PRELIMINARY RESULTS TO ESTABLISH THE DAAO SYSTEM AS AN ALTERNATIVE SELECTION STRATEGY ON APPLE

Selection after transformation is an essential step to detect transgenic plant cells effectively.
A new selection system is available using a D-amino acid oxidase (DAAO) encoded by the marker gene dao1. This selection system is based on selection with D-amino acids and allows the selection as well as the elimination of transgenic plants by the same marker.
In this study we tested the DAAO system in apple transformation.
Based on regeneration experiments on apple leaf explants using different concentrations of D-serine and D-alanine, it was found that concentrations higher than 2 mM are toxic for plant cells and inhibit shoot regeneration of non-transgenic cells.
Using a binary vector that allows selection both with D-serine and kanamycin, we obtained only regenerates on medium containing kanamycin, although leaf explants were vital and covered by callus on medium containing 2 mM D-serine.

XII EUCARPIA Symposium on Fruit Breeding and Genetics

C. Hättasch, M.-V. Hanke, H. Flachowsky

transgenic apple, D-amino acid oxidase, dao1, D-amino acids, Agrobacterium tumefaciens, selection, elimination

https://doi.org/10.17660/ActaHortic.2009.814.39