Interspecific aggression causes negative selection on sexual characters

Interspecific aggression originating from mistaken species recognition may cause selection on secondary sexual characters, but this hypothesis has remained untested. Here we report a field experiment designed to test directly whether interspecific aggression causes selection on secondary sexual characters, wing spots, in wild damselfly populations. Males of Calopteryx virgo are more aggressive toward males of C. splendens with large than with small wing spots. This differential interspecific aggression may cause negative selection on wing spot size. Indeed, our results show that directional survival selection on wing spot size of C. splendens males was changed by experimental removal of C. virgo males. Without removal, directional selection went from positive to negative with increasing relative abundance of C. virgo males. In populations where C. virgo males were removed, this relationship disappeared. These results verify that interspecific aggression can cause negative selection on sexual characters. Thus, interspecific aggression has the potential to cause divergence on these characters between two species offering an alternative explanation for reinforcement for generating character displacement in secondary sexual characters.     

Selective predation on wing morphology in sympatric damselflies

Although predation is thought to affect species divergence, the effects of predator-mediated natural selection on species divergence and in nonadaptive radiations have seldom been studied. Wing melanization in Calopteryx damselflies has important functions in sexual selection and interspecific interactions and in species recognition. The genus Calopteryx and other damselfly genera have also been put forward as examples of radiations driven by sexual selection. We show that avian predation strongly affects natural selection on wing morphology and male wing melanization in two congeneric and sympatric species of this genus (Calopteryx splendens and Calopteryx virgo). Predation risk was almost three times higher for C. virgo, which has an exaggerated degree of wing melanization, than it was for the less exaggerated, sympatric congener C. splendens. Selective predation on the exaggerated species C. virgo favored a reduction and redistribution of the wing melanin patch. There was evidence for nonlinear selection involving wing patch size, wing patch darkness, and wing length and width in C. splendens but weaker nonlinear selection on the same trait combinations in C. virgo. Selective predation could interfere with species divergence by sexual selection and may thus indirectly affect male interspecific interactions, reproductive isolation, and species coexistence in this genus.     

Sympatric shift in a male sexual ornament in the damselfly Calopteryx splendens

Character displacement is a process by which interactions between two species that exhibit similar traits, results in geographical patterns of trait divergence in one or both species. These traits evolve to reduce costs of interspecific interactions in sympatry and thus differ from their condition in allopatry. In male damselflies Calopteryx splendens, large wing spots are sexually selected. However, in sympatric populations with Calopteryx virgo, wing spot size decreases as C. virgo abundance increases. The stability of this pattern is unclear, because previous studies have focused on sympatric populations with potentially fluctuating relative abundances. We studied the wing spot sizes of C. splendens in both sympatric and allopatric populations. Our data show that male C. splendens’ wing spots are larger in allopatry than in sympatry with C. virgo. We suggest that both interspecific aggression and avoidance of interspecific reproductive interactions may result in this pattern, although their relative importance remains unclear.     

Gender differences in species recognition and the evolution of asymmetric sexual isolation

Closely related sympatric species are expected to evolve strong species discrimination because of the reinforcement of mate preferences. Fitness costs of heterospecific matings are thought to be higher in females than in males, and females are therefore expected to show stronger species discrimination than males. Here, we investigated gender and species differences in sexual isolation in a sympatric species pair of Calopteryx damselflies. The genus Calopteryx is one of the classic examples of reproductive character displacement in evolutionary biology, with exaggerated interspecific differences in the amount of dark wing coloration when species become sympatric. Experimental manipulation of the extent of dark wing coloration revealed that sexual isolation results from both female and male mate discrimination and that wing melanization functions as a species recognition character. Female choice of conspecific males is entirely based on wing coloration, whereas males in one species also use other species recognition cues in addition to wing color. Stronger species discrimination ability in males is presumably an evolutionary response to an elevated male predation risk caused by conspicuous wing coloration. Gender differences in species discrimination and fitness costs of male courtship can thus shed new light on the evolution of asymmetric sexual isolation and the reinforcement of mate preferences.     

