Assessment of rampant genitalic variation in the spider genus Homalonychus (Araneae, Homalonychidae)

Abstract. Animal genitalia are often complex and thought to vary little within species but differ between closely related species making them useful as primary characters in species diagnosis. Spiders are no exception, with nearly all of the 40,462 (at the time of this writing) described species differentiated by genitalic characteristics. However, in some cases, the genitalia of putative species are not uniform, but rather vary within species. When intraspecific variation overlaps interspecific variation, it can be difficult (if not impossible) to place a name on a specimen. The quantification of shape variation in genitalia has not often been attempted, probably because until recently it was not a methodologically and computationally simple process. In the two currently recognized species of the spider genus Homalonychus , genitalic variation is rampant in both male and female structures, with some parts of the genitalia (e.g., the retrolateral tibial apophysis) differing in each specimen examined. In this study, geometric morphometric analysis employing landmark data is used to quantify both intra- and interspecific variation in this genus. The large amount of variation is condensed into two or three groups depending on the structures examined, and these groups correspond to either the two species or to previously established mitochondrial DNA clades within one of the species. The results also show that analyses of female structures do not separate the groups as readily as the analyses of the male structures. The large amount of variation present in some structures is not correlated with geography or population genetic structure.     

Hierarchical comparative analysis of genetic and genitalic geographical structure: testing patterns of male and female genital evolution in the scarab beetle Phyllophaga hirticula (Coleoptera: Scarabaeidae)

It is generally accepted that genitalia are among the fastest evolving characters in insects and that selection on these structures may increase speciation rates in groups with polygamous mating systems. If selection is causing genitalic divergence between or among populations of a species, one prediction is that geographical structure of genitalic morphology would be in place before genetic structure of a rapidly evolving neutral marker. The current study tests this hypothesis in the geographically widespread scarab beetle Phyllophaga hirticula by evaluating whether standing variation in male and female genitalia is more or less geographically structured than a mitochondrial genetic marker. Geographical structure of mitochondrial (mt)DNA and male and female genitalic shape were analysed using analysis of variance, multivariate analysis of variance, Mantel tests, and tests of spatial autocorrelation. The results show that, although female genitalia are more geographically structured than mtDNA, male genitalia are not. This pattern suggests that selection on female genitalic variation may be causing divergence of these structures among populations.  © 2009 The Linnean Society of London, Biological Journal of the Linnean Society , 2009, 96 , 135–149.     

SPECIES ISOLATION,GENITAL MECHANICS,AND THE EVOLUTION OF SPECIES-SPECIFIC GENITALIA IN THREE SPECIES OF MACRODACTYLUS BEETLES (COLEOPTERA,SCARABEIDAE, MELOLONTHINAE)

The question asked was why male genitalic structures have diverged in three syntopic species of Macrodactylus beetles. Four hypotheses were evaluated: 1. The ways in which male genitalia mesh with internal female structures indicate that selection for species isolation via mechanical exclusion (“lock and key”) is unlikely to explain the genitalic differences. 2. The specific mate recognition hypothesis also clearly fails to explain genitalic differences due to the implausibility of postulated environmental effects on genitalia, and lack of postulated coevolution of male and female morphologies. 3. Selection for species isolation via differences in genitalic stimulation (sensory lock and key) is unlikely due to relatively infrequent cross-specific pair formation and intromission in the field, and “excessive” numbers of species-specific genitalic structures and male courtship behavior patterns which nevertheless occasionally fail. It also fails to explain the frequent failure of intraspecific copulations to result in sperm transfer. This hypothesis cannot, however, be rejected as confidently as the previous hypotheses. 4. Conditions under which sexual selection by cryptic female choice could take place are common. Females frequently exercise their ability to prevent sperm transfer by conspecific males even after intromission has occurred, and females generally mate repeatedly, probably with different males. Males behave as if cryptic female choice is occurring, courting assiduously while their genitalia are within the female. Sexual selection by female choice could thus contribute to the divergence in genitalic structures.     

