Beneficial rhizobacteria can inhibit foliar pathogen infection by activation of defense responses, yet it the mechanisms of rhizobacteria-induced disease resistance remain largely unknown. Here, inoculation of susceptible maize plants with Pseudomonas fluorescens MZ05 significantly reduced disease occurrence caused by the leaf pathogen Setosphaeria turcica. Gene expression profiles of MZ05-inoculated plants were investigated by RNA-sequencing analyses, showing that several differentially expressed genes were positively associated with the metabolic processes of benzoxazinoids. Accordantly, the inoculation with P. fluorescens MZ05 resulted in a significant increase in the levels of 2,4-dihydroxy-7-methoxy-2H-1,4-benzoxazin-3(4H)-one (DIMBOA) in the maize leaves. Furthermore, pre-inoculation with P. fluorescens MZ05 enhanced the transcription of two defense-related marked genes PAL and PR2a, as well as BX2 and GLU2, which are involved in DIMBOA biosynthesis, in pathogen-infected leaves. Defense responses in the inoculated plants were also greatly stronger and quicker than that in non-inoculated plants after pathogen attacks. However, virus-mediated silencing of BX2 or GLU2 remarkably attenuated the MZ05-induced effects, as evidenced by more disease occurrence and lower transcription of PAL and PR2a. Collectively, these findings indicated that the MZ05-induced increases of DIMBOA levels participated in the mediation of priming, which was the key mechanism in the rhizobacteria-induced host resistance.