LAUSR

Upon inoculation, common beans immune to Pseudomonas savastanoi pv phaseolicola race 5 (R5) accumulate resveratrol, a phytoalexin. How resveratrol acts upon on this bacterium is not known, although in animal pathogenic bacteria in vitro resveratrol reduces ATPase (ATP = adenosine triphosphate) activity, cellular motility, quorum sensing, and biofilm formation. In this study, mass spectrometry was used to monitor the effects of resveratrol on R5. R5 responded by producing multidrug efflux proteins to pump resveratrol out of cells. Changes in R5 enzyme abundances were consistent with a slowed tricarboxylic acid cycle, the consequence of which likely impeded ATP production by oxidative phosphorylation. There also were enzymatic shifts consistent with decreased amounts of flagellar proteins and decreased pools of purines. A motility assay confirmed a reduction in R5 flagellar movement in resveratrol, and mass spectrometry of metabolite extracts confirmed decreased pools of guanosine 5'-monophosphate and adenosine 5'-monophosphate. Mass spectrometry also detected the accumulation of a reactive aldehyde byproduct of resveratrol catabolism. Overall, the study reveals that resveratrol likely imparts its antibiotic activity during plant immunity by disturbing the bacterial tricarboxylic acid cycle, interfering with ATP biosynthesis at the electron transport chain, and by decreasing bacterial proteins needed for pathogenicity and leaf colonization. Mass spectrometry data files for this study can be retrieved from massive.ucsd.edu (MSV000090171 and MSV000090172).