Contamination of agricultural soils with heavy metal leads to a decrease in the crop quality and yield, as well as increases in public health risks. In this study, we aimed to evaluate the impact of soil contamination with lead (Pb) on the growth, photosynthesis, respiration, and coupling between these physiological processes, as well as temporal dynamics of Pb uptake and accumulation by lettuce (Lactuca sativa L.) plants. For this 46-day pot experiment, Pb(NO3)2 was mixed with loamy Retisol soil with the rate of 0, 50, and 250 mg kg−1. No significant differences in plant biomass accumulation were found between plants grown on Pb-free and Pb-rich soil, but root-weight ratio, root-to-shoot weight ratio, and leaf area were lower, and the number of leaves and leaf weight per unit area were significantly higher in plants grown on soil contaminated with Pb than in their counterparts grown on Pb-free soil. The concentration of Pb in plant root and shoot followed the increase in soil Pb, with Pb content in the roots being higher than in the shoots. Soil Pb decreased chlorophyll content, net CO2 assimilation rate and photosynthetic light use efficiency, but caused an increase in the leaf respiration rate regardless of whether respiration was performed in the light or in darkness. Increased ratio of respiration to photosynthesis reflects the shift in the carbon balance of lettuce plants toward carbon losses under stress conditions of soil contamination with Pb.