Rice (Oryza sativa L.) as the staple food is widely grown, but the current high tropospheric ozone (O₃) pollution has seriously threatened the rice production. We performed an experiment on four rice cultivars widely grown in the Yangtze River Delta region of China, which were exposed to five O₃ treatments under open top chambers. The effects of O₃ on leaf physiology, growth, yield and grain quality of rice were comprehensively explored, and the results showed that (i) elevated O₃ reduced the light-saturated rate of CO₂ assimilation (A_sat) and stomatal conductance (g_s), and the O₃-induced decrease in A_sat was mainly attributed to stomatal limitation under low O₃ treatment whereas stomatal and biochemical co-limitation under high O₃ treatment; (ii) elevated O₃ did not affected the harvest index since the inhibition of shoot biomass and yield was the same extent, which indicated that O₃ does not alter the remobilization of carbon assimilates from vegetative organs to grains; (iii) only ambient air plus 60 ppb O₃ treatment (NF60) significantly reduced the yield of rice, which was caused by the combined reduction of individual grain weight, grain number, and panicle number; (iv) elevated O₃ did not significantly affect the concentration of mineral nutrients in grains, while the effect of O₃ on the concentration of amino acids was consistent with biphasic O₃-response profile. NF60 had a greater reduction in the concentration of essential amino acids than non-essential amino acids; (v) NF60 significantly reduced the accumulation of mineral nutrients and amino acid in grain due to a decrease in grain yield. These results are conducive to a comprehensive understanding of the impacts of O₃ on rice, and also indicated that the risk assessment of O₃ on crops would need to consider both crop yield and grain quality.