Study on Protoplast Isolation conditions of Rosa Chinensis

Plasmolysis and frozen methods were used to pre-treat young leaves of Rosa chinensis. Protoplast was isolated by mixed enzyme solution of cellulase, macerozyme and mannitol. The factors of protoplast yield and viability were investigated. The results showed that Cellulase R-10 had extremely significant influence on the isolation of proto- plasts, and the suitable enzyme liquid combination was: cellulase R-10 1.5%, macerozyme 0.5%, mannitol 0.6 mol/L, with 12 h of enzymolysis time. Plasmolysis enhanced protoplast yield but reduced protoplast viability. The frozen pre-treatment also led to an increment of protoplast yield, and protoplast viability reached the maximum after 12 h frozen.