The role of AHK5 in abiotic and biotic stress signalling

Abstract

In Arabidopsis thaliana, eight histidine kinases (HKs) have been identified which function in hormone signalling, stimuli perception, and plant development. To better elucidate HK roles in signalling, the function of the least characterised HK, AHK5, in stress tolerance was investigated using a T-DNA insertion knockout line (ahk5-1). Reduced inhibition of seedling root growth was seen in ahk5-1 in response to salinity when compared to wild-type Col-0 in tissue culture assays. In mature plants, ahk5-1 showed greater fresh weight gain under either salinity or drought stress. Loss of AHK5 function did not alter cold stress tolerance, nor basal and acquired heat stress tolerance in terms of seedling root elongation. Infection with the biotrophic pathogen Pseudomonas syringae pv. tomato DC3000 revealed ahk5-1 is compromised in disease resistance, exhibiting increased chlorosis and in planta bacterial growth. Levels of the plant hormones salicylic acid, jasmonic acid, and abscisic acid, alongside the bacterial phytotoxin coronatine, were lower in pathogen challenged ahk5-1 mutants compared to wild-type plants. The ahk5-1 mutant was also more susceptible to the necrotrophic pathogen Botrytis cinerea, supporting more fungal growth and displaying accelerated symptom development. Hydrogen peroxide production has been linked with both resistance and susceptibility towards B. cinerea; in ahk5-1, 3,3-diaminobenzidene (DAB) staining suggested reduced hydrogen peroxide production in response to infection. Complementation and expression of AHK5 with either full-length genomic AHK5 under the 35S CaMV promoter or full-length AHK5 cDNA under the native promoter rescued the ahk5-1 mutant stress response phenotypes. In summary, AHK5 was found to negatively regulate abiotic stress tolerance whilst positively contributing towards resistance against pathogens employing different lifestyles. To begin to establish an AHK5 signalling network, tandem affinity purification coupled with LC-MS/MS was employed for identification of possible AHK5 interacting proteins. Suggestions for further optimisation of the purification method are presented. The role of AHK5 in regulation of plant stress responses through modulation of reactive oxygen species and hormone signalling and through protein-protein interactions are reviewed. Suggestions for further investigation are also discussed.