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Protoplasts prepared from the aseptic leaves of Rose Supreme in Gladiolus hybridus Hort. were cultured. The yield of protoplasts with vigour was the highest under the following conditions: 1.5% cellulase Onzuka RS, 0.5% pectinase Y-23, 0.6 mol· L~(-1) mannitol, pH 5.8, digestion time 2 h, and temperature 27℃. After isolation and purification, protoplasts were cultured in solid and liquid combined medium, which included KM8P, 0.6 mol· L~(-1) mannitol, 1.0 mg· L~(-1) 2, 4-D, 0.2 mg· L~(-1) NAA and 0.2 mg· L~(-1) KT. The division of the protoplasts first emerged in medium after cultured 4 days, and emerged only a few times.
The yield of protoplasts with vigour was the highest under the following conditions: 1.5% cellulase Onzuka RS, 0.5% pectinase Y-23, 0.6 mol · L - ( -1) mannitol, pH 5.8, digestion time 2 h, and temperature 27 ° C. After isolation and purification, protoplasts were cultured in solid and liquid combined medium, which included KM8P, 0.6 mol · L -1 mannitol, 1.0 mg · L -1,2,4-D, 0.2 mg · L -1 NAA and 0.2 mg · L -1 KT. The division of the protoplasts first emerged in medium after cultured 4 days, and Viewed only a few times.