How climate and rising carbon dioxide concentrations (pCO2) have influenced competition between C3 and C4 plants over the last 50 years is a critical uncertainty in climate change research. Here we used carbon isotope (δ13C) values of the saprotrophic lawn fungus Amanita thiersii to integrate the signal of C3 and C4 carbon in samples collected between 1982 and 2009 from the Midwestern USA. We then calculated 13C fractionation (Δ) to assess the balance between C3 and C4 photosynthesis as influenced by mean annual temperature (MAT), mean annual precipitation over a 30 year period (MAP-30), and pCO2. Sporocarp Δ correlated negatively with MAT (−1.74‰ °C−1, 79% of variance) and positively with MAP (9.52‰ m−1, 15% of variance), reflecting the relative productivity of C3 and C4 grasses in lawns. In addition, Δ values correlated positively with pCO2 (0.072‰ ppm−1, 5% of variance). Reduced photorespiration with rising pCO2 accounted for 20% of this increased Δ, but the remaining 80% is consistent with increased assimilation of C3-derived carbon by Amanita thiersii resulting from increased productivity of C3 grasses with rising pCO2. Between 1982 and 2009, pCO2 rose by 46 ppm and the relative contribution of C3 photosynthesis to Amanita thiersii carbon increased 18.5%. The δ13C value of Amanita thiersii may integrate both lawn maintenance practices and the physiological responses of turf grasses to rising CO2 concentrations.