Domestication has dramatically increased crop size and biomass, reflecting the enhanced accumulation of photosynthates. However, we still lack solid empirical data on the impacts of domestication on photosynthetic rates at different light intensities and on leaf anatomy, and of the relationships of photosynthesis with aboveground biomass. In this study, we measured the photosynthetic rate at three photosynthetic photon flux densities of 2000 (high), 1000 (moderate) and 400 μmol m-2 sec-1 (low light intensity), dark respiration, relative chlorophyll content (SPAD), leaf morphology, and aboveground biomass in 40 wild, 91 semiwild, and 42 domesticated cotton genotypes. The study was replicated for two years (growing years 2018 and 2019). During the first domestication stage (transition from wild to semiwild genotypes), domestication led to higher photosynthetic rates measured under high light intensity, higher SPAD, larger leaf area (LA), and lower leaf mass per unit area (LMA), contributing to greater aboveground biomass accumulation in both study years. During the second domestication stage (transition from semiwild to domesticated genotypes), domestication significantly enhanced photosynthesis under low light intensity and reduced LMA, which were associated with increased aboveground biomass in both study years. In conclusion, photosynthesis improvement at different light intensities has been a gradual domestication phase specific process with the rate of photosynthesis enhanced under high light during the first domestication stage, and under low light during the second domestication stage. We argue that these differences reflect a higher proportion of LA photosynthesizing under low light due to enhanced canopy expansion at the second domestication stage.