In the evolution of parthenogenesis (female reproduction without male fertilization), males and females may be in conflict over genetic transmission to the next generation. In the termite Reticulitermes speratus, queens produce their replacements parthenogenetically while using normal sexual reproduction by mating with kings to produce other colony members. Here, we report that termite queens produce parthenogenetic offspring in the presence of kings by closing micropyles (tiny openings for sperm entry) of unfertilized eggs. Our field survey revealed that there is a large variation in the number of micropyles in the eggs, and that some of the eggs have no micropyle. Microsatellite analysis indicated that micropyleless eggs develope parthenogenetically, whereas eggs with micropyles get fertilized and develope sexually. Comparison of the number of micropyles between the eggs laid by old- and young-queens showed that only old-queens produce micropyleless eggs. Our results demonstrate that the queens control egg fertilization without interference from the kings and produce their replacements parthenogenetically as needed. This suggests that parthenogenesis can evolve in favour of the females’ own interests independently of males’ interests.