Copulation behaviour of Glossina pallidipes (Diptera: Muscidae) outside and inside the female, with a discussion of genitalic evolution

If species-specific male genitalia are courtship devices under sexual selection by cryptic female choice, then species-specific aspects of the morphology and behaviour of male genitalia should often function to stimulate the female during copulation. The morphology and behaviour of the complex, species-specific male genitalia of the tsetse fly, Glossina pallidipes Austen, were determined from both direct observations and dissections of flash-frozen copulating pairs; we found that some male genitalic traits probably function to stimulate the female, while others function to restrain her. The male clamps the ventral surface of the female's abdomen tightly with his powerful cerci. Clamping does not always result in intromission. Clamping bends the female's body wall and her internal reproductive tract sharply, posteriorly and dorsally, and pinches them tightly. The male performed sustained, complex, stereotyped, rhythmic squeezing movements with his cerci that were not necessary to mechanically restrain the female and appeared instead to have a stimulatory function. Six different groups of modified setae on and near the male's genitalia rub directly against particular sites on the female during squeezing. The designs of these setae correlate with the force with which they press on the female and the probable sensitivity of the female surfaces that they contact. As expected under the hypothesis that these structures are under sexual selection by female choice, several traits suspected to have stimulatory functions have diverged in G. pallidipes and its close relative, G. longipalpis. Additional male non-genitalic behaviour during copulation, redescribed more precisely than in previous publications, is also likely to have a courtship function. The elaborate copulatory courtship behaviour and male genitalia may provide the stimuli that previous studies showed to induce female ovulation and resistance to remating.     

Selection of a conspicuous male genitalic trait in the seedbug Lygaeus simulans

Phenotypic selection of a male genitalic trait based on insemination success in the seedbug Lygaeus simulans, Deckert 1985, was demonstrated in this study. In a laboratory experiment, 97 pairs of virgin animals copulated once. The females consisted of six groups of full sisters, all sisters of a group having been raised in the same box. Related females were used in order to lower the variance of female preferences and to thereby increase the chances of detecting selection of male traits by female choice. The males were independent, i.e. each male descended from a unique pair of parents. Multiple regressions of insemination success on linear and nonlinear terms of a male genitalic trait (processus gonopori length) and male weight revealed stabilizing selection on the genitalic trait. According to functional considerations this selection is most probably the result of female choice of male processus length.     

Sexual selection,phenotypic variation,and allometry in genitalic and non‐genitalic traits in the sexually size‐dimorphic stick insect Micrarchus hystriculeus

The mobility hypothesis could explain the evolution of female‐biased size dimorphism if males with a smaller body size and longer legs have an advantage in scramble competition for mates. This hypothesis is tested by performing a selection analysis in the wild on Micrarchus hystriculeus (Westwood) (Phasmatodea), a sexually size dimorphic stick insect endemic to New Zealand. This analysis examined the form and strength of sexual selection on body size, leg lengths (front, mid and hind), and clasper size (a genitalic trait), and also quantified the degree of phenotypic variation and the allometric scaling pattern of these traits. By contrast to the mobility hypothesis, three lines of evidence were found to support significant stabilizing sexual selection on male hind leg length: a significant nonlinear selection gradient, negative static allometry, and a low degree of phenotypic variation. Hind leg length might be under stabilizing selection in males if having average‐sized legs facilitates female mounting or improves a male's ability to achieve the appropriate copulation position. As predicted, a negative allometric scaling pattern and low phenotypic variation of clasper size is suggestive of stabilizing selection and supports the ‘one‐size‐fits‐all’ hypothesis. Opposite to males, the mid and hind leg lengths of females showed positive static allometry. Relatively longer mid and hind leg lengths in larger females might benefit individuals via the better support of their larger abdomens. © 2014 The Linnean Society of London, Biological Journal of the Linnean Society, 2014, 113 , 471–484.     

