Juvenile hormone (JH) is an important regulator of development and physiology in insects. While in many insect species, including bumble bees, JH functions as gonadotropin in adults, in some highly eusocial insects its role has shifted to regulate social behavior including division of labor, dominance and aggression. Studying JH functions across social insect species is important for understanding the evolution of sociality; however, these studies have been limited due to inability to reduce JH levels without surgically removing its glandular source, the corpora allata. Precocene is known to inhibit JH biosynthesis in several non-social insects, but was poorly studied in social insects. Here, we tested if precocene-I can effectively reduce JH levels in Bombus terrestris workers, and examined its effects on their physiology and behavior. Precocene-I treatment of 3-worker groups decreased JH titer and ovarian activation, irrespective of the bees' dominance rank within the group, and was remedied by JH replacement therapy. Precocene-I also decreased aggressiveness and increased ester-sterility-signal production. These changes were rank-dependent, affected mainly in the most reproductive and the least aggressive workers, respectively, and could not be remedied by JH replacement therapy. These results clearly confirm the role of JH as a gonadotropin and mediator of aggression in B. terrestris, and that JH effects are associated with worker dominance rank. The ability to chemically reduce JH titer provides us with a non-intrusive method to probe the evolutionary changes associated with JH and the hormonal mechanisms that are associated with reproduction and behavior in social insects.