SIMULATING RANGE EXPANSION: MALE SPECIES RECOGNITION AND LOSS OF PREMATING ISOLATION IN DAMSELFLIES

Prolonged periods of allopatry might result in loss of the ability to discriminate against other formerly sympatric species, and can lead to heterospecific matings and hybridization upon secondary contact. Loss of premating isolation during prolonged allopatry can operate in the opposite direction of reinforcement, but has until now been little explored. We investigated how premating isolation between two closely related damselfly species, Calopteryx splendens and C. virgo , might be affected by the expected future northward range expansion of C. splendens into the allopatric zone of C. virgo in northern Scandinavia. We simulated the expected secondary contact by presenting C. splendens females to C. virgo males in the northern allopatric populations in Finland. Premating isolation toward C. splendens in northern allopatric populations was compared to sympatric populations in southern Finland and southern Sweden. Male courtship responses of C. virgo toward conspecific females showed limited geographic variation, however, courtship attempts toward heterospecific C. splendens females increased significantly from sympatry to allopatry. Our results suggest that allopatric C. virgo males have partly lost their ability to discriminate against heterospecific females. Reduced premating isolation in allopatry might lead to increased heterospecific matings between taxa that are currently expanding and shifting their ranges in response to climate change.     

Wing pigmentation in Calopteryx damselflies: a role in thermoregulation?

Body melanization may show adaptive variation related to thermoregulation ability, and it is to be expected that the degree of melanization will change among populations or closely related species across environmental gradients of solar radiation and/or environmental temperature. Some melanized secondary sexual traits may also play a role in sexual selection, leading to interpopulation variation, which would not be predicted by thermoregulation pressures alone. We studied the relationships between the interpopulation variation in wing pigmentation level (i.e. melanized secondary sexual trait) of two closely related species of Calopteryx damselfly, and both solar radiation and maximum environmental temperature estimates. Wing pigmentation differs between these species, is gender specific and is used in species' discrimination. Only Calopteryx virgo meridionalis males showed a significant negative partial correlation between wing pigmentation degree and temperature. However, C. virgo meridionalis females showed a positive significant partial correlation between wing pigmentation degree and solar radiation. Wing pigmentation in Calopteryx xanthostoma males was not related to solar radiation or temperature. Thus, thermoregulation pressures poorly explained the observed variations in wing pigmentation between populations, although they might have an adaptive significance at the species' level. As wing pigmentation showed important latitudinal variation, several other selection pressures which might act on melanized traits are briefly discussed. © 2011 The Linnean Society of London, Biological Journal of the Linnean Society, 2011, 103 , 36–44.     

Character displacement in the fighting colours of Hetaerina damselflies

Aggression between species is a seldom-considered but potentially widespread mechanism of character displacement in secondary sexual characters. Based on previous research showing that similarity in wing coloration directly influences interspecific territorial aggression in Hetaerina damselflies, we predicted that wing coloration would show a pattern of character displacement (divergence in sympatry). A geographical survey of four Hetaerina damselfly species in Mexico and Texas showed evidence for character displacement in both species pairs that regularly occurs sympatrically. Hetaerina titia, a species that typically has large black wing spots and small red wing spots, shifted to having even larger black spots and smaller red wing spots at sites where a congener with large red wing spots is numerically dominant (Hetaerina americana or Hetaerina occisa). Hetaerina americana showed the reverse pattern, shifting towards larger red wing spots where H. titia is numerically dominant. This pattern is consistent with the process of agonistic character displacement, but the ontogenetic basis of the shift remains to be demonstrated.     

