Phomopsis blight, caused by Phomopsis vexans (P. vexans) in brinjal plants is one of the main diseases in India. In order to manage this malady, we initially screened seed samples of 14 cultivars obtained from seed suppliers for the disease incidence. Based on the disease incidence and severity, the samples were categorized as resistant and susceptible cultivars of brinjal. It was established that MEBH-9' was a sensitive cultivar, while the Kolar local was resistant to the disease. Two ideal strains of Rhizosphere colonizing bacteria (RCB), Pseudomonas putida strain Has-1/c (HM229805), and Phylloplane Colonizing Bacteria (PCB), Bacillus subtilis strain Br/ph-33 (KJ867501), previously characterized as potential plant growth-promoting rhizobacteria, were tested for their potential biocontrol of this disease. Both susceptible and resistant cultivars' seedlings were treated with different combinations of PCB and RCB, PCB alone, RCB alone, and challenge inoculated with P. vexans. In order to ascertain whether there was signalling between defense enzyme activation and plant protection after PCB and RCB treatment, defense enzymes were quantified. Brinjal plants treated with PCB+RCB combination and challenge inoculated with P. vexans showed a significant increase in the activity of Phenylalanine ammonia-lyase (PAL), Peroxidase (POX), Lipoxygenase (LOX), Polyphenol oxidase (PPO), Catalase, Chitinase, β -1,3-glucanase, and Hydrogen peroxide (H2O2) content. The current study reveals that the defense enzymes and PR-proteins are gradually induced and accumulated, which enhance the resistance in brinjal plants against P. vexans-causing fruit rot disease.