微型结球白菜原生质体游离与纯化研究

Isolation and Purification of Proplast of the Micro-heading Chinese Cabbage

以微型结球白菜无菌苗子叶为材料,探讨酶液浓度、酶解时间、酶液渗透压、离心转数对其原生质体分离效果的影响以及蔗糖浓度对纯化结果的影响。结果表明:以1.0%CellulaseR-10,0.08%Pectinase,0.4 mol/L甘露醇,酶解6 h,第1次离心转速1 200 r/min,24%蔗糖的条件组合最佳,原生质体产量为2.10×107个/g(FW),活力达91.20%。

Using the micro-heading Chinese cabbage cotyledon-free vaccine as material, the enzyme concentration, hydrol- ysis time, enzyme solution osmotic pressure, centrifugal rotation of its protoplast isolation as well as the effect of sucrose concentration on the purification of the outcome were explored. The results showed that：the best combination was the 1. 0% Cellulase R-10, 0. 08o/oo Pectinase, 0. 4 mol/L rnannitol, hydrolysis 6 h, the first centrifugal speed of 1 200 r/min, and 24% sucrose,the production of protoplast was 2. 10×10^7 Unit/g（FW）,activity was 91.20%.