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Seedlings and old trees of Scots pine in self-regenerating, old, virgin boreal forest in northern Sweden were compared in terms of the species composition of their associated ectomycorrhizal (EM) fungi. The natural regeneration of seedlings was mimicked by annual seeding for 10 yr (1986–1995) in intact field vegetation at three sites. In 1995, all seedlings were harvested, and 171 were examined for mycorrhizas. Twenty-five soil cores were taken in order to study the mycorrhizas on the established trees. Most short roots were mycorrhizal. Using mycorrhizal morphology and restriction fragment length polymorphisms (RFLP) analysis, we were able to distinguish 43 ectomycorrhizal taxa. Fourteen of these taxa were identified using a sporocarp and mycelial culture based internal transcribed spacer (ITS)-RFLP reference database, and another three were identified by mycorrhizal morphology. Cenococcum geophilum …