Project description:The objectives of this study encompassed a thorough exploration of the potential implications of protein profiling in hairy roots, specifically focusing on optimizing and enhancing C. asiatica organ cell biofactories. In this pursuit, we categorized established C. asiatica hairy root lines according to their capacity for centelloside production, classifying them into HIGH, MID, or LOW categories. For comparative analysis, wild adventitious (Adv) roots were extracted from in vitro C. asiatica seedlings and cultivated in solid MS medium at 25°C in complete darkness, serving as control specimens. This meticulous, label-free proteomic analysis enabled the successful identification of several proteins. Our research substantially builds upon and extends the findings presented by Alcalde et al. (2022) (DOI: 10.3389/fpls.2022.1001023). In their study, distinctive morphological and metabolic variations were noted among different C. asiatica hairy root lines. Such differences are presumably attributable to the random incorporation of a selective set of genes from the T-DNA, with particular emphasis on the rol and aux genes.
Project description:Centella asiatica is an important medicinal plant with a wide range of bioactivities associated with its secondary metabolites. Using two extraction procedures, metabolomic approaches were used to investigate changes in the metabolome of C. asiatica cells treated with exogenous MeJA. GC–MS and LC–MS platforms were employed for semi-targeted and untargeted analyses, respectively. Multivariate data analyses indicated concentration-dependent changes in the metabolite profiles, indicative of the cellular response to MeJA. Annotation of biomarkers correlated with the treatment indicate differential responses in flavonoid-, phenylpropanoid (cinnamates)- and terpenoid pathways and changes in fatty acid profiles. MeJA treatment triggered the accumulation of bicyclic sesquiterpenoids (aristolochene, deoxy-capsidiol, 15-hydroxysolavetivone, solavetivone, 3-hydroxylubimin) and a tricyclic sesquiterpenoid (phytuberin), indicating the stimulatory effect of MeJA on this branch of the terpenoid pathways. In contrast, flavonoids were mostly negatively correlated with the treatment. The presence of the sesquiterpenoids in MeJA-elicited cells and other tentatively identified metabolites (abscisic acid, fatty acids, phytosterols and metabolites of shikimate–phenylpropanoid pathways) indicates that the changes in the metabolome are associated with a defensive function in response to elicitation by MeJA, rather than just the amplification of existing terpene pathways. These results provide a detailed and comprehensive picture of metabolic changes occurring in C. asiatica cells in response to MeJA elicitation and contribute to the understanding of flexible and controllable aspects of metabolic manipulation.