Abstract
The following protocol describes the genetic transformation of wheat using the BioRad PDS/1000-He particle delivery system. Immature embryos are isolated 12–16 days post-anthesis, the embryonic axis is removed, and the immature scutella are precultured for 1–2 days prior to particle bombardment. Gold particles are coated with plasmid DNA containing the gene(s) of interest plus a selectable marker gene, in this instance bar (bialaphos resistance), and are fired into the cells to deliver the DNA. Subsequent tissue culture and regeneration steps allow recovery of plantlets, assisted by the inclusion of PPT (phosphinothricin tripeptide), the active ingredient of glufosinate-ammonium containing herbicides, to help select transformants. This updated method introduces selection earlier in the regeneration process which provides a shortened protocol while maintaining high transformation efficiencies.
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Acknowledgments
Caroline Sparks and Angela Doherty (Rothamsted Research) receive grant-aided support from the Biotechnology and Biological Sciences Research Council (BBSRC) Designing Future Wheat project [BB/P016855/1].
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Sparks, C.A., Doherty, A. (2020). Genetic Transformation of Common Wheat (Triticum aestivum L.) Using Biolistics. In: Rustgi, S., Luo, H. (eds) Biolistic DNA Delivery in Plants. Methods in Molecular Biology, vol 2124. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0356-7_12
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DOI: https://doi.org/10.1007/978-1-0716-0356-7_12
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Publisher Name: Humana, New York, NY
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Online ISBN: 978-1-0716-0356-7
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