Biological and geochemical CO2 proxies provide critical constraints on understanding the role of atmospheric CO2 in driving climate change during Earth history. As no single existing CO2 proxy is without its limitations, there is a clear need for new approaches to reconstructing past CO2 concentrations. Here we develop a new pre-Quaternary CO2 proxy based on the stable carbon isotope composition (δ13C) of astomatous land plants. In a series of CO2-controlled laboratory experiments, we show that the carbon isotope discrimination (Δ13C) of a range of bryophyte (liverwort and moss) species increases with atmospheric CO2 across the range 375 to 6000 ppm. Separate experiments establish that variations in growth temperature, water content and substrate type have minor impacts on the Δ13C of liverworts but not mosses, indicating the greater potential of liverworts to faithfully record past variations in CO2. A mechanistic model for calculating past CO2 concentrations from bryophyte Δ13C (White et al., 1994) is extended and calibrated using our experimental results. The potential for fossil liverworts to record past CO2 changes is investigated by analyzing the δ13C of specimens collected from Alexander Island, Antarctica dating to the greenhouse world of the mid-Cretaceous. Our analysis and isotopic model yield mid-Cretaceous CO2 concentrations of 1000--1400 ppm, in general agreement with independent proxy data and long-term carbon cycle models. The exceptionally long evolutionary history of bryophytes offers the possibility of reconstructing CO2 concentrations back to the mid-Ordovician, pre-dating all currently used quantitative CO2 proxies.