Twelve-year-old Norway spruce (Picea abies [L.] Karst.) trees were exposed to ambient (AC) or elevated (EC) [ambient + 350 µmol(CO2) mol-1] CO2 concentrations in open-top-chamber (OTC) experiment under the field conditions of a mountain stand. Short-term (4 weeks, beginning of the vegetation season) and long-term (4 growing seasons, end of the vegetation season) effects of this treatment on biochemical parameters of CO2 assimilation were evaluated. A combination of gas exchange, fluorescence of chlorophyll a, and application of a mathematical model of ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBPCO) activity was used. The analysis showed that the depression of photosynthetic activity by long-term impact of elevated CO2 was mainly caused by decreased RuBPCO carboxylation rate. The electron transport rate as well as the rate of ribulose-1,5-bisphosphate (RuBP) formation were also modified. These modifications to photosynthetic assimilation depended on time during the growing season. Changes in the spring were caused mainly by local deficiency of nitrogen in the assimilating tissue. However, the strong depression of assimilation observed in the autumn months was the result of insufficient carbon sink capacity. and O. Urban, M. V. Marek.