Soybean, Glycine max, is a relatively new crop in Europe gaining increasing interest for its potential to fix atmospheric nitrogen and thus increase sustainability of modern agricultural production systems. Under temperate conditions, root-lesion nematodes (Pratylenchus spp.) are one of the most prominent plant-parasitic nematode taxa of economic concern in soybean. Here, we investigated the susceptibility of soybean cultivars towards naturally occurring populations of Pratylenchus spp. and their effects on soybean yield. At the site Bundesallee the cultivars ‘Abelina’, ‘Primus’ and ‘Taifun3’ supported the highest infestation and multiplication of a mixed population of P. neglectus and P. crenatus, while ‘Sultana’ and ‘Solena’ were least susceptible. At the field site Groß Lüsewitz, ‘Primus’ and ‘Merlin’ cultivars were significantly more susceptible to P. neglectus than ‘Sultana’. Soybean yield was reduced by high initial densities of Pratylenchus spp. Results are expected to contribute to a better nematode management.