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A comparison of transgenic barley lines produced by particle bombardment and Agrobacterium-mediated techniques

  • Genetic Transformation and Hybridization
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Abstract

Two barley transformation systems, Agrobacterium-mediated and particle bombardment, were compared in terms of transformation efficiency, transgene copy number, expression, inheritance and physical structure of the transgenic loci using fluorescence in situ hybridisation (FISH). The efficiency of Agrobacterium-mediated transformation was double that obtained with particle bombardment. While 100% of the Agrobacterium-derived lines integrated between one and three copies of the transgene, 60% of the transgenic lines derived by particle bombardment integrated more than eight copies of the transgene. In most of the Agrobacterium-derived lines, the integrated T-DNA was stable and inherited as a simple Mendelian trait. Transgene silencing was frequently observed in the T1 populations of the bombardment-derived lines. The FISH technique was able to reveal additional details of the transgene integration site. For the efficient production of transgenic barley plants, with stable transgene expression and reduced silencing, the Agrobacterium-mediated method appears to offer significant advantages over particle bombardment.

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Acknowledgements

We gratefully acknowledge Dr. H. Salvo-Garrido for helpful discussions. Funding for this research was provided by the European Commission (4th program FAIR), the Biotechnology and Biological Sciences Research Council and the Department of the Environment, Food and Rural Affairs (DEFRA)

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Authors and Affiliations

  1. John Innes Centre, Norwich Research Park, Colney, Norwich, NR4 7UH, UK

    S. Travella, S. M. Ross, J. Harden, C. Everett, J. W. Snape & W. A. Harwood

  2. Present address: Institute of Plant Biology, University of Zürich, 8008, Zürich, Switzerland

    S. Travella

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  1. S. Travella
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  2. S. M. Ross
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  3. J. Harden
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  4. C. Everett
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  5. J. W. Snape
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  6. W. A. Harwood
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Correspondence to W. A. Harwood.

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Communicated by L. Peña

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Travella, S., Ross, S.M., Harden, J. et al. A comparison of transgenic barley lines produced by particle bombardment and Agrobacterium-mediated techniques. Plant Cell Rep 23, 780–789 (2005). https://doi.org/10.1007/s00299-004-0892-x

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  • DOI: https://doi.org/10.1007/s00299-004-0892-x

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