Pseudomonas putida is widely used as a biocontrol agent, however, mechanisms by which it initiates the plants' defense response remains obscure. To gain an insight into the molecular changes that occur in plants upon plant growth-promoting rhizobacteria colonization, root transcriptome analysis by using a microarray was performed in rice using P. putida RRF3 (a rice rhizosphere isolate). Data analysis revealed a differential regulation of 61 transcripts (48 h post-treatment), of which, majority corresponded to defense response, cell wall modification and secondary metabolism. Seven genes encoding salicylic acid (SA) responsive pathogenesis-related proteins were up-regulated significantly (fold change ranges from 1 to 4), which suggests that RRF3 has a profound impact on a SA-mediated defense signaling mechanism in rice. Investigations performed at later stages of RRF3 colonization by real-time polymerase chain reaction and high-performance liquid chromatography (HPLC) analysis confirmed the above results, demonstrating RRF3 as a potent biocontrol agent. Further, the impact of RRF3 colonization on root exudation, in particular, exudation of SA was investigated by HPLC. However, analysis revealed RRF3 to have a negative impact on root exudation of SA. Overall, this study shows that P. putida RRF3 immunizes the rice plants by re-organizing the root transcriptome to stimulate plant defense responses ('priming'), and simultaneously protects itself from the primed plants by altering the rhizosphere chemical constituents.