DNA sequence analysis of a 1.4-kb SalI-HindIII segment located approximately 2 kb upstream of the Rhizobium leguminosarum bv. phaseoli syrM gene revealed the presence of an open reading frame (ORF3) encoding a putative 295-amino acid polypeptide with a molecular mass of 33,401 Da. ORF3 is homologous to a R. fredii host-inducible gene. The proteins encoded by R. l. bv. phaseoli ORF3 and by the R. fredii host-inducible gene share 37% sequence identity. In contrast to the R. fredii host-inducible gene, expression of ORF3 is not induced in the presence of Phaseolus vulgaris root exudates or by specific flavonoids, able to induce nodulation genes in R. l. bv. phaseoli. A R. l. bv. phaseoli ORF3 mutant was constructed by site-directed deletion/replacement mutagenesis. This mutant strain is not affected in symbiotic nitrogen fixation but exhibits a delay in nodulation on Phaseolus vulgaris. Moreover, this mutant was shown to be defective in competition for nodulation.