Involvement of tonB-exbBD1D2 operon in infection of Xanthomonas campestris phage phi L7

phi L7 is a lytic bacteriophage infecting Xanthomonas campestris pv. campestris, a Gram-negative bacterium producing xanthan gum and causing black rot in crucifers. A mutant resistant to phi L7 was isolated by Tn5 mutagenesis. Sequence analysis indicated that the gene responsible for the mutation is tonB encoding an inner membrane protein previously shown to be required for iron uptake and pathogenesis. This gene is clustered with three other genes, tonB-exbB-exbD1-exbD2. Results of insertional mutations, DNA and protein sequence analyses, phage sensitivity tests, transfection tests, complementation tests, and phage adsorption assays together with the cellular location of the proteins indicate that TonB, ExbB, and ExbD1 are essential for penetration of phage phi L7. The genome organization, structural features of the tonB-exb region, and transcriptional analyses including Northern hybridization, reporter assays, and primer extension together indicate that the four genes are organized into an operon.