SUMMARYEffects of mildew on nitrate reductase (NaR) in leaves of barley were investigated throughout the second dark period and into the third photoperiod after inoculation using both in vivo and in vitro assays of the enzyme activity. Initially, infection depressed NaR activity, but as the dark period progressed the depression disappeared, only to reappear in the following photoperiod.An altered flux of nitrate from the roots was not responsible for decreased NaR activity, since infection stimulated this flux 2 d after inoculation. Similarly, a comparison of activities measured using an in vivo assay, with and without added nitrate, showed that the supply of the other substrate, NADH, was not limiting the rate of reduction.Although NaR activity was depressed during the second and third photoperiods, the rate of in vivo nitrate reduction, calculated from the difference between the flux of nitrate delivered to the leaves and the nitrate content of the leaves, was stimulated by infection over the initial 2 d after inoculation. Consequently, inferences made elsewhere which concern effects of mildew infection on nitrate metabolism, and which depend solely on measurements of NaR activity, may be unwarranted because even the infected plant contains an excess of active NaR.