Abstract
This chapter details a reproducible method for the transformation of spring wheat using Agrobacterium tumefaciens via the direct inoculation of bacteria into immature seeds in planta as described in patent WO 00/63398(1. Transformation efficiencies from 1 to 30% have been obtained and average efficiencies of at least 5% are routinely achieved. Regenerated plants are phenotypically normal with 30–50% of transformation events carrying introduced genes at single insertion sites, a higher rate than is typically reported for transgenic plants produced using biolistic transformation methods.
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References
Christou, P. (1992). Genetic transformation of crop plants using microprojectile bombardment. Plant J. 2,275–281.
Taylor, N. J. and Fauquet, C. M. (2002) Microparticle bombardment as a tool in plant science and agricultural biotechnology. DNA Cell Biol. 21,963–977.
Rasco-GauntS. Riley, A., Cannell, M., Barcelo, P. and Lazzeri, P. A. (2001) Procedures allowing the transformation of a range of European elite wheat (Triticum aestivum L.) varieties via particle bombardment. J. Exp. Bot. 52,865–874.
Barsby, T., Power, J. B., Freeman, J., Ingram, H. M., Livesey, N. L., Risacher, T. and Davey, M. R. (2001) Transformation of wheat, in The World Wheat Book – A History of Wheat Breeding .Lavoisier Publishing, France and USA, and Intercept Ltd., UK, pp. 1081–1103.
Bhalla, P. L., Ottenhof, H. H. and Singh, M. B. (2006). Wheat transformation – an update of recent progress. Euphytica 149,353–366.
Risacher, T. and Craze, M. (1992) WO 00/63398. Plant Transformation Method.
Cheng, M., Fry, J. E., Pang, S., Zhou, H., Hironaka, C. M., Duncan, D. R., Conner, T. W. and Wan, Y. (1997) Genetic transformation of wheat mediated by Agrobacterium tumefaciens . Plant Physiol. 115,971–980.
Cheng, M., Hu, T. C., Layton, J., Liu, C. N. and Fry, J. E. (2003) Desiccation of plant tissues post-Agrobacterium infection enhances T-DNA delivery and increases stable transformation efficiency in wheat. In Vitro Cell Dev. Biol. 39,595–604.
Uze, M., Wunn, J., Puonti-Kaerlas, J., Potrykus, I. and Sautter, C. (1997) Plasmolysis of precultured immature embryos improves Agrobacterium. -mediated gene transfer to rice (Oryza sativa L.) Plant Sci. 130,87–95.
Hood, E. E., Gelvin, S. B., Melchers, L. S. and Hoekema, A. (1993) New Agrobacterium vectors for plant transformation. Transgen. Res. 2,208–218.
Firek, S., Ozcan, S., Warner, S. A. and Draper, J. (1993) A wound-induced promoter driving npt-II expression limited to dedifferentiated cells at wound sites is sufficient to allow selection of transgenic shoots. Plant Mol. Biol. 22,129–42.
McElroy, D., Zhang, W., Cao, J. and Wu, R. (1990) Isolation of an efficient actin promoter for use in rice transformation. Plant Cell, 2,163–171.
Jefferson, R. A. (1987) Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol. Biol. Rep. 5,387–405.
Schünmann, P. H. D., Surin, B. and Waterhouse, P. M. (2003) A suite of novel promoters and terminators for plant biotechnology. II. The pPLEX series for use in monocots. Funct. Plant. Biol. 30453–460.
Herrera-Estrella, L., De Block, M., Messens, E., Hernalsteens, J.-P., Van Montagu, M. and Schell, J. (1983) Chimeric genes as dominant selectable markers in plant cells. EMBO J. 2,987–995.
Sambrook, J., Fritsch, E. F. and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, New York.
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Risacher, T., Craze, M., Bowden, S., Paul*, W., Barsby, T. (2009). Highly Efficient Agrobacterium-Mediated Transformation of Wheat Via In Planta Inoculation. In: Jones, H., Shewry, P. (eds) Transgenic Wheat, Barley and Oats. Methods in Molecular Biology™, vol 478. Humana Press. https://doi.org/10.1007/978-1-59745-379-0_7
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DOI: https://doi.org/10.1007/978-1-59745-379-0_7
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