介质钙离子对白芷悬浮培养细胞及其原生质体增殖的影响

本工作首先利用《复杂络合平衡体系》计算并配制了对自由钙离子浓度具有络合平衡缓冲能力的MS液体培养基,并用电极法验证了其可靠性。在精确控制Ca~(2 )浓度条件下,利用计数法和~3H-TdR标记DNA合成的方法系统研究了不同钙离子浓度对白芷悬浮细胞及原生质体细胞增殖的影响。原生质体第一次细胞分裂所需Ca~(2 )浓度(10mmol/L)比细胞增殖所需Ca~(2 )浓度(1mmol/L)为高;不同钙离子浓度对原生质体壁再生、活力及第一次细胞分裂的作用也不一样,壁再生所需最适Ca~(2 )浓度为50mmol/L,原生质体存活以及第一次细胞分裂所需最适Ca~(2 )浓度为5—10 mmol/L,当Ca~(2 )浓度小于10~(-4)mol/L时细胞及原生质体的增殖受到很大程度的抑制,细胞死亡数目较多。结果表明介质钙离子浓度与细胞及原生质的增殖密切相关。

THE EFFECTS OF EXTRACELLULAR CALCIUM ON PROLIFERATION OF ANGELICA DAHURICA CELLS AND ITS PROTOPLASTS

We exploited the microcomputer software "MAX CHELATOR V5.6 1992" and utilized it to prepare the MS liquid medium in which the expected cocentration of free Ca2 was obtained when certain concentration of Ca2 chelator existed and proved by using Caz sensitive electrode (modle 402). The effects of various concentrations of Ca2 on the proliferation of Angelica dahurica cells and its protoplasts were studied by the methods of cell counting and 3H-TdR labeled DNA synthesis under the conditions of precise control of Ca2 concentrations. It was found that the concentration of Ca2 favorite with the proliferation of protoplasts was higher than that favorite with the proliferation of cell, the former was 10 mmol/L, where- as the later was 1 mmol/L. It was also found that various concentrations of Ca2 produced different effects on the cell wall regeneration, valibility and the first cell division of protoplasts, the favorite concentration of Ca2 for cell wall regeneration, valibility and the first cell division of protoplasts were 50 mmol/L, 10 mmol/L and 5 mmol/L, respectively. When the concentration of Ca2 was lower than 10-4 mol/L, the proliferations of cells and protoplasts were inhibited obviously and the number of died cell were increased. Those results suggested that there were close relationship between the concentration of Ca2 in the medium and the proliferations of cells and protoplasts.