O3-FACE (Ozone-free air control enrichment) platform has been established for observing the effect of elevated tropospheric ozone concentration on the gas exchange and chlorophyll fluorescence of two rice varieties (Wuyunjing 21 and Liangyoupeijiu). The results showed that high ozone concentration decreased the net photosynthetic rate (Pn), stomatal conductance (Gs), and transpiration rate (Tr) of rice leaves. After 76d fumigation the decline in them for Wuyunjing 21 was as follows: 21.7%, 26.64% and 24.74% respectively, and that for Liangyoupeijiu was as follows: 25.53%, 30.31% and 25.48% respectively; however, no significant impact on leaf intercellular CO2 concentration was observed. Chlorophyll fluorescence kinetics parameters changed as can be seen by the decrease in F0 (initial fluorescence in the dark), ETR (The apparent electron transfer rate) and psiPSII (actual photochemical efficiency of PS II in the light), and the increase in NPQ (non-photochemical quenching). After 76 days of O3 treatment, the NPQ of Wuyunjing 21 and Liangyoupeijiu was enhanced by 16.37% and 11.77%, respectively. The impact of ozone on rice was a cumulative effect, and the extent of variation in the above parameters and the differences between the two varieties were enlarged as the O3 treatment time increased; At the same time because the rice leaf intercellular CO2 concentration did not significantly reduce, the inferred decrease in net photosynthetic rate was restricted by non-stomatal factors. The results of this experiment indicated that Liangyoupeijiu was more susceptible to ozone than Wuyunjing 21.