Earth history was punctuated during the Permo-Carboniferous [300 -250 million years (Myr) ago] by the longest and most severe glaciation of the entire Phanerozoic Eon. But significant uncertainty surrounds the concentration of CO 2 in the atmosphere through this time interval and therefore its role in the evolution of this major prePleistocene glaciation. Here, I derive 24 Late Paleozoic CO 2 estimates from the fossil cuticle record of arborsecent lycopsids of the equatorial Carboniferous and Permian swamp communities. Quantitative calibration of Late Carboniferous (330 -300 Myr ago) and Permian (270 -260 Myr ago) lycopsid stomatal indices yield average atmospheric CO 2 concentrations of 344 ppm and 313 ppm, respectively. The reconstructions show a high degree of self-consistency and a degree of precision an order of magnitude greater than other approaches. Low CO 2 levels during the PermoCarboniferous glaciation are in agreement with glaciological evidence for the presence of continental ice and coupled models of climate and ice-sheet growth on Pangea. Moreover, the Permian data indicate atmospheric CO 2 levels were low 260 Myr ago, by which time continental deglaciation was already underway. Positive biotic feedbacks on climate, and geotectonic events, therefore are implicated as mechanisms underlying deglaciation.fossil plants ͉ paleoclimates ͉ stomata ͉ carbon isotopes P aleobotanical approaches for quantitatively estimating the pre-Quaternary CO 2 content of the Earth's atmosphere exploit the inverse relationship between leaf stomatal index (SI, fraction of epidermal cells that are stomata) and CO 2 shown by modern trees (1). However, their application has focused on the Cretaceous and Early Tertiary when extant species are represented in the fossil record (2). Attempts to extend this approach further back into the Paleozoic era with the genus Ginkgo are much less secure because of the need to switch to related taxa, such as pteridosperm genera, whose SIs lie outside the range of the calibration dataset (3). These considerations severely limit the utility of the plant fossil record for evaluating the role of CO 2 as a greenhouse gas during the Late Paleozoic glaciation (4), a time when quite large discrepancies are evident between geochemical modeling studies of the long-term carbon cycle (5), and paleobiological proxy CO 2 measurements (3, 6).Here, I report 24 quantitative Carboniferous and Permian atmospheric CO 2 estimates derived from the stomatal characteristics of fossil arborscent lycopsids, the spore-bearing vascular plant group dominating equatorial Euramerican and Cathaysian coal swamp floras during the Late Palaeozoic (7). The fossil lycopsid SIs were calibrated against the response to atmospheric CO 2 variations of three geographically distinct contemporary tropical Lycopodium cernuum populations, taken to be a reasonable nearest living equivalent (NLE) taxon among extant lycopsid species (8). Phylogenetically, Isoetes represents the nearest living relative to members of the Lepidodendron, Bothr...