Ralstonia solanacearum AmpD is required for wild-type bacterial wilt virulence

Abstract A gene resembling enterobacterial ampD was identified in the bacterial wilt pathogen, Ralstonia solanacearum. The gene lies 13 bp 3' of pehSR, a two-component positive regulator of virulence factors such as plant cell wall-degrading polygalacturonases and bacterial motility. AmpD, an N-acetylmuramyl-l-alanine amidase, degrades and recycles bacterial cell wall components and also plays a role in the induction of beta-lactamase, which confers ampicillin resistance. AmpD is probably not involved in beta-lactamase regulation in R. solanacearum, because the species produces no detectable beta-lactamase activity and is not ampicillin resistant. However, the R. solanacearum ampD gene restores inducible beta-lactamase activity to an Escherichia coli ampD mutant, demonstrating that the gene encodes an AmpD protein that can function in a heterologous background. An R. solanacearumampD chromosomal mutant was motile, produced wild-type levels of polygalacturonase activity and had wild-type cell and colony morphology. This mutant also grew normally in minimal medium and in plant tissue. Nonetheless, the ampD mutant was significantly reduced in bacterial wilt virulence on eggplant and tomato, suggesting a previously unsuspected role for N-acetylmuramyl-l-alanine amidase in plant pathogenesis.