N2 fixation rates were measured in the 0–1000 m layer at 13 stations located in the open western and central Mediterranean Sea (MS) during the PEACETIME cruise (late spring 2017). While the spatial variability of N2 fixation was not related to Fe, P nor N stocks, the surface composition of the diazotrophic community indicated a strong eastward increasing longitudinal gradient for the relative abundance of non-cyanobacterial diazotrophs (NCD) (mainly γ-Proteobacteria) and conversely eastward decreasing for UCYN-A (mainly -A1 and -A3) as did N2 fixation rates. UCYN-A4 and A3 were identified for the first time in the MS. The westernmost station influenced by Atlantic waters, and characterized by highest stocks of N and P, displayed a patchy distribution of diazotrophic activity with an exceptionally high rate in the euphotic layer of 72.1 nmol N L−1 d−1, which could support up to 19 % of primary production. At this station at 1 %PAR depth, UCYN-A4 represented up to 94 % of the diazotrophic community. These in situ observations of higher UCYN-A relative abundance in nutrient rich stations while NCD increased in the more oligotrophic stations, suggest that the nutrient conditions could determine the composition of the diazotrophic communities and in turn the N2 fixation rates. The impact of Saharan dust deposition on N2 fixation and diazotrophic communities was also investigated, under present and future projected conditions of temperature and pH during short term (3–4 days) experiments at three stations. New nutrients from simulated dust deposition triggered a significant stimulation of N2 fixation (from 41 % to 565 %). The strongest increase in N2 fixation was observed at the stations dominated by NCD and did not lead on this short time scale to change in the diazotrophic community composition. Under projected future conditions, N2 fixation was either exacerbated or unchanged, in that later case this was probably due to a too low nutrient bioavailability or an increased grazing pressure. The future warming and acidification likely benefited NCD (Pseudomonas) and UCYN-A2 while disadvantaged UCYN-A3 without knowing which effect (alone or in combination) is the driver, especially since we do not know the temperature optima of these species not yet cultivated as well as the effect of acidification.