杠柳细胞悬浮培养体系中蔗糖浓度与氮源的考察

Studies on Sucrose Concentration and Nitrogen Source in the System of Cell Suspension Culture of Periploca sepium Bunge

目的考察MS培养基中蔗糖浓度和氨态氮与硝态氮的比例(NH4+∶NO3-)以及初始总氮量对杠柳悬浮细胞生长和杠柳毒苷与4-甲氧基水杨醛积累的影响。方法以3%蔗糖浓度为对照,设置1%、2%、4%、5%浓度梯度;设置NH4+-NO3-分别为0∶1、1∶0、2∶1、1∶1、1∶2共5个处理;总氮含量以MS基本培养基(总氮含量60mmol·L-1)为对照,实验组总氮含量分别为对照组的1/4、1/2和2倍。结果 4%的蔗糖最有利于杠柳细胞生长,5%的蔗糖有助于杠柳毒苷的合成,3%的蔗糖有利于4-甲氧基水杨醛的合成,4%的蔗糖最有利于杠柳毒苷和4-甲氧基水杨醛产量的提高。NH4+-NO3-为1∶2有助于细胞生长和杠柳毒苷和4-甲氧基水杨醛的合成。总氮含量为N(60mmol·L-1)时细胞生长最好;总氮含量为1/2N(30mmol·L-1)时杠柳毒苷和4-甲氧基水杨醛含量最高,且产量也最高。结论蔗糖浓度、氮源中氨态氮与硝态氮比例以及总氮含量的改变均可显著影响杠柳细胞生长和杠柳毒苷与4-甲氧基水杨醛的积累。

OBJECTIVE To study the effects of sucrose concentration,the proportion of NH＋4and NO-3 and initial total nitrogen content in the MS culture medium on the growth of suspension cell and accumulation of periplocin and 4-methoxysalicylaldehyde.METHODS The sucrose concentration gradient was set up as 1%,2%,4% and 5%,with 3% sucrose as control.The proportions of NH＋4 and NO-3 were 0∶1,1∶0,2∶1,1∶1 and 1∶2 in five groups.Total nitrogen content of control group was 60 mmol·L-1 in MS minimal medium.The total nitrogen contents of experimental groups were 1/4,1/2 and 2 times as that of the control.RESULTS 4% sucrose was in favor of the growth of cell.5% sucrose benefited synthetizing the ingredient of periplocin and 3% sucrose benefited synthetizing the ingredient of 4-methoxysalicylaldehyde.4% sucrose was in favor of the increase of the output of periplocin and 4-methoxysalicylaldehyde.It was beneficial to increase the growth of cell and compose the ingredients of periplocin and 4-methoxysalicylaldehyde while the proportion of NH＋4 and NO-3 was 1 to 2.60 mmol·L-1 total nitrogen content was good for the growth of cell.30 mmol·L-1 total nitrogen content improved the content and output of the ingredient of periplocin and 4-methoxysalicylaldehyde.CONCLUSION The sucrose concentration,the proportion of NH＋4 and NO-3,and total nitrogen content can affect on the growth of cell and the accumulation of the ingredients of periplocin and 4-methoxysalicylaldehyde.