Genetic transformation of wheat via Agrobacterium-mediated DNA delivery

B - Book chapters etc edited externally

Sparks, C. A., Doherty, A. and Jones, H. D. 2014. Genetic transformation of wheat via Agrobacterium-mediated DNA delivery. in: Henry, R. J. and Furtado, A. (ed.) Cereal genomics: methods and protocols. Methods in Molecular Biology, vol 1099 Springer, New York. pp. 235-250

AuthorsSparks, C. A., Doherty, A. and Jones, H. D.
EditorsHenry, R. J. and Furtado, A.
Abstract

The method described involves an initial incubation of wheat immature embryos in a liquid culture of Agrobacterium tumefaciens. The Agrobacterium strain is engineered to contain a binary vector with a gene of interest and a selectable marker gene placed between the T-DNA borders; the T-DNA is the region transferred to the plant cells, thus harnessing the bacterium’s natural ability to deliver specific DNA into host cells. Following the initial inoculation with the Agrobacterium, the embryos are co-cultivated for several days after which the Agrobacterium is selectively destroyed using an antibiotic. Tissue culture of the embryos on plant media with a correct balance of hormones allows embryogenic callus formation followed by regeneration of plantlets, and in the later stages of tissue culture a selectable marker (herbicide) is included to minimize the incidence of non-transformed plants. This protocol has been used successfully to generate transformed plants of a wide range of wheat varieties, both spring and winter bread wheats (T. aestivum L.) and durum wheats (T. turgidum L.).

KeywordsWheat ; Agrobacterium tumefaciens; Transformation; T-DNA; Cocultivation ; Immature embryo ; Transgenic plants
Page range235-250
Year of Publication2014
Book titleCereal genomics: methods and protocols. Methods in Molecular Biology, vol 1099
PublisherSpringer, New York
ISBN9781627037150
Digital Object Identifier (DOI)doi:10.1007/978-1-62703-715-0_19
Open accessPublished as non-open access
Funder project or codeWheat
Other filePages from 2014_Book_CerealGenomics.pdf
Output statusPublished
Publication dates
Online31 Oct 2013
Copyright licensePublisher copyright

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