Net photosynthetic rates and mesophyll conductances were measured under standardized conditions for leaves of two C3 and one C4 annual species grown at temperatures of 20 to 32°C. Plants were grown with varying day and night temperatures, and also at constant temperatures equal to all the day and night temperatures used. Plants were grown with 8, 12, and 16 hours of light per day. This design allowed determination of whether photosynthetic characteristics were best correlated with day, night, mean, or time-weighted mean temperatures, The results showed that for Glycine max (L.) Merr. (C3) night temperature was most important in determining photosynthetic characteristics, while in Helianthus annuus L. (C3) and Amaranthus hypochondriacus L. (C4) the time-weighted mean temperature was most important. The results for all species were consistent with the hypothesis that development of photosynthetic characteristics is related to a balance between the rate of leaf expansion and the rate of photosynthesis under the growth conditions.