Both GacS-regulated lipopeptides and the type three secretion system contribute to Pseudomonas cichorii induced necrosis in lettuce and chicory

Pseudomonas cichorii SF1-54, the causal agent of lettuce midrib rot disease, produces lipopeptides cichofactins and cichopeptins which are important virulence factors. The GacS/GacA two-component system is well known to regulate production of lipopeptides in pseudomonads. Additionally, the functions of the type three secretion system (T3SS) in P. cichorii-plant interactions are not clarified. In this study, we investigated the role of the GacS-regulated lipopeptides and the T3SS in pathogenicity of P. cichorii SF1-54 on two host plants, chicory and lettuce, by constructing mutants in hrpL, which encodes the key sigma factor to control T3SS expression, and gacS. Compared with the wildtype, the hrpL mutant produced lipopeptides at a similar level but the gacS mutant was strongly impaired in lipopeptide production. The mutant deficient in hrpL did not significantly differ from the wildtype in virulence on chicory and lettuce. The gacS mutant exhibited significantly less symptoms on both host plants compared to the wildtype and the hrpL mutant. Intriguingly, the gacS hrpL-double mutant no longer produced lipopeptides, lost virulence and showed impaired colonization on chicory, but was still weakly virulent on lettuce. Thus, contribution of both the GacS-regulated lipopeptides and T3SS to virulence of P. cichorii SF1-54 is host plant dependent.