Regeneration of transgenic plants from the commercial apple cultivar Royal Gala

Abstract

A transformation system was developed for the commercial apple (Malus X domestica Borkh.) cultivar Royal Gala. Leaf pieces from in vitro-grown shoots were cocultivated for 2 days with Agrobacterium tumefaciens strain LBA4404 containing the binary vectors pKIWI105 or pKIWI110. Shoots were produced on a shooting medium containing kanamycin (50 mg·L(-1)). A 2-day incubation period on kanamycin-free medium prior to antibiotic selection enhanced the regeneration of kanamycin-resistant shoots. The majority of the kanamycin-resistant shoots also expressed GUS (β-glucuronidase) activity. The putatively transformed shoots were rooted on a medium containing kanamycin (50 mg·L(-1)). Rooted plants were established in a greenhouse, and plants transformed with pKIWI110, which contains a mutant Arabidopsis acetolactate synthase gene, were shown to be resistant to the herbicide Glean(™). Integration of T-DNA into the apple genome was confirmed by PCR and Southern hybridization analyses.