Sperm Precedence Research Articles

Patterns of sperm use in the scorpionfly Panorpa germanica L. (Mecoptera: Panorpidae)

Sperm competition can be a powerful selective force in the evolution of reproductive strategies and mating systems. In studies on sperm competition, patterns of sperm use are typically reported as the mean species value of P 2, determined as the proportion of offspring sired by the second male to copulate with a doubly mated female. However, the within-species variance in P 2 has mostly been ignored, although taking this variance into account may be crucial for understanding the underlying mechanisms of sperm competition. Paternity analysis among the offspring of doubly mated females of Panorpa germanica (Mecoptera, Panorpidae) revealed a relationship between relative copulation durations of both males and the proportion of offspring each male will sire. This correlation between proportional copulation durations and paternity suggests mixing of sperm from different males inside the female’s spermatheca. Yet, sperm mixing appears to be incomplete, as paternity was overall slightly shifted towards the second male on average fathering a higher proportion of the offspring than its relative copulation duration would predict in case of complete sperm mixing. For individual males, however, the outcome of sperm competition is rather unpredictable as the intraspecific variance in P 2 was found to be very high, irrespective of copulation durations. Possible causes of the observed variance in P 2 and the partial last male sperm precedence are discussed.

On the functional significance of juvenile hormone in the accessory sex glands of male Heliothis virescens

The storage of large quantities of juvenile hormone (JH) in male abdomens is a phenomenon known from some species of moths. Juvenile hormone, stored in male accessory sex glands (ASG), may be transferred to the female during copulation, but the physiological significance of the JH transfer remains unclear. Here, using the moth Heliothis virescens as a model, we show that JH transferred from male to the promiscuous female promotes JH synthesis and egg development in the female. We propose that this explains the functional significance of JH transfer in species that exhibit last male sperm precedence, and that this hormone acts as a bioactive substance which the first male to mate uses for co-opting and regulating the female's gonadotropic mechanisms, thereby ensuring that despite last male sperm precedence he will sire a significant number of viable offspring.

The snail's love-dart delivers mucus to increase paternity.

Many of the seemingly bizarre animal behaviours can be understood only by acknowledging the power of sex to shape evolution. A case in point is the so-called love-dart that some terrestrial molluscs shoot at their prospective sexual partners. Given that the likelihood of copulation is not different after solid hits than after complete misses, why do these suitors act so violently towards their chosen mates? Previously, it was shown that successful dart shooting enhances paternity. We conducted an experiment to determine whether the dart achieves its effect by a purely mechanical action or by transferring a bioactive substance. We found that injections of mucus from a gland associated with the dart more than doubled paternity relative to injections of saline. These results support the hypothesis that the dart transfers a substance capable of reconfiguring the spermatophore-receiving organs. While dart shooting probably evolved as the result of sperm competition, a role for cryptic female choice cannot be excluded. Our results imply that if cryptic female choice is operating in this system, it is likely to be based on the properties of the mucus and not on properties of the dart itself. Since we also found evidence of early-male sperm precedence, we conclude that snails can optimize their reproductive success by mating with virgins and shooting their darts accurately.

The effects of copulation duration in the bruchid beetle Callosobruchus maculatus

The importance of cryptic female choice, i.e. female post-copulatory influence over male reproductive success, in driving the evolution of male traits remains controversial. The main aim of this thesis was to understand the post-copulatory consequences of sexual interactions and the importance of cryptic female choice in two species of beetle.Males of the red flour beetle Tribolium castaneum use their legs to rub the lateral edges of the female elytra during mating. When manipulating female perception of this behaviour, I found that females preferentially use the sperm of males with vigorous leg rubbing when they mate with more than one male. Leg rubbing also appeared to increase female rate of oviposition. Females do not seem to gain any indirect benefits by preferring males with an intense leg rubbing behaviour since this behaviour was found to have very low narrow sense heritability and did not appear to be condition dependent in its expression.Males of the bruchid beetle Callosobruchus maculatus have spiny genitalia that harm their mates. Females kick males during copulation and when prevented from kicking, suffered reduced lifetime offspring production as a consequence of more extensive injuries. Males were not able to delay female remating, increase rate of oviposition or increase sperm precedence by inflicting relatively severe injuries to non-kicking females. Hence, the injuries appear to be side effects of male efforts to remain in copula. When copulation duration was manipulated, ejaculate size and female lifetime offspring production increased with the length of copulation. Females reduced their mating rate when they had access to water, suggesting that they obtain water from the large ejaculates and trade-off their need for additional water against the costs of mating. Males may then reduce the benefits of remating by providing their mates with a large amount of water. Females did not increase their remating propensity to avoid inbreeding when they had mated to brothers. Together, these studies reveal the complexity of sexual interactions and the importance of post-copulatory processes for the fitness of both males and females.

