In forest ecosystems, species use different strategies to increase their competitive ability for nitrogen (N) acquisition. The acquisition of N by trees is regulated by tree internal and environmental factors including mycorrhizae. In this study, we investigated the N uptake strategies of three co-occurring tree species [European beech (Fagus sylvatica L.), sycamore maple (Acer pseudoplatanus L.) and Norway maple (Acer platanoides L.)] in the understorey of a beech-dominated, N-limited forest on calcareous soil over two consecutive seasons. For this purpose, we studied 15N uptake capacity as well as the allocation to N pools in the fine roots. Our results show that European beech had a higher capacity for both inorganic and organic N acquisition throughout the whole growing season compared with sycamore maple and Norway maple. The higher capacity of N acquisition in beech indicates a better adaption of beech to the understorey conditions of beech forests compared with the seedlings of other tree competitors under N-limited conditions. Despite these differences, all three species preferred organic over inorganic N sources throughout the growing season and showed similar seasonal patterns of N acquisition with an increased N uptake capacity in summer. However, this pattern varied with N source and year indicating that other environmental factors not assessed in this study further influenced N acquisition by the seedlings of the three tree species.