摘要
研究从烟草叶片分离原生质体的若干影响因素.通过酶解法降解细胞壁释放出原生质体,利用离心和蔗糖漂浮法从粗提取液中纯化出原生质体,并通过血球计数板计数来比较制备的效果.结果表明:酶的种类、质膜稳定剂、渗透压稳定剂、pH及酶解时间是影响原生质体制备的重要因素.纤维素酶含量2%,果胶酶含量1%,2-N-吗啉乙烷磺酸(MES)浓度50mmol/L,Ca2+与甘露醇作为渗透压稳定剂,pH6左右,温度28℃,酶解时间4h时原生质体的分离效果最佳.
The factors affecting isolation and purification of protoplasts from tobacco leaves were studied. The protoplasts were released by digesting cell walls with cellulase and pectolase, and were purificated from the crude extracts by centrifugation and suspending in sucrose solution. The isolation efficiency of the protoplasts （in different condition） was decided by counting their number with globulimeter. The results showed that enzyme, cell member stabilitizer, osmotic pressure stabilizer, pH, enzymolysis time significantly affected isolation of protoplasts from leaves and in this experiment temperature pH and enzymolysis time are the most important influence factors.
出处
《安徽大学学报（自然科学版）》
CAS
北大核心
2006年第6期91-94,共4页
Journal of Anhui University(Natural Science Edition)
关键词
烟草
原生质体
分离纯化
影响因素
tobacco
protoplast
isolation and purification
influence factor
