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Published on October 26, 2007; 10.1104/pp.107.105353

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Received July 11, 2007
Accepted October 16, 2007

Induction of a Small Heat Shock Protein and its Functional Roles in Nicotiana Plants in the Defense Response against Ralstonia solanacearum

Milimo Maimbo , Kouhei Ohnishi , Yasufumi Hikichi , Hirofumi Yoshioka , and Akinori Kiba *

Laboratory of Plant Pathology and Biotechnology, Faculty of Agriculture, Kochi University, Nankoku 783-8502, Japan; Research Institute of Molecular Genetics, Kochi University, Nankoku 783-8502, Japan; Laboratory of Defense in Plant-Pathogen Interactions, Graduate School of Bioagricultural Sciences, Nagoya University, Chikusa-ku, Nagoya 464-8601, Japan

* Corresponding author; email: akiba{at}cc.kochi-u.ac.jp.

In tobacco, Ralstonia solanacearum OE1-1 (RsOE1-1) is pathogenic, whereas R. solanacearum 8107 (Rs8107) is non-pathogenic and induces the hypersensitive response (HR). To elucidate the molecular mechanisms of plant–R. solanacearum interactions, we used differential display to isolate a cDNA fragment, A6, regulated in tobacco by inoculation with RsOE1-1. The deduced amino acid sequence predicted from full length A6-cDNA showed similarity to small heat shock proteins from Arabidopsis thaliana (hypothetical protein), Medicago truncatula and Cucumis melo; we therefore designated A6 to correspond to Ntshsp17 (Nicotiana tabacum small heat shock protein 17). Recombinant Ntshsp17 overproduced in Escherichia coli exhibited molecular chaperone function. Expression of Ntshsp17 was increased in tobacco leaves inoculated with both RsOE1-1 and Rs8107. Expression was induced by heat treatment, and by treatment with aminocyclopropane carboxylic acid, H2O2, methyl jasmonate and salicylic acid. Ntshsp17 expression was induced by inoculation with a hypersensitive response- and pathogenicity (hrp)-gene mutant of Rs8107 that does not induce the HR, but not by Agrobacterium-mediated transient expression of INF1, an HR elicitor. In Nbshsp17 (an Ntshsp17 ortholog in N. benthamiana)-silenced plants, expression of EREBP, PR-1a and PR-4 genes was compromised, but the expression of EF1-{alpha} was scarcely affected. Appearance of the HR was not affected in the silenced plants. In the silenced plants, growth of Rs8107 was accelerated. Bacterial growth and wilt symptoms elicited by RsOE1-1 were also accelerated in the silenced plants. These results indicate that this small heat shock protein might have a role in the HR-independent defenses in Nicotiana plants.






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