Nitrogen (N) fixation is the main source of ‘new’ N for N-limited ecosystems like subarctic and arctic tundra. This crucial ecosystem function is performed by a wide range of N2 fixer (diazotroph) associations that could differ fundamentally in their timing and amount of N release to the soil. To assess the importance of different associative N2 fixers for ecosystem N cycling, we tracked 15N-N2 into four N2-fixer associations (with a legume, lichen, free-living, moss) and into soil, microbial biomass and non-diazotroph-associated plants 3 days and 5 weeks after in situ labelling. In addition, we tracked 13C from 13CO2 labelling to assess if N and C fixation are linked. Three days after labelling, half of the fixed 15N was recovered in the legume soils, indicating a fast release of fixed N2. Within 5 weeks, the free-living N2 fixers released two-thirds of the fixed 15N into the soil, whereas the lichen and moss retained the fixed 15N. Carbon and N2 fixation were linked in the lichen shortly after labelling, in free-living N2 fixers 5 weeks after labelling, and in the moss at both sampling times. The four investigated N2-fixer associations released fixed N2 at different rates into the soil, and non-diazotroph-associated plants have no access to ‘new’ N within several weeks after N2 fixation. Although legumes and free-living N2 fixers are immediate sources of ‘new’ N for N-limited tundra ecosystems, lichens and especially mosses, do not contribute to increase the N pool via N2 fixation in the short term.