Summary• The effects of zinc (Zn) toxicity on photosynthesis and respiration were investigated in sugar beet (Beta vulgaris) plants grown hydroponically with 1.2, 100 and 300 lM Zn.• A photosynthesis limitation analysis was used to assess the stomatal, mesophyll, photochemical and biochemical contributions to the reduced photosynthesis observed under Zn toxicity.• The main limitation to photosynthesis was attributable to stomata, with stomatal conductances decreasing by 76% under Zn excess and stomata being unable to respond to physiological and chemical stimuli. The effects of excess Zn on photochemistry were minor. Scanning electron microscopy showed morphological changes in stomata and mesophyll tissue. Stomatal size and density were smaller, and stomatal slits were sealed in plants grown under high Zn. Moreover, the mesophyll conductance to CO 2 decreased by 48% under Zn excess, despite a marked increase in carbonic anhydrase activity. Respiration, including that through both cytochrome and alternative pathways, was doubled by high Zn.• It can be concluded that, in sugar beet plants grown in the presence of excess Zn, photosynthesis is impaired due to a depletion of CO 2 at the Rubisco carboxylation site, as a consequence of major decreases in stomatal and mesophyll conductances to CO 2 .Abbreviations: A N , net photosynthesis; AOX, alternative oxidase; APX, ascorbate peroxidase; BL, DL, MCL, NSL and SL, biochemical conductance, diffusional conductance, mesophyll conductance, and nonstomatal and stomatal limitations, respectively; C a , C C and C i, atmospheric, chloroplastic and substomatal CO 2 concentrations, respectively; CA, carbonic anhydrase; Chl, chlorophyll; ETR, electron transport rate; F PSII, actual photosystem II efficiency; FAAS, flame atomic absorption spectroscopy; F o , minimal Chl fluorescence in the dark; F v ⁄ F m , maximum potential photosystem II efficiency; g m and g s , CO 2 mesophyll and stomatal conductances, respectively; J max , in vivo maximum rate of electron transport driving regeneration of RuBP; Mn-SOD, manganese-superoxide dismutase; PPFD, photosynthetic photon flux density; PSII and PSI, photosystems II and I, respectively; V c,max , in vivo maximum rate of Rubisco carboxylation; WC, water content.