Offensive and defensive sperm competition roles in the dung beetle Onthophagus taurus (Coleoptera: Scarabaeidae)

Sperm competition is predicted to generate opposing selection pressures on males. On one hand, selection should favour ‘defensive’ adaptations that protect a male’s ejaculate from displacement, while, on the other hand, selection should favour ‘offensive’ adaptations that overcome paternity assurance mechanisms of rivals. Here, we use the sterile male technique to assess sperm precedence when a male dung beetle Onthophagus taurus mates in both a defensive (first male) and an offensive (second male) role. Significant variation in a male’s sperm precedence (both P1 and P2) was detected, and an individual’s defensive (P1) and offensive (P2) abilities were positively correlated. Thus, it appears that sexual selection simultaneously selects for ‘defensive’ and ‘offensive’ adaptations in O. taurus. We discuss a variety of male traits in O. taurus that potentially contribute to a male’s ability to be successful when mating in an ‘offensive’ and a ‘defensive’ role.

A strain of the brown-winged green bug, Plautia crossota stali (Heteroptera: Pentatomidae) with blue-green body colouration: Genetic control and sexual competitivity

A blue-green (BG) strain was established from a laboratory colony of Plautia crossota stali. This strain was characterized by a unique blue-green body colouration and pale green egg colouration with grey forewings, compound eyes and ocelli. The testicular epithelium for male adults was whitish in contrast to the orange-yellow colour for normal males. Reciprocal crosses between normal and BG strains indicated that the traits specific to the BG strain were recessive against normal phenotype and controlled by a single Mendelian unit. Double mating experiments in which BG females were mated with a normal and a BG male in this or the other order suggested that the sperm of BG males were as competitive as those of normal males. It was also found that the sperm of two males were mixed throughly soon after the second mating, although the precedence of sperm from a second male varied greatly for each female adult. The BG strain may be used as a genetic marker for behavioural, ecological and genetic studies in P. c. stali.

RELATIVE ABUNDANCE AND THE SPECIES-SPECIFIC REINFORCEMENT OF MALE MATING PREFERENCE IN THE CHRYSOCHUS (COLEOPTERA: CHRYSOMELIDAE) HYBRID ZONE

Most studies of reinforcement have focused on the evolution of either female choice or male mating cues, following the long-held view in sexual selection theory that mating mistakes are typically more costly for females than for males. However, factors such as conspecific sperm precedence can buffer females against the cost of mating mistakes, suggesting that in some hybrid zones mating mistakes may be more costly for males than for females. Thus, the historical bias in reinforcement research may underestimate its frequency. In this study, we present evidence that reinforcement has driven the evolution of male choice in a hybrid zone between the highly promiscuous leaf beetles Chrysochus cobaltinus and C. auratus, the hybrids of which have extremely low fitness. In addition, there is evidence for male choice in these beetles and that male mating mistakes may be costly, due to reduced opportunities to mate with conspecific females. The present study combines laboratory and field methods to quantify the strength of sexual isolation, test the hypothesis of reproductive character displacement, and assess the link between relative abundance and the strength of selection against hybridization. We document that, while sexual isolation is weak, it is sufficient to produce positive assortative mating. In addition, reproductive character displacement was only detected in the relatively rare species. The strong postzygotic barriers in this system are sufficient to generate the bimodality that characterizes this hybrid zone, but the weak sexual isolation is not, calling into question whether strong prezygotic isolation is necessary for the maintenance of bimodality. Growing evidence that the cost of mating mistakes is sufficient to shape the evolution of male mate choice suggests that the reinforcement of male mate choice may prove to be a widespread occurrence.

