Genital damage in the orb-web spider Argiope bruennichi (Araneae: Araneidae) increases paternity success

The morphology of male genitalia often suggests functions besidessperm transfer that may have evolved under natural or sexualselection. In several species of sexually cannibalistic spiders,males damage their paired genitalia during mating, limitingthem to one copulation per pedipalp. Using a triple-mating experiment,we tested if genital damage in the orb-web spider Argiope bruennichiincreases male fitness either through facilitating his escapefrom an aggressive female or by obstructing the female's inseminationducts against future copulation attempts from other males. Wefound no survival advantage for males damaging their pedipalps;however, copulations into a previously used insemination ductwere significantly shorter when the previous male had left partsof his genitalia inside the insemination duct. Because copulationduration determines paternity in this species, our result suggeststhat male genital damage in A. bruennichi is sexually selected.By breaking off parts of their intromittent organs inside avirgin female, males can reduce sperm competition and therebyincrease their paternity success.     

Female behaviour and the interaction of male and female genital traits mediate sperm transfer during mating

Natural selection and post‐copulatory sexual selection, including sexual conflict, contribute to genital diversification. Fundamental first steps in understanding how these processes shape the evolution of specific genital traits are to determine their function experimentally and to understand the interactions between female and male genitalia during copulation. Our experimental manipulations of male and female genitalia in red‐sided garter snakes (Thamnophis sirtalis parietalis) reveal that copulation duration and copulatory plug deposition, as well as total and oviductal/vaginal sperm counts, are influenced by the interaction between male and female genital traits and female behaviour during copulation. By mating females with anesthetized cloacae to males with spine‐ablated hemipenes using a fully factorial design, we identified significant female–male copulatory trait interactions and found that females prevent sperm from entering their oviducts by contracting their vaginal pouch. Furthermore, these muscular contractions limit copulatory plug size, whereas the basal spine of the male hemipene aids in sperm and plug transfer. Our results are consistent with a role of sexual conflict in mating interactions and highlight the evolutionary importance of female resistance to reproductive outcomes.     

Dermacentor variabilis and Dermacentor andersoni: Genital sex pheromones

There is evidence for the existence of a previously undescribed sex pheromone (or pheromones) in the ticks Dermacentor variabilis and D. andersoni. In addition to 2,6-dichlorophenol, which attracts mate-seeking males, a pheromone released on the cuticle of the female genital area enables the sexually excited male to locate the gonopore. The compound (or compounds) appears to act as a contact pheromone; male copulation responses are greatly reduced when the female genital surface is washed with solvents, especially hexane. It is also a potent excitant; males will puncture or dislodge barriers placed over the gonopore to copulate. However, the response is eliminated if the genital area is washed (hexane or acetone) prior to sealing the gonopore, suggesting the reproductive system as the source of the pheromone. A species specific copulation-eliciting pheromone appears necessary to excite the male to form and implant its spermatophore in the vulva of a conspecific female. Males encountering trans-specific females probe their gonopores, but mating attempts are almost always aborted within 5–10 min. The copulation-eliciting pheromone may be the same, or similar, to that used to locate the gonopore. Physical differences in the shape of the female gonopore in the two species, although slight, may contribute to the male's ability to identify conspecific females. Using this pheromone-guided process of attraction and identification, females present in mixed species populations will almost always be distinguished and inseminated by conspecific mate-seeking males.     

How the length of genital parts affects copulation performance in a carabid beetle: implications for correlated genital evolution between the sexes

To identify factors leading to the correlated evolution of exaggerated male and female genitalia, we studied the effects of the variable dimensions of corresponding functional genital parts (male copulatory piece and female vaginal appendix) on copulatory performance in the polygamous carabid beetle Carabus (Ohomopterus) maiyasanus. We used mating pairs of individuals from two populations to increase the variances in genital dimensions and determined the copulation performance (insemination and spermatophore replacement, and copulation time) in single‐ and double‐mating situations. In single mating, insemination success was not affected by genital dimensions, although the copulation time was significantly shorter when the male aedeagus was longer. In the double‐mating experiment, insemination and replacement of spermatophores by the second male succeeded more frequently when the copulatory piece was shorter and the vaginal appendix was longer, and when the difference between the length of the copulatory piece and the vaginal appendix was smaller. Thus, a matching of the corresponding genital parts between the sexes increases the male's reproductive success in sperm competition, but elongation of the copulatory piece cannot be explained simply by the improvement in male reproductive success. We discuss possible factors for the elongation of genital parts in terms of sexual conflict and reproductive interference through interspecific copulation.     

