Little is known about ozone (O3) effects on adult trees in the field, where ecophysiological parameters control pollutant uptake. It was the goal of this study to examine how ambient and above ambient O3 concentrations affect gas exchange of mature Norway spruce (Picea abies (L.) Karst.). Therefore, twigs were enclosed in chambers and exposed to different O3 concentrations for one and two seasons over three years. During winter periods twigs were maintained under ambient conditions. Data from the shade crown of spruce trees at 1000 m a.s.l. are presented. After one and two fumigation periods no clear treatment effects on gas exchange were observed in twigs fumigated with O3 concentrations ranging from zero up to ambient (A) + 60 ppb. However, O3 at 90 ppb reduced photosynthesis and conductance. CO2 response curves indicated that in A + 90 twigs the efficiency of CO2 uptake was diminished. Observed losses in Pn of A + 90 twigs were greater than reductions in conductance indicating that stomatal closure alone did not limit CO2 uptake. We conclude that ambient and slightly above ambient O3 concentrations do not alter gas exchange of mature Norway spruce. Therefore, suppositions on O3 damage on mature spruce trees should be critically questioned.