In plants, pollen donor composition can differ during the early phases of reproduction through various selection mechanisms favouring self, related or nonrelated pollen donors, but such differences have not been examined under natural conditions because paternity is difficult to analyse in a natural setting. Here, we performed paternity analyses based on microsatellite genotyping of individual pollen grains deposited on female flowers (n = 773) and seeds (n = 304) to evaluate pollen donor composition from three individuals of the insect-pollinated monoecious tree Castanea crenata in a natural forest. Spatial genetic structure was also investigated. A mean self-pollen rate of 90.2% was observed at the pollination stage, but a low selfing rate of 0.3% was observed at the seed stage. In outcross events, however, pairwise distance and relatedness between maternal and paternal parents were not different between pollination and seed stages. We also observed significant positive relatedness, based on clear fine-scale genetic structure of individual trees within 80 m of one another, and 71% of seeds were derived using pollen grains of related trees within 80 m. The results suggest that the mechanism of self-incompatibility strongly avoids self-pollen before seed production. However, the avoidance of biparental inbreeding was not obvious between pollination and seed stages.