B - Book chapters etc edited externally
Sparks, C. A. and Doherty, A. 2020. Genetic transformation of common wheat (Triticum aestivum L.) using biolistics. in: Rustgi, S. and Luo, H. (ed.) Biolistic DNA Delivery in Plants New York Humana Press Inc.
Authors | Sparks, C. A. and Doherty, A. |
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Editors | Rustgi, S. and Luo, H. |
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 pre-cultured 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. |
Keywords | Biolistics; Gene gun; Particle bombardment; Wheat; Genetic transformation; Immature embryo; Transgenic plants; DNA delivery; Tissue culture; Regeneration |
Year of Publication | 2020 |
Book title | Biolistic DNA Delivery in Plants |
Publisher | Humana Press Inc |
Place of publication | New York |
Series | Methods in Molecular Biology, vol 2124 |
ISBN | 978-1-0716-0355-0 |
Digital Object Identifier (DOI) | https://doi.org/10.1007/978-1-0716-0356-7_12 |
Funder | Biotechnology and Biological Sciences Research Council |
Funder project or code | Designing Future Wheat (DFW) [ISPG] |
Open access | Published as non-open access |
Output status | Published |
Publication dates | |
Online | 11 Apr 2020 |
Copyright license | Publisher copyright |
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