The Maritimes Basin contains some of Earth's best-studied records of late Palaeozoic terrestrial environments, which have informed much understanding of ichnology and plant and tetrapod evolution during this interval. However, less attention has been paid to the basin's only fluvio-aeolian constituent - the Cap-aux-Meules Formation (Iles-de-la-Madeleine, Quebec, Canada) - which has previously been considered unfossiliferous and of uncertain age. In this paper, we report plants and diverse trace fossils from the Cap-aux-Meules Formation and describe their sedimentological context. Fluvial facies record deposition in a dryland river system with ephemeral floodplain lakes. The fluvial fossil assemblage includes 9 invertebrate ichnogenera characteristic of the Scoyenia ichnofacies and a fossil stump of a probable woody seed plant, which expands globally sparse records of upright fossil trees in dryland fluvial facies. The presence of dryland vegetation is further evidenced by rhizoliths and vegetation-induced sedimentary structures, comprising accretionary mounds and decay-related forms. In aeolian facies, interdune deposits contain 8 invertebrate ichnogenera of the Scoyenia ichnofacies, in addition to three forms of tetrapod aestivation burrow which constitute the earliest examples of vertebrate burrows from aeolian strata. Dune facies host abundant tetrapod trackways, resembling Erpetopus and Varanopus curvidactylus, that are characteristic of the late Artinskian-Kungurian (late early Permian) Erpetopus biochron and provide the first direct biostratigraphic age constraint for the Cap-aux-Meules Formation. The formation represents the culmination of long-term aridification in Euramerica, which began in the late Carboniferous, and witnessed the expansion of dryland habitats, the rise to dominance of coniferous seed plants, and the infaunalization of vertebrates and invertebrates. The trace fossils are archetypal of lower Permian aeolian strata, where the earliest widespread evidence for faunal colonisation of desert dune-fields appears globally.