Hindsight by genitalia: photo-guided copulation in butterflies

1. Butterflies have two pairs of extraocular photoreceptive neurons on the genitalia. Here we report that the genital photoreceptors have a crucial role in achieving copulation. 2. We first investigated mating behavior of the butterfly Papilio xuthus in an outdoor cage. The mating behavior consists of six steps: the male approaches the female (1), maneuvers into a ventor-to-ventor position with the female (2), searches the female's genitalia (3), clasps the genitalia (4), the pair copulates (5), and finally separates (6). 3. We bilaterally ablated the P1 pair of photoreceptors, in the males, and observed mating behavior with virgin females. Of the intact males 66% copulated successfully, whereas only about 25% of the treated males could copulate. About 40% of P1 ablated males stopped the mating behavior during step 3: the males could not locate the female's genitalia. P1 ablation in females did not have a clear effect in this study. 4. Electrophysiological measurements showed that the P1 response of a male drops sharply when it correctly locates the female's genitalia. We hypothesize that the sharp drop in the P1 response informs the male that the female's vagina is correctly positioned for penis insertion. The P1 ablated males never experience such␣a␣response drop, as there is no P1 response to begin with. Accepted: 18 October 1996     

The Functional Significance of Chiral Genitalia: Patterns of Asymmetry,Functional Morphology and Mating Success in the Praying Mantis Ciulfina baldersoni

Genital asymmetry is relatively common and widespread throughout the animal kingdom. The functional significance of genital asymmetry is however, poorly understood for most species. Male praying mantids of the genus Ciulfina are remarkable in possessing complex and directionally asymmetric genital phallomeres in some species, and chirally dimorphic/antisymmetric genitalia in others. Here we explore the chiral dimorphism in male genitalia of Ciulfina baldersoni which appear to exhibit genital antisymmetry. We test whether genital orientation influences mating success, copulation duration and the attachment duration of spermatophores. Additionally we investigate genital interactions between male and females using x-ray micro-computed tomography (micro-CT) and scanning electron microscopy (SEM). Lastly we assess whether genital asymmetry is associated with non-genital morphological asymmetry of a range of traits. Our results highlight the complex functional morphology of genitalia in this praying mantis species and yet demonstrate no functional difference between dextral and sinistral morphs other than the direction of attachment with both morphs enjoying equal levels of mating success. Chiral morphs also did not strongly associate with any other forms of asymmetry. We therefore conclude that genital chirality in Ciulfina baldersoni is a likely case of antisymmetry with no functional significance to genital orientation, and is likely to be selectively neutral.     

Mating behavior and the function of the male genital spine in the ground beetle Carabus clathratus

The morphologies of male genitalia often appear harmful or aggressive, as if they may inflict physical damage upon females during copulation. Such male genitalia are often thought to function in intra- and intersexual interactions during mating. In the carabid genus Carabus, division Spinulati, males possess a spine (spinula) on the intromittent organ, of which function is unknown. To reveal the function of the spinula, we studied the mating behavior and genital coupling of a Spinulati species, Carabus (Limnocarabus) clathratus. The males positioned the spinula along the inner wall of the vaginal opening throughout copulation. This placement created a small dent and subsequently a melanized patch (wound) on the vaginal wall, but the spinula rarely penetrated the vaginal wall. The spinula did not reach the innermost part of the vagina where the spermatophore is deposited. These results suggest that the spinula is not used for inflicting damage on female genitalia or manipulating spermatophores of rival males. During spermatophore formation, the male partially withdrew the aedeagus, and only the aedeagal tip and endophallus remained within the female. By placing the spinula against the vaginal wall, the male could hold the endophallus within the vaginal chamber in the unstable copulatory posture. Thus, our observations suggest that the spinula primarily functions as an "anchor" to maintain the coupling of the male and female genitalia and thereby ensure insemination.     

Significance of constraints on genital coevolution: Why do female Drosophila appear to cooperate with males by accepting harmful matings?

