To study aspects of the ecology of grassland species, in a comparative experiment, plants of P. lanceolata and P. major were grown in pots in a greenhouse, and subjected to a gradual nitrate depletion for several weeks. Control plants were weekly supplied with nitrate. Growth, leaf appearance and disappearance, concentrations of cations and inorganic anions, soluble and insoluble reduced nitrogen concentrations, in vivo nitrate reductase activity (NRA) and the concentration of non-structural carbohydrates in several parts of the plants were followed. Depletion of nitrate caused a reduction of shoot growth, both in biomass and number of leaves. Withering of leaves increased. Accumulation of root dry matter was little (P. lanceolata), or not (P. major) affected. The concentration of reduced nitrogen in all tissues also decreased, both that of the soluble and that of the insoluble fraction. As a result, nitrogen use efficiency (NUE, g dry matter produced per mmol N incorporated) increased by nitrate depletion. NRA was higher in the roots than in the leaves, and decreased with increasing nitrate depletion.In control plants, nitrate became also limiting. This resulted in decreasing nitrate concentrations in leaves and roots. In the leaves, the decrease in nitrate concentration was preceded by a decrease in NRA. The decrease of the nitrate concentration was parallelled by an increase in the concentration of soluble sugar.No major differences in the response towards nitrate depletion were observed between the two species.