Size, operational sex ratio, and mate-guarding success of the carrion beetle, Necrophila americana

When male insects guard females until oviposition, the benefits from last-male sperm precedence must outweigh the costs of relinquishing additional fertilizations. The profitability of guarding is increased when males guard large, fecund females and when females are scarce because fewer fertilizations are sacrificed. However, the male reproductive success is not only determined by the profitability of guarding but also by his ability to maintain guarding. In this study, we used male carrion beetles (Necrophila americana) to examine the effects of sex ratio, male relative size, and female quality on the ability to guard. First, we present a model of mate guarding that explores factors, such as sperm precedence, sex ratio, male size, and female quality, that influence the profitability of postcopulatory riding. Our model predicts that large N. americana males should preferentially guard the largest female only when the sex ratio is male biased and sperm precedence is above 80%. In contrast, small males gain little from guarding because they are not likely to maintain it and be the last male to mate. Then, we tested these predictions by manipulating sex ratio, relative male size, and female quality. All males in equal sex ratio and large males in male-biased sex ratio guarded females significantly longer than did males in female-biased sex ratio. In male-biased sex ratio, large males guarded significantly longer and achieved more takeovers than small males. Large females were guarded longer. The success of guarding males in this beetle depends on their size relative to other males and the operational sex ratio.

Sperm Precedence in Overwintered Colorado Potato Beetle (Coleoptera: Chrysomelidae) and Its Implications for Insecticide Resistance Management

Colorado potato beetle, Leptinotarsa decemlineata (Say), is the most important insect defoliator of potatoes and is infamous for its ability to develop insecticide resistance. Sperm precedence is an important consideration in developing resistance management plans. We determined the precedence of sperm from a postdiapause spring mating by using irradiated sterile males. We also investigated whether spring mating (with or without fertilization) affects reproduction of overwintered females. Precedence of sperm from spring matings versus overwintered sperm from fall matings was almost complete. Some fertilization from fall matings did take place in this experiment, but it exceeded 10% in only one of the 22 pairings. Females mated only the previous fall laid fewer eggs than spring-mated females, but spring mating did not seem to improve the hatch rate of deposited eggs. On average, mated females started laying 1.6 d sooner than fall-mated females, but this difference was not significant. Hatch rate did not change with time. Precedence of spring matings over stored sperm can help compensate for assortative summer mating among resistant individuals due to differences in developmental time between refuges and treated areas.

The effect of female polyandry and sperm precedence on the evolution of sexual difference in dispersal timing

Predispersal copulation and unpredictable environment facilitate the evolution of female-biased dispersal in species, where females are functionally monandrous. Females should migrate and reproduce over different habitats to spread their risks due to environmental fluctuation. On the other hand, males do not have to disperse because their risks are spread by their mating partners who produce their offspring in different habitats. However, when females are functionally polyandrous, it is expected that they will not contribute to spreading the male's risk extensively. Therefore, by simulation with the individual based model, the present study evaluated how female polyandry influences the sexual difference in dispersal timing. This model revealed that when females are polyandrous, the timing of female remating and sperm priority patterns have an important influence on the evolution of sex-biased dispersal. Particularly when female remating is not synchronized with dispersal or when last-male sperm precedence does not exist, female-biased dispersal is evolved.

Gamete Compatibility and Sperm Competition Affect Paternity and Hybridization Between SympatricAsteriasSea Stars

Gamete interactions may strongly influence speciation and hybridization in sympatric broadcast-spawning marine invertebrates. We examined the role of gamete compatibility in species integrity using cross-fertilization studies between sympatric Asterias sea stars from a secondary contact zone in the northwest Atlantic. In crosses between single males and single females, gametes of both species were compatible and produced viable, fertile hybrid offspring, but with considerable variation in the receptivity of eggs to heterospecific sperm. Differential compatibility of heterospecific gametes was detected in sperm competition studies in which we used a nuclear DNA marker to assign paternity to larval offspring. Several families showed conspecific sperm precedence in A. forbesi eggs, and one family showed competitive superiority of A. forbesi sperm fertilizing A. rubens eggs. Gametic interactions are an important component of prezygotic reproductive isolation in sympatric Asterias. The interaction between gametes of these closely related sea stars is consistent with the function of gamete recognition systems that are known to mediate fertilization success and speciation in other marine invertebrates.

