在24和72小时暴露期间,使用香芹酚以5、10、25、50、100、250和500μM的浓度进行激发过程,在西洋参毛状根培养物中确定了人参皂苷(三萜皂苷)的积累。这项研究是第一个将香芹酚用作激发子的研究。八种人参皂苷的含量,Rb1,Rb2,Rb3,Rc,Rd,Rg1、Rg2和Re,使用HPLC分析确定。此外,应用定量RT-PCR方法评估法尼基二磷酸合成酶的相对表达水平,角鲨烯合成酶,和在研究的培养物中的丹麦尼二醇合成酶基因。添加香芹酚(100μM)是增加人参皂苷产量的有效方法。所有检测到的皂苷含量和生产率最高的是,分别,在72小时激发后,20.01毫克·g-1d.w.和5.74毫克·L-1·day-1。在香芹酚的影响下,西洋参培养物中单个代谢物的产生谱发生了变化。在香芹酚处理下,大多数检查的原人参二醇衍生物的生物合成减少。相比之下,属于Rg组的人参皂苷水平升高。香芹酚对Re代谢物的作用最强,与对照相比实现7.72倍的增加。皂素Rg2,未在未经处理的样品中检测到,是在香芹酚刺激后积累的,暴露于10μM激发子72小时后达到最大浓度。
The accumulation of ginsenosides (triterpenic saponins) was determined in Panax quinquefolium hairy root cultures subjected to an elicitation process using carvacrol at 5, 10, 25, 50, 100, 250, and 500 μM concentrations during 24 and 72 h exposure. This study was the first one in which carvacrol was applied as an elicitor. The content of eight ginsenosides, Rb1, Rb2, Rb3, Rc, Rd, Rg1, Rg2, and Re, was determined using HPLC analysis. Moreover, the quantitative RT-PCR method was applied to assess the relative expression level of farnesyl diphosphate synthase, squalene synthase, and dammarenediol synthase genes in the studied cultures. The addition of carvacrol (100 μM) was an effective approach to increase the production of ginsenosides. The highest content and productivity of all detected saponins were, respectively, 20.01 mg∙g-1 d.w. and 5.74 mg∙L-1∙day-1 after 72 h elicitation. The production profile of individual metabolites in P. quinquefolium cultures changed under the influence of carvacrol. The biosynthesis of most examined protopanaxadiol derivatives was reduced under carvacrol treatment. In contrast, the levels of ginsenosides belonging to the Rg group increased. The strongest effect of carvacrol was noticed for Re metabolites, achieving a 7.72-fold increase in comparison to the control. Saponin Rg2, not detected in untreated samples, was accumulated after carvacrol stimulation, reaching its maximum concentration after 72 h exposure to 10 μM elicitor.