Fossil wood is described from the Late Permian Middleton Formation, Adelaide Subgroup, Beaufort Group of the Karoo Supergroup in the Easter Cape Province, South Africa. The wood consists of in situ tree trunks up to 9 m long, partially enclosed in fine-grained sandstone surrounded by argillaceous mudstone and siltstone. The strata are poorly exposed due to the eroded and denuded land surface. All are lying horizontally and none are in upright growth positions. Comparisons of the study site lithologies with known surrounding Middleton Formation lithostratigraphy supports the interpretation that the palaeoenvironment responsible for the transport and deposition of the fossil wood was fluvial, with sandstone representing in-channel deposits and the mudstone-siltstone flood plain sequences where the trees were originally located before being uprooted and transported within the channels. Thin sections of twenty-nine wood samples revealed the presence of four existing taxa, Agathoxylon africanum, Agathoxylon karooensis, Australoxylon natalense and Australoxylon teixeirae. Growth rings of all the samples show a wide range of ring width from 1 mm to 11 mm, indicating growth under suitable climatic regimes with adequate water supply interspersed by very wet and warm periods. Crushed earlywood, s-shaped cell walls and torn cell walls suggest felling of trees during storm and/or flood events followed by sedimentary transport in palaeochannels prior deposition within the channel sands.