Induction of promising antibacterial prenylated isoflavonoids from different subclasses by sequential elicitation of...

Elicited soybean (Glycine max (L.) Merrill, Leguminosae) seedlings can produce prenylated isoflavonoids from different subclasses, namely pterocarpans (glyceollins), isoflavones and coumestans. These prenylated isoflavonoids serve as defence compounds and can possess antimicrobial activity. Recently, we showed that priming with reactive oxygen species (ROS) specifically stimulated the production of glyceollins in Rhizopus spp.-elicited soybean seedlings (ROS + R). In this study, we achieved diversification of the inducible subclasses of prenylated isoflavonoids in soybean, by additional stimulation of two prenylated isoflavones and one prenylated coumestan. This was achieved by using a combination of the relatively long-lived ROS representative, H₂O₂, with AgNO₃ prior to microbial elicitation. Microbial elicitation was performed with a live preparation of either a phytopathogenic fungus, Rhizopus spp. or a symbiotic bacterium, Bacillus subtilis. B. subtilis induced 30% more prenylated isoflavones than Rhizopus spp. in (H₂O₂ + AgNO₃)-treated seedlings, without significantly compromising the total levels of glyceollins, compared to (ROS + R)-treated seedlings. The most abundant prenylated isoflavone induced was 6-prenyl daidzein, which constituted 60% of the total isoflavones. The prenylated coumestan, phaseol, was also induced in the (H₂O₂ + AgNO₃)-treated and microbially elicited seedlings. Based on previously developed quantitative structure-activity relationship (QSAR) models, 6-prenyl daidzein and phaseol were predicted to be promising antibacterials. Overall, we show that treatment with H₂O₂ and AgNO₃ prior to microbial elicitation leads to the production of promising antibacterial isoflavonoids from different subclasses. Extracts rich in prenylated isoflavonoids may potentially be applied as natural antimicrobial agents.