Abstract: Taking Cymbidium hybridum leaves and roots as material, the effects of enzyme combination, shaking speed, enzymolysis time, enzymolysis temperature, and the ratio of materials enzyme on protoplast isolation were studied. The results showed that：after 90 min pretreatment for plasmolysis, protoplast yield from leaves was the highest using mixed enzyme (2.0% cellulose + 0.1% pectinase + 0.2% macerozyme + 0.2% driselase) with shaking speed 50 r·min^-1 at 28 ℃ for 6 h. And protoplast yield from roots was the highest using mixed enzyme (2.0% cellulose + 0.3% pectinase + 0.1% macerozyme + 0.1% driselase) under the same conditions.

Key words: Cymbidium hybridum, protoplasts, isolation

摘要： 以大花蕙兰的叶片、根为试材，分析不同酶液组合、摇床转速、酶解温度、材料酶液比、酶解时间等因素对其原生质体分离的影响。大花蕙兰叶片、根经过90 min质壁分离预处理后，在温度28 ℃，于摇床上50 r·min^-1振荡酶解6 h，叶、根分别在酶解液为纤维素酶2.0%＋果胶酶0.1%＋离析酶0.2%＋崩溃酶0.2%和纤维素酶2.0%＋果胶酶0.3%＋离析酶0.1%＋崩溃酶0.1%中得到最高产量的原生质体。

关键词: 大花蕙兰, 原生质体, 分离