Local adaptation of plants to mycorrhizal fungi helps determine the outcome of mycorrhizal interactions. However, there is comparatively little work exploring the potential for evolution in interactions with ectomycorrhizal fungi, and fewer studies have explored the heritability of mycorrhizal responsiveness, which is required for local adaptation to occur. We set up a reciprocal inoculation experiment using seedlings and soil from four populations of Scots pine (Pinus sylvestris) from Scotland, measuring seedling response to mycorrhizal inoculation after 4 months. We estimated heritability for the response traits and tested for genotype × environment interactions. While we found that ectomycorrhizal responsiveness was highly heritable, we found no evidence that pine populations were locally adapted to fungal communities. Instead, we found a complex suite of interactions between pine population and soil inoculum. Our results suggest that, while Scots pine has the potential to evolve in response to mycorrhizal fungi, evolution in Scotland has not resulted in local adaptation. Long generation times and potential for rapid shifts in fungal communities in response to environmental change may preclude the opportunity for such adaptation in this species, and selection for other factors such as resistance to fungal pathogens may explain the pattern of interactions found.