TRANSFORMATION OF HAPLOID DATURA-INNOXIA PROTOPLASTS AND ANALYSIS OF THE PLASMID INTEGRATION PATTERN IN REGENERATED TRANSGENIC PLANTS

We developed a highly efficient transformation protocol for the PEG-me diated direct transfer of plasmid DNA into protoplasts of haploid Datu ra innoxia. Vectors harbouring a neomycin phosphotransferase II gene o r a hygromycin B phosphotransferase gene under the control of differen t promoters were used in the transformation experiments. Various amoun ts of plasmid DNA were applied without any carrier DNA to show the dir ect influence of the plasmid DNA concentration on the transformation e fficiency. Approximately 95 % of the selected calli were regenerated t o plants; 20 % of them remained haploid. Total DNA of different transg enic plants was analysed with regard to the integration pattern of the plasmid DNA. Plants carrying only one or two copies of the vector DNA were observed as well as individuals with multi-copy integration (up to ten or more copies).