Sperm Precedence Research Articles

Fertilization by proxy: rival sperm removal and translocation in a beetle

Competition between diierent males’ sperm for the fertilization of ova has led to the evolution of a diversity of characters in male reproductive behaviour, physiology and morphology. Males may increase sperm competition success either by enhancing the success of their own sperm or by negating or eliminating the success of rival sperm. Here, we ¢nd that in the £our beetleTribolium castaneum, the second male to mate gains fertilization precedence over previous males’ sperm and fertilizes approximately two-thirds of the eggs. It is not known what mechanism underlies this pattern of last-male sperm precedence; however, the elongate tubules of the female sperm storage organ may encourage a‘last-in, ¢rst-out’sperm use sequence. Here we present an additional or alternative mechanism of sperm precedence whereby previously deposited sperm are removed from the female tract by the mating male’s genitalia. In addition to providing evidence for sperm removal inT. castaneum, we also show that removed, non-self sperm may be translocated back into the reproductive tracts of new, previously unmated females, where the translocated sperm go on to gain signi¢cant fertilization success.We found that, in 45 out of 204 crosses, sperm translocation occurred and in these 45 crosses over half of the oispring were sired by spermatozoa which had been translocated between females on the male genitalia. In the natural environment of stored food, reproductively activeT. castaneum adults aggregate in dense mating populations where copulation is frequent (we show in three naturally occurring population densities that copula duration and intermating intervals across three subsequent matings average 1^2min). Selection upon males to remove rival sperm may have resulted in counter-selection upon spermatozoa to survive removal and be translocated into new females where they go on to fertilize in signi¢cant numbers.

Sperm competition in the absence of fertilization in Caenorhabditis elegans.

Hermaphrodite self-fertilization is the primary mode of reproduction in the nematode Caenorhabditis elegans. However, when a hermaphrodite is crossed with a male, nearly all of the oocytes are fertilized by male-derived sperm. This sperm precedence during reproduction is due to the competitive superiority of male-derived sperm and results in a functional suppression of hermaphrodite self-fertility. In this study, mutant males that inseminate fertilization-defective sperm were used to reveal that sperm competition within a hermaphrodite does not require successful fertilization. However, sperm competition does require normal sperm motility. Additionally, sperm competition is not an absolute process because oocytes not fertilized by male-derived sperm can sometimes be fertilized by hermaphrodite-derived sperm. These results indicate that outcrossed progeny result from a wild-type cross because male-derived sperm are competitively superior and hermaphrodite-derived sperm become unavailable to oocytes. The sperm competition assays described in this study will be useful in further classifying the large number of currently identified mutations that alter sperm function and development in C. elegans.

Paternal investment in Poecilimon veluchianus bushcrickets: beneficial effects of nuptial feeding on offspring viability

In most bushcrickets, males transfer a large spermatophore during copulation that is afterwards consumed by the female. In some species this nuptial gift enhances offspring fitness and is therefore believed to function as paternal investment. To determine whether this is the case, I examined whether a male's own offspring benefit from spermatophore consumption in the bushcricket Poecilimon veluchianus. Females that consumed a spermatophore produced offspring with increased residuals of dry weight compared to females that were prevented from feeding on the spermatophore. This beneficial effect of spermatophore consumption occurred within the first 4 days after copulation. An increased dry weight indicates higher energy reserves because offspring dry weight correlates significantly with the lifespan of starved larvae and because spermatophore consumption increased the lifespan of starved offspring. During egg-laying, females apply a liquid substance to the soil that hardens and probably serves as protection for the egg clutch. The amount of this substance correlated with the number of eggs laid but did not differ between spermatophore treatments. In P. veluchianus, females mate frequently and there is last-male sperm precedence. The spermatophore thus only constitutes paternal investment when offspring produced before female remating benefit from spermatophore consumption. The dry weight of offspring increased during the first 4 days after spermatophore consumption and thus within the natural remating interval. This shows that the spermatophore has a paternal investment function in addition to its already known sperm protection function.

