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Abstract Nitrogen fertilization affect growth and yield of timothy (Phleum Pratense L.), but there are conflicting report on its effect on nutritive value. Two experiments were conducted to describe changes in nutritive value of field-grown timothy fertilized with four rates of N fertilizer during spring and summer growth cycles, and to analyse relationships between parameters of nutritive value and the leaf to weight ratio (LWR). Early in the regrowth cycles when the harvestabe shoot biomass was approximately 200 g dry matter (DM) m−2 and under non-limiting N and water conditions, the in vitro true digestibility (IVTD) of DM was greater in spring than in summer because the concentration of neutral-detergent fibre (NDF) was lower and its in vitro cell wall digestibility (IVCWD) was greater. However, the rates of decline in IVTD and IVCWD and the rate of increase in NDF were greater in spring than in summer. Nitrogen deficiency decresed IVTD and IVCWD, and increased NDF concentration early in the regrowth cycles. Nitrogen deficiency, however, reduced the rate of change in the three parameters of nutritive value. By the end of growth cycles, IVTD was 59 and 42 g kg−1 DM greater when no N was applied than with non-limiting N conditions in spring and summer respectively. When no N was applied, the IVCWD was 76 and 52 g kg−1 NDF greater, whereas the NDF concentration was 35 and 42 g kg−1 DM less than under non-limiting N conditions in spring and summer respectively. The N deficiency increased the proportion of leaves in the biomass, which can explain for the most part the positve effect of the N deficiency on the nutritive value of timothy. This positive effect of the N deficiency on nutritive value though the proportion of leaves in the biomass, however, was reduced by the negative effect of N deficiency on the nutritive value of leaves or stems or both.