported in this publication.

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Author notes

  1. M. Dutt

    Present address: Citrus Research and Education Center, University of Florida/IFAS, 700 Experiment Station Road, Lake Alfred, FL, 33850, USA

Authors and Affiliations

  1. Mid-Florida Research and Education Center, University of Florida/IFAS, 2725 Binion Road, Apopka, FL, 32703-8504, USA

    Z. T. Li, S. A. Dhekney & D. J. Gray

Authors
  1. M. Dutt
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  2. Z. T. Li
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  3. S. A. Dhekney
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  4. D. J. Gray
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Corresponding author

Correspondence to D. J. Gray.

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Communicated by K.K. Kamo.

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Dutt, M., Li, Z.T., Dhekney, S.A. et al. Transgenic plants from shoot apical meristems of Vitis vinifera L. “Thompson Seedless” via Agrobacterium-mediated transformation. Plant Cell Rep 26, 2101–2110 (2007). https://doi.org/10.1007/s00299-007-0424-6

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  • DOI: https://doi.org/10.1007/s00299-007-0424-6

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