Evidence for lock-and-key mechanisms in the internal genitalia of the Apamea moths (Lepidoptera, Noctuidae)

Abstract. Fifty of the fifty-six species of the genus Apamea known from North America and three Palaearctic species were analysed for lock-and-key characters in their internal genitalia, mainly in the male vesica and the female bursa copulatrix. There were an average of 4.5 such characters per species, structurally corresponding in the two sexes. Anatomically they form a postcopulatory but prezygotic isolation mechanism. In some closely related species, the internal genitalia are very similar, but these species have a precopulatory isolation mechanism in the presence or absence of male abdominal coremata. Closely related species did not have more lock-and-key characters than unrelated species, which is taken to indicate absence of character displacement. The anatomical distribution of the lock-and-key characters was examined and the organs of eight species are illustrated.
The lock-and-key hypothesis has been abandoned by several earlier authors but mainly on consideration of external genitalia. In Apamea the invariable functional correspondence between sexes in the sperm transferring organs, and the overall species-specifity of characters but non-existence of interspecific differences under a precopulatory mechanism indicate that (a) lock-and-keys are functioning and (b) they act as isolation mechanisms. Alternative hypotheses of genitalic evolution are reviewed.     

HAVE MALE AND FEMALE GENITALIA COEVOLVED? A PHYLOGENETIC ANALYSIS OF GENITALIC MORPHOLOGY AND SEXUAL SIZE DIMORPHISM IN WEB‐BUILDING SPIDERS (ARANEAE: ARANEOIDEA)

Sexual size dimorphism (SSD) can strongly influence the evolution of reproductive strategies and life history. If SSD is extreme, and other characters (e.g., genitalic size) also increase with size, then functional conflicts may arise between the sexes. Spiders offer an excellent opportunity to investigate this issue because of their wide range of SSD. By using modern phylogenetic methods with 16 species of orb-weaving spiders, we provide strong evidence for the "positive genitalic divergence" model, implying that sexual genitalic dimorphism (SGD) increases as SSD increases. This pattern is supported by an evolutionary mismatch between the absolute sizes of male and female genitalia across species. Indeed, our findings reveal a dramatic reversal from male genitalia that are up to 87x larger than female genitalia in size-monomorphic species to female genitalia that are up to 2.8x larger in extremely size-dimorphic species. We infer that divergence in SGD could limit SSD both in spiders, and potentially in other taxa as well. Further, male and female body size, as well as male and female genitalia size, are decoupled evolutionarily. Finally, we show a negative scaling (hypoallometry) of male and female genitalic morphology within sexes. Evolutionary forces specific to each sex, such as larger female size (increased fecundity) or smaller male size (enhanced mate-searching ability), may be balanced by stabilizing selection on relative genitalic size.     

The function and evolution of male and female genitalia in Phyllophaga Harris scarab beetles (Coleoptera: Scarabaeidae)

Genitalia diversity in insects continues to fuel investigation of the function and evolution of these dynamic structures. Whereas most studies have focused on variation in male genitalia, an increasing number of studies on female genitalia have uncovered comparable diversity among females, but often at a much finer morphological scale. In this study, we analysed the function and evolution of male and female genitalia in Phyllophaga scarab beetles, a group in which both sexes exhibit genitalic diversity. To document the interaction between male and female structures during mating, we dissected flash‐frozen mating pairs from three Phyllophaga species and investigated fine‐scale morphology using SEM. We then reconstructed ancestral character states using a species tree inferred from mitochondrial and nuclear loci to elucidate and compare the evolutionary history of male and female genitalia. Our dissections revealed an interlocking mechanism of the female pubic process and male parameres that appears to improve the mechanical fit of the copulatory position. The comparative analyses, however, did not support coevolution of male and female structures and showed more erratic evolution of the female genitalia relative to males. By studying a group that exhibits obvious female genitalic diversity, we were able to demonstrate the relevance of female reproductive morphology in studies of male genital diversity.     

