An increase in global demand for crop‐based products necessitates an increased crop yield. Optimizing photosynthesis, which is sensitive to environmental fluctuations, offers a promising strategy to improve crop yield and resilience. Photosynthetic responses often lag behind changes in irradiance, resulting in the loss of potential carbon gain. Additionally, global warming is accompanied by unexpected chilling spells, further affecting photosynthesis. Thus, developing chilling‐resilient crops and optimizing photosynthetic responses to fluctuating light is critical. This can be achieved by identifying genetic markers associated with desirable photosynthetic traits in plant populations. However, the combined effect of chilling and photosynthetic responses to fluctuating light remains unexplored, and there is a lack in populations designed to explore these responses. Thus, exploration needs to be done in pre‐existing populations where there is phenotypic variation in photosynthetic responses and in how chilling affects these responses within parental lines. This study examined the variation in photosynthetic responses of the parental lines of a Multi‐parent‐Advanced‐Generation Inter‐Cross (MAGIC) population of tomato ( Solanum lycopersicum ) under fluctuating light and suboptimal temperatures. Photosynthetic responses to step increases and step decreases in irradiance were measured using modulated chlorophyll fluorescence and the effect of lowered temperature (14°C) on these responses was investigated. The results showed variation in the kinetics of the response of quantum efficiency of PSII (Φ PSII ) to step changes in irradiance under control and chilling conditions. Chilling had a minimal effect on the photosynthetic responses of some parental lines, indicating resilience to chilling. These findings highlight the potential of exploring genetic components to breed climate‐resilient crops.