The mechanisms driving the coevolution of male and female genital morphologies are still debated. Female genitalia in Drosophila species bear membranous “pouches” or hardened “shields,” which the male genital armature contact during copulation. Although shield‐like structures likely serve to mitigate the effects of harmful mating, some authors have suggested that soft pouches, which do not prevent male genitalia from inflicting wounds, represent a congruent sensory organ. To elucidate the evolutionary forces responsible for the development of such organs, I examined the effects of artificial damage to various genital parts of female Drosophila erecta on reproductive success. Despite a high survival rate among females, damage to the ovipositor plate resulted in frequent failure of insemination and in the embedment of eggs into the substrate. Damage to the vaginal shield resulted in increased mortality and frequent failure of egg embedment, with an egg blocking the vagina under the damaged shield in some females. Wounding of the pouch had less of an effect on both mating and oviposition success, suggesting that the structure “lures” the male trauma‐causing organs to areas where the resultant wounds do not interfere with insemination or oviposition. These data show that the dual functions of female genitalia (mating and oviposition) mediate genital coevolution.     

Experimental modifications imply a stimulatory function for male tsetse fly genitalia,supporting cryptic female choice theory

One of the most sweeping of all patterns in morphological evolution is that animal genitalia tend to diverge more rapidly than do other structures. Abundant indirect evidence supports the cryptic female choice (CFC) explanation of this pattern, which supposes that male genitalia often function to court females during copulation; but direct experimental demonstrations of a stimulatory function have been lacking. In this study, we altered the form of two male genital structures that squeeze the female’s abdomen rhythmically in Glossina pallidipes flies. As predicted by theory, this induced CFC against the male: ovulation and sperm storage decreased, while female remating increased. Further experiments showed that these effects were due to changes in tactile stimuli received by the female from the male’s altered genitalia, and were not due to other possible changes in the males due to alteration of their genital form. Stimulation from male genital structures also induces females to permit copulation to occur. Together with previous studies of tsetse reproductive physiology, these data constitute the most complete experimental confirmation that sexual selection (probably by CFC) acts on the stimulatory properties of male genitalia.     

The Mating Behaviour of the Velvet Ant, <Emphasis Type="Italic">Nemka viduata</Emphasis> (Hymenoptera: Mutillidae)

In the present study, the mating behaviour of the velvet ant Nemka viduata (Pallas) (Mutillidae) is described both from field and laboratory observations. The whole pairing interaction, lasting around two hours, includes several behavioural phases. During pre-copula, the male seizes the female’s neck with his mandibles, and then starts to rhythmically stroke the prothorax of the female with his forelegs (this behaviour is also resumed after copulation) before curving his abdomen in order to couple the genital parts, including genital armatures (the male parameres remaining outside the female body); just prior to copulation, the female extrudes the sting, and immediately after copulation begins, she stridulates for 7–10 s, this behaviour is repeated when the pair separates. During copulation (lasting around two minutes), the male moves his antennae rhythmically, hitting the back of the female’s head with the scape. Generally, recently-mated males become aggressive towards females, but more tolerant after a few days. During the whole pairing act, females are held by the males’ mandibles, and in the field they are carried off in flight or by walking to a safe place to copulate. This would suggest that larger males, which can lift a wider range of female sizes, have a reproductive advantage, as indicated by data obtained on their load-lifting capacity with respect to the size distribution of females. A review of mating behaviour in mutillid wasps and comparisons with other lineages of aculeate and non-aculeate Hymenoptera are also given.     

Mating behavior of the parasitic honeybee mite Tropilaelaps clareae

The mating behavior of adult Tropilaelaps clareae males and females was observed in glass test tubes. The male jumped on the dorsum of the female, stretched legs I forward, and hooked their distal ends on the frontal margin of the female dorsum. He then slipped sideways to the female venter. In the venter-to-venter position, the male clasped the female between her legs I and II, with his legs I. The male then moved backwards until his gnathosoma reached about 1/2 way along the epigynial plate of the female. The male vibrated and probably pushed the spermatophore out of his body. Next he moved laterally and placed the gnathosoma between coxae III and IV where the gonopore is located. Probably the sperm was introduced into the gonopore by means of the spermatodactyl. Subsequently the male moved to one side and stroked the anterior part of the epigynial plate and the gonopore area with his leg II. He repeated these contacts on average 282 times (variation 100 up to 522). The male then moved on the other side of the female venter and repeated the whole procedure. Finally, the male withdrew himself backwards from the female. The total mating process lasted between 3 and 42 min, with an average of 23 min. Multiple matings of both, males and females were observed.     