Fitness benefits of multiple mating versus female mate choice in the cellar spider (Pholcus phalangioides)

Recent studies have demonstrated that mating with multiple males can be beneficial for females and her offspring even if males contribute nothing but sperm. This was mainly established for species in which sperm from several males mix in the reproductive tract of the female, thus allowing sperm competition and/or female sperm choice. However, in species with last male sperm precedence, female re-mating decides against the previous male by strongly limiting his reproductive success. We tested the effect of female re-mating behaviour using the cellar spider Pholcus phalangioides, which shows strong last males sperm precedence and moderate levels of polyandry under natural situations. We predicted that females prevented from remating even though they are receptive would show reduced reproductive success compared to females that accept two copulations and females that reject a second male, since the latter two treatments were allowed to behave according to their decisions. However, if the number of matings per se had an effect on oviposition or on offspring performance, double-mated females should perform better compared to both treatments of once-mated females. We measured female fecundity and fertility over a period of 140 days, comparable to the species' natural reproductive peak season. Two thousand one hundred and fifty-two offspring from 67 first egg sacs were reared under two feeding levels. We registered development time and survival, and measured offspring adult size and mass. We found a positive effect of double mating, as in this treatment, oviposition probability was higher compared to the other treatments. Interestingly, adult female offspring of the DM treatment that were raised under low food level had a higher condition index compared to those from FS and RM, but development time, size and mass at adulthood were not affected by mating treatment. Female choice only seemed to affect hatching latency of the offspring. Overall, the main predictor of female reproductive output and success was female body size.

SEXUAL SELECTION WHEN FERTILIZATION IS NOT GUARANTEED

Much of the theory of sexual selection assumes that females do not generally experience difficulties getting their eggs fertilized, yet sperm limitation is occasionally documented. How often does male limitation form a selection for female traits that improve their mating rate? The question is difficult to test, because if such traits evolve to be efficient, sperm limitation will no longer appear to be a problem to females. Here, we suggest that changes in choosiness between populations, and in particular between virgin and mated females, offer an efficient way to test this hypothesis. We model the "wallflower effect," that is, changes in female preferences due to time and mortality costs of remaining unmated (for at least some time). We show that these costs cause adaptive reductions of female choice, even if mate encounter rates appear high and females only rarely end their lives unfertilized. We also consider the population consequences of plastic or fixed mate preferences at different mate encounter rates. If mate choice is plastic, we confirm earlier verbal models that virgins should mate relatively indiscriminately, but plastic increase of choosiness in later matings can compensate and intensify sexual selection on the male trait, particularly if there is last male sperm precedence. Plastic populations will cope well with unusual conditions: eagerness of virgins leads to high reproductive output and a relaxation of sexual selection at low population densities. If females lack such plasticity, however, population-wide reproductive output may be severely reduced, whereas sexual selection on male traits remains strong.

Sperm competition and evidence of sperm fertilization patterns in the carrion ball-roller beetle Canthon cyanellus cyanellus LeConte (Scarabaeidae: Scarabaeinae)

Field observation and laboratory experiments were conducted to investigate sperm competition and patterns of sperm fertilization under different experimental conditions in the carrion ball-roller beetle Canthon cyanellus cyanellus. Males in nature can mate with females whose spermathecae contains fertile sperm from other males. Sperm precedence was investigated using a visible genetic marker. The progeny of red (homozygous recessive) virgin females mated once with a red male and afterward, once with a green beetle (homozygous dominant) and vice versa, revealed that regardless of its color, the last male to mate fertilized c.a. 50% of the eggs, suggesting strong sperm competition. Males were able to achieve higher levels of paternity (more than 80%) when mated ad libitum with previously mated females, although large amount of variance in paternity does not exclude the possibility of first male sperm precedence or female cryptic choice. These results suggest that repeated mating and sperm replacement are the mechanisms by which last males achieve sperm precedence.

