The function of insect external genitalia has played a significant role in exploring insect mating mechanisms and male fertilization strategies. However, due to the privacy of genital coupling, insect copulatory mechanisms have only been investigated in a few insect groups. In this study, we observed the mating behavior using freeze-fixated pairs in copula to reveal the copulatory mechanism of the longhorn beetle Moechotypa diphysis (Pascoe, 1871). At the beginning stage of mating, the male M. diphysis usually takes 30 min to control the female and then extends its median lobe and endophallus. Approximately 80% of males (19/24) of M. diphysis exhibit multiple expansions (the membranous endophallus expands and enters into the female genital tract), ranging from two to five times. There are two types of expansions: short ones lasting for 1.4 to 49 s and long ones ranging from 1.03 to 7.23 min. During copulation, male tarsi continuously grasped the female elytra, thorax, and abdomen to help the male to initiate and maintain copulation. Male genital structures are closely connected to female genital structures: the apical phallomere and flagellum on the male endophallus contacting the bursa copulatrix duct and the spermathecal duct of the female, and the abundant microstructures on the surface of the everted male endophallus directly anchoring the female genital tract. Finally, we discuss the possible reasons for the evolution of their complex mating-related structures. Our research will help to explore the evolutionary mechanisms of insect genital structures.
