Dill (Anethum graveolens L.) has been used in traditional Uighur medicine for its various pharmacological activities. Previous studies have suggested that dill seed essential oil (DSEO) has anti-Candida potential and the mechanism of its action also has been studied. Our study examined whether DSEO induces apoptosis in the human pathogen Candida albicans ATCC 64550. Our results indicate that C. albicans ATCC 64550 cells treated with DSEO show some typical apoptosis characters, such as decrease in adenosine triphosphatase (ATPase) activity, chromatin condensation, nuclear fragmentation, and phosphatidylserine (PS) exposure. The DSEO promoted cytochrome c (cyt c) release and metacaspase activation, which resulted in C. albicans ATCC 64550 apoptosis. l-cysteine prevented the DSEO-induced nuclear fragmentation, PS externalization, and metacaspase activation, thus indicating that the reactive oxygen species (ROS) is an important mediator of DSEO-induced apoptosis. To our knowledge, this study is the first to report the induction of apoptosis of this pathogen with concomitant metacaspase activation by DSEO.