积雪草(L.)Urban是一种重要的药典植物,不仅用于医学,而且用于美容。建立了积雪草搅动的枝条培养物以研究乙烯利的影响,茉莉酸甲酯,l-苯丙氨酸(Eth50µM,MeJa50µM,L-Phe2.4g/L培养基,分别;补充的七个变体)对次生代谢产物的积累:主要的积雪草苷(积雪草苷和羟基积雪草苷)和选定的酚酸,和生物质中的类黄酮。在补充后2天和6天收获微芽。通过UPLC-MS/MS(centellosides)和HPLC-DAD(酚类)分析甲醇提取物中的次级代谢物。与参考文化相比,单个次生代谢产物的浓度增加如下:centellosides高达5.6倍(积雪草苷),酚酸高达122倍(对香豆酸)和类黄酮高达22.4倍(kaempherol)。对于补充的不同变体,观察到单个化合物的最高产量增加。变体C(50µMMeJa),最适合百叶皂甙和类黄酮积累,选择用于生物反应器的实验。生物反应器Plantform™,与RITA®系统和搅拌培养物相比,在积雪草芽培养物中,次生代谢产物的生产似乎是最有利的。酚酸,类黄酮,Centelloside,Plantform™系统中的总次级代谢产物生产力是1.8倍,1.7倍,2.8折,2.1折,分别,高于MeJa激发的激越文化,4.3倍,7.3折,12.2折,7.2倍,分别,高于对照搅动培养物。
Centella asiatica (L.) Urban is an important pharmacopoeial plant used not only in medicine but also in cosmetology. C. asiatica agitated shoot cultures were established to study the influence of ethephon, methyl jasmonate, l-phenylalanine (Eth 50 µM, MeJa 50 µM, L-Phe 2.4 g/L of medium, respectively; seven variants of the supplementation) on the accumulation of secondary metabolites: the main centellosides (asiaticoside and madecassoside) and selected phenolic acids, and flavonoids in the biomass. Microshoots were harvested two and six days after the supplementation. Secondary metabolites were analyzed in methanolic extracts by UPLC-MS/MS (centellosides) and by HPLC-DAD (phenolics). In comparison with the reference cultures, the concentrations of individual secondary metabolites increased as follows: centellosides up to 5.6-fold (asiaticoside), phenolic acids up to 122-fold (p-coumaric acid) and flavonoids up to 22.4-fold (kaempherol). The highest production increase of individual compounds was observed for different variants of supplementation. Variant C (50 µM MeJa), the most optimal for centellosides and flavonoid accumulation, was selected for the experiment with bioreactors. Bioreactor Plantform™, compared to RITA® system and agitated cultures, appeared to be the most advantageous for secondary metabolites production in C. asiatica shoot cultures. The phenolic acid, flavonoid, centelloside, and total secondary metabolite productivity in Plantform™ system is 1.8-fold, 1.7-fold, 2.8-fold, 2.1-fold, respectively, higher than in MeJa elicitated agitated cultures, and 4.3-fold, 7.3-fold, 12.2-fold, 7.2-fold, respectively, higher than in control agitated cultures.