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目的研究一氧化氮(NO)、茉莉酸甲酯(methyl jasmonate,MJ)与水杨酸(salicylic acid,SA)对肉苁蓉悬浮细胞生长和苯乙醇苷(PeGs)生物合成的影响。方法肉苁蓉细胞悬浮培养体系中加入硝普钠(sodium nitroprusside,SNP)、MJ和SA诱导子。结果低浓度外源性NO有利于悬浮细胞的生长,但对PeGs的积累作用不明显,MJ和SA有利于PeGs的积累,但抑制悬浮细胞的生长,且三者的最适作用浓度和作用时间存在差异。在细胞培养初期(0 d)加入0.05 mmol·L-1SNP,细胞对数生长初期(12 d)加入50μmol·L-1SA,对数生长中期(16 d)加入20μmol·L-1MJ,细胞干重可达对照组的1.42倍,PeGs总量可达373.52 mg·L-1,约为对照的4.17倍;松果菊苷可达21.892 mg·g-1,为对照的4.02倍。结论 3种诱导子协同作用能够显著提高肉苁蓉细胞悬浮培养体系中次生代谢产物的含量。
Objective To investigate the effects of nitric oxide (NO), methyl jasmonate (MJ) and salicylic acid (SA) on the growth of suspension cells and the biosynthesis of Petidiposide (PGE) in Cistanche deserticola. Methods Cistanche tubulosa cell suspension culture system was added sodium nitroprusside (SNP), MJ and SA inducer. Results Low concentration of exogenous NO was beneficial to the growth of suspension cells, but not obvious to PeGs accumulation. MJ and SA were beneficial to the accumulation of PeGs, but inhibited the growth of suspension cells. The optimum concentration and duration has a difference. 0.05 mmol·L-1SNP was added into the initial stage of cell culture (0 d), 50 μmol·L-1 SA was added to the logarithmic growth phase (12 d), and 20 μmol·L-1MJ was added to the logarithmic growth phase (16 d) Up to 1.42 times that of the control group. The total amount of PeGs was 373.52 mg · L-1, which was about 4.17 times that of the control, and that of echinacoside was 21.892 mg · g-1, which was 4.02 times of the control. Conclusion The synergistic effects of three kinds of elicitors can significantly increase the content of secondary metabolites in Cistanche tubulosa cell suspension culture system.