Models predicting ecosystem carbon dioxide (CO2) exchange under future climate change rely on relatively few real world tests of their assumptions and outputs. Here, we demonstrate a rapid and cost-effective method to estimate CO2 exchange from intact vegetation patches under varying atmospheric CO2 concentrations. We ﬁnd that net ecosystem CO2 uptake (NEE) in a boreal forest rose linearly by 4.7 0.2% of the current ambient rate for every 10 ppm CO2 increase, with no detectable inﬂuence of foliar biomass, season, or nitrogen (N) fertilization. The lack of any clear short-term NEE response to fertilization in such an N-limited system is inconsistent with the instantaneous downregulation of photosynthesis formalized in many global models. Incorporating an alternative mechanism with considerable empirical support – diversion of excess carbon to storage compounds – into an existing earth system model brings the model output into closer agreement with our ﬁeld measurements. A global simulation incorporating this modiﬁed model reduces a long-standing mismatch between the modeled and observed seasonal amplitude of atmospheric CO2. Wider application of this chamber approach would provide critical data needed to further improve modeled projections of biosphere–atmosphere CO2 exchange in a changing climate.