Multiplication and Virulence in Plant Tissues of Escherichia coli Clones Producing Pectate Lyase Isozymes PLb and PLe at High Levels and of an Erwinia chrysanthemi Mutant Deficient in PLe.

The phytopathogenic enterobacterium Erwinia chrysanthemi strain EC16 produces four isozymes of pectate lyase (PL), an extracellular enzyme that macerates parenchymatous plant tissues and kills plant cells. A 1.8-kilobase EcoRI DNA fragment containing the entire pelE gene was deleted from the E. chrysanthemi chromosome by marker exchange of a cloned fragment that had been modified in vitro. The resulting mutant, UM1001, produced the isozymes PLa, PLb, and PLc, but not PLe. Mutant UM1001 was compared with wild-type E. chrysanthemi, with Escherichia coli JA221, and with JA221 containing expression vectors with cloned pel genes producing high levels of PLe (pPEL748) or PLb (pPEL343) for the ability to multiply and cause symptoms in intact potato tubers. Tubers were injected with less than 100 bacteria per inoculation site and incubated aerobically or anaerobically. While maceration occurred only in anaerobically incubated tubers, all of the bacteria, including nonpectolytic E. coli controls, multiplied substantially under all conditions. E. coli JA221(pPEL748) caused significantly more maceration than E. coli JA221(pPEL343) or wild-type E. chrysanthemi. Mutant UM1001 caused significantly less maceration than the wild-type E. chrysanthemi. The results establish the importance of PLe in the pectolytic arsenal of E. chrysanthemi by demonstrating that production of PLe can enable E. coli to aggressively macerate tuber tissue and that deletion of pelE significantly diminishes the virulence of E. chrysanthemi.