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目的:研究外源茉莉酸甲酯诱导后丹参毛状根中酚酸类有效成分形成的次生代谢分子机制。方法:用茉莉酸甲酯(MJ,100μmol.L-1)对丹参毛状根进行诱导实验,分别在0,6,12,24,36 h收获材料,运用实时定量PCR对丹参中迷迭香酸次生代谢途径上的关键酶基因的mRNA表达水平进行检测。同时,采用LC-MS测定各个时间点毛状根材料中迷迭香酸(RA)、丹酚酸B(LAB)及咖啡酸(CA)的含量。结果与结论:外源性信号分子MJ作用于丹参毛状根,酚酸类成分迅速积累,表现为CA,RA均在诱导后24 h达到最大值,LAB的含量36 h后达到最高。其诱导机制可能与不同程度的激活RA合成中苯丙氨酸途径上关键酶基因PAL,4CL,C4H与酪氨酸途径中TAT,HPPR基因的表达有关,基因表达在时间呈现先后性,其中,4CL与PAL的贡献度较大。这为今后更深一步的研究丹参水溶性酚酸类成分的次生代谢途径机制提供了依据。
Objective: To study the molecular mechanism of secondary metabolism of phenolic acids in hairy root of Salvia miltiorrhiza induced by methyl jasmonate. Methods: Hairy roots of Salvia miltiorrhiza Bunge were induced with methyl jasmonate (MJ, 100μmol·L-1). The materials were harvested at 0, 6, 12, 24 and 36 h respectively. Real- Acid secondary metabolic pathway of the key enzyme gene mRNA expression levels were detected. At the same time, the contents of rosmarinic acid (RA), salvianolic acid B (LAB) and caffeic acid (CA) in hairy root materials at various time points were determined by LC-MS. RESULTS AND CONCLUSION: Exogenous signaling molecule MJ acts on the hairy roots of Salvia miltiorrhiza. The contents of phenolic acids rapidly accumulate. The contents of CA and RA reached the maximum at 24 h after induction. The content of LAB reached the peak at 36 h. The induction mechanism may be related to the activation of PAL, 4CL, C4H and TAT, HPPR genes in the pathway of phenylalanine pathway in RA synthesis to varying degrees, and the expression of gene in time appears in sequence. Among them, 4CL and PAL contribute more. This provides a basis for further study on the secondary metabolic pathway of Salvia miltiorrhiza water-soluble phenolic acids.