Abstract: Plasma membrane H⁺-ATPases belong to a family of proton pumps and are widely found in the plasma membranes of plants and fungi; they play important roles in maintaining the transmembrane electrochemical proton gradient required for nutrient uptake and regulating intracellular pH value. Plasma membrane H⁺-ATPases are required for the life activities of fungi. When the plasma membrane ATPase is lacking, the mutants exhibit obvious growth defects or even fail to grow, indicating the potential of plasma membrane ATPase as a target of fungicides. Moreover, due to the low homology between plasma membrane H⁺-ATPases of fungi and plants, fungicides developed targeting plasma membrane ATPase are biosafety. Recently, the structure elucidation cryo-electron microscopy structures of plasma membrane H⁺-ATPases in Saccharomyces cerevisiae and Neurospora crassa has revealed that they are hexamer, elucidating the mechanism of action of plasma membrane H⁺-ATPases and providing a theoretical basis for structure-based drug design. This paper systematically elaborates the structure and function of plasma membrane H⁺-ATPases in fungi and reviews the research status of research on plasma membrane H⁺-ATPases as targets of novel fungicides, which may provide a theoretical basis for the discovery and application of novel fungicides.