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A spontaneousXanthomonas campestris pv. phaseoli H2 O2 -resistant mutant emerged upon selection with 1 mM H2 O2 . In this report, we show that growth of this mutant under noninducing conditions gave high levels of catalase, alkyl hydroperoxide reductase (AhpC and AhpF), and OxyR. The H2 O2 resistance phenotype was abolished inoxyR -minus derivatives of the mutant, suggesting that elevated levels and mutations inoxyR were responsible for the phenotype. Nucleotide sequence analysis of theoxyR mutant showed three nucleotide changes. These changes resulted in one silent mutation and two amino acid changes, one at a highly conserved location (G197 to D197) and the other at a nonconserved location (L301 to R301) in OxyR. Furthermore, these mutations inoxyR affected expression of genes in theoxyR regulon. Expression of anoxyR -regulated gene,ahpC , was used to monitor the redox state of OxyR. In the parental strain, a high level of wild-type OxyR repressedahpC expression. By contrast, expression ofoxyR5 from theX. campestris pv. phaseoli H2 O2 -resistant mutant and its derivativeoxyR5 G197D with a single-amino-acid change on expression vectors activatedahpC expression in the absence of inducer. The other single-amino-acid mutant derivative ofoxyR5 L301R had effects onahpC expression similar to those of the wild-typeoxyR . However, when the two single mutations were combined, as inoxyR5 , these mutations had an additive effect on activation ofahpC expression.

Mutations in oxyR Resulting in Peroxide Resistance in Xanthomonas campestris