Male-female conflict and genitalia: failure to confirm predictions in insects and spiders

Some recent models suggest a new role for evolutionary arms races between males and females in sexual selection. Female resistance to males is proposed to be driven by the direct advantage to the female of avoiding male-imposed reductions in the number of offspring she can produce, rather than by the indirect advantage of selecting among possible sires for her offspring, as in some traditional models of sexual selection by female choice. This article uses the massive but hitherto under-utilized taxonomic literature on genitalic evolution to test, in a two-step process, whether such new models of arms races between males and females have been responsible for rapid divergent evolution of male genitalia. The test revolves around the prediction that 'new arms races' are less likely to occur in species in which females are largely or completely protected from unwanted sexual attentions from males (e.g. species which mate in leks or in male swarms, in which males attract females from a distance, or in which females initiate contact by attracting males from a distance). The multiple possible mechanical functions of male genitalia are summarized, and functions of male genitalic structures in 43 species in 21 families of Diptera are compiled. Functions associated with intromission and insemination (e.g. seizing and positioning the female appropriately, pushing past possible barriers within the female, orienting within the female to achieve sperm transfer), which are unlikely to be involved in new arms races when females are protected, are shown to be common (> 50 % of documented cases). This information is then used to generate the new arms race prediction: differences in genitalic form among congeneric species in which females are protected should be less common than differences among congeneric species in which females are vulnerable to harassment by males. This prediction was tested using a sample of 361 genera of insects and spiders. The prediction clearly failed, even when the data were adjusted to take into account several possible biases. Comparative analyses within particular taxonomic groups also failed to show the predicted trends, as did less extensive data on other non-genitalic male display traits. Arms races, as defined in some recent models, seem to have been less important in male-female coevolution of genitalic structures than has been suggested. By elimination, alternative interpretations, such as traditional female choice, which do not predict associations between female protection from harassment and rapid divergent evolution, are strengthened.     

A Test for Female Attraction to Male Orange Coloration in Poecilia picta

Examining the distribution of female preference and attractive male secondary sexual characters in closely related species can help evaluate the various models for the evolution of female preference. We examined whether orange coloration in males was attractive to females of Poecilia picta, a species in the same subgenus as the guppy, P. reticulata. Females did not respond differently to males with or without a conspicuous orange stripe on the dorsum of the tail, a characteristic ornamentation of the male of this species. This lack of preference in a taxon that possesses a conspicuous, secondary sexual character directly contradicts the predictions of the sensory exploitation hypothesis for the evolution of female preference, but may support a model of antagonistic sexual selection proposed by Rice (1996).     

Mechanical reproductive isolation via divergent genital morphology between Carabus insulicola and C. esakii with implications in species coexistence

The mechanical isolation hypothesis predicts that physical incompatibility between divergent reproductive morphologies hinders hybridization between populations. However, evidence for this hypothesis remains scarce. We examined this hypothesis using two parapatric carabid beetles, Carabus insulicola and C. esakii, which are of the subgenus Ohomopterus and exhibit a species-specific genital lock-and-key system. Our interspecific crossing experiment revealed that incompatibility of genital morphologies served as a strong postmating-prezygotic isolation barrier. This isolation was asymmetric: a decrease in female fitness was more costly in the cross with greater genitalic incompatibility between a C. esakii female and a C. insulicola male. These two species share a limited sympatric area, but the mechanism responsible for their coexistence is unclear given no evidence of premating isolation via male mate choice. A comparison of the present results with those of previous studies that quantified reproductive isolation between Ohomopterus species suggest that strong mechanical isolation via genitalic incompatibility plays a major role in species isolation, but that it may be less important in species coexistence.     

Divergent patterns of diversification in courtship and genitalic characters of Timema walking‐sticks

Understanding the patterns of diversification in sexual traits and the selection underlying such diversification represents a major unresolved question in evolutionary biology. We examined the phylogenetic diversification for courtship and external genitalic characters across ten species of Timema walking‐sticks, to infer the tempos and modes of character change in these sexual traits and to draw inferences regarding the selective pressures underlying speciation and diversification in this clade. Rates of inferred change in male courtship behaviours were proportional to speciation events, but male external genitalic structures showed a pattern of continuous change across evolutionary time, with divergence proportional to branch lengths. These findings suggest that diversification of courtship behaviour is mediated by processes that occur in association with speciation, whereas diversification of genitalia occurs more or less continuously, most likely driven by forces of sexual selection.     

