The aim of this study was to assess genetic diversity and spatial distribution of 123 strains of the root-rot pathogen Neonectria macrodidyma isolated from fine living roots of bare-root nursery cultivated Pinus sylvestris. We found that ITS rDNA sequences were 100% identical for all strains. Arbitrary primed PCR fragment analysis showed the presence of two distinct N. macrodidyma types (Type 1 and Type 2), which included respectively 11 and 14 genotypes composed of 1–29 strains. Results of this study indicate that N. macrodidyma is commonly associated with fine living roots of pine seedlings, is largely disseminated by asexual means of local genotypes and has even distribution in the forest nursery soils. In living roots, N. macrodidyma is likely present as dormant propagules but under favourable conditions, it may develop rapidly and have a significant negative effect on plant health and productivity.