一氧化氮、茉莉酸甲酯与水杨酸对肉苁蓉悬浮细胞生长及苯乙醇苷生物合成的影响

Effects of Nitric Oxide,Methyl Jasmonate and Salicylic Acid on the Phenylethanoid Glycosides Production and Cell Growth in Suspension Cultures of Cistanche deserticola

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 study the effects of nitric oxide（NO）,methyl jasmonate（MJ） and salicylic acid（SA） on the phenylethanoid glycoside formation and growth of suspension cultures of Cistanche deserticola.METHODS Suspension cell culture of C.deserticola was established and treatment using sodium nitroprusside（As NO donor）,MJ and SA was carried out.Content determination of echinacoside and total phenylethanoid glycosides（PeGs） was performed using RP-HPLC and UV-spectrophotometer,respectively.RESULTS NO could not enhance the accumulation of PeGs,but low concentration can promote cells growth.MJ and SA greatly enhanced the accumulation of PeGs and restricted the growth of cells.Besides,their optimal elicitation dosage and addition time were different.Maximal PeGs production was achieved with combined treatment using three elicitors at different cell growth stages（0.05 mmol·L-1 sodium nitroprusside added on day 0,50 μmol·L-1 SA on 12nd day and 20 μmol·L-1 MJ added on 16th day）.Under this combined treatment,dry weight,total PeGs and echinacoside yield was 1.42,4.17 and 4.02 times of that in untreated cells.CONCLUSION Three elicitors had different effects on the accumulation and growth of suspension cell culture,and the combined treatment of three elicitors resulted in significant increase of the yield of secondary metabolites in C.deserticola suspension cells.