SYNERGISTIC ENHANCEMENT OF PROTOPLAST GROWTH BY OXYGENATED PERFLUOROCARBON AND PLURONIC F-68

Cell suspension-derived protoplasts of albino Petunia hybrida were gro wn for 10 d at the interface between aqueous culture medium (KM8P) and an oxygenated (10 mbar for 15 min) perfluorocarbon liquid, perfluorod ecalin. Protoplasts synthesised new cell walls and divided normally at the perfluorodecalin/culture medium interface, with a mean viability after 10 d of > 92.0%. The mean plating efficiency of protoplasts was elevated by 37% (P<0.05) following culture at the perfluorodecalin/med ium interface, but was unaltered by perfluorodecalin or oxygen separat ely. The mean plating efficiency of protoplasts cultured at the interf ace was further increased to a maximum of 52% above control, in the pr esence of oxygenated perfluorodecalin and KM8P medium supplemented wit h the non-ionic, co-polymer surfactant, Pluronic F-68 at 0.01% (w/v). These findings demonstrate the effectiveness of oxygenated perfluorode calin for promoting protoplast growth, by facilitating oxygen delivery . The finding that Pluronic F-68 further increased the plating efficie ncy of protoplasts cultured at the perfluorocarbon/aqueous interface s uggests that these agents improve growth through separate, but cumulat ive, mechanisms.