Abstract:

The basic work for the system establishment about protoplast culture and somatic hybrids is to formulate efficient protoplast isolation and protoplast-culturing system. The conditions of isolation and purification of protoplast from somatic callus of Allium cepa were studied. The results showed that the optimal enzyme solution for protoplast isolation was 2.0% cellulase Onozuka R-10+ 0.1% pectolgase Y-23+0.5% macerozyme R-10+0.60 mol/L mannitol +0.1% MES and CPW solution. The enzyme mixture was for 5 h at (25±2)℃. High yield and viability protoplasts were obtained when purified by centrifugation at 600 r/min for 5 min and the callus for 12 days.