The ferric iron absorption is one of the most important limiting factors of bacterial growth of Xanthomonas oryzae pv. oryzae. It has been previously speculated that rpfFxoo might be involved in the ferric iron metabolism of the pathogen. In the present study, ΔrpfFxoo, a gene deletion mutant, was generated from the wild-type strain PXO99A of Xoo through the homologous recombination, and Fe content was assayed using flame atomic absorption in PXO99A and ΔrpfFxoo. The results indicated that the recovery was 99.7% and the relative standard deviation was 1.89 under optimized AAS operating conditions. The increase in Fe absorption in PXO99A and ΔrpfFxoo was observed with the increasing time. However, the ferric content of ΔrpfFxoo was significantly lower than that of PXO99A (P<0.05). It is suggested that rpfFxoo is involved in iron metabolism in Xanthomonas oryzae pv. oryzae.