中药材2024，Vol.47Issue(9)：2146-2153,8.DOI:10.13863/j.issn1001-4454.2024.09.002
茉莉酸甲酯诱导刺人参不定根三萜皂苷合成及其关键酶基因表达研究
Biosynthesis of Triterpenoid Saponins and Key Enzymes Gene Expression from Adventitious Root of Oplopanax elatus Induced by Methyl Jasmonate
摘要
Abstract
Objective:To establish a propagation system for the tissue culture of Oplopanax elatus adventitious roots using liquid fermentation in a bioreactor,to investigate the effects of methyl jasmonate(MeJA)on the accumulation of triterpenoid total saponins and the expression of key enzyme gene involved in biosynthesis.Methods:The optimum medium hormone ratio for inducing adventitious root was selected by taking Oplopanax elatus adventitious roots as explants.The adventitious root was expanded in 10 L airlift bioreactor by suspension culture of liquid medium.The Oplopanax elatus adventitious roots was induced by 0,50,100 and 200 μmol/L methyl jas-monate(MeJA),the content of total saponins in adventitious roots was determined by vanillin and glacial acetic acid method.The ex-pression levels of key enzyme genes SQS,SQE,BAS,P450 and UGT in the triterpene saponin synthesis pathway of Oplopanax elatus adven-titious roots were determined by qPCR after 0,6 and 12 hours induction by MeJA.Results:Adding 3 mg/L IBA in the solid MS medium could induce Oplopanax elatus adventitious roots,adding 3 mg/L IBA in the liquid MS medium could achieve the suspension culture pro-liferation of Oplopanax elatus adventitious roots.The biomass increased by about 40 times after 30 days of adventitious roots cultivation with 10 L airlift bioreactor.Under the induction of 100 μmol/L MeJA,the content of triterpenoid saponins of Oplopanax elatus adventitious roots was 2.3 times that of the control group.The expression levels of SQS,SQE,BAS,P450 and UGT,which were the key genes of triter-pene saponin synthesis pathway,increased to 1.3～58.6 times of the control.Conclusion:The liquid fermentation culture and propagation system of Oplopanax elatus adventitious roots is successfully established.Exogenous addition of MeJA can induce the up-regulation of key enzyme gene expression in the pathway of triterpene saponins synthesis,thereby increasing the accumulation of triterpene saponins.This study provides a basis for elucidating the biosynthesis mechanism of triterpenoid saponins in Oplopanax elatus.关键词
刺人参/不定根/茉莉酸甲酯(MeJA)/三萜皂苷/气升式生物反应器Key words
Oplopanax elatus Nakai/Adventitious roots/Methyl jasmonate(MeJA)/Triterpenoid saponins/Airlift bioreactor分类
中医学引用本文复制引用
向文珍,张贺,王鑫,康孟莹,王宇,李玉花,吴昊..茉莉酸甲酯诱导刺人参不定根三萜皂苷合成及其关键酶基因表达研究[J].中药材,2024,47(9):2146-2153,8.基金项目
国家自然科学基金项目(U21A20243,32271823) （U21A20243,32271823）
黑龙江省自然基金项目(TD2022C001,YQ2021C003) （TD2022C001,YQ2021C003）
