The inverse relationship between the number of stomata on a leaf surface and the atmospheric carbon dioxide concentration ([CO(2)]) in which the leaf developed allows plants to optimise water-use efficiency (WUE), but it also permits the use of fossil plants as proxies of palaeoatmospheric [CO(2)]. The ancient conifer family Araucariaceae is often represented in fossil floras and may act as a suitable proxy of palaeo-[CO(2)], yet little is known regarding the stomatal index (SI) responses of extant Araucariaceae to [CO(2)]. Four Araucaria species (Araucaria columnaris, A. heterophylla, A. angustifolia and A. bidwillii) and Agathis australis displayed no significant relationship in SI to [CO(2)] below current ambient levels (~380 ppm). However, representatives of the three extant genera within the Araucariaceae (A. bidwillii, A. australis and Wollemia nobilis) all exhibited significant reductions in SI when grown in atmospheres of elevated [CO(2)] (1,500 ppm). Stomatal conductance was reduced and WUE increased when grown under elevated [CO(2)]. Stomatal pore length did not increase alongside reduced stomatal density (SD) and SI in the three araucariacean conifers when grown at elevated [CO(2)]. These pronounced SD and SI reductions occur at higher [CO(2)] levels than in other species with more recent evolutionary origins, and may reflect an evolutionary legacy of the Araucariaceae in the high [CO(2)] world of the Mesozoic Era. Araucariacean conifers may therefore be suitable stomatal proxies of palaeo-[CO(2)] during periods of "greenhouse" climates and high [CO(2)] in the Earth's history.