Interspecific aggression,not interspecific mating,drives character displacement in the wing coloration of male rubyspot damselflies (Hetaerina)

Traits that mediate intraspecific social interactions may overlap in closely related sympatric species, resulting in costly between-species interactions. Such interactions have principally interested investigators studying the evolution of reproductive isolation via reproductive character displacement (RCD) or reinforcement, yet in addition to reproductive interference, interspecific trait overlap can lead to costly between-species aggression. Previous research on rubyspot damselflies (Hetaerina spp.) demonstrated that sympatric shifts in male wing colour patterns and competitor recognition reduce interspecific aggression, supporting the hypothesis that agonistic character displacement (ACD) drove trait shifts. However, a recent theoretical model shows that RCD overshadows ACD if the same male trait is used for both female mate recognition and male competitor recognition. To determine whether female mate recognition is based on male wing coloration in Hetaerina, we conducted a phenotype manipulation experiment. Compared to control males, male H. americana with wings manipulated to resemble a sympatric congener (H. titia) suffered no reduction in mating success. Thus, female mate recognition is not based on species differences in male wing coloration. Experimental males did, however, experience higher interspecific fighting rates and reduced survival compared to controls. These results greatly strengthen the case for ACD and highlight the mechanistic distinction between ACD and RCD.     

Molecular population divergence and sexual selection on morphology in the banded demoiselle (Calopteryx splendens)

The importance of sexual selection in population divergence is of much interest, mainly because it is thought to cause reproductive isolation and hence could lead to speciation. Sexually selected traits have been hypothesized to diverge faster between populations than other traits, presumably because of differences in the strength, mechanism or dynamics of selection. We investigated this by quantifying population divergence in eight morphological characters in 12 south Swedish populations of a sexually dimorphic damselfly, the banded demoiselle (Calopteryx splendens). The morphological characters included a secondary sexual character, the male melanized wing spot, which has an important function in both inter- and intrasexual selection. In addition, we investigated molecular population divergence, revealed by amplified fragment length polymorphism (AFLP) analysis. Molecular population divergence was highly significant among these Northern European populations (overall F(st)=0.054; pairwise population F(st)'s ranged from approximately 0 to 0.13). We found evidence for isolation-by-distance (r=0.70) for the molecular markers and a significant correlation between molecular and phenotypic population divergence (r=0.39). One interpretation is that population divergence for the AFLP loci are affected by genetic drift, but is also indirectly influenced by selection, due to linkage with loci for the phenotypic traits. Field estimates of sexual and natural selection from two of the populations revealed fairly strong sexual selection on wing spot length, indicating that this trait has the potential to rapidly diverge, provided that variation is heritable and the observed selection is chronic.     

The evolution of species recognition in competitive and mating contexts: the relative efficacy of alternative mechanisms of character displacement

Sympatric divergence in traits affecting species recognition can result from selection against cross‐species mating (reproductive character displacement, RCD) or interspecific aggression (agonistic character displacement, ACD). When the same traits are used for species recognition in both contexts, empirically disentangling the relative contributions of RCD and ACD to observed character shifts may be impossible. Here, we develop a theoretical framework for partitioning the effects of these processes. We show that when both mate and competitor recognition depend on the same trait, RCD sets the pace of character shifts. Moreover, RCD can cause divergence in competitor recognition, but ACD cannot cause divergence in mate recognition. This asymmetry arises because males with divergent recognition traits may avoid needless interspecific conflicts, but suffer reduced attractiveness to conspecific females. Therefore, the key empirical issue is whether the same or different traits are used for mate recognition and competitor recognition.     

Does character displacement demonstrate density-dependent expression in females? A test on the wing shape of two species of European damselflies

Character displacement (CD) is the evolutionary process which leads to the divergence in trait expression of closely related species in regions where species co-occur, compared to allopatric populations. In Europe CD has been investigated in males of Calopteryx splendens and C. virgo and has been related to species recognition. If species recognition is relevant for males, also females should benefit from CD. The most obvious differences between females of these two species are wing profile and colour. We sampled females from allopatric and from sympatric populations with different relative abundances of these species. Wing shape and pigmentation were evaluated for each damselfly. CD was found in wing profile but not in wing transparency. The relative abundance of species significantly affected CD, but with a different pattern in each species. The prediction that wing shape become more different from the allopatric state when the species was relatively rare, but more similar to the allopatric state when the species was common was evident only for C. splendens. Wing shape changes might increase differences in flying patterns making males more effective to discriminate between heterospecific females. So, CD we observed may be the result of a selection directed to reduce interspecific reproductive interference.     

