The role of N03-and N02-in the induction of nitrite reductase (NiR) activity in detached leaves of 8-day-old barley (Hordeum vulgare L.) seedlings was investigated. Barley leaves contained 6 to 8 micromoles N02-/gram fresh weight x hour of endogenous NiR activity when grown in N-free solutions. Supply of both N02-and NO3-induced the enzyme activity above the endogenous levels (5 and 10 times, respectively at 10 millimolar NO2-and N03-over a 24 hour period). In N03-supplied leaves, NiR induction occurred at an ambient N03-concentration of as low as 0.05 millimolar; however, no NiR induction was found in leaves supplied with NO2-until the ambient NO2-concentration was 0.5 millimolar. Nitrate accumulated in N02-fed leaves. The amount of NO3-accumulating in N02-fed leaves induced similar levels of NiR as did equivalent amounts of NO3-accumulafing in N03--fed leaves. Induction of NiR in N02--fed leaves was not seen until N03-was detectable (30 nanomoles/gram fresh weight) in the leaves. The internal concentrations of N03-, irrespective of N source, were highly correlated with the levels of NiR induced.When the reduction of N03-to NO2-was inhibited by W042-, the induction of NiR was inhibited only partially. The results indicate that in barley leaves NiR is induced by N03-directly, i.e. without being reduced to N02-, and that absorbed NO2-induces the enzyme activity indirectly after being oxidized to N03-within the leaf.