Erwinia chrysanthemi pv. zeae is one of the Erwinia chrysanthemi pathovars that infects on both dicotyledons and monocotyledons. However, little is known about the molecular basis and regulatory mechanisms of its virulence. By using a transposon mutagenesis approach, we cloned the genes coding for an E. chrysanthemi pv. zeae synthase of acyl-homoserine lactone (AHL) quorum-sensing signals (expI Ecz ) and a cognate response regulator (expR Ecz ). Chromatography analysis showed that expI Ecz encoded production of the AHL signal N-(3-oxo-hexanoyl)-homoserine lactone (OHHL). Null mutation of expI Ecz in the E. chrysanthemi pv. zeae strain EC1 abolished AHL production, increased bacterial swimming and swarming motility, disabled formation of multicell aggregates, and attenuated virulence of the pathogen on potato tubers. The mutation also marginally reduced the inhibitory activity of E. chrysanthemi pv. zeae on rice seed germination. The mutant phenotypes were rescued by either exogenous addition of AHL signal or in trans expression of expI Ecz . These data demonstrate that the AHL-type QS signal plays an essential role in modulation of E. chrysanthemi pv. zeae cell motility and the ability to form multicell aggregates and is involved in regulation of bacterial virulence.Erwinia chrysanthemi pv. zeae is the major pathogen responsible for bacterial stalk rot of maize around the world (20,29,31). The pathogen was also found to cause rice foot rot in Asian countries, including Japan, China, India, Indonesia, and South Korea (11,17). E. chrysanthemi pv. zeae is a member of the pathovars of the gram-negative bacterium Erwinia chrysanthemi, which has been divided, on the basis of its pathogenicity on host plants and certain biochemical and physiological differences, into six pathovars: pv. chrysanthemi, pv. dianthicola, pv. dieffenbachiae, pv. paradisiaca, pv. parthenii, and pv. zeae (7). The variations among these pathovars were subsequently verified by molecular genetic analysis (20,21). While the rationality of this pathovar classification is still under debate and a new scheme of classification has been proposed (29), in which E. chrysanthemi pv. zeae was delineated as a novel species, Dickeya zeae, the distinct status of E. chrysanthemi pv. zeae in different taxonomy schemes may reflect its unshakeable differences from other pathovars, which will hereby be collectively referred to as E. chrysanthemi strains for convenience and to be consistent with numerous previous studies. Recent constant outbreaks of rice foot rot disease caused by E. chrysanthemi pv. zeae have stirred serious concern (17); however, little is known about the molecular bases of its host specificity and the pathogenic mechanisms of this important pathovar of E. chrysanthemi.Among the closely related bacterial pathogens, a few E. crysanthemi strains that infect dicotyledonous plants, such as strains EC3937 and EC16, have been characterized extensively at biochemical and genetic levels. They are known to cause the soft rot disease that is characterized...