Sensory trap as the mechanism of sexual selection in a damselfly genitalic trait (Insecta: Calopterygidae)

During copulation, males of some calopterygid damselfly species displace the sperm stored in the spermatheca: the male genital appendages enter into the spermathecal ducts and physically remove sperm. In Calopteryx haemorrhoidalis, the genital appendages are too wide to penetrate the spermathecae, but males use a different mechanism in which the aedeagus stimulates the vaginal sensilla that control spermathecal sperm release. Since these sensilla are used during egg fertilization and oviposition, it was hypothesized that this function evolved before the male stimulatory ability. I investigated this using Hetaerina cruentata, a species whose position in the Calopterygidae phylogeny is more basal than Calopteryx. Given this position and having determined that males of this species are not able to displace sperm of their conspecific females during copulation, it was expected that H. cruentata females would eject sperm when stimulated with the aedeagi of C. haemorrhoidalis but not when stimulated with the aedeagi of their conspecifics. This prediction was confirmed. In order to investigate the widespread nature of this result, some other Calopteryx species-Calopteryx xanthostoma and Calopteryx virgo-were investigated. The results were similar to those of H. cruentata: conspecific males were unable to stimulate their females, but females ejected sperm when stimulated with C. haemorrhoidalis aedeagi. Morphometric analysis suggests that the mechanistic explanation for the stimulatory ability of C. haemorrhoidalis genitalia is that the aedeagal region that makes contact with the vaginal sensilla is wider in C. haemorrhoidalis than in the other species. These results suggest that the sensory "bias" shown and shared by H. cruentata, Calopteryx splendens, C. virgo, and C. haemorrhoidalis females represents an ancestral condition and that the male stimulatory ability is absent in the evolutionary history of the clade. These pieces of evidence as well as another one presented elsewhere, which indicates that C. haemorrhoidalis males vary in their stimulatory ability, constitute the three criteria for a case of sexual selection via exploitation of a female sensory bias. These results also provide support to the sensory trap hypothesis that indicates that the female bias-in this case, egg fertilization and oviposition-evolved in a context different from sexual selection. Considering that the male genital appendages responsible for physically removing spermathecal sperm in other calopterygids are present in C. haemorrhoidalis, I suggest that males were once able to displace spermathecal sperm physically. Such ability may have been later impeded by a reduction in size of the spermathecal ducts. Possibly, one of the latest events in this sequence is the male's stimulatory ability. This hypothetical series of events suggests a coevolutionary scenario in which the central actor is the sperm stored in the spermathecae.     

Permanent genetic resources added to Molecular Ecology Resources Database 1 February 2011-31 March 2011

This article documents the addition of 111 microsatellite marker loci to the Molecular Ecology Resources Database. Loci were developed for the following species: Acipenser oxyrinchus desotoi, Anopheles nuneztovari sensu lato, Asellus aquaticus, Calopteryx splendens, Calopteryx virgo, Centaurea aspera, Centaurea seridis, Chilina dombeyana, Proctoeces cf. lintoni and Pyrenophora teres f. teres.     

Phenotypic integration and conserved covariance structure in calopterygid damselflies

By comparing the phenotypic (P) variance-covariance matrices between closely related taxa or conspecific populations, one can study the outcome of the interplay between selection and developmental constraints in phenotypic evolution. Shared patterns of phenotypic integration are also of interest and might result from similarities in either selection or developmental pathways. We compared P-matrices and phenotypic integration indices between populations and species of the damselfly genus Calopteryx. P(max)-comparisons between parapatric C. splendens populations revealed stronger conserved phenotypic covariance structure than P(max)-comparisons between species, suggesting that divergence in its early stages proceeds along phenotypic lines of least resistance. Within- and among-population correlations in C. splendens were highly concordant, in further support of initial divergence along P(max). Despite some similarities in overall phenotypic integration between C. splendens and C. virgo, these two species only had several P-matrix eigenvectors in common, indicating that after reproductive isolation, divergence has proceeded against P(max).     

