The interior regions of South Africa have had less attention devoted to archaeological research than coastal regions, and palaeoenvironmental studies are also more limited. As such, little is known about the interaction between human behaviours and past environments in these semi-arid regions. Here, we present an archaeobotanical and mineralogical study from the Middle Stone Age site of Mwulu's Cave, Limpopo Province. Our study shows the importance of using taphonomical approaches prior to interpreting archaeobotanical assemblages, while provides with novel information on the plants used by ancient inhabitants of Mwulu's. The grass phytolith composition is of environmental significance, where a shift from C 4 Panicoideae to C 3 grasses is observed in the last occupation event. This tentatively suggests a shift in rainfall regime, from summer rainfall conditions to an increase in winter rain, during Marine Isotope Stage 5b in the Polokwane region, or a decrease in rainfall seasonality. Although we are unable to chronostratigraphically associate this change in the plant composition, our study adds evidence in support of previous propositions for an expansion of the winter rainfall zone into the interior regions of South Africa.