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We studied mycorrhizal associations of North American Carya laciniosa and Carya cordiformis trees, successfully acclimated to local habitat conditions of the historic Kórnik Arboretum in Poland, in order to better understand mycorrhizal host range extensions in new environments. The root systems of Carya seedlings (1–3 years old), regenerated under a canopy of mature hickory trees, were analyzed using microscopic, morphological, and molecular techniques. Our results, for the first time, indicate that C. laciniosa and C. cordiformis have both arbuscular and ectomycorrhizal associations. In the cleared and stained roots of both Carya species, typical structures of arbuscular mycorrhizae (vesicles, arbuscules, hyphal coils, and intercellular nonseptate hyphae) were detected. On the basis of ITS rDNA sequencing, 40 ectomycorrhizal fungal taxa were revealed, with 25 on C. laciniosa and 19 on C. cordiformis …