Advances in Fertilization. In flowering plants, fertilization is initiated by the delivery of immotile sperm cells to the boundary between two female gametes, the egg cell and the central cell. During female gamete development in Arabidopsis, the nucleus of the central cell becomes positioned toward this boundary. How this specific polarized nuclear position is maintained is unclear and whether it plays a role in successful fertilization remains to be determined. By disrupting and manipulating F-actin dynamics in the central cell, we identified that the balance of F-actin dynamics is important for nuclear positioning in the central cell and that the presence of intact F-actin cables in the central cell correlates with successful karyogamy regardless of the central cell nuclear position. We also report that the surface of the central cell nucleus is enriched in F-actin. Thus, the central cell nucleus might also serve as a site that organizes F-actin cytoskeleton to promote sperm cell nucleus movement and karyogamy.