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


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.


Non-Agrobacterium strains Genetic transformation Solanumtuberosum Potato Transbacter Rhizobia 



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)


  1. An G, Watson BD, Chiang CC (1986) Transformation of tobacco, tomato, potato and Arabidopsis thaliana using a binary Ti vector system. Plant Physiol 81:301–305PubMedCrossRefGoogle Scholar
  2. Broothaerts W, Mitchell HJ, Weir B, Kaines S, Smith LMA, Yang W, Mayer JE, Rodriguez CR, Jefferson RA (2005) Gene transfer to plants by diverse bacteria. Nature 433:629–633PubMedCrossRefGoogle Scholar
  3. Chilton M-D (2005) Adding diversity to plant transformation. Nat Biotechnol 23:309–310PubMedCrossRefGoogle Scholar
  4. Craig W, Gargano D, Scotti N, Nguyen TT, Lao NT, Kavanagh TA, Dix PJ, Cardi T (2005) Direct gene transfer in potato: a comparison of particle bombardment of leaf explants and PEG-mediated transformation of protoplasts. Plant Cell Rep 24:603–611PubMedCrossRefGoogle Scholar
  5. Crowell E, McGrath J, Douches D (2008) Accumulation of vitamin E in potato (Solanum tuberosum) tubers. Transgenic Res 17:205–217PubMedCrossRefGoogle Scholar
  6. Giddings G, Allison G, Brooks D, Carter A (2000) Transgenic plants as factories for biopharmaceuticals. Nat Biotechnol 18:1151–1155PubMedCrossRefGoogle Scholar
  7. Herres P, Schippers-Rozenboom M, Jacobsen E, Visser R (2002) Transformation of a large number of potato varieties: genotype-dependent variation in efficiency and somaclonal variability. Euphytica 124:13–22CrossRefGoogle Scholar
  8. Jefferson RA, Kavanagh TA, Bevan MW (1987) GUS fusions—beta-glucuronidase as a sensitive and versatile gene fusion marker in higher-plants. EMBO J 6:3901–3907PubMedGoogle Scholar
  9. Kok-Jacon G, Vincken J-P, Suurs L, Wang D, Liu S, Visser R (2007) Expression of alternansucrase in potato plants. Biotechnol Lett 29:1135–1142PubMedCrossRefGoogle Scholar
  10. Kramer LC, Choudoir MJ, Wielgus SM, Bhaskar PB, Jiang JM (2009) Correlation between transcript abundance of the RB Gene and the level of the RB-mediated late blight resistance in potato. Mol Plant-Microbe Interact 22:447–455PubMedCrossRefGoogle Scholar
  11. Lacroix B, Citovsky V (2009) Agrobacterium aiming for the host chromatin: host and bacterial proteins involved in interactions between T-DNA and plant nucleosomes. Commun Integr Biol 2:42–45PubMedCrossRefGoogle Scholar
  12. Lacroix B, Loyter A, Citovsky V (2008) Association of the Agrobacterium T-DNA-protein complex with plant nucleosomes. Proc Natl Acad Sci 105:15429–15434PubMedCrossRefGoogle Scholar
  13. Li M, Li H, Jiang H, Wu G (2008) Establishment of a highly efficient Agrobacterium tumefaciens-mediated leaf disc transformation method for Broussonetia papyrifera. Plant Cell Tissue Organ Culture 93:249–255CrossRefGoogle Scholar
  14. Mullins E, Milbourne D, Petti C, Doyle-Prestwich BM, Meade C (2006) Potato in the age of biotechnology. Trends Plant Sci 11:254–260PubMedCrossRefGoogle Scholar
  15. Murashige TF, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plantarum 15:473–476CrossRefGoogle Scholar
  16. Neumann K, Stephan DP, Ziegler K, Huhns M, Broer I, Lockau W, Pistorius EK (2005) Production of cyanophycin, a suitable source for the biodegradable polymer polyaspartate, in transgenic plants. Plant Biotechnol J 3:249–258PubMedCrossRefGoogle Scholar
  17. Nottenburg C, Rodriguez CR (eds) (2007) Agrobacterium-mediated gene transfer: a lawyer’s perspective. Agrobacterium. Springer, New York, 750Google Scholar
  18. Pandian A, Hurlstone C, Liu Q, Singh S, Salisbury P, Green A (2006) Agrobacterium-mediated transformation protocol to overcome necrosis in elite Australian Brassica juncea lines. Plant Mol Biol Reporter 24:103–103CrossRefGoogle Scholar
  19. Petti C (2007) Elucidating the propensity to genetically transform Solanum tuberosum L. and investigating the consequences for subsequent risk assessment studies. Biology. Maynooth, National University of Ireland, Maynooth, 384Google Scholar
  20. Petti C, Kumar S (eds) (2010) The potato (Solanum tuberosum L.): an overview of genetic transformation and related biosafety issues. Applications of plant biotechnology: in vitro propagation, plant transformations and secondary metabolite production IK international Pvt. Ltd, pp 336–376Google Scholar
  21. Richter LJ, Thanavala Y, Arntzen CJ, Mason HS (2000) Production of hepatitis B surface antigen in transgenic plants for oral immunization. Nat Biotechnol 18:1167–1171PubMedCrossRefGoogle Scholar
  22. Schünmann PHD, Coia G, Waterhouse PM (2002) Biopharming the SimpliRED™ HIV diagnostic reagent in barley, potato and tobacco. Mol Breed 9:113–121CrossRefGoogle Scholar
  23. Shahin EA, Simpson RB (1986) Gene transfer system for potato. Hortscience 21:1199–1201Google Scholar
  24. Song J, Bradeen JM, Naess SK, Raash JA, Wielgus SM, Haberlach GT, Liu J, Kuang H, Austin-Phillips S, Buell CR, Helgeson JP, Jiang J (2003) Gene RB cloned from Solanum bulbocastanum confers broad spectrum resistance to potato late blight. Proc Natl Acad Sci USA 100:9128–9133PubMedCrossRefGoogle Scholar
  25. Sparrow P, Irwin J, Dale P, Twyman R, Ma J (2007) Pharma-planta: road testing the developing regulatory guidelines for plant-made pharmaceuticals. Transgenic Res 16:147–161PubMedCrossRefGoogle Scholar
  26. U.N. Secretariat (2007) World population prospects, the 2006 revision. New York, The Department of Economic and Social Affairs, United NationsGoogle Scholar
  27. van der Vossen E, Gros J, Sikkema A, Muskens M, Wouters D, Wolters P, Pereira A, Allefs S (2005) The Rpi-blb2 gene from Solanum bulbocastanum is an Mi-1 homolog conferring broad-spectrum late blight resistance in potato. Plant J 44:208PubMedCrossRefGoogle Scholar
  28. Yu J, Langridge WHR (2001) A plant-based multicomponent vaccine protects mice from enteric diseases. Nat Biotechnol 19:548–552PubMedCrossRefGoogle Scholar

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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