A novel A -C i curve (net CO 2 assimilation rate of a leaf -A n -as a function of its intercellular CO 2 concentration -C i ) analysis method (Plant, Cell & Environment 27, 137-153, 2004) was used to estimate the CO 2 transfer conductance ( g i ) and the maximal carboxylation ( V cmax ) and electron transport ( J max ) potentials of ageing, non-senescing Pseudotsuga menziesii leaves in relation to their nitrogen (N) content and protein and pigment composition. Both g i and the stomatal conductance ( g sc ) of leaves were closely coupled to V cmax , J max and A n with all variables decreasing with increasing leaf age. Consequently, both C i and C c (chloroplastic CO 2 concentration) remained largely conserved through successive growing seasons. The N content of leaves, as well as the amount of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) and other sodium dodecyl sulfate-soluble proteins, increased during the first three growing seasons, then stabilized or decreased only slightly afterwards. Thus, the age-related photosynthetic nitrogen use efficiency (PNUE) decline of leaves was not a consequence of decreased allocation of N towards Rubisco and other proteins involved in bioenergetics and light harvesting. Rather, loss of photosynthetic capacity was the result of the decreased activation state of Rubisco and proportional down-regulation of electron transport towards the photosynthetic carbon reduction (PCR) and photorespiratory (PCO) cycles in response to a reduction of CO 2 supply to the chloroplasts' stroma. This study emphasizes the regulatory potential and homeostaticity of C c -rather than photosynthetic metabolites or C i -in relation to the commonly observed correlation between photosynthesis and g sc .Key-words : A -C i curves; CO 2 transfer conductance; leaf ageing; photosynthesis model Pseudotsuga menziesii ; Rubisco activity.Abbreviations : A c , RuBP-saturated CO 2 assimilation rate; A j , RuBP-limited CO 2 assimilation rate; A n , net CO 2 assimilation rate; α , leaf absorptance; δ 13 C, stable carbon isotope composition; ∆ , carbon isotope discrimination; C c and C i , chloroplastic and intercellular CO 2 concentration, respectively; ELISA, enzyme-linked immunosorbtion assay; Γ *, chloroplastic CO 2 photocompensation point; Γ , CO 2 compensation point; g i , CO 2 transfer conductance; g sc , stomatal conductance to CO 2 ; I , incident irradiance; J , photochemical electron transport rate; J max , maximal photochemical electron transport rate; Θ , curvature factor of the non-rectangular hyperbola describing the light response of J ; Φ , quantum yield of photochemical electron flow; K c and K o , Michaelis-Menten constants for RuBP carboxylation and oxygenation, respectively; LMA, leaf dry mass allocated per unit area; N, nitrogen; PCR, photosynthetic carbon reduction; PCO, photosynthetic carbon oxidation; PNUE, photosynthetic nitrogen use efficiency; PPFD, photosynthetic photon flux density; R d , mitochondrial respiration in the light; RuBP, ribulose-1,5-bisphosphate; Rubisco, r...