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Nat Protoc. 2009;4(2):117-24. doi: 10.1038/nprot.2008.224.

Modeling Pseudomonas aeruginosa pathogenesis in plant hosts.

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  • 1Department of Surgery at Massachusetts General Hospital, 50 Blossom St., Boston, Massachusetts 02114, USA.

Abstract

A pathogenic model in which both the pathogen and its host are amenable to genetic manipulation can greatly facilitate the understanding of bacterial pathogenesis. Plants are genetically tractable and can be used as experimental models for human microbial pathogenesis. We present protocols for both lettuce and Arabidopsis leaf infection models using the opportunistic human bacterial pathogen, Pseudomonas aeruginosa. The lettuce model allows for high-throughput qualitative analysis of virulence and is suitable for screening large numbers of bacterial strains, whereas the Arabidopsis model provides a quantitative approach and permits the tracking of bacterial cell proliferation in planta. The lettuce model takes approximately 24 h including bacterial growth using store-bought lettuce, and the Arabidopsis model takes 4-6 weeks to grow the plants and a similar time as with lettuce to infect the plants. Both models are monitored for up to 5 d post-infection. These methodologies can and have been used to identify novel and critical P. aeruginosa pathogenicity agents, as virulence factors are often conserved across phylogeny.

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