Influence of Fusarium oxysporum f. sp. cubense (E.F. Smith) Snyder and Hansen on 2,4-diacetylphloroglucinol (DAPG) production in the rhizosphere of banana cultivar Rasthali by Pseudomonas fluorescens was investigated. The purified extracts of Pfm strain of P. fluorescens isolated from banana rhizosphere inhibited the growth and spore germination of F. oxysporum f. sp. cubense under laboratory conditions. DAPG extracted from the cultures of the strain was observed as distinct spots in thin layer chromatographic plates at Rf value of 0.88. The extracts of soil inoculated with P. fluorescens and challenge inoculated with F. oxysporum f. sp. cubense eluted at retention time ranges from 20.00 min to 21.30 min. The quantity of DPAG production was less in the extracts of soil inoculated with P. fluorescens and challenge inoculated with F. oxysporum f. sp. cubense as compared to P. fluorescens alone inoculated soil. The talc formulation of Pfm strain also reduced vascular discolouration due to the pathogen in banana plants when inoculated at 15 g/plant.