Abstract
Brachypodium distachyon is an attractive genomics and biological model system for grass research. Recently, the complete annotated genome sequence of the diploid line Bd21 has been released. Genetic transformation technologies are critical for the discovery and validation of gene function in Brachypodium. Here, we describe an efficient procedure enabling the Agrobacterium-mediated transformation of a range of diploid and polyploid genotypes of Brachypodium. The procedure relies on the transformation of compact embryogenic calli derived from immature embryos using either chemical selection alone or a combination of chemical and visual screening of transformed tissues and plants. Transformation efficiencies of around 20% can routinely be achieved using this protocol. In the context of the BrachyTAG programme (BrachyTAG.org), this procedure made possible the mass production of Bd21T-DNA mutant plant lines.
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Acknowledgements
The authors would like to thank Barbara Worland (John Innes Centre, UK) for valuable technical assistance as well as John Doonan (John Innes Centre, UK), Magda Opanowicz (John Innes Centre, UK), and John Vogel (USDA-ARS, Albany, CA, USA) for kindly providing Brachypodium polyploid seed stocks. This work was supported by the UK Biotechnology and Biological Sciences Research Council (BBSRC).
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Thole, V., Vain, P. (2012). Agrobacterium-Mediated Transformation of Brachypodium distachyon . In: Dunwell, J., Wetten, A. (eds) Transgenic Plants. Methods in Molecular Biology, vol 847. Humana Press. https://doi.org/10.1007/978-1-61779-558-9_12
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DOI: https://doi.org/10.1007/978-1-61779-558-9_12
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