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Journal of Bacteriology, December 2001, p. 7126-7134, Vol. 183, No. 24
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.24.7126-7134.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Differential Roles of the Pseudomonas aeruginosa PA14 rpoN Gene in Pathogenicity in Plants, Nematodes, Insects, and Mice

Erik L. Hendrickson,1,2,dagger Joulia Plotnikova,1,2 Shalina Mahajan-Miklos,1,2,Dagger Laurence G. Rahme,3,4 and Frederick M. Ausubel1,2,*

Department of Genetics1 and Department of Surgery,3 Harvard Medical School, and Department of Molecular Biology2 and Shriner's Burns Institute,4 Massachusetts General Hospital, Boston, Massachusetts 02114

Received 4 June 2001/Accepted 18 September 2001

We cloned the rpoN (ntrA, glnF) gene encoding the alternate sigma factor sigma 54 from the opportunistic multihost pathogen Pseudomonas aeruginosa strain PA14. A marker exchange protocol was used to construct the PA14 rpoN insertional mutation rpoN::Genr. PA14 rpoN::Genr synthesized reduced levels of pyocyanin and displayed a variety of phenotypes typical of rpoN mutants, including a lack of motility and the failure to grow on nitrate, glutamate, or histidine as the sole nitrogen source. Compared to wild-type PA14, rpoN::Genr was ca. 100-fold less virulent in a mouse thermal injury model and was significantly impaired in its ability to kill the nematode Caenorhabditis elegans. In an Arabidopsis thaliana leaf infectivity assay, although rpoN::Genr exhibited significantly reduced attachment to trichomes, stomata, and the epidermal cell surface, did not attach perpendicularly to or perforate mesophyll cell walls, and proliferated less rapidly in Arabidopsis leaves, it nevertheless elicited similar disease symptoms to wild-type P. aeruginosa PA14 at later stages of infection. rpoN::Genr was not impaired in virulence in a Galleria mellonella (greater wax moth) pathogenicity model. These data indicate that rpoN does not regulate the expression of any genes that encode virulence factors universally required for P. aeruginosa pathogenicity in diverse hosts.

* Corresponding author. Mailing address: Department of Molecular Biology, Wellman 10, Massachusetts General Hospital, Boston, MA 02114. Phone: (617) 726-5969. Fax: (617) 726-5949. E-mail: ausubel{at}molbio.mgh.harvard.edu.

dagger Present address: Department of Microbiology, University of Washington, Seattle, WA 98195.

Dagger Present address: Microbia, Inc., Cambridge, MA 02139.

Journal of Bacteriology, December 2001, p. 7126-7134, Vol. 183, No. 24
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.24.7126-7134.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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