咖啡叶片原生质体分离条件的研究

摘要

Hydrolysis parameters of protoplast with high yield and activity were determined for C canephora The effects of enzyme combinations, mannitol concentration in the mixed enzyme solution, enzymolysis time and pretreatment time on protoplast isolations were studied. The results are as follow: Leaves grown for 10?5 days were cut and then treated with enzymes consisting of 2% cellulase, 0.5% pectinase, 0.3% macerozyme and 0.7 mol/L mannitol and are sustainied for 20 hours after 24-hour dark pretreatment, its protoplast yield and activity was the highest.%以中粒种咖啡(Coffea canephora)叶片组织为材料,研究酶液组合、酶解时间、酶液渗透压及预处理措施对其原生质体分离效果的影响,以期确定能够酶解出高数量且高活力的原生质体的酶解条件.结果表明:获得有活力原生质体的最佳酶液组合为2％纤维素酶+0.5％果胶酶+0.3％离析酶,同时酶解时间为20h、酶液渗透压即甘露醇浓度为0.7 mol/L,预处理措施为黑暗24 h的咖啡叶片组织得到8.1×105个/g、活力达到73％的原生质体.