Articles
TRANSFORMATION OF APPLE CULTIVARS WITH T4-LYSOZYME-GENE TO INCREASE FIRE BLIGHT RESISTANCE
Genes encoding lytic proteins are a potential source of antibacterial resistance genes for use in plants.
Transgenic potato plants expressing a bacteriophage T4 lysozyme gene have increased resistance to the bacterial pathogen Erwinia carotovora subsp. atroseptica (Düring et al., 1993) and T4 lysozyme has in vitro activity against E. amylovora.
A transformation system was developed for several apple cultivars: ‘Pinova’, ‘Pirol’, ‘Pilot’, ‘Pingo’, ‘Remo’, ‘Elstar’ and ‘Liberty’. Leaf pieces of in vitro grown microshoots were inoculated with Agrobacterium tumefaciens strain EHA 105 containing the plasmid binary vector pSR 8–36 bearing a chimeric lysozyme gene naturally occurring in bacteriophage T4 and previously used for transformation of potato for increased resistance to Erwinia carotovora atroseptica (Porsch et al., 1998). 8 weeks after inoculation, regenerants were produced on leaf explants cultivated on a selective medium containing kanamycin and cefotaxime.
Regenerants were placed on several media amended with 0, 50 or 100 mg/l paromomycin.
Transformation was confirmed by an ELISA for the NPT II protein and by PCR tests to confirm the presence of the lysozyme gene.
