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Agrobacterium-Mediated Transformation of Bread and Durum Wheat Using Freshly Isolated Immature Embryos

  • Huixia Wu
  • Angela Doherty
  • Huw D. Jones
Protocol
Part of the Methods in Molecular Biology™ book series (MIMB, volume 478)

Abstract

Agrobacterium-mediated transformation of wheat is becoming a viable alternative to the more established biolistic protocols. It offers advantages in terms of simple, low-copy-number integrations and can be applied with similar efficiencies to specific durum wheat and spring and winter bread wheat types varieties.

Keywords:

Agrobacterium tumefaciens bread wheat durum wheat AGL1 inoculation immature embryo 

Notes

Acknowledgements

Rothamsted Research receives grant-aided support from the Biotechnology and Biological Sciences Research Council (BBSRC) of the UK. CIMMYT receives grant-aided support for wheat genetic transformation from the European Commission, the Japanese Government, and the Molecular Plant Breeding Cooperative Research Center.

References

  1. 1.
    Broothaerts, W., Mitchell, H. J., Weir, B., Kaines, S., Smith, L. M. A., Yang, W., Mayer, J. E., Roa-Rodriguez, C. and Jefferson, R. A. (2005) Gene transfer to plants by diverse species of bacteria.Nature 433,629–633.CrossRefGoogle Scholar
  2. 2.
    Jones, H. D., Doherty, A. and Wu, H. (2005) Review of methodologies and a protocol for the Agrobacterium-mediated transformation of wheat.Plant Methods 1,5.CrossRefGoogle Scholar
  3. 3.
    Lazo, G. R., Stein, P. A. and Ludwig, R. A. (1991) A DNA Transformation-competent Arabidopsis genomic library in Agrobacterium Bio-Technology 9,963–967.Google Scholar
  4. 4.
    Wu, H., Sparks, C., Amoah, B. and Jones, H. D. (2003) Factors influencing successful Agrobacterium-mediated genetic transformation of wheat Plant Cell Rep. 21,659–668.Google Scholar
  5. 5.
    Hellens, R. P., Edwards, E. A., Leyland, N. R., Bean, S. and Mullineaux, P. M. (2000) pGreen: a versatile and flexible binary Ti vector for Agrobacterium-mediated plant transformation Plant Mol. Biol. 42,819–832.CrossRefGoogle Scholar
  6. 6.
    Amoah, B. K., Wu, H., Sparks, C. and Jones, H. D. (2001) Factors influencing Agrobacterium-mediated transient expression of uidA in wheat inflorescence tissue J. Exp. Bot. 52,1135–1142.CrossRefGoogle Scholar
  7. 7.
    Wu, H., Doherty, A. and Jones, H. D. (2008) Efficient and rapid Agrobacterium-mediated transformation of durum wheat (Triticum turgidum L. ssp durum) using additional virulence genes. Transgen. Res. 17, 425–436.CrossRefGoogle Scholar
  8. 8.
    Sparks, C. A. and Jones, H. D. (2004) Transformation of wheat by biolistics, in Curtis, I. P., ed.), Transgenic Crops of the World Essential Protocols, (Kluwer, Dordrecht, pp. 19–35.Google Scholar
  9. 9.
    Sambrook, J., Fritsch, E. F. and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory, New York.Google Scholar
  10. 10.
    Tingay, S., McElroy, D., Kalla, R., Fieg, S., Wang, M. B., Thornton, S. and Brettell, R. (1997) Agrobacterium tumefaciens-mediated barley transformation Plant J. 11,1369–1376.CrossRefGoogle Scholar
  11. 11.
    Wu, H., Sparks, C. A. and Jones, H. D. (2006) Characterisation of T-DNA loci and vector backbone sequences in transgenic wheat produced by Agrobacterium-mediated transformation Mol. Breed. 18,195–208.CrossRefGoogle Scholar

Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Huixia Wu
    • 1
  • Angela Doherty
    • 2
  • Huw D. Jones
    • 2
  1. 1.Genetic Resources and Enhancement UnitCIMMYTMexico DFMexico
  2. 2.Centre for Crop Genetic ImprovementDepartment of Plant Sciences, Rothamsted ResearchHarpendenUK

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