The sensitivity of plant carbon isotope fractionation (13Δleaf) to changes in atmospheric CO2 concentrations (Ca) is the subject of heavy debate, with some studies finding no sensitivity, while others show a strong dependency. We tested the hypothesis of photosynthetic homeostasis by using δ13C of n-alkanes, cuticles, and bulk organic matter of gymnosperm-rich rocks (Arundel Clay) from two sites deposited during the Aptian, a time that experienced signifi-cant Ca variations. Our results show no effect of Ca on 13Δleaf, and a relatively constant Ci/Ca (0.64 ± 0.04, 1σ; i—intercellular space), a value that is similar to that of modern gymnosperms. These results suggest that Aptian gymnosperms used homeostatic adjustments with rising Ca, probably involving increased carbon assimilation and/or stomatal closure, a response also found in modern gymnosperms. The similarity between Aptian and modern gymnosperms suggests that the processes responsible for regulating CO2 and water vapor exchange during photosynthesis have remained unaltered in gymnosperms for the past 128 m.y.