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PLANTS INTERACTING WITH OTHER ORGANISMS

Tomato Mitogen-Activated Protein Kinases LeMPK1, LeMPK2, and LeMPK3 Are Activated during the Cf-4/Avr4-Induced Hypersensitive Response and Have Distinct Phosphorylation Specificities^1,[C],[W]

Laboratory of Phytopathology, Wageningen University, Wageningen, The Netherlands (I.J.E.S., M.H.A.J.J.); and Department of Biological Sciences, University of South Carolina, Columbia, South Carolina (J.W.S.)

Tomato (Solanum lycopersicum) plants with the Cf-4 resistance gene recognize strains of the pathogenic fungus Cladosporium fulvum that secrete the avirulence protein Avr4. Transgenic tomato seedlings coexpressing Cf-4 and Avr4 mount a hypersensitive response (HR) at 20°C, which is suppressed at 33°C. Within 120 min after a shift from 33°C to 20°C, tomato mitogen-activated protein (MAP) kinase (LeMPK) activity increases in Cf-4/Avr4 seedlings. Searching tomato genome databases revealed at least 16 LeMPK sequences, including the sequence of LeMPK1, LeMPK2, and LeMPK3 that cluster with biotic stress-related MAP kinase orthologs from Arabidopsis (Arabidopsis thaliana) and tobacco (Nicotiana tabacum). LeMPK1, LeMPK2, and LeMPK3 are simultaneously activated in Cf-4/Avr4 seedlings, and, to reveal whether they are functionally redundant or not, recombinant LeMPKs were incubated on PepChip Kinomics slides carrying peptides with potential phosphorylation sites. Phosphorylated peptides and motifs present in them discriminated between the phosphorylation specificities of LeMPK1, LeMPK2, and LeMPK3. LeMPK1, LeMPK2, or LeMPK3 activity was specifically suppressed in Cf-4-tomato by virus-induced gene silencing and leaflets were either injected with Avr4 or challenged with C. fulvum-secreting Avr4. The results of these experiments suggested that the LeMPKs have different but also overlapping roles with regard to HR and full resistance in tomato.

The author responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (www.plantphysiol.org) is: Matthieu H.A.J. Joosten (matthieu.joosten{at}wur.nl).

^[C] Some figures in this article are displayed in color online but in black and white in the print edition.

^[W] The online version of this article contains Web-only data.

www.plantphysiol.org/cgi/doi/10.1104/pp.107.101063

^* Corresponding author; e-mail matthieu.joosten{at}wur.nl; fax 31–317483412.

Received April 13, 2007; accepted April 27, 2007; published May 3, 2007.

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