The Fur protein is a global regulator of iron metabolism in many bacterial species. However, Fur homologs from some rhizobia appear not to mediate iron-dependent gene expression in vivo. Here, transcriptional profiling analysis showed that more than one-fourth of the genes within the iron stimulon of Bradyrhizobium japonicum were aberrantly controlled by iron in a fur mutant. However, Fur has only a modest role in regulating iron transport genes. Quantitative real time reverse transcriptase PCR measurements confirmed abnormal gene expression in iron-limited cells of the fur strain, thereby demonstrating that Fur must function under those conditions. The findings show that B. japonicum Fur is involved in iron-dependent gene expression, and support the conclusion that rhizobial Fur proteins have novel functions compared with well studied model systems.