大蒜原生质体游离和纯化的研究

Studies on the Protoplast Isolation and Purification of Allium sativum L.

以大蒜胚性悬浮细胞系为研究试材,对原生质体分离纯化技术进行了研究。结果表明:2.0%纤维素酶OnozukaR-10+0.2%果胶酶Pectolgase+0.55mol/L甘露醇+5mmol/LCaCl2+5mmol/LMES的酶液,酶解5h,大蒜原生质体分离效果最好。纯化时,适当提高离心力的短时间离心有利于原生质体纯化过程中产量和活力的保持,以1000r/min离心5min,效果为好。

The conditions of isolation and purification of protoplast from somatic embryogenic cell suspension of Allium sativum L.were studied.The results showed that the optimal enzyme solution for protoplast isolation was 2.0%Cellulase Onozuka R-10＋0.2%Pectolgase＋0.55 mol/L mannitol＋5 mmol/L MES.The enzyme mixture was shaken（90 r/min）for 5 h at（25±2）℃.High yield and viability protoplasts were obtained when purified by centrifugation at 1000 r/min for 5min.