Articles
THE USE OF PHENOTYPIC MARKERS TO IDENTIFY BRASSICA GENOTYPES FOR ROUTINE HIGH THROUGHPUT AGROBACTERIUM-MEDIATED TRANSFORMATION
Article number
706_28
Pages
239 – 246
Language
English
Abstract
Plant transformation is an important research tool for testing gene function in addition to producing commercial GM crops.
For gene testing to become a routine procedure it is important to identify easy-to-transform genotypes, with reproducible and reliable transformation efficiencies that respond well when handled by different users.
In Brassica oleracea we have identified phenotypic markers associated with transformation potential.
This led to the identification of a high throughput B. oleracea genotype, now used by a number of research groups for routine transformation.
This paper describes the successful application of these phenotypic markers in the screening of 81 genotypes of Brassica napus, to identify candidates for routine transformation.
Genotypes with high shoot regeneration potential that regenerate via a callus phase, critically in the absence of tissue culture blackening, and with high Agrobacterium susceptibility resulted in higher transformation efficiencies.
Transformation rates of up to 17% were observed (based on the percentage of explants giving rise to independent transgenic plantlets). This study is being carried out as part of BRACT (Biotechnology Resources for Arable Crop Transformation, www.bract.org).
For gene testing to become a routine procedure it is important to identify easy-to-transform genotypes, with reproducible and reliable transformation efficiencies that respond well when handled by different users.
In Brassica oleracea we have identified phenotypic markers associated with transformation potential.
This led to the identification of a high throughput B. oleracea genotype, now used by a number of research groups for routine transformation.
This paper describes the successful application of these phenotypic markers in the screening of 81 genotypes of Brassica napus, to identify candidates for routine transformation.
Genotypes with high shoot regeneration potential that regenerate via a callus phase, critically in the absence of tissue culture blackening, and with high Agrobacterium susceptibility resulted in higher transformation efficiencies.
Transformation rates of up to 17% were observed (based on the percentage of explants giving rise to independent transgenic plantlets). This study is being carried out as part of BRACT (Biotechnology Resources for Arable Crop Transformation, www.bract.org).
Authors
P.A.C. Sparrow, P.J. Dale, J.W. Snape, J.A. Irwin
Keywords
callus, tissue culture blackening, shoot regeneration, susceptibility, oil seed rape, canola
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