Abstract
Plant cell cultures represent important model systems to understand metabolism and its modulation by regulatory factors. Even in controlled conditions, cell metabolism is highly dynamic and can be fully characterized only by time course experiments. Here, we show that statistical analysis of this type of data gains power if it moves to approaches able to compare the ‘trends’ of the different metabolites. In particular, we show how generalized additive models can be used to model the time-dependent profile of anthocyanin synthesis in grapevine cell suspension cultures (Vitis vinifera cv. Gamay), following treatment with 100 μm methyl jasmonate. The sampling was performed daily for 20 days of culturing following elicitation at day 5. All samples were analyzed by UPLC-MS/MS for the identification and quantification of fifteen anthocyanin compounds. The models confirmed the separation in the anthocyanin biosynthetic pathway between delphinidin-based and cyanidin-based compounds, showing that methyl jasmonate modulates the anthocyanin concentration profiles. Our results clearly indicate that the combination of high-throughput metabolomics and state of the art statistical modeling is a powerful approach to study plant metabolism. This approach is expected to gain popularity due to the growing availability of low-cost high-throughput ‘omic’ assays.
Highlights
Plant cell cultures are model systems to dissect plant biosynthetic pathways and to investigate which factors influence metabolism
The treatment of plant cell cultures with biotic or abiotic elicitors often induces the production of secondary metabolites, which in natural conditions are synthesized in response to pathogen attack or environmental stimuli.[1]
A variety of molecules can act as elicitor,[2] among them, jasmonic acid (JA) and methyl jasmonate (MeJA) own a place of merit
Summary
Plant cell cultures are model systems to dissect plant biosynthetic pathways and to investigate which factors influence metabolism. JA and MeJA are fatty acid derived plant hormones that occur ubiquitously in the plant kingdom and act as regulators of defense responses and other plant processes They trigger a signal transduction chain, which in turn activates multiple secondary biosynthetic pathways.[3] When exogenously applied to plant cell cultures in the concentration range from 10 to 200, MeJA μM leads to an increased production of secondary metabolites including alkaloids, volatile terpenes and anthocyanins.[4,5,6] Anthocyanins are a class of phenylpropanoids responsible of the red, blue or purple color of grapes and of wine. Water was used as weak eluting solvent (A) and methanol as strong eluting solvent (B); formic acid 5% v/v was used as additive in both
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