Mating strategies in two species of dart-poison frogs: a comparative study

Comparative field studies of species of dart-poison frogs in the genus Dendrobates were carried out to test predictions from two hypotheses that attempt to explain female-female competition for mates in species of Dendrobates with male parental care. The sex role reversal hypothesis proposes that males invest so much time and energy in parental care that receptive males are rare relative to receptive females, and females compete to find and mate with receptive males. The parental quality hypothesis proposes that females compete to monopolize the parental effort of particular males, because they potentially suffer a cost when their mates care for the offspring of other females. Comparisons between species with male parental care (Dendrobates leucomelas) and female parental care (Dendrobates histrionicus) contradicted prediction of the sex role reversal hypothesis, but were consistent with predictions of the parental quality hypothesis. Male D. histrionicus did not compete for mates more aggressively than male D. leucomelas, and male D. leucomelas were not more selective about mating than male D. histrionicus. Female D. leucomelas and D. histrionicus were both selective about mating; female D. leucomelas associated with and competed for particular males, whereas female D. histrionicus did not.     

The exoskeleton of the female genitalic region in Petrobiellus takunagae (Insecta: Archaeognatha): Insect-wide terminology,homologies, and functional interpretations

The exoskeleton of the female genitalic region (abdominal venters 7–9) in Petrobiellus takunagae (Machilidae-Petrobiellinae) is studied using light microscopy and SEM. Sclerites are distinguished from membrane by the degree of cuticular flexibility. However, the microsculpture of the cuticle is shown to be useful in characterising the heterogeneity of the cuticle and in detecting weak sclerotisations. The morphology of Petrobiellus is compared with that in Trigoniophthalmus alternatus (Machilidae-Machilinae) described previously. While venter 7 is similar, venters 8 and 9 show many differences in the presence/absence or fusion/separation of particular sclerites. This suggests female genitalic morphology to be a valuable character system for phylogenetic and taxonomic work in Archaeognatha. Comparison with other insect orders is aimed at detecting homologous structures and conditions. Important points are: (1) Petrobiellus has a sclerotised genital lobe posteriorly on venter 7, similar to Zygentoma and Dictyoptera; it bears the gonopore. (2) Petrobiellus has a posterior sclerite on venter 9 that is very similar to a sclerite of Odonata. (3) The morphology of the coxal lobes of venter 9 (gonoplacs) suggests their function as a sheath of the ovipositor. From female genitalic morphology we deduce the process of oviposition, describing an external egg transportation tract.     

Correlated evolution of multivariate traits: detecting co-divergence across multiple dimensions

Tests of correlated evolution typically treat phenotypic characters as univariate variables, even though different trait attributes may contribute to their association with other traits. In this study, patterns of character covariation among species are analysed in a multivariate framework to test for both correlated rates and directions of evolutionary change in traits forming the genitalic complex of male grasshoppers. Although the covariation structure differs among traits, and among the constituent species of two grasshopper clades, significant co-divergence was detected among the most closely interacting genitalic traits (i.e. intromittent characters) in both clades. Co-divergence across shape space is not accompanied by similar rates of evolution among species, although the intromittent characters tend to show accelerated evolution (relative to nonintromittent characters). Differences in the evolutionary trajectories among traits may relate to their varied roles during mating. The study emphasizes the importance of a multivariate framework for detecting macroevolutionary patterns of correlated change.     

