In a 10-year (1996-2005) biodiversity experiment, the mechanisms underlying the increasingly positive effect of biodiversity on plant biomass production shifted from sampling to complementarity over time. The effect of diversity on plant biomass was associated primarily with the accumulation of higher total plant nitrogen pools (N g m-2) and secondarily with more efficient N use at higher diversity. The accumulation of N in living plant biomass was significantly increased by the presence of legumes, C4 grasses, and their combined presence. Thus, these results provide clear evidence for the increasing effects of complementarity through time and suggest a mechanism whereby diversity increases complementarity through the increased input and retention of N, a commonly limiting nutrient.