Courtship behavior of the mosquitoSabethes cyaneus (Diptera: Culicidae)

Unlike any other mosquito reported, Sabethes cyaneus(Fabricius) displays an elaborate courtship before and during copulation. A male approaches a female suspended from a horizontal stick, suspends himself in front of her as he grasps her folded wings, and proceeds with a series of discrete stereotyped behaviors that involve proboscis vibration and movement of iridescent blue paddles on his midlegs. The sequence of these behaviors is as follows: freeleg waving, swinging, copulation attempt, superficial coupling, waving, genital shift, waggling, and release. Insemination occurs after genital shift. The only overt reciprocation by the female is abdomen lowering during the male's swinging. Courtship is often unsuccessful, and males are usually rejected during freeleg waving. The relation between male performance and mating success remains obscure.     

Female control of mate plugging in a female-cannibalistic spider (Micaria sociabilis)

Background

Sperm competition imposes a strong selective pressure on males, leading to the evolution of various physiological, morphological and behavioral traits. Sperm competition can be prevented by blocking or impeding the access to female genitalia by means of a mating plug. We investigated the factors responsible for plug production and function in the promiscuous female-cannibalistic spider Micaria sociabilis (Gnaphosidae).

Results

We performed mating trials using females with and without a plug that consists of an amorphous mass. The mating trials demonstrated that the probability of male plugging increased non-linearly with the duration of copulation. Copulation duration and plug production seem to be controlled by the female. We found that females terminated matings later if males were fast at genital coupling. Whereas incomplete plugs had disappeared on the day following copulation, complete plugs persisted (40%). In matings with females with complete plugs, only a small proportion of males (7%) were able to remove the plug, indicating the high effectiveness of plugging. Moreover, males ceased attempts to copulate with plugged females with higher probability. 3D X-ray microscopy of the female and male genitalia showed that the plug material can extend far into the female genital tract and that the plug material is produced by a massive gland inside the palpal organ of the modified male pedipalps.

Conclusions

Our study demonstrates that the mating plug in M. sociabilis constitutes an effective male strategy to avoid sperm competition that seems to be under female control.

Electronic supplementary material

The online version of this article (doi:10.1186/s12862-014-0278-9) contains supplementary material, which is available to authorized users.     

Mating behavior and genital damage during copulation in the leaf beetle Lema coronata (Chrysomelidae: Criocerinae)

The criocerine leaf beetle Lema coronata Baly has extremely long genitalia that reach more than twice the body length in both sexes. We observed mating behavior of this species in the laboratory and inspected the male genital morphology using a scanning electron microscope. The males did not perform pre-copulatory courtship and post-copulatory guarding of the mates, and copulation lasted only for about 30 min. The surface of male genitalia is smooth without any special structures at the tip. A fragment of broken male genitalia was detected in the spermathecal duct of one female. We discuss the adaptive significance of male genital damage and the selective factor of elongated genitalia.     

Copulatory mechanism in a sexually cannibalistic spider with genital mutilation (Araneae: Araneidae: Argiope bruennichi)

Genitalia are among the fastest evolving morphological traits as evidenced by their common function as diagnostic traits in species identification. Even though the main function of genitalia is the successful transfer of spermatozoa, the presence of diverse structures that are obviously not necessary for this suggests that genitalia are a target of sexual selection. The male genitalia of many spider species are extremely complex and equipped with numerous sclerites, plates and spines whose functions are largely unknown. Selection on male genitalia may be particularly strong in sexually cannibalistic spiders, where mating success of males is restricted to a single female. We investigated the copulatory mechanism of the sexually cannibalistic orb weaving spider Argiope bruennichi by shock freezing mating pairs and revealed a complicated interaction between the appendices and sclerites that make up the male gonopods (paired pedipalps). The plate that covers the female genital opening (scape) is secured between two appendices of the male genital bulb, while three sclerites that bear the sperm duct are unfolded and extended into the female copulatory opening. During copulation, females attack and cannibalise the male and males mutilate their genitalia in about 80% of cases. Our study demonstrates that (i) genital coupling is largely accomplished on the external part of the female genitalia, (ii) that the mechanism requires an interaction between several non-sperm-transferring structures and (iii) that there are two predetermined breaking points in the male genitalia. Further comparative work on the genus Argiope will test if the copulatory mechanism with genital mutilation indeed is an adaptation to sexual cannibalism or if cannibalism is a female counter adaptation to male monopolisation through genital plugging.     

