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Analysis of Erwinia chrysanthemi EC16 pelEuidA, pelLuidA, and hrpNuidA Mutants Reveals Strain-Specific Atypical Regulation of the Hrp Type III Secretion System

February 2004 , Volume 17 , Number  2
Pages  184 - 194

Jong Hyun Ham , 1 Yaya Cui , 2 James R. Alfano , 1 Pablo Rodríguez-Palenzuela , 3 Clemencia M. Rojas , 1 Arun K. Chatterjee , 2 and Alan Collmer 1

1Department of Plant Pathology, Cornell University, Ithaca, NY 14853-4203, U.S.A.; 2Department of Plant Pathology, University of Missouri, Columbia, MO 65211, U.S.A.; 3Departamento de Biotecnología, E.T.S. Ingenieros Agrónomos, Ciudad Universitaria s/n 28040 Madrid, Spain

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Accepted 23 September 2003.

The plant pathogen Erwinia chrysanthemi produces a variety of factors that have been implicated in its ability to cause soft-rot diseases in various hosts. These include HrpN, a harpin secreted by the Hrp type III secretion system; PelE, one of several major pectate lyase isozymes secreted by the type II system; and PelL, one of several secondary Pels secreted by the type II system. We investigated these factors in E. chrysanthemi EC16 with respect to the effects of medium composition and growth phase on gene expression (as determined with uidA fusions and Northern analyses) and effects on virulence. pelE was induced by polygalacturonic acid, but pelL was not, and hrpN was expressed unexpectedly in nutrient-rich King's medium B and in minimal salts medium at neutral pH. In contrast, the effect of medium composition on hrp expression in E. chrysanthemi CUCPB1237 and 3937 was like that of many other phytopathogenic bacteria in being repressed in complex media and induced in acidic pH minimal medium. Northern blot analysis of hrpN and hrpL expression by the wild-type and hrpL∷ΩCmr and hrpS∷ΩCmr mutants revealed that hrpN expression was dependent on the HrpL alternative sigma factor, whose expression, in turn, was dependent on the HrpS putative σ54 enhancer binding protein. The expression of pelE and hrpN increased strongly in late logarithmic growth phase. To test the possible role of quorum sensing in this expression pattern, the expI/expR locus was cloned in Escherichia coli on the basis of its ability to direct production of acyl-homoserine lactone and then used to construct expI mutations in pelEuidA, pelLuidA, and hrpNuidA Erwinia chrysanthemi strains. Mutation of expI had no apparent effect on the growth-phase-dependent expression of hrpN and pelE, or on the virulence of E. chrysanthemi in witloof chicory leaves. Overexpression of hrpN in E. chrysanthemi resulted in approximately 50% reduction of lesion size on chicory leaves without an effect on infection initiation.

© 2004 The American Phytopathological Society