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林业科学 ›› 2001, Vol. 37 ›› Issue (1): 23-27.doi: 10.11707/j.1001-7488.20010104

• 论文及研究报告 • 上一篇    下一篇

水双相法分离泡桐幼苗根细胞质膜的研究

洪剑明 贾慧君 郑槐明   

  1. 首都师范大学生物系,北京100037;中国林业科学研究院,北京100091;中国林学会,北京100091
  • 收稿日期:2000-03-16 修回日期:1900-01-01 出版日期:2001-01-25 发布日期:2001-01-25

STUDIES ON SEPARATING PLASMA MEMBRANE FROM ROOTS OF PAULOWNIA SEEDLING BY AQUEOUS TWO-PHASE PARTITIONING

Hong Jianming,Jia Huijun,Zheng Huaiming   

  1. Department of Biology, Capital Normal University Beijing100037;The Research Institute of Forestry, CAF Beijing100091;The Chinese Society of Forestry Beijing100091
  • Received:2000-03-16 Revised:1900-01-01 Online:2001-01-25 Published:2001-01-25

摘要:

通过对草本植物水双相法分离细胞质膜技术的改进,建立起木本植物泡桐根细胞质膜的分离纯化方法。其改进主要有:在匀浆介质中添加了抗坏血酸,加大了聚乙烯聚吡咯烷酮的用量,由草本植物一般用量0.05%~0.1%增至0.6%;对第1次双相分离后下相液中剩余的质膜进行了再回收。通过对质膜和细胞器各项标记酶指标的测定,确定了6.3%的葡聚糖(Dextran)T-500和6.3%的聚乙二醇(PEG)4000为主组成的两相系统分离的根细胞质膜具有较高的收率和纯度。该方法的建立为在细胞和分子水平上研究林木根系对离子吸收机理与生长发育的关系打下了基础。

关键词: 泡桐, 水双相分离法, 细胞质膜, 标记酶, 氧化还原活性

Abstract:

This paper reports the method of partitioning and purifying plasma membrane from roots of woody plants of Paulownia by improving the method of preparing plasma membrane by aqueous two-phase partitioning in herbaceous plants. The main improvements are adding ascorbic acid and increasing an amount of insoluble pvpp(polyvinylpolypyrrolidone) from 0.05%~0.1% of herbaceous plants to 0.6% in this homogenization medium, and then again collecting the plasma membrane from the lower phase after the first aqueous two-phase partitioning. Through determining the different kinds of marker enzymes of plasma membranes and organelles, that plasma membrane preparations of high yield and purity were easily obtained in aqueous polymer two-phase systems mainly constituted by 6.3% Dextran T-500 and 6.3% polyethylene glycol 4000. This method lays the foundations for the studies on the relation between the principles of roots absorption ions and growth, development of a woody plant at cellular and molecular level.

Key words: Paulownia, Aqueous two-phase partitioning, Plasma membrane, Marker enzyme, Activity of oxidation-reduction