Sexual selection research on spiders: progress and biases

The renaissance of interest in sexual selection during the last decades has fuelled an extraordinary increase of scientific papers on the subject in spiders. Research has focused both on the process of sexual selection itself, for example on the signals and various modalities involved, and on the patterns, that is the outcome of mate choice and competition depending on certain parameters. Sexual selection has most clearly been demonstrated in cases involving visual and acoustical signals but most spiders are myopic and mute, relying rather on vibrations, chemical and tactile stimuli. This review argues that research has been biased towards modalities that are relatively easily accessible to the human observer. Circumstantial and comparative evidence indicates that sexual selection working via substrate-borne vibrations and tactile as well as chemical stimuli may be common and widespread in spiders. Pattern-oriented research has focused on several phenomena for which spiders offer excellent model objects, like sexual size dimorphism, nuptial feeding, sexual cannibalism, and sperm competition. The accumulating evidence argues for a highly complex set of explanations for seemingly uniform patterns like size dimorphism and sexual cannibalism. Sexual selection appears involved as well as natural selection and mechanisms that are adaptive in other contexts only. Sperm competition has resulted in a plethora of morphological and behavioural adaptations, and simplistic models like those linking reproductive morphology with behaviour and sperm priority patterns in a straightforward way are being replaced by complex models involving an array of parameters. Male mating costs are increasingly being documented in spiders, and sexual selection by male mate choice is discussed as a potential result. Research on sexual selection in spiders has come a long way since Darwin, whose spider examples are reanalysed in the context of contemporary knowledge, but the same biases and methodological constraints have persisted almost unchanged through the current boom of research.

The effect of an inversion system and the time interval between matings on postcopulatory sexual selection in the seaweed fly, Coelopa frigida

The seaweed fly, Coelopa frigida, exhibits LMSP. A large chromosomal inversion system affects many traits including egg-to-adult viability via heterosis. Consequently, there is also considerable potential for cryptic female mate choice to operate on the basis of sperm karyotype. Here, we investigated the effect of time interval and chromosomal inversion karyotype on postcopulatory sexual selection. Homokaryotypic females were mated with a male of the same and a male of the opposite homokaryotype. The order of the matings was varied so cryptic female mate choice could operate either in concert or antagonistically with LMSP. LMSP was found when there was a 24 h time interval between matings, irrespective of the order in which the males were mated. However, when the males were mated in quick succession the order of mating was important. When LMSP and cryptic female mate choice work in concert a high level of LMSP was found. However, when the male of opposite homokaryotype mated first, then first male sperm precedence was observed. This suggests that polyandrous females might be able to bias paternity but only when matings occur in quick succession. Consequently, population density is likely to affect the operation of postcopulatory sexual selection.

Effect of Seminal Fluids in Mating Between M and S Forms ofAnopheles gambiae

Previous studies have shown that sympatric populations of M and S molecular forms of Anopheles gambiae sensu stricto exhibit strong assortative mating. In the few documented cases of cross-mating between M and S forms, females that mated with amale of the alternative form were often also mated with a male of their own form. A potential explanation for the association between cross-mating and double mating could be that male accessory gland or sperm proteins that are responsible for inducing refractoriness to further mating by females have diverged between the M and S forms. This mechanism of postmating reproductive isolation would have important implications for our understanding of the speciation processes in the An. gambiae complex. We tested for this mechanism, by comparing the likelihood of mating, feeding, and laying eggs, as well as the fertility of females presented with males of their own form or the alternate form in the laboratory. We also compared the likelihood of remating in cross-mated and assortatively-mated females, and we analyzed their progeny to unravel patterns of sperm precedence. We found that cross-mated females differed from assortatively-mated females only in terms of egg-hatching rate and larval survival but that these effects could be attributed to hybrid vigor rather than differential response to seminal products. Cross-mating between forms was not associated with remating behavior. These results indicate that the sex proteins responsible for inhibiting further insemination and triggering the gonotrophic cycle in females have not diverged between these M and S populations. We discuss alternative explanations for the patterns of cross-mating and multiple mating observed in the field.

