Production of kanamycin resistant rice tissues following DNA uptake into protoplasts

Rice protoplasts (Oryza sativa L. v Taipei 309) have been transformed to kanamycin resistance following uptake of pCaMVNEO induced by electroporation, PEG and PEG combined with electroporation. Protoplast-derived colonies selected on medium containing 100 g/ml of kanamycin expressed NPTII activity, and contained DNA that hybridised to a 1.0 Kb BamHI fragment of pCaMVNEO carrying the NPTII gene. Expression of the transformation frequency in relative terms (number of kanamycin resistant colonies compared to the number of colonies on kanamycin free medium) gave frequencies of 26.1%, 8.5% and 2.9% following electroporation, PEG and PEG with electroporation respectively. In absolute terms (number of kanamycin resistant colonies compared to the number of protoplasts plated) these represent frequencies of 19.9×10^–5, 9.0×10^–5 and 2.7×10^–5 for the three procedures.