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Immunogold Labeling of Hrp Pili of Pseudomonas syringae pv. tomato Assembled in Minimal Medium and In Planta

February 2001 , Volume 14 , Number  2
Pages  234 - 241

Wenqi Hu , 1 , 2 Jing Yuan , 1 Qiao-Ling Jin , 1 , 2 Patrick Hart , 2 and Sheng Yang He 1 , 2

1Department of Energy Plant Research Laboratory, Michigan State University, East Lansing 48824, U.S.A.; 2Department of Botany and Plant Pathology, Michigan State University, East Lansing 48824, U.S.A.

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Accepted 1 November 2000.

Hypersensitive reaction and pathogenicity (hrp) genes are required for Pseudomonas syringae pv. tomato (Pst) DC3000 to cause disease in susceptible tomato and Arabidopsis thaliana plants and to elicit the hypersensitive response in resistant plants. The hrp genes encode a type III protein secretion system known as the Hrp system, which in Pst DC3000 secretes HrpA, HrpZ, HrpW, and AvrPto and assembles a surface appendage, named the Hrp pilus, in hrp-gene-inducing minimal medium. HrpA has been suggested to be the Hrp pilus structural protein on the basis of copurification and mutational analyses. In this study, we show that an antibody against HrpA efficiently labeled Hrp pili, whereas antibodies against HrpW and HrpZ did not. Immunogold labeling of bacteria-infected Arabidopsis thaliana leaf tissue with an Hrp pilus antibody revealed a characteristic lineup of gold particles around bacteria and/or at the bacterium-plant contact site. These results confirm that HrpA is the major structural protein of the Hrp pilus and provide evidence that Hrp pili are assembled in vitro and in planta.

Additional keywords: avirulence gene, disease resistance, EPEC, Salmonella spp., Shigella spp., Yersinia spp.

© 2001 The American Phytopathological Society