Ordering Matters: Combinatorial Song Structure Governs Male Responses in a Cicada Species

ABSTRACT The composition of complex acoustic sequences can significantly influence receiver responses. In Meimuna opalifera , a cicada species, males produce a complex calling song whose main section comprises two components: the former and latter parts. A previous study suggested that these components play distinct roles in eliciting male responses. We investigated whether the arrangement of these components influences male response frequency. By preparing stimuli with a normal order, reversed order, elongated versions of the latter part, and conducting playback experiments, we found that male responses significantly declined during reversed‐order playback. Furthermore, in normal‐order songs, the majority of male responses occurred during the latter part. These results indicate that the arrangement of acoustic units is crucial for eliciting response calls (RCs) in M. opalifera . This is the first evidence in insects that the ordering of air‐borne acoustic signal units alters receiver responses, highlighting the importance of signal combinations in male–male communication.