Effect of Seminal Fluids in Mating Between M and S Forms of <I>Anopheles gambiae</I>

Previous studies have shown that sympatric populations of M and S molecular forms of Anopheles gambiae sensu stricto exhibit strong assortative mating. In the few documented cases of cross-mating between M and S forms, females that mated with amale of the alternative form were often also mated with a male of their own form. A potential explanation for the association between cross-mating and double mating could be that male accessory gland or sperm proteins that are responsible for inducing refractoriness to further mating by females have diverged between the M and S forms. This mechanism of postmating reproductive isolation would have important implications for our understanding of the speciation processes in the An. gambiae complex. We tested for this mechanism, by comparing the likelihood of mating, feeding, and laying eggs, as well as the fertility of females presented with males of their own form or the alternate form in the laboratory. We also compared the likelihood of remating in cross-mated and assortatively-mated females, and we analyzed their progeny to unravel patterns of sperm precedence. We found that cross-mated females differed from assortatively-mated females only in terms of egg-hatching rate and larval survival but that these effects could be attributed to hybrid vigor rather than differential response to seminal products. Cross-mating between forms was not associated with remating behavior. These results indicate that the sex proteins responsible for inhibiting further insemination and triggering the gonotrophic cycle in females have not diverged between these M and S populations. We discuss alternative explanations for the patterns of cross-mating and multiple mating observed in the field.

Multiple sperm storage organs facilitate female control of paternity

It has been proposed that multiple sperm storage organs (spermathecae) could allow polyandrous females to control paternity. There is little conclusive evidence for this since insemination of individual spermathecae is generally not experimentally manipulable. Here, we examined sperm use patterns in the Australian redback spider (Latrodectus hasselti), which has paired, independent spermathecae. We assessed paternity when two rivals were forced to inseminate a single storage organ or opposite storage organs. When males inseminated a single spermatheca, mean paternity of the female's first mate was 79.8% (median 89.4%), and 38% of first mates achieved 100% paternity. In contrast, when males inseminated opposite organs, the mean paternity of the first mate was 49.3% (median 49.9%), only 10% of males achieved complete precedence, and paternity was normally distributed, suggesting sperm mixing. Males responded to this difference by avoiding previously inseminated female reproductive tracts. Complete sperm precedence can only be achieved if females permit males to copulate with both reproductive tracts. Females often cannibalize smaller males during their first copulation, thus limiting their paternity to 50%. These data show that multiple sperm storage organs can increase female control of paternity.

Why do male Callosobruchus maculatus harm their mates?

The importance of cryptic female choice, i.e. female post-copulatory influence over male reproductive success, in driving the evolution of male traits remains controversial. The main aim of this thesis was to understand the post-copulatory consequences of sexual interactions and the importance of cryptic female choice in two species of beetle.Males of the red flour beetle Tribolium castaneum use their legs to rub the lateral edges of the female elytra during mating. When manipulating female perception of this behaviour, I found that females preferentially use the sperm of males with vigorous leg rubbing when they mate with more than one male. Leg rubbing also appeared to increase female rate of oviposition. Females do not seem to gain any indirect benefits by preferring males with an intense leg rubbing behaviour since this behaviour was found to have very low narrow sense heritability and did not appear to be condition dependent in its expression.Males of the bruchid beetle Callosobruchus maculatus have spiny genitalia that harm their mates. Females kick males during copulation and when prevented from kicking, suffered reduced lifetime offspring production as a consequence of more extensive injuries. Males were not able to delay female remating, increase rate of oviposition or increase sperm precedence by inflicting relatively severe injuries to non-kicking females. Hence, the injuries appear to be side effects of male efforts to remain in copula. When copulation duration was manipulated, ejaculate size and female lifetime offspring production increased with the length of copulation. Females reduced their mating rate when they had access to water, suggesting that they obtain water from the large ejaculates and trade-off their need for additional water against the costs of mating. Males may then reduce the benefits of remating by providing their mates with a large amount of water. Females did not increase their remating propensity to avoid inbreeding when they had mated to brothers. Together, these studies reveal the complexity of sexual interactions and the importance of post-copulatory processes for the fitness of both males and females.