水双相法分离泡桐幼苗根细胞质膜的研究

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

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 0 5%～ 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 50 0和 6 3%的聚乙二醇 (PEG) 4 0 0 0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。

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 005%～01% of herbaceous plants to 06% 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.