A single amino acid insertion in the WRKY domain of the Arabidopsis TIR–NBS–LRR–WRKY‐type disease resistance protein SLH1 (sensitive to low humidity 1) causes activation of defense responses and hypersensitive cell death

Abstract

Summary: In this study we characterized the sensitive to low humidity 1 (slh1) mutant of Arabidopsis ecotype No‐0 which exhibits normal growth on agar plate medium but which on transfer to soil shows growth arrest and development of necrotic lesions. cDNA microarray hybridization and RNA gel blot analysis revealed that genes associated with activation of disease resistance were upregulated in the slh1 mutants in response to conditions of low humidity. Furthermore, the slh1 mutants accumulate callose, autofluorescent compounds and salicylic acid (SA). We demonstrate that SA is required for the slh1 phenotype but not PAD4 or NPR1. SLH1 was isolated by map‐based cloning and it encodes a resistance (R)‐like protein consisting of a domain with Toll and interleukin‐1 receptor homology (TIR), a nucleotide‐binding domain (NB), leucine‐rich repeats (LRR) and a carboxy‐terminal WRKY domain. SLH1 is identical to the R gene RRS1‐R of the Arabidopsis ecotype Nd‐1, a gene which confers resistance to the bacterial pathogen Ralstonia solanacearum GMI1000 and also functions as an R gene to this pathogen in No‐0. We identified a 3 bp insertion mutation in slh1 that results in the addition of a single amino acid in the WRKY domain; thereby impairing its DNA‐binding activity. Our data suggest that SLH1 disease resistance signaling may be negatively regulated by its WRKY domain in the R protein and that the constitutive defense activation conferred by the slh1 mutation is inhibited by conditions of high humidity.