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The ΔfliD mutant of Pseudomonas syringae pv. tabaci, which secretes flagellin monomers, induces a strong hypersensitive reaction (HR) in non-host tomato cells

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Abstract

To investigate the role of flagella and monomer flagellin in the interaction between Pseudomonas syringae pv. tabaci and plants, non-polar fliC and fliD mutants were produced. The ORFs for fliC and fliD are deleted in the ΔfliC and ΔfliD mutants, respectively. Both mutants lost all flagella and were non-motile. The ΔfliC mutant did not produce flagellin, whereas the ΔfliD mutant, which lacks the HAP2 protein, secreted large amounts of monomer flagellin into the culture medium. Inoculation of non-host tomato leaves with wild-type P. syringae pv. tabaci or the ΔfliD mutant induced a hypersensitive reaction (HR), whereas the ΔfliC mutant propagated and caused characteristic symptom-like changes. In tomato cells in suspension culture, wild-type P. syringae pv. tabaci induced slight, visible HR-like changes. The ΔfliC mutant did not induce HR, but the ΔfliD mutant induced a remarkably strong HR. Expression of the hsr203J gene was rapidly and strongly induced by inoculation with the ΔfliD mutant, compared to inoculation with wild-type P. syringae pv. tabaci. Furthermore, introduction of the fliC gene into the ΔfliC mutant restored motility and HR-inducing ability in tomato. These results, together with our previous study, suggest that the flagellin monomer of pv. tabaci acts as a strong elicitor to induce HR-associated cell death in non-host tomato cells.

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Acknowledgments

We thank Dr. A. Collmer (Cornell University, Ithaca, N.Y.) and the Leaf Tobacco Research Laboratory of Japan Tobacco Inc. for providing P. syringae pv. tomato and pv. tabaci, respectively. We are grateful to the Plant Cell Bank of The Institute of Physical and Chemical Research (RIKEN) and Dr. Y. Hikichi (Kochi University, Japan) for providing the tomato cell culture Sly-1 and the hsr203J cDNA clone, respectively. We are also grateful to Dr. K. Kamimura (Okayama University, Japan) for technical assistance in the observation of bacteria by transmission electron microscopy. This work was supported in part by a Grant-in-Aid for Scientific Research on Priority Areas (A) (No. 12052215) from the Ministry of Education, Culture, Sports, Science and Technology in Japan

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Authors and Affiliations

  1. Laboratory of Plant Pathology and Genetic Engineering, Faculty of Agriculture, Okayama University, 1-1-1 Tsushima-naka, 700-8530, Okayama , Japan

    R. Shimizu, F. Taguchi, M. Marutani, Y. Inagaki, K. Toyoda, T. Shiraishi & Y. Ichinose

  2. Research Institute for Biological Sciences (RIBS) Okayama, Yoshikawa 7459-1, 716-1241, Kayo-cho, Okayama , Japan

    T. Mukaihara

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  1. R. Shimizu
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  2. F. Taguchi
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  3. M. Marutani
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  4. T. Mukaihara
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  5. Y. Inagaki
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  6. K. Toyoda
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  7. T. Shiraishi
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  8. Y. Ichinose
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Correspondence to Y. Ichinose.

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Communicated by A. Kondorosi

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Shimizu, R., Taguchi, F., Marutani, M. et al. The ΔfliD mutant of Pseudomonas syringae pv. tabaci, which secretes flagellin monomers, induces a strong hypersensitive reaction (HR) in non-host tomato cells. Mol Gen Genomics 269, 21–30 (2003). https://doi.org/10.1007/s00438-003-0817-3

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