The introduction of alien plants can influence biodiversity and ecosystems. However, its consequences for soil microbial communities remain poorly understood. We addressed the impact of alien ectomycorrhizal (EcM) pines on local arbuscular mycorrhizal (AM) fungal communities in two regions with contrasting biogeographic histories: in South Africa, where no native EcM plant species are present; and in Argentina, where EcM trees occur naturally. The effect of alien pines on AM fungal communities differed between these regions. In South Africa, plantations of alien EcM pines exhibited lower AM fungal richness and significantly altered community composition, compared with native fynbos. In Argentina, the richness and composition of local AM fungal communities were similar in plantations of alien EcM pines and native forest. However, the presence of alien pines resulted in slight changes to the phylogenetic structure of root AM fungal communities in both regions. In pine clearcut areas in South Africa, the richness and composition of AM fungal communities were intermediate between the native fynbos and the alien pine plantation, which is consistent with natural regeneration of former AM fungal communities following pine removal. We conclude that the response of local AM fungal communities to alien EcM pines differs between biogeographic regions with different histories of species coexistence.