The fungi associated with diseased and healthy shoots of young, planted Betula pendula were studied at five sites in Scotland where crown die-back of birch is occurring. At each site, 30 diseased and 30 healthy shoots were sampled twice, with 1 yr-old shoots collected in May 2002, and 4–5-month-old shoots collected in September/October 2002. Overall, 34 different fungal species were recorded, with 92% and 60% of all diseased and healthy shoot pieces, respectively, colonised by at least one fungal species. The main colonisers of both diseased and healthy shoots were Melanconium bicolor, Discula betulina and Godronia urceolus, and the fungi most frequently found on diseased shoots only included Marssonina betulae and Fusarium avenaceum. The pathogenicity of these fungi was tested on seedlings of B. pendula and B. pubescens. Melanconium bicolor required a wound for lesion development on both birch species and caused larger lesions on B. pubescens than on B. pendula. Discula betulina caused lesions on stems and shoots of both birch species, including non-wounded current shoots. Marssonina betulae caused lesions and die-back of wounded and non-wounded current shoots of B. pendula seedlings. This fungus was not pathogenic on B. pubescens. Fusarium avenaceum caused restricted lesions on the main stems of both birch species. Godronia urceolus was not pathogenic on either birch species. Further work needs to focus on the role of these fungi in causing crown die-back of young, planted B. pendula in Scotland.