The temperature dependence of planktonic metabolism in the subtropical North Atlantic Ocean was assessed on the basis of measurements of gross primary production (GPP), community respiration (CR) and net community production (NCP), as well as experimental assessments of the response of CR to temperature manipulations. Metabolic rates were measured at 68 stations along three consecutive longitudinal transects completed during the Malaspina 2010 Expedition, in three different seasons. Temperature gradients were observed in depth and at basin and seasonal scale. The results showed seasonal variability in the metabolic rates, the highest rates being observed during the spring transect. The overall mean integrated GPP / CR ratio was 1.39 ± 0.27 decreasing from winter to summer, and the NCP for the subtropical North Atlantic Ocean during the cruises exhibits net autotrophy (NCP > 0) in about two-thirds (66%) of the total sampled communities. Also, we reported the activation energies describing the temperature dependence of planktonic community metabolism, which was generally higher for CR than for GPP in the subtropical North Atlantic Ocean, as the metabolic theory of ecology predicts. Furthermore, we made a comparison of activation energies describing the responses to in situ temperature in the field (EaCR Combining double low line 1.64 ± 0.36 eV) and those derived experimentally by temperature manipulations (EaCR Combining double low line 1.45 ± 0.6 eV), which showed great consistency. © 2014 Author(s).