Organization and environmental regulation of the Pseudomonas syringae pv. syringae 61 hrp cluster

Abstract

The ability of Pseudomonas syringae pv. syringae 61 to elicit the hypersensitive response in nonhost plant species has been linked to a cluster of hrp/hrm genes whose expression appears to be environmentally regulated. To understand the genetic organization of this hrp/hrm gene cluster and its expression during the interaction with nonhost plant species better, we constructed a set of chromosomal hrp-uidA fusions in P. syringae pv. syringae 61 by Tn5-gusA1 mutagenesis of the cloned hrp/hrm gene cluster and transferred them into the genome by marker exchange mutagenesis. Complementation analysis employing plasmid-borne Tn5-gusA1 insertions and previously characterized chromosomal TnphoA mutations defined at least eight apparent transcriptional units within the hrp/hrm cluster, several of which were multicistronic. The expression of hrp-uidA fusions in seven of these apparent hrp transcriptional units increased following inoculation into tobacco leaves. Enhanced expression from a representative fusion was detected 1 h after inoculation of tobacco leaves. The induction observed in planta was similar to the levels detected following culture of the bacteria in minimal-salts medium: irrespective of the carbon source. Complex amino acid sources, such as peptone, repressed the expression of P. syringae pv. syringae 61 hrp genes at levels exceeding 0.028%. The results indicate that enhanced expression of hrp genes occurs early in the interaction with nonhost plant species in an apparent response to altered nutritional conditions.