Abstract
Centella asiatica is a perrenial herb that grows in tropical regions with numerous medicinal properties mostly attributed to the presence of pentacyclic triterpenoids. Interestingly, this plant also possess a significant amount of phenylpropanoid-derived chlorogenic acids (CGAs) that have recently been reported to confer neuroprotective properties. In a biotechnological attempt to increase the biosynthesis of CGA-derivatives in cultured Centella cells, acibenzolar-S-methyl was applied as a xenobiotic inducer in combination with quinic acid and shikimic acid as precursor molecules. Applying a semi-targeted metabolomics-based approach, time and concentration studies were undertaken to evaluate the effect of the manipulation on cellular metabolism leading to CGA production. Phytochemical extracts were prepared using methanol and analyzed using a UHPLC-qTOF-MS platform. Data was processed and analyzed using multivariate data models. A total of four CGA-derivatives, annotated as trans-5-feruloylquinic acid, 3,5 di-caffeoylquinic acid, 3,5-O-dicaffeoyl-4-O-malonylquinic acid (irbic acid) and 3-caffeoyl, 5-feruloylquinic acid, were found to be upregulated by the acibenzolar-S-methyl treatment. To the best of our knowledge, this is the first report on the induction of CGA derivatives in this species. Contrary to expectations, the effects of precursor molecules on the levels of the CGAs were insignificant. However, a total of 16 metabolites, including CGA derivatives, were up-regulated by precursor treatment. Therefore, this study shows potential to biotechnologically manipulate C. asiatica cells to increase the production of these health beneficial CGAs.
Highlights
In order to enhance secondary metabolite production, the biotechnological manipulation of cultured cells with plant signaling molecules and elicitors is becoming widespread in plant research (Bourgaud et al, 2001; Hellwig et al, 2004)
A semi-targeted LC-Murashige and Skoog (MS) based metabolomic approach was employed to determine the metabolic response of C. asiatica cells to ASM as an inducer, in combination with precursors QA and ShA as well as to the latter individual precursors without a pre-induction step
The plant metabolomics approach, based on an UHPLC-qTOFMS insource collision induced dissociation (ISCID) platform, was effective to investigate the effect of exogenously applied inducer of the phenylpropanoid pathway to C. asiatica cells along with precursor molecules of chlorogenic acids (CGAs), i.e., QA and ShA
Summary
In order to enhance secondary metabolite production, the biotechnological manipulation of cultured cells with plant signaling molecules and elicitors is becoming widespread in plant research (Bourgaud et al, 2001; Hellwig et al, 2004). These systems allow for a number of plant bioactive constituents to be rapidly biosynthesized, extracted and analyzed. Exogenous application of such elicitors has been shown to induce metabolic changes by upregulating or downregulating certain defense-associated pathways in plants (Hayat et al, 2010). An attempt to increase the biosynthesis of CGAs in Centella asiatica with the exogenous application of SA proved ineffective (Ncube et al, 2016)
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