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Biolistic- and Agrobacterium-Mediated Transformation Protocols for Wheat

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Plant Cell Culture Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 877))

Abstract

After rice, wheat is considered to be the most important world food crop, and the demand for high-quality wheat flour is increasing. Although there are no GM varieties currently grown, wheat is an important target for biotechnology, and we anticipate that GM wheat will be commercially available in 10–15 years. In this chapter, we summarize the main features and challenges of wheat transformation and then describe detailed protocols for the production of transgenic wheat plants both by biolistic and Agrobacterium-mediated DNA-delivery. Although these methods are used mainly for bread wheat (Triticum aestivum L.), they can also be successfully applied, with slight modifications, to tetraploid durum wheat (T. turgidum L. var. durum). The appropriate size and developmental stage of explants (immature embryo-derived scutella), the conditions to produce embryogenic callus tissues, and the methods to regenerate transgenic plants under increasing selection pressure are provided in the protocol. To illustrate the application of herbicide selection system, we have chosen to describe the use of the plasmid pAHC25 for biolistic transformation, while for Agrobacterium-mediated transformation the binary vector pAL156 (incorporating both the bar gene and the uidA gene) has been chosen. Beside the step-by-step methodology for obtaining stably transformed and normal fertile plants, procedures for screening and testing transgenic wheat plants are also discussed.

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Acknowledgements

László Tamás is in receipt of grants from the Hungarian Scientific Research Fund (OTKA T 46703 and 67844) and of the Bilateral Intergovernmental Science and Technology Cooperation (KR-1/2007). Rothamsted Research receives grant-aided support from the Biotechnology and Biological Sciences Research Council of the United Kingdom.

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Authors and Affiliations

  1. Centre for Agricultural Research, Hungarina Academy of Sciences, Martonvásár, Hungary

    Cecília Tamás-Nyitrai

  2. Óbuda University, Budapest, Hungary

    Cecília Tamás-Nyitrai

  3. Cereal Transformation Group, Centre for Crop Genetic Improvement, Plant Sciences Department, Rothamsted Research, Harpenden, Hertfordshire, UK

    Huw D. Jones (Research Group Leader)

  4. Department of Plant Physiology, Eötvös Loránd University, Budapest, Hungary

    László Tamás

Authors
  1. Cecília Tamás-Nyitrai
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  2. Huw D. Jones
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  3. László Tamás
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Corresponding author

Correspondence to László Tamás .

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Editors and Affiliations

  1. , Unidad de Bioquímica y Biología Molecula, Centro de Investigación Científica de Yu, Apartado Postal 87, Cordemex, Mérida, Mexico

    Víctor M. Loyola-Vargas

  2. , Departamento de Ingeniería Genética de P, Ctr Investigación, Estudios Avanzados In, Apartado Postal 629, Irapuato, 36821, Mexico

    Neftalí Ochoa-Alejo

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Tamás-Nyitrai, C., Jones, H.D., Tamás, L. (2012). Biolistic- and Agrobacterium-Mediated Transformation Protocols for Wheat. In: Loyola-Vargas, V., Ochoa-Alejo, N. (eds) Plant Cell Culture Protocols. Methods in Molecular Biology, vol 877. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-818-4_27

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  • DOI: https://doi.org/10.1007/978-1-61779-818-4_27

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-817-7

  • Online ISBN: 978-1-61779-818-4

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