Functional morphology and evolution of the hyper-elongated intromittent organ in Cassida leaf beetles (Coleoptera: Chrysomelidae: Cassidinae)

The peculiar phenomenon of hyper-elongation of intromittent organs is well known in a number of insect groups. However, the unresolved questions of how and why such a phenomenon originated independently many times continue to attract biologists' attention. To be able to detect the evolutionary driving mechanisms that enabled insects to repeatedly acquire such a peculiarity, first of all the structural key features and the mechanics of these organs have to be determined. In the present study, the morphology of the reproductive organs of two species of the beetle genus Cassida, with a special focus on the male structures, was scrutinised in detail during copulation and at rest using different microscopy techniques. We found that the hyper-elongated structure of the intromittent organ, called flagellum, is part of the male ejaculatory duct. When the flagellum is inserted into the female spermathecal duct, longitudinal muscles of the ejaculatory duct, but not the flagellum, are shortened. These results strongly suggest that the contraction of the longitudinal muscles of the ejaculatory duct causes propulsion of the flagellum into the highly spiralled spermathecal duct of the female. The tip of the cuticular flagellum is curled up, which can suggest that its physical properties differ from those of the rest of the flagellum. Considering the preceding modelling studies, this property aids the flagellum in penetrating within the highly spiralled and very variable female duct. Based on our morphological results and in comparison with the morphology of intromittent organs of other beetles, we discuss the evolutionary origin of the hyper-elongation in the Cassida species and propose a hypothesis that explains the independent origin of the hyper-elongation of intromittent organs.