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Transgenic Research

, Volume 20, Issue 2, pp 377–386 | Cite as

Gene transfer into Solanum tuberosum via Rhizobium spp.

  • Toni Wendt
  • Fiona Doohan
  • Dominik Winckelmann
  • Ewen Mullins
Original Paper

Abstract

Agrobacterium tumefaciens-mediated transformation (ATMT) is the preferred technique for gene transfer into crops. A major disadvantage of the technology remains the complexity of the patent landscape that surrounds ATMT which restricts its use for commercial applications. An alternative system has been described (Broothaerts et al. in Nature 433:629-633, 2005) detailing the propensity of three rhizobia to transform the model crop Arabidopsis thaliana, the non-food crop Nicotiana tabacum and, at a very low frequency, the monocotyledonous crop Oryza sativa. In this report we describe for the first time the genetic transformation of Solanumtuberosum using the non-Agrobacterium species Sinorhizobium meliloti, Rhizobium sp. NGR234 and Mesorhizobiumloti. This was achieved by combining an optimal bacterium and host co-cultivation period with a low antibiotic regime during the callus and shoot induction stages. Using this optimized protocol the transformation frequency (calculated as % of shoots equipped with root systems with the ability to grow in rooting media supplemented with 25 μg/ml hygromycin) of the rhizobia strains was calculated at 4.72, 5.85 and 1.86% for S. meliloti, R. sp. NGR234 and M. loti respectively, compared to 47.6% for the A. tumefaciens control. Stable transgene integration and expression was confirmed via southern hybridisation, quantitative PCR analysis and histochemical screening of both leaf and/or tuber tissue. In light of the rapid advances in potato genomics, combined with the sequencing of the potato genome, the ability of alternative bacteria species to genetically transform this major food crop will provide a novel resource to the Solanaceae community as it continues to develop potato as both a food and non-food crop.

Keywords

Non-Agrobacterium strains Genetic transformation Solanumtuberosum Potato Transbacter Rhizobia 

Notes

Acknowledgments

We wish to acknowledge the Teagasc Core Research Programme for funding this research and the Teagasc Walsh Fellowship for supporting Toni Wendt.

Supplementary material

11248_2010_9423_MOESM1_ESM.doc (45 kb)
Supplementary material 1 (doc 45 kb)
11248_2010_9423_MOESM2_ESM.pdf (222 kb)
Supplementary material 2 (PDF 222 kb)

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Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • Toni Wendt
    • 1
    • 2
  • Fiona Doohan
    • 2
  • Dominik Winckelmann
    • 1
  • Ewen Mullins
    • 1
  1. 1.Teagasc Crops Research CentrePlant Biotechnology UnitCarlowIreland
  2. 2.School of Biology and Environmental SciencesUniversity College DublinDublin 4Ireland

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