Abstract: Nitrogen (N) transfer from legume trees to associated crops is a key factor for the N economy of low-input agroforestry systems. In this work, we presented a new approach to estimate N transfer based on the N-15 content of root exudates and N released by root turnover of the donor plant (Gliricidia sepium) and the temporal change of the N-15 content of the receiver plant (Dichanthium aristatum). The study was carried out in greenhouse using two isotopic methods: N-15 leaf feeding (LF) and the natural N-15 abundance (NA). Measurements of exudate N-15 were made at several dates before and after tree pruning. A time-dependent box model was devised to quantify N transfer in time and to make comparisons between the isotopic methods. In NA, although tree roots and exudates presented a similar N-15 signature before tree pruning, exudates were strongly depleted in N-15 after pruning. In LF, exudates were always depleted in N-15 in relation to tree roots. Hence, the current assumption used in N transfer studies concerning the equal N-15/N-14 distribution in tissues of the donor plant and in its excreted N was not confirmed in our study. Before pruning, N transfer functioned as a two-N-source system (soil N and exudates N) and both isotopic methods provided similar estimates: 11-12% for LF and 10-15% for NA. Calculations per-formed with the model indicated that N transfer occurred with small or nil fractionation of N-15 in exudates. After pruning, there was a third N source associated with N released from tree root turnover. During this period, the isotopic signature of the receiver plant showed a transient state due to the progressive decrease of N-15 content of that N source. The amount of N derived from the tree represented 65% of the total N content of the. grass at the end of the experiments. (c) 2007 Elsevier B.V. All rights reserved.