Fertile transgenic plants from direct gene transfer to protoplasts of Lolium perenne L. and Lolium multiflorum Lam.

摘要: Summary We adapted and applied a method of direct gene transfer to protoplasts the monocots Lolium perenne L. multiflorum Lam. A total 20 clones, which produced green transgenic plants expressing transgenes GUS ( uid A) neomycin phosphotransferase II (NPT II), have been obtained in four independent transformation experiments. mild selection at early stage protoplast culture step-by-step callus strategy were employed recover transformed clones. frequency nearly 5×10 −6 (antibiotic-resistant clones/total protoplasts) was achieved, varied with plant species, cell suspension lines, plasmids used. The proportion albino regenerants antibiotic-resistant clones higher than non-transformed from control integration foreign DNA into genomes confirmed by Southern hybridization. expression reporter detected histochemical staining fluorometric assay plants, transmitted expressed progeny plants. These results demonstrate high efficiency species.