In Flammulina velutipes the origin and development of the dikaryon (as indicated by the formation of clamp connexions) has been studied in colonies derived either from a multi-basidiospore inoculum or from a paired inoculum of two compatible monokaryons. Occasional clamped hyphae arose, but not until 3–4 d after inoculation, and no further hyphae of this kind were initiated once the growing fringe of the colony had become established. In colonies with diameters of 40–70 mm, both clamped and unclamped leading hyphae were present in the growing fringe. The eventual dominance of the clamped hyphae may be due to their greater growth rate. No anastomoses were seen in the monokaryon. When compatible monokaryons from separate inocula met and clamped hyphae were formed, no development of the clamped condition arose in advance of the growing, clamped hyphae. There was no evidence that extensive nuclear migration plays a part in dikaryotization as it does in Coprinus cinereus and Schizophyllum commune.