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Bacteria of genus Methylobacterium have been found to stimulate plant growth and regulate the level of ethylene in crop plants. This work aimed to test the induction of defense responses in red pepper against bacterial spot by stress ethylene level reduction mediated by the ACC deaminase activity of Methylobacterium strains. In greenhouse experiments, the disease index value observed in Methylobacterium strains treated red pepper plants were lower than that of uninoculated treatment. Plants treated with Methylobacterium strains challenge inoculated with Xanthomonas campestris pv. vesicatoria (XCV) showed significantly reduced disease symptom and lowered ethylene emission under greenhouse condition. The ACC and ACO (1-aminocyclopropane-1- carboxylate oxidase) accumulated in the red pepper leaves were significantly reduced with Methylobacterium strains inoculation compared to treatment with pathogen alone. In addition PR proteins related to ISR (Induced systemic resistance) including β-1,3-glucanase, PAL, PO and PPO were increased in Methylobacterium strains inoculated red pepper plants. To confirm the localization of the gfp tagged Methylobacterium spp., red pepper leaf surfaces were visualized by CLSM. The leaves of pathogen alone treatments showed no fluorescent bacterial cells. Fluorescent bacterial cells were observed on all the treatments containing gfp tagged Methylobacterium spp. inoculated red pepper leaves. This study supports that the Methylobacterium strains might increase the activity of defense enzymes by modulating the ethylene biosynthesis pathway, suggesting the possibility of using Methylotrophic bacteria as potential bio-control agents.