Genetic transformation of wheat via Agrobacterium-mediated DNA delivery

Caroline A. Sparks, Angela Doherty, Huw D Jones

Research output: Chapter in Book/Report/Conference proceedingChapter

25 Citations (SciVal)

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.).

Original languageEnglish
Title of host publicationCereal Genomics
Subtitle of host publicationMethods and Protocols
EditorsRobert J. Henry, Agnelo Furtado
PublisherSpringer Nature
Pages235-250
Number of pages16
Volume1099
ISBN (Electronic)978-1-62703-715-0
ISBN (Print)978-1-62703-714-3
DOIs
Publication statusPublished - 2014

Publication series

NameMethods in Molecular Biology
PublisherHumana press
Volume1099
ISSN (Print)1064-3745

Keywords

  • Agrobacterium
  • Agrobacterium tumefaciens
  • DNA, Bacterial
  • Gene Transfer Techniques
  • Plants, Genetically Modified
  • Seeds
  • Transformation, Genetic
  • Triticum

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