The promotion of Agrobacterium mediated transformation in Atropa belladonna L. by acetosyringone

Abstract

Leaf segments from young seedlings of Atropa belladona were infected with a disarmed strain of Agrobacterium tumefaciens C 58 C 1, harboring the plasmid pGV 3850::1103 containing the coding sequences of neomycin phosphotransferase which confers resistance to the antibiotic kanamycin. Explants were infected with A. tumefaciens, and cultured overnight with and without acetosyringone (AS). The mean free quencies of explants producing calli on the two kanamycin-containing media were 21% and 71% when the explants were infected with ba/cteria cultured in the presence of AS compared with 2% in those infected with bacteria grown without AS. The transformed calli, when subcultured on kanamycin-containing media, produced transgenic plants with roots in frequencies ranging between 57% and 83%. The transgenic plants were successfully transferred to soil. Southern hybridization showed the integration of foreign DNA into the plant genome. Three plants were progeny tested, out of which two showed monogenic dominant segregation for kan^R.