084 Response of Transgenic `Royal Gala' Apple (Malus ×domestica Borkh.) Shoots, Containing the Modified Cecropin MB39 Gene, to Erwinia amylovora

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  • 1USDA-ARS, BARC-W, Bldg.010A, Beltsville, MD 20705; 2USDA-ARS, HCRL, 3420 NW Orchard Ave., Corvallis, OR 97330

Transgenic `Royal Gala' apple (Malus × domestica Borkh.) shoots were obtained by Agrobacterium-mediated gene transfer using the plasmid binary vector pGV-osm-AC with a T-DNA encoding a chimeric gene consisting of a secretory sequence from barley-amylase joined to the modified cecropin MB39 coding sequence. Shoots were placed under the control of a wound-inducible, osmotin promoter from tobacco. The integration of the cecropin MB39 gene into apple was confirmed by Southern blot analysis. The transformation efficiency was 1.5% when internodes from etiolated shoots were used as explants and 2% when leaf explants were used. Both non- and transgenic tetraploid plants were produced by treatment of leaf explants with colchicine at 25 mg·L-1, and polyploidy was confirmed by flow cytometry. Of the diploid transgenics, three of seven were significantly more resistant to Erwinia amylovora than the non-transgenic `Royal Gala' control. Also, in one instance, a tetraploid transgenic was significantly more resistant than the diploid shoot from which it was derived.

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