Filamentous fungi from Plantago lanceolata L. leaves: contribution to the pattern and stability of bioactive metabolites.

The aim of this study was to test contribution of plant-associated microorganism (PAMs) to metabolite stability/instability in a medicinal plant matrix. Therefore, PAM strains were isolated and identified based on relevant DNA sequences from Plantago lanceolata leaves. Sterile water extracts of P. lanceolata were incubated with the isolated strains and antioxidants (ascorbic acid (AA), and EDTA) for 15 days, and changes in the concentrations of chief bioactive constituents (aucubin, catalpol, acteoside (=verbascoside)) were quantified by capillary electrophoresis. Phenolic breakdown-products were identified by GC-MS. PAMs were identified from the genera Epicoccum, Bipolaris, Cladosporium, Leptosphaerulina, Aspergillus, Eurotium and Penicillium (pathongens, endophytes, and other species). Some fungi caused significant decomposition of the chief constituents (p<0.001). Surprisingly, some strains inhibited breakdown of acteoside (p<0.001). Meanwhile, concentration of several phenolic acids increased in fungi-infested extracts (p<0.001). Gentisic acid, 4-hydroxyphenyl acetic acid, 4-hydroxybenzoic acid and hydroxytyrosol were only present when the extract was infested with a PAM. The products are powerful antioxidants and chelators. Concentrations of phenolic acids influenced acteoside stability significantly (p<0.01), as shown by basic data-mining techniques. AA and EDTA also significantly inhibited acteoside breakdown in sterile model solutions (p<0.05). Our results suggest that the phenolic acid mixture (produced during the fungal proliferation) protected acteoside from breakdown, possibly via its antioxidant activity and metal complexing ability. It was shown that PAMs can increase or decrease the stability of chief metabolites in herbal matrices, and can significantly alter the chemical pattern of the plant matrix.