Gas exchange measurements were carried out on ash and oak trees in a forest plantation during three whole growing seasons characterized by different water availability (2001, 2002 and 2003). A quantitative limitation analysis was applied to estimate the effects of drought and leaf ontogeny on stomatal (SL) and non-stomatal limitations (NSL) to light-saturated net photosynthesis (Amax), relative to the seasonal maximum rates obtained under conditions of optimal soil water content. Furthermore, based on combined gas exchange and chlorophyll fluorescence measurements, NSL was partitioned into a diffusive (due to a decrease in mesophyll conductance, MCL) and a biochemical component (due to a decrease in carboxylation capacity, BL). During the wettest year (2002), the seasonal pattern of both Amax and stomatal conductance (gsw) was characterized in both species by a rapid increase during spring and a slight decline over the summer. However, with a moderate (year 2001) or a severe (year 2003) water stress, the summer decline of Amax and gsw was more pronounced and increased with drought intensity (30–40% in 2001, 60–75% in 2003). The limitation analysis showed that during the spring and the autumn periods SL, MCL and BL were of similar magnitude. By contrast, from the summer data it emerged that all the limitations increased with drought intensity, but their relative contribution changed. At mild to moderate water stress (corresponding to values of gsw > 100 mmol H2O m−2 s−1) about two-thirds of the decline in Amax was attributable to SL. However, with increasing drought intensity, NSL increased more than SL and nearly equalled it when the stress was very severe (i.e. with gsw < 60 mmol H2O m−2 s−1). Within NSL, MCL represented the main component, except at the most severe water stress levels when it was equalled by BL. It is concluded that diffusional limitations (i.e. SL + MCL) largely affect net assimilation during most of the year, whereas biochemical limitations are quantitatively important only during leaf development and senescence or with severe droughts.