Antibacterial mode of action of trans-cinnamaldehyde derived from cinnamon bark (Cinnamomum verum) essential oil against Agrobacterium tumefaciens

The fumigant antibacterial activities of 50 plant essential oils belonging to 10 families were investigated against Agrobacterium tumefaciens. Among the test plant essential oils, Cinnamomum verum (cinnamon bark) essential oil showed the most potent fumigant antibacterial activity. When we investigated the antibacterial activities of compounds identified from cinnamon bark essential oil and 9 congeners of trans-cinnamaldehyde, lengths of inhibition zone of trans-cinnamaldehyde, salicylaldehyde and hydrocinnamaldehyde were 1.28, 1.73, and 1.24 cm at 0.625 mg/paper disc concentration, respectively. To determine the mode of action of trans-cinnamaldehyde and salicylaldehyde, intercellular reactive oxygen species (ROS) generation and cell membrane integrity were determined using a confocal laser scanning microscopy. Furthermore, we compared the up- and down-regulated gene expression of A. tumefaciens treated with trans-cinnamaldehyde and salicylaldehyde with that of untreated A. tumefaciens. With cutoffs of vertical bar log2FC vertical bar > 1 and FDR < 0.05, 29 and 43 down-regulated genes and 27 and 117 up-regulated genes were found in the treatment of trans-cinnamaldehyde and salicylaldehyde, respectively. Based on the ROS generation results, cell membrane integrity assay, and gene expression, we conclude that the antibacterial mode of action of trans-cinnamaldehyde and salicylaldehyde is ROS generation by the Fenton reaction caused by the down-regulation of an ATP synthesis-related gene cluster, corrupted iron ion homeostasis, and a corrupted ROS defense mechanism. The high concentration of ROS damaged the A. tumefaciens cell membrane, which caused cell death.