The chloride channel family gene CLCd negatively regulates pathogen-associated molecular pattern (PAMP)-triggered immunity in Arabidopsis

Chloride channel (CLC) family genes are ubiquitous from prokaryotes to eukaryotes and encode proteins with both channel and transporter activities. The Arabidopsis thaliana genome encodes seven CLC genes, and their products are found in a variety of cellular compartments and have various physiological functions. However, a role for AtCLCs in plant innate immunity has not previously been demonstrated. Here it is reported that AtCLCd is a negative regulator of pathogen-associated molecular pattern (PAMP)-triggered immunity (PTI). T-DNA insertion mutants of AtCLCd exhibited enhanced responses to the elicitor, flg22. The PTI phenotypes of the clcd mutants were rescued by expression of AtCLCd. Overexpression of AtCLCd led to impaired flg22-induced responses. In line with a role for AtCLCd in PTI, the clcd mutants were more resistant to a virulent strain of the bacterial pathogen Pseudomonas syringae pv. tomato DC3000 when spray inoculated, while AtCLCd-overexpressing lines displayed increased susceptibility to this pathogen. Interestingly, flg22 treatment was found to repress the expression of AtCLCd. In addition, its expression was elevated in mutants of the flg22 pattern recognition receptor (PRR) FLS2 and the PRR regulatory proteins BAK1 and BKK1, and reduced in an FLS2-overexpressing line. These latter findings indicate that FLS2 complexes regulate the expression of AtCLCd, further supporting a role for AtCLCd in PTI.