Increased emergence of drug resistance and DON pollution pose a severe problem in Fusarium head blight (FHB) control. While the H⁺ antiporter (DHA) family plays crucial roles in drug resistance, the characterization of DHA transporters has not been systematically studied in pathogenetic fungi. In this study, a systematic gene deletion analysis of all putative DHA transporter genes was carried out in Fusarium graminearum, and one DHA1 transporter FgQdr2 was found to be involved in multiple drug resistance, ion homeostasis, and virulence. Further exploration showed that FgQdr2 is mainly localized in the cell membrane; its expression under normal growth conditions is comparatively low, but sufficient for the regulation of drug efflux. Additionally, investigation of its physiological substrates demonstrated that FgQdr2 is essential for the transport of K⁺, Na⁺, Cu²⁺, and the regulation of the membrane proton gradient. For its roles in the FHB disease cycle, FgQdr2 is associated with fungal infection via regulating the biosynthesis of virulence factor deoxynivalenol (DON), the scavenging of the phytoalexin, as well as both asexual and sexual reproduction in F. graminearum. Overall, the results of this study reveal the crucial roles of FgQdr2 in multiple drug resistance, ion homeostasis, and pathogenicity, which advance the understanding of the DHA transporters in pathogenetic fungi.

Maintaining cellular calcium (Ca²⁺) homeostasis is essential for many aspects of cellular life. The high-osmolarity glycerol (HOG) mitogen-activated protein kinase (MAPK) pathway responsible for signal integration and transduction plays crucial roles in environmental adaptation, especially in the response to osmotic stress. Hog1 is activated by transient Ca²⁺ increase in yeast, but the functions of the HOG pathway in Ca²⁺ homeostasis are largely unknown. We found that the HOG pathway was involved in the regulation of Ca²⁺ homeostasis in Fusarium graminearum, a devastating fungal pathogen of cereal crops. The deletion mutants of HOG pathway displayed increased sensitivity to Ca²⁺ and FK506, and elevated intracellular Ca²⁺ content. Ca²⁺ treatment induced the phosphorylation of FgHog1, and the phosphorylated FgHog1 was transported into the nucleus by importin β FgNmd5. Moreover, the increased phosphorylation and nuclear accumulation of FgHog1 upon Ca²⁺ treatment is independent of the calcineurin pathway that is conserved and downstream of the Ca²⁺ signal. Taken together, this study reported the novel function of FgHog1 in the regulation of Ca²⁺ homeostasis in F. graminearum, which advance the understanding of the HOG pathway and the association between the HOG and calcineurin pathways in fungi.