The genetics of somatic incompatibility in tetrapolar Collybia fusipes was studied using eight dikaryotic isolates collected from the wild and their experimentally derived progeny. Monokaryons from each isolate were all paired with the same unrelated monokaryon and also paired together in all combinations. The somatic compatibility of the two resulting sets of dikaryons was studied. Two different types of somatic incompatible interaction were observed, lightly or heavily pigmented lines developing between the two isolates. The dikaryons that had one nuclear type in common and one coming from sibling monokaryons were compatible in 7–27% of the cases, incompatible with a lightly pigmented interaction in 30–93% and incompatible with a heavily pigmented interaction in 0–53%. The results suggest that at least three to four loci control the somatic incompatibility in C. fusipes, one of them alone controlling the heavily pigmented interaction.