Immunocompetence, developmental stability and wingspot size in the damselfly Calopteryx splendens L

Calopteryx splendens males exhibit a remarkable variation in wing pigmentation both within and between populations. In this study, we examined whether the wingspots of male C. splendens are related to male quality. We measured the nylon implant encapsulation rate for 85 males and found that males with larger wingspots had a faster encapsulation rate, indicating a better immunocompetence. We also found that the encapsulation rate was positively correlated with the density of haemocytes in the haemolymph. Another measurement of male quality, fluctuating asymmetry of wingspots, correlated negatively with the size of the wingspots. Males with asymmetrical wingspots also had lower encapsulation rates than more symmetrical males. Our results suggest that the size of wingspot is an indicator of male quality in C. splendens.     

The Role of Wing Coloration in Sex Recognition and Competitor Recognition in Rubyspot Damselflies (Hetaerina spp.)

The decision rules that animals use for distinguishing between conspecifics of different age and sex classes are relevant for understanding how closely related species interact in sympatry. In rubyspot damselflies (Hetaerina spp.), the red wing coloration of mature males is hypothesized to be a key trait for sex recognition and competitor recognition within species and the proximate trigger for interspecific male–male aggression. We tested this hypothesis by manipulating the wing coloration of tethered conspecific intruders and measuring the responses of territory holders of three species in the field. As predicted, covering the red spots of mature males with black ink nearly eliminated territorial responses, and in some cases, territorial holders clasped the blackened males as if they were females. Adding red spots to female wings triggered territorial responses and nearly eliminated sexual responses. Immature males with artificial red spots were attacked at the same rate as mature male intruders, and much more frequently than were immature male controls. The results varied somewhat by species. In H. titia, the only species of Hetaerina with substantial black wing pigmentation, the effects of blackening the red spots of intruders varied both geographically and seasonally. But even when blackening the red spots of male intruders did not reduce the aggressive response of H. titia territory holders, adding artificial red spots to female wings elicited aggressive responses and nearly eliminated sexual responses. The results of this study further strengthen the evidence that interspecific aggression in Hetaerina results from overlap in territorial signals and that the derived black wing pigmentation of H. titia reduces interspecific aggression.     

Wing pigmentation in territorial male damselflies, Calopteryx haemorrhoidalis: a possible relation to sexual selection

One striking characteristic in adult males of some odonate species is the presence of wing pigmentation. In Calopteryx species, males show a series of pre- and postcopulatory behavioural displays during which they face females while showing their pigmented wings. One hypothesis to explain the precopulatory flying displays and the associated wing pigmentation is that they may serve a sexual selection function. I investigated this in the territorial damselfly Calopteryx haemorrhoidalis. Males of this species defend aquatic substrates that females use for oviposition. Observational evidence indicated that males with a higher proportion of wing pigmentation were more likely to defend a territory, obtained more matings, had fewer gut parasites, survived in the study site and stayed in territories for longer. Experimental evidence suggested that the relationship mating success and wing pigmentation still held when controlling for the size of the substrate defended by territorial males. Similar to other studies in the Calopterygidae, these results suggest that wing pigmentation may be favoured by sexual selection. I discuss, however, whether an alternative function for male copulatory courtship displays and wing pigmentation, as sexual and/or species recognition, may also explain the evolution of these traits. Copyright 2002 The Association for the Study of Animal Behaviour. Published by Elsevier Science Ltd. All rights reserved.     

