Long-term fire exclusion may weaken ecosystem resistance to the return of fire. We investigated how a surface wildfire that occurred after several decades of fire exclusion affected a southern Appalachian forest transitioning from a fire-adapted to a fire-intolerant state. Tree traits associated with fire adaptation often co-occur with traits for nutrient conservation, including the ectomycorrhizal (ECM) association. In the absence of fire, the ECM association may facilitate the accumulation of organic matter, which becomes colonized by fine roots that then become vulnerable to consumption or damage by fire. Therefore, a deeper organic horizon might make stands of fire-adapted, ECM trees less resistant to a surface wildfire than stands of arbuscular mycorrhizal (AM), fire-intolerant trees. To test this hypothesis, we established plots in stands that fall along a gradient of mycorrhizal tree relative abundance both inside and outside the perimeter of the 2016 Rock Mountain wildfire. With increasing relative abundance of ECM trees, we found increasing organic horizon depth and mass and slower rates of decay, even for litter of ECM tree species. We calculated a major (73-83%) reduction in fine root biomass and length in the organic horizon following the wildfire. Over three years post-fire, we observed a higher probability of crown decline, basal sprouting and aboveground biomass mortality with increasing abundance of ECM trees. We propose that the biogeochemistry of mycorrhizal associations can help explain why fire exclusion makes stands of fire-adapted trees less resistant to a surface wildfire than those with fire-intolerant trees.