Abstract
This investigation cultured Cecropia obtusifolia cells in suspension to evaluate the effect of nitrate deficiency on the growth and production of chlorogenic acid (CGA), a secondary metabolite with hypoglycemic and hypolipidemic activity that acts directly on type 2 diabetes mellitus. Using cell cultures in suspension, a kinetics time course was established with six time points and four total nitrate concentrations. The metabolites of interest were quantified by high-performance liquid chromatography (HPLC), and the metabolome was analyzed using directed and nondirected approaches. Finally, using RNA-seq methodology, the first transcript collection for C. obtusifolia was generated. HPLC analysis detected CGA at all sampling points, while metabolomic analysis confirmed the identity of CGA and of precursors involved in its biosynthesis. Transcriptome analysis identified differentially expressed genes and enzymes involved in the biosynthetic pathway of CGA. C. obtusifolia probably expresses a key enzyme with bifunctional activity, the hydroxycinnamoyl-CoA quinate hydroxycinnamoyl transferase and hydroxycinnamoyl-CoA shikimate/quinate hydroxycinnamoyl transferase (HQT/HCT), which recognizes shikimic acid or quinic acid as a substrate and incorporates either into one of the two routes responsible for CGA biosynthesis.
Highlights
Plant secondary metabolites (SMe) are compounds with a restricted presence in taxonomic groups that participate in the interaction of cells with the environment to ensure the survival of the organism and provide an evolutionary advantage in its ecosystem [1,2]
The results show that chlorogenic acid (CGA) is detectable throughout the entire kinetics time course and in all nitrate concentrations evaluated, see Supplementary Figure S2; the CGA identity was corroborated by enrichment analysis of one of the samples using the standard to perform a coelution, see Supplementary Figure S3a,b
Our results show that CGA biosynthesis increased throughout the kinetics time course
Summary
Plant secondary metabolites (SMe) are compounds with a restricted presence in taxonomic groups that participate in the interaction of cells with the environment to ensure the survival of the organism and provide an evolutionary advantage in its ecosystem [1,2]. A wide range of biological activities are exhibited, such as antimicrobial, antigerminative, toxic, and attractant properties or symbiont interaction. These features have allowed researchers to identify more than 200,000 plant. SMe, largely represented by terpenes, alkaloids, and polyphenols [3,4]. Polyphenols have received considerable attention due to their pharmacological and biomedical potential in chronic human degenerative diseases such as cataracts, macular degeneration, endothelial dysfunction, arthritis, cancer, and diabetes mellitus [5].
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