INTERSEXUAL ARMS RACE? GENITAL COEVOLUTION IN NEPHILID SPIDERS (ARANEAE,NEPHILIDAE)

Genital morphology is informative phylogenetically and strongly selected sexually. We use a recent species-level phylogeny of nephilid spiders to synthesize phylogenetic patterns in nephilid genital evolution that document generalized conflict between male and female interests. Specifically, we test the intersexual coevolution hypothesis by defining gender-specific indices of genital complexity that summarize all relevant and phylogenetically informative traits. We then use independent contrasts to show that male and female genital complexity indices correlate significantly and positively across the phylogeny rather than among sympatric sister species, as predicted by reproductive character displacement. In effect, as females respond to selection for fecundity-driven fitness via giantism and polyandry (perhaps responding to male-biased effective sex ratios), male mechanisms evolve to monopolize females (male monogamy) via opportunistic mating, pre- and postcopulatory mate guarding, and/or plugging of female genitalia to exclude subsequent suitors. In males morphological symptoms of these phenomena range from self-mutilated genitalia to total castration. Although the results are compatible with both recently favored sexual selection hypotheses, sexually antagonistic coevolution, and cryptic female choice, the evidence of strong intersexual conflict and genitalic damage in both sexes is more easily explained as sexually antagonistic coevolution due to an evolutionary arms race.     

Evolution of an ornament,the dewlap,in females of the lizard genus Anolis

Male ornaments have been the subject of numerous studies on sexual selection and communication, although female ornaments have garnered substantially less study, even though female ornaments are well developed in some species. The factors that have propelled the evolution of elaborate ornaments in females are poorly understood but may include genetic correlations between the sexes, social selection, sensory drive or species recognition. We used simulation‐based comparative methods and a newly estimated phylogeny to test these four hypotheses to explain female ornamentation within the diverse neotropical lizard genus Anolis. We found support for the sensory drive hypothesis and the social selection hypothesis; the female dewlap was larger in species that use more arboreal habitats, as well as in species where the sexes were less dimorphic. We did not find support for the genetic correlation hypothesis or the species recognition hypothesis. We propose that the size of the female dewlap may evolve in response to sensory drive differentially affecting species in different habitats, as well as social selection such as male mate choice or intrasexual competition for territory among females. © 2012 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, 106 , 191–201.     

Sexual selection and the evolution of genital shape and complexity in water striders

Animal genitalia show two striking but incompletely understood evolutionary trends: a great evolutionary divergence in the shape of genitalic structures, and characteristic structural complexity. Both features are thought to result from sexual selection, but explicit comparative tests are hampered by the fact that it is difficult to quantify both morphological complexity and divergence in shape. We undertake a comparative study of multiple nongenitalic and male genital traits in a clade of 15 water strider species to quantify complexity and shape divergence. We show that genital structures are more complex and their shape more divergent among species than nongenital traits. Further, intromittent genital traits are more complex and have evolved more divergently than nonintromittent genital traits. More importantly, shape and complexity of nonintromittent genital traits show correlated evolution with indices of premating sexual selection and intromittent genital traits with postmating sexual selection, suggesting that the evolution of different components of genital morphology are shaped independently by distinct forms of sexual selection. Our quantitative results provide direct comparative support for the hypothesis that sexual selection is associated with morphological complexity in genitalic traits and highlight the importance of quantifying morphological shape and complexity, rather than size in studies of genital evolution.     

A time-sequence functional analysis of mating behaviour and genital coupling in Drosophila: role of cryptic female choice and male sex-drive in the evolution of male genitalia

Male genitalia in Drosophila exemplify strikingly rapid and divergent evolution, whereas female genitalia are relatively invariable. Whereas precopulatory and post-copulatory sexual selection has been invoked to explain this trend, the functional significance of genital structures during copulation remains obscure. We used time-sequence analysis to study the functional significance of external genitalic structures during the course of copulation, between D. melanogaster and D. simulans. This functional analysis has provided new information that reveals the importance of male-driven copulatory mechanics and strategies in the rapid diversification of genitalia. The posterior process, which is a recently evolved sexual character and present only in males of the melanogaster clade, plays a crucial role in mounting as well as in genital coupling. Whereas there is ample evidence for precopulatory and/or post-copulatory female choice, we show here that during copulation there is little or no physical female choice, consequently, males determine copulation duration. We also found subtle differences in copulatory mechanics between very closely related species. We propose that variation in male usage of novel genitalic structures and shifts in copulatory behaviour have played an important role in the diversification of genitalia in species of the Drosophila subgroup.