Reduction in the photosynthetic capacity of plants is presumed to negatively affect their fungal symbionts. To test this hypothesis under natural conditions, we artificially removed 100% of previous year needles in two successive years on Scots pine trees (Pinus sylvestris L.) to simulate pine sawfly attack. Despite a decline in the shoot growth of defoliated trees, root biomass did not differ from control trees. The ergosterol (fungal biomass) and starch concentration of fine roots, however, slightly declined in defoliated trees. Percent ectomycorrhizal colonization of fine root tips remained high in both defoliated and control trees. The dominant tubercle morphotypes were slightly more abundant in the control than in defoliated trees. In contrast to the relatively weak effects on vegetative ectomycorrhizae, reproduction declined near the defoliated pines. Average sporocarp numbers and, consequently, the relative fungal investment to reproduction of the estimated total fungal biomass were more than three times higher near controls than defoliated trees in the first treatment year. Defoliation also reduced the diversity of ectomycorrhizal species producing sporocarps. Mutualistic fungal symbionts may thus alter their reproductive investment in response to restrictions on host resources. Because fungal biomass in the roots as well as colonization percentage remained unchanged, Scots pine evidently continues to invest in the maintenance of the symbiosis despite the reduced photosynthetic capacity due to defoliation.