The current concentrations of O3 have been shown to cause significant negative effects on crop yield. The present levels of ozone may not induce visible symptoms in most of plants, but can result in substantial losses in reproductive output. This paper considers the impact of ambient O3 on gas exchange, photosynthetic pigments, chlorophyll (Chl) fluorescence and carbohydrate levels in the flag leaf of wheat plants during various stages of reproductive development using open-top chambers. Mean O3 concentration was 45.7 ppb during wheat growth and 50.2 ppb after flag leaf development. Reproductive stage showed higher exceedence of O3 above 40 ppb compared to the vegetative stage. Diurnal variations in net photosynthetic rate (PN) and stomatal conductance (gs), intercellular CO2 concentration (Ci), Fv/Fm ratio, photosynthetic pigments, soluble sugars, and starch were measured at 10, 30, and 50 days after flag leaf expansion (DAFE). The results showed reductions in PN, gs, Fv/Fm ratio, photosynthetic pigments and starch, and increases in Ci, F0, and soluble sugars in nonfiltered chambers (NFCs) compared to filtered chambers (FCs). Maximum changes in measured parameters were observed at 50 DAFE (i.e. grain filling and setting phase). Diurnal variation in PN showed double peaked curve in both FCs and NFCs, but delayed peak and early depression in NFCs. Stomatal conductance was significantly lower in NFCs. The study suggests that higher prevalence of ambient O3 during reproductive development led to significant alteration in physiological vitality of wheat having potential negative influence on yield. and R. Rai, M. Agrawal, S. B. Agrawal.