Nitrogen (N) is one of the major nutrients limiting plant growth in terrestrial ecosystems. To avoid plant-microbe competition, previous studies on plant N uptake preference often used hydroponic experiments on fine roots of seedlings and demonstrated ammonium preference for conifer species; however, we lack information about N uptake and translocation in the field. In this paper, we described a method of in situ paired N-15 labeling and reported the rates and time course of N uptake and translocation by mature trees in situ. We added N-15-enriched ammonium or nitrate, together with the nitrification inhibitor dicyandiamide, to paired Larix kaempferi (Lamb.) Carr (larch) trees from 30-, 40- and 50-year-old plantations. Fine roots, coarse roots, leaves and small branches were collected 2, 4, 7, 14 and 30 days after labeling. Nitrate uptake and translocation averaged 1.59 +/- 0.16 mu g N-15 g(-1) day(-1), which is slightly higher than ammonium (1.08 +/- 0.10 mu g N-15 g(-1) day(-1)), in all tree organs. Nitrate contributed 50-78% to N uptake and translocation, indicating efficient nitrate use by larch in situ. We observed no age effect. We suggest that sampling leaves after 4 days of N-15 labeling is sufficient to detect mature tree N uptake preference in situ. Whole-tree N-15-ammonium recovery equaled that of N-15-nitrate 30 days after N-15 addition, implying the importance of both ammonium and nitrate to mature larch N use in the long run. We conclude that our method is promising for studying mature tree N uptake preference in situ and can be applied to other conifer and broadleaf species. We suggest using highly enriched N-15 tracer to overcome soil dilution and a nitrification inhibitor to minimize ammonium transformation to nitrate. Our study revealed mature tree N preference in situ and demonstrated the strong contribution of nitrate toward mature larch growth on soils rich in nitrate.