Frequent Failure of Male Monopolization Strategies as a Cost of Female Choice in the Black Widow Spider Latrodectus tredecimguttatus

In many mating systems, males strive for securing paternity through monopolizing females. As male monopolization attempts often contradict female interests, this conflict may fuel an evolutionary arms race. In the widow spider genus Latrodectus, females are commonly polyandrous, whereas males are monogynous, hence restricted to mate with a single female, making paternity protection particularly important. Potential mating plugs (specialized embolus sclerites of male copulatory organs) have been discovered in the complex female genital tracts of several Latrodectus species. In this study, we investigated mating strategies in the Mediterranean black widow spider Latrodectus tredecimguttatus and tested the adaptive value of female attacks against male monopolization efforts. In a double mating experiment, we manipulated the number of insertions (=copulations) for first and second males to assess female behaviour and male embolus sclerite placement success. Our results indicate that first males′ embolus sclerites inside the females′ sperm stores physically block sclerites of subsequent males. While female attacks did not affect the deposition of potential mating plugs, they significantly reduced copulation duration. Irrespective of female aggression, male sclerite placement failure occurred frequently, but large males were more successful than smaller competitors. We suggest that the complex genital morphology in both sexes may have co‐evolved antagonistically and female morphology could serve to favour large males for fertilization.     

No evidence for external genital morphology affecting cryptic female choice and reproductive isolation in Drosophila

Genitalia are one of the most rapidly diverging morphological features in animals. The evolution of genital morphology is proposed to be driven by sexual selection via cryptic female choice, whereby a female selectively uptakes and uses a particular male's sperm on the basis of male genital morphology. The resulting shifts in genital morphology within a species can lead to divergence in genitalia between species, and consequently to reproductive isolation and speciation. Although this conceptual framework is supported by correlative data, there is little direct empirical evidence. Here, we used a microdissection laser to alter the morphology of the external male genitalia in Drosophila, a widely used genetic model for both genital shape and cryptic female choice. We evaluate the effect of precision alterations to lobe morphology on both interspecific and intraspecific mating, and demonstrate experimentally that the male genital lobes do not affect copulation duration or cryptic female choice, contrary to long‐standing assumptions regarding the role of the lobes in this model system. Rather, we demonstrate that the lobes are essential for copulation to occur. Moreover, slight alterations to the lobes significantly reduced copulatory success only in competitive environments, identifying precopulatory sexual selection as a potential contributing force behind genital diversification.     

Female wing spreading as acceptance signal in theDrosophila virilis group of species

Females of manyDrosophila species spread apart their wings prior to copulation. In the present study we found female wing spreading to provoke male copulation attempts inDrosophila virilis-group species, helping the males to attempt copulation when the female is ready to mate. The males of most species, however, rarely responded to female wing spreading by copulation attempt without licking the female genitalia before and/or after female wing spreading bout. Blocking the female genitalia (D. virilis, D. novamexicana) reduces males' tendency to attempt copulation after female wing spreading. In these, and most other species of the group, female wing spreading seems to be an efficient signal only when combined with stimuli from female genitalia.     

Sexual coercion does not exclude luring behavior in the climbing camel-spider <Emphasis Type="Italic">Oltacola chacoensis</Emphasis> (Arachnida,Solifugae, Ammotrechidae)

Sexual coercion in the form of forced copulation has been used as a typical example to illustrate the conflict of interests between females and males. Among arthropods, forced copulation has been reported for some groups of insects and crustaceans, but not for arachnids. In the present work, we analyse and describe the behavioral patterns of mating behavior of the climbing camel-spider, Oltacola chacoensis, relating it to relevant morphological features, In this species, the male forcefully clasps the female’s genital region with his chelicerae and locks her fourth pair of legs with his pedipalps. In some cases, the cuticle of the female’s abdomen was damaged by this cheliceral clasping. In contrast to other camel-spiders, the female O. chacoensis never remained motionless during mating, but continuously shook her body, opening her chelicerae notably towards the male. Despite this coercive context, males performed copulatory courtship (tapping with pedipalps) and females showed an apparent cooperative behavior (they remained still during a short period of the sperm transfer phase). These results strengthen the idea that sexual coercion (in the form of forced copulation) and luring behavior (in the form of copulatory courtship) are not two mutually-exclusive male’s strategies during a single copulation.