Isolation of Protoplast and Ion Channel Recording in Plasma Membrane of Suspension Cells of Populus euphratica

The authors used suspension cells of Populus euphratica to isolate protoplast in the present study. Protoplasts were successfully obtained after 4 hours incubation in enzyme solution containing 1.0% cellulase "onozuka" R-10, 0.01% pectolyase Y-23, 0.15% macerozyme R-10 and 0.1% hemicellulase at 25℃. Outward and inward single channels in plasma membrane were observed using cell-attached recording of patch-clamp technique. In this study, single channel records showed that more than one species of channel were obtained. These attempts in protoplast isolation and ion channel recording offers the opportunity to characterize cellular mechanisms of salt tolerance in tree species.