Fluorescent Pseudomonas (FP) is a major group of plant growth promoting rhizobacteria and a well-known synthesizer of siderophores, which imparts a selective advantage on rhizosphere competence and their biocontrol traits. The present study was aimed at examining the factors affecting the production of siderophores and their potential biocontrol traits. Sixteen FP isolates were shortlisted based on their siderophore-producing ability in chrome azural S medium. The isolates were checked for variations in siderophore production under varying incubation times, temperatures, pH, iron (Fe3+) concentrations and mutagens. In addition, the iron binding affinity of siderophores, mycelial inhibition assay and plant growth promotion traits were assessed. Results showed that the siderophore production was highly influenced by the time of incubation, changes in pH, temperature and iron concentration. Chemical characterization showed that the produced siderophores were hydroxamates. Maximum siderophore production was observed at pH 7 whereas UV and EtBr exposure invariably suppressed siderophore production drastically in all isolates. All FPs from maize rhizosphere showed excellent siderophore production which could be due to the competence in strategy-II of the plant rhizosphere and significant growth inhibition on Fusarium oxysporum. Our results suggest the inclination of siderophores to iron, in terms of various criteria affecting production and the possible role of environmental mutations that affect the natural iron harvesting mechanism.