The effects of prochloraz on membrane permeability and germination of uredospores of Puccinia recondita were investigated to determine its potential mode of action on wheat leaf rust control activity. Disease control activity of ergosterol biosynthesis inhibitors (EBIs) and their activities on uredospore membrane permeability and germination were examined with wheat leaf rust pathogen, both in vitro and in vivo. While wheat leaf rust was not controlled by prochloraz, electrolytic leakage and spore germination of P. recondita uredospore was the highest with the use of prochloraz among the eight fungicides tested. Prochloraz stimulated uredospore of P. recondita to germinate at a higher ratio. Although certain EBIs, such as hexaconazole, showed excellent control activity, their effects on uredospore membrane permeability and germination was much inferior to prochloraz. Therefore, results of this study suggest that effects of EBIs on membrane permeability and germination of uredospore are not always correlated with their disease control activity.