In present investigation total 10 isolates were obtained from rhizospheric and non-rhizospheric soil sample of Vadodara locality. All the isolates were capable of restricting mycelial growth of soil-borne wilt pathogen Fusarium oxysporum and Fusarium udum. Among all the isolates, PR3 showed maximum antipathogenic activity i.e. 42.23 and 45.80 % against F. oxysporum and F. udum and hence, this isolate was further characterized on the basis of morphological, biochemical and molecular characteristics as P. fluorescens AAU PR3 (NCBI Accn. No.: KJ161327). It also showed siderophore production and cell wall degrading enzyme lipase production which represents a possible mechanism of biocontrol of fungal pathogens. P. fluorescens AAU PR3 was found highly resistant to major antibiotics tested. Isolate P. fluorescens AAU PR3 was also capable of reducing wilt symptoms in greengram and also enhanced root and shoot length, fresh as well as dry biomass of greengram in presence and absence of F. udum under laboratory and pot trial conditions. From the present research, native rhizospheric fluorescent P. fluorescens AAU PR3 (NCBI Accn No: KJ161327) has emerged out as a potential biocontrol strain against soil-borne fungal pathogen F. udum.