Nitrogen fertilization is known to have a limited effect on the digestibility of grasses. In a previous paper, however, we reported that N deficiency increased the proportion of leaves in the shoot biomass, and hence, indirectly increased the digestibility of timothy (Phleum pratense L). This was mitigated by the direct negative effect of N deficiency on the digestibility of leaves or stems or both early in the regrowth. The objective of this study was to determine the direct effect of N deficiency on three parameters of nutritive value of leaves and stems of timothy cv. Champ. The evolution of leaf and stem in vitro true digestibility (IVTD), in vitro cell wall digestibility (IVCWD), and neutral detergent fiber (NDF) concentration of field-grown timothy fertilized with four rates of N was studied during a spring and summer regrowth cycle by sampling at weekly intervals. The rate of decline in leaf nutritive value in spring was greater than that in summer, and was less than the rate of decline in stem nutritive value during spring. Nitrogen deficiency consistently increased the NDF concentration of leaves but had a lesser effect on the NDF concentration of stems. Nitrogen deficiency decreased leaf IVCWD when the leaf-to-weight ratio (LWR) was high early in the spring and summer regrowth cycles. At the end of regrowth when LWR values were low, leaf IVCWD was greatest when no N was applied. As a result, the negative effect of N deficiency on the leaf IVTD during the early part of the spring and summer regrowth cycles diminished as the regrowth cycles progressed. Nitrogen deficiency had a limited effect on stem IVCWD. We concluded that the direct negative effect of N deficiency on the digestibility of the timothy shoot biomass can be attributed primarily to its effect on the NDF concentration and cell wall digestibility of leaves. Key words: Phleum pratense L., grasses, digestibility, cell wall, nitrogen