| 烟草原生质体的分离纯化 |
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| 引用本文： | 江力,孔小卫,吴晓杰,李少松.烟草原生质体的分离纯化[J].安徽大学学报(自然科学版),2006,30(6):91-94. |
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| 作者姓名： | 江力 孔小卫 吴晓杰 李少松 |
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| 作者单位： | 合肥工业大学,生物与食品工程学院,安徽,合肥,230009;安徽大学,生命科学学院,安徽,合肥,230039 |
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| 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.
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| 关 键 词： | 烟草 原生质体 分离纯化 影响因素 |
| 文章编号： | 1000-2162（2006）06-0091-04 |
| 收稿时间： | 2006-07-13 |
| 修稿时间： | 2006-07-13 |
Isolation and purification of protoplasts from tobacco leaves |
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| JIANG Li,KONG Xiao-wei,WU Xiao-jie,LI Shao-song.Isolation and purification of protoplasts from tobacco leaves[J].Journal of Anhui University(Natural Sciences),2006,30(6):91-94. |
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| Authors: | JIANG Li KONG Xiao-wei WU Xiao-jie LI Shao-song |
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| Institution: | 1. School of Biotechnology and Food Engineering, Hefei University of Technology, Hefei 230009, China; 2. School of Life Science, Anhui University, Hefei 230039, China |
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| Abstract: | 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. |
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| Keywords: | tobacco protoplast isolation and purification influence factor |
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