Biological dinitrogen (N2) fixation provides the major source of new nitrogen (N) to the open ocean, contributing more than atmospheric and riverine inputs to the N supply. Yet the fate of the diazotroph-derived N (DDN) in the planktonic food web is poorly understood due to technical limitations. The main goals of this study were to (i) quantify how much of DDN is released to the dissolved pool during N2 fixation and how much is transferred to bacteria, phytoplankton and zooplankton, (ii) to compare the DDN release and transfer efficiencies under contrasting N2 fixation activity and diversity the oligotrophic waters of the Western Tropical South Pacific (WTSP) Ocean. We used nanometer scale secondary ion mass spectrometry (nanoSIMS) coupled with 15N2 isotopic labelling and flow cytometry cell sorting to track the DDN transfer to plankton, in regions were the diazotroph community was either dominated by Trichodesmium or by UCYN-B. After 48 h, ~ 20–40 % of the N2 fixed during the experiment was released to the dissolved pool when Trichodesmium dominated, while the DDN release was not quantifiable when UCYN-B dominated. ~ 7–15 % of the total fixed N (net N2 fixation + release) was transferred to non-diazotrophic plankton within 48 h, with higher transfer efficiencies (15 ± 3 %) when UCYN-B dominated as compared to when Trichodesmium dominated (9 ± 3 %). Most of the DDN (> 90 %) was transferred to picoplankton (Synechococcus, Prochlorococcus and bacteria) in all experiments. The cyanobacteria Synechococcus and Prochlorococcus were the primary beneficiaries (~ 65–70 % of the DDN transfer), followed by heterotrophic bacteria (~ 23–34 % of the DDN transfer). The DDN transfer in bacteria was the highest (34 ± 7 %) when UCYN-B were dominating the diazotroph community. Regarding higher trophic level, the DDN transfer to the dominant zooplankton species was more efficient when the diazotroph community was dominated by Trichodesmium (~ 5–9 % of the DDN transfer) than when it is dominated by UCYN-B (~ 28 ± 13 % of the DDN transfer). To our knowledge, this study provides the first quantification of DDN release and transfer to phytoplankton, bacteria and zooplankton communities in open ocean waters. It reveals that despite UCYN-B fix N2 at lower rates compared to Trichodesmium in the WTSP, the DDN from UCYN-B is much more available and efficiently transferred to the planktonic food web than the DDN coming from Trichodesmium.