The role of interspecific interference competition in character displacement and the evolution of competitor recognition

The extent to which interspecific interference competition has contributed to character evolution is one of the most neglected problems in evolutionary biology. When formerly allopatric species come into secondary contact, aggressive interactions between the species can cause selection on traits that affect interspecific encounter rates (e.g. habitat preferences, activity schedules), competitor recognition (e.g. colouration, song), and fighting ability (e.g. weaponry, body size). We define agonistic character displacement (ACD) as the process of phenotypic evolution in a population caused by interference competition with one or more sympatric species and which results in shifts in traits that affect the rate, intensity or outcome of interspecific aggression. After clarifying the relationships between ACD and other evolutionary processes that may occur when species come into secondary contact, we develop an individual‐based, quantitative genetic model to examine how traits involved in competitor recognition would be expected to evolve under different secondary contact scenarios. Our simulation results show that both divergence and convergence are possible outcomes, depending on the intensity of interspecific exploitative competition, the costs associated with mutual versus unilateral recognition, and the extent of phenotypic differences prior to secondary contact. We then devise a set of eight criteria for evaluating putative examples of ACD and review the empirical literature to assess the strength of existing evidence and to identify promising avenues for future research. Our literature search revealed 33 putative examples of ACD across insects, fishes, bats, birds, lizards, and amphibians (15 divergence examples; 18 convergence examples). Only one example satisfies all eight criteria for demonstrating ACD, but most case studies satisfy four or more criteria. The current state of the evidence for ACD is similar to the state of the evidence for ecological character displacement just 10 years ago. We conclude by offering suggestions for further theoretical and empirical research on ACD.     

Interspecific interactions and learning variability jointly drive geographic differences in mate preferences

Co‐occurrence of closely related species can cause behavioral interference in mating and increase hybridization risk. Theoretically, this could lead to the evolution of more species‐specific mate preferences and sexual signaling traits. Alternatively, females can learn to reject heterospecific males, to avoid male sexual interference from closely related species. Such learned mate discrimination could also affect conspecific mate preferences if females generalize from between species differences to prefer more species‐specific mating signals. Female damselflies of the banded demoiselle (Calopteryx splendens) learn to reject heterospecific males of the beautiful demoiselle (C. virgo) through direct premating interactions. These two species co‐occur in a geographic mosaic of sympatric and microallopatric populations. Whereas C. virgo males have fully melanized wings, male C. splendens wings are partly melanized. We show that C. splendens females in sympatry with C. virgo prefer smaller male wing patches in conspecific males after learning to reject heterospecific males. In contrast, allopatric C. splendens females with experimentally induced experience with C. virgo males did not discriminate against larger male wing patches. Wing patch size might indicate conspecific male quality in allopatry. Co‐occurrence with C. virgo therefore causes females to prefer conspecific male traits that are more species specific, contributing to population divergence and geographic variation in female mate preferences.     

Rapid evolution of sexual signals in sympatric Calopteryx damselflies: reinforcement or ‘noisy‐neighbour’ ecological character displacement?

Enhanced prezygotic isolation in sympatry is one of the most intriguing patterns in evolutionary biology and has frequently been interpreted as evidence for reinforcement. However, the frequency with which reinforcement actually completes speciation remains unclear. The Jewelwing damselflies (Calopteryx aequabilis and C. maculata) have served as one of the few classic examples of speciation via reinforcement outside of Drosophila. Although evidence for wing pattern displacement and increased mate discrimination in this system have been demonstrated, the degree of hybridization and gene flow in nature are unknown. Here, we show that sympatric populations of these two species are the result of recent secondary contact, as predicted under a model of speciation via reinforcement. However, we found no phenotypic evidence of hybridization in natural populations and a complete association between species-specific haplotypes at two different loci (mitochondrial CO I and nuclear EF1-alpha), suggesting little or no contemporary gene flow. Moreover, genealogical and coalescent-based estimates of divergence times and migration rates indicate that, speciation occurred in the distant past. The rapid evolution of wing colour in sympatry is recent, therefore, relative to speciation and seems to be better explained by selection against wasting mating effort and/or interspecific aggression resulting from a 'noisy neighbour' signalling environment.