Specialized and shared functions of the histidine kinase- and HOG1 MAP kinase-mediated signaling pathways in Alternaria alternata, a filamentous fungal pathogen of citrus

Signal transduction pathways are critical for the coordination of complex cellular processes in cells. In Alternaria alternata, a necrotrophic fungal pathogen of citrus, cloning and characterization of a gene coding a Group III histidine kinase (AaHSK1) and the yeast HOG1 ortholog (AaHOG1) showed the two genes to operate, both uniquely and synergistically, in a number of physiological and pathological functions. Systemic loss-of-function genetics in A. alternata revealed that AaHSK1 is a primary regulator for cellular resistance to sugar osmotic stress and for sensitivity to dicarboximide or phenylpyrrole fungicides. These functions were likely modulated by unknown mechanisms rather than solely by the AaHOG1-mediated pathway. AaHOG1, which conferred cellular resistance to salts and oxidative stress, also bypassed AaHSK1, even though deletion of AaHSK1 affected AaHOG1 phosphorylation. Phosphorylation of AaHOG1 was increased when the fungus was treated with osmotic stress, fungicides or H2O2. Fungal mutants impaired in AaHSK1, AaHOG1, AaAP1 (encoding a redox-responsive transcription factor) or AaFUS3 (encoding a MAP kinase) were all hypersensitive to 2-chloro-5-hydroxypyridine (CHP) or 2,3,5-triiodobenzoic acid (TIBA). An AaHOG1::sGFP (synthetic green fluorescent protein) fusion protein became localized in the nucleus in response to H2O2, CHP, TIBA, fungicides, but not glucose. Glucose, however, enhanced AaHOG1 phosphorylation and nuclear localization in the AaHSK1 deficient background. Accumulation of the AaHSK1 gene transcript was negatively regulated by AaHOG1, AaAP1 or AaFUS3. AaHOG1 was necessary for fungal pathogenicity, yet AaHSK1 was completely dispensable for pathogenicity. Our results highlight a dramatic flexibility and uniqueness in the signaling pathways that are involved in responding to diverse environmental stimuli in A. alternata.