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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.).
