Reproductive character displacement explains strengthening of mechanical barriers in damselflies

ABSTRACT Reinforcing natural selection against maladaptive hybrids can favor the strengthening of premating reproductive isolation driving a pattern of Reproductive Character Displacement (RCD). In a recent study conducted in North-West (NW) (older) Spanish hybrid zone, was detected an asymmetric reinforcement of the mechanical isolation in the reciprocal cross direction between I. graellsii males and I. elegans females. Furthermore, in the North-Central and Mediterranean (NCM) (younger) hybrid zone was also detected a similar strengthening of the mechanical isolation, consistent with a pattern of asymmetric reinforcement in this hybrid zone as well. In this study, we did geometric morphometrics analyses, shape, and Centroid Size (CS), on male and female secondary sexual traits to investigate whether reinforcement has generated a pattern of RCD of these traits in both hybrid zones. We detected, in the NW hybrid zone, unidirectional RCD (CS) of the male caudal appendages of I. graellsii , and bidirectional RCD (shape) of the female prothorax. Consistently with the prediction that the signal of reinforcement may diminish rapidly once reinforcement ceases to operate, a stronger signal of RCD was detected in the NCM than in the NW hybrid region. In this region, was detected unidirectional RCD (CS) of the male caudal appendages which was consistent with the lock-and-key mechanism of genital coevolution, as well as RCD (shape) of the female prothorax of I. elegans. Interestingly, our study highlights the importance of using geometric morphometrics to deal with the complexity of reproductive structures and controlling for environmental and geographic factors to investigate RCD.