不同质膜稳定剂对烟草原生质体细胞壁再生的影响

The Effect of Different Cell Membrane Stabilizer on Reformation of Cell Wall from Tobacco Protoplasts

质膜稳定剂能增加完整原生质体数量,防止质膜破坏,促进原生质体细胞壁再生和细胞分裂形成细胞团。本研究拟在原生质体酶解液中分别加入CaCl2.2H2O、KH2PO4、MES 3种质膜稳定剂,或3种质膜稳定剂组合分离、提纯、培养原生质体,探讨3种质膜稳定剂对烟草原生质体细胞壁再生的影响。结果表明:3种质膜稳定剂对于解离出的原生质体的质量及原生质体细胞壁再生和原生质体细胞分裂都有一定影响,最佳浓度分别为CaCl2.2H2O 10 mM、KH2PO40.7 mM、MES 5 mM;3种质膜稳定剂组合以CaCl2.2H2O 5 mM、KH2PO40.35 mM与MES 2.5 mM的组合最佳,原生质体壁再生率达88.5%,细胞分裂率达77.5%,表明能最快促进烟草原生质体细胞壁再生及细胞分裂生长。

The cell membrane stabilizer can increase the number of intact protoplasts, so as to prevent the dis- ruption of the cell membrane, and promote the cell wall reformation from protoplast and formation of mitotic cell clusters. To investigate the effect of membrane stabilizers on the reformation of tobacco cell wall from pro- toplasts, three kinds of cell membrane stabilizers including either CaC12 .2H20, or KH2PO4 or MES was added to the enzyme solution of protoplast, or their combination was supplemented in the solution, and proto- plast was isolated, purified and cultrued. The results showed that three kinds of membrane stabilizers had certain effects on the protoplast quality, cell wall reformation and protoplast division. The optimum concentra- tion was CaC12.2H20 10 mM, KH2PO4 0.7 mM, and MES 5 mM. Among the tested membrane stabilizer combinations, the best one was CaC12.2H20 5 mM, KH2PQ 0.35 mM and MES 2.5 mM, and the cell wall reformation rate and cell division rate were as high as 88.5% and 77.5% , respectively.