Genetic transformation of common wheat (Triticum aestivum L.) using biolistics

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.

AuthorsSparks, C. A. and Doherty, A.
EditorsRustgi, 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.

KeywordsBiolistics; Gene gun; Particle bombardment; Wheat; Genetic transformation; Immature embryo; Transgenic plants; DNA delivery; Tissue culture; Regeneration
Year of Publication2020
Book titleBiolistic DNA Delivery in Plants
PublisherHumana Press Inc
Place of publicationNew York
SeriesMethods in Molecular Biology, vol 2124
ISBN978-1-0716-0355-0
Digital Object Identifier (DOI)https://doi.org/10.1007/978-1-0716-0356-7_12
FunderBiotechnology and Biological Sciences Research Council
Funder project or codeDesigning Future Wheat
Open accessPublished as non-open access
Output statusPublished
Publication dates
Online11 Apr 2020
Copyright licensePublisher copyright

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