Electrophoretic Confirmation of Sperm Mixing in Mated Colorado Potato Beetles (Coleoptera: Chrysomelidae)

Sperm precedence in the Colorado potato beetle, Leptinotarsa decemlineata (Say), was investigated using phosphoglucomutase (Pgm) allozymes as genetic markers. Females were mated to 2 males carrying different Pgm alleles, and the proportion of offspring sired by each male was determined by cellulose acetate gel electrophoresis. Our results indicate that Pgm alleles in the Colorado potato beetle are inherited in the Mendelian fashion, and there is no change in sperm utilization over time. We also were able to confirm that sperm precedence in this insect is incomplete, with approximately equal to 72% of the larvae fathered by males at the 2nd mating. Possible implications of incomplete sperm precedence for insecticide resistance management are discussed.

COPULATORY BEHAVIOR, GENITAL MORPHOLOGY, AND MALE FERTILIZATION SUCCESS IN WATER STRIDERS.

Recent theoretical and empirical interest in postmating processes have generated a need for increasing our understanding of the sources of variance in fertilization success among males. Of particular importance is whether such postmating sexual selection merely reinforces the effects of premating sexual selection or whether other types of male traits are involved. In the current study, we document large intraspecific variation in last male sperm precedence in the water strider Gerris lateralis. Male relative paternity success was repeatable across replicate females, showing that males differ consistently in their ability to achieve fertilizations. By analyzing shape variation in male genital morphology, we were able to demonstrate that the shape of male intromittent genitalia was related to relative paternity success. This is the first direct experimental support for the suggestion that male genitalia evolve by postmating sexual selection. A detailed analysis revealed that different components of male genitalia had different effects, some affecting male ability to achieve sperm precedence and others affecting male ability to avoid sperm precedence by subsequent males. Further, the effects of the shape of the male genitalia on paternity success was in part dependent on female morphology, suggesting that selection on male genitalia will depend on the frequency distribution of female phenotypes. We failed to find any effects of other morphological traits, such as male body size or the degree of asymmetry in leg length, on fertilization success. Although males differed consistently in their copulatory behavior, copulation duration was the only behavioral trait that had any significant effect on paternity.

Sperm Competition in a Nuptial Feeding Spider, Pisaura Mirabilis

Abstract The paper tests two hypotheses about sperm priority in the spider Pisaura mirabilis. A 'phylogenetic constraints hypothesis' states that since the females have conduit spermathecae, first male priority should prevail. On the other hand, males offer nuptial prey to the females and females mate with multiple males. The evolution of these traits is most easily understood if late mating males also have a substantial fertilization success. The results indicate a compromise solution. Sterile-male technique with double-mated females indicated a first male priority pattern (P1 = ca 70%, after adjusting for sterilization damage and experimental mortality). However, the success of the fourth male of quadroublemated females was unexpectedly high (adjusted P4 = ca 24%, not different from P of 2 two-male matings). This lends support to a supplementary hypothesis of constant last male success, which may turn an initial first-male advantage into a last-male advantage, when the number of males mating with a female raises above a certain number. Independent of mating order, males may increase their share of fertilizations by long copulation times. It was tested whether female choice for symmetric males might account for differential male success. However, fluctuating asymmetry (FA) in male leg length showed no relationships with fertilization success or copulation duration.

Last-Male Sperm Priority and the Mating System of the Haplogyne Spider Tetragnatha extensa (Araneae: Tetragnathidae)

Austad (1984) proposed a relation between female spermathecal anatomy and sperm priority, such that in a haplogyne spider with “cul-de-sac” spermathecae a last-male sperm priority was expected. A laboratory experiment using both normal and irradiated males (sterile-male technique) confirmed this prediction; last males fertilized ca. 70% of eggs in the first egg sac of double-mated females. Mating series with up to four males indicate an antichronological order of male success. Thus, when females laid multiple egg sacs, earlier males fertilized an increasing proportion of eggs in successive sacs. In nature this effect may be counteracted by the fact that females remate between sacs. These statistical patterns emerged in spite of considerable randomness in paternity order. Field observations showed that males survived as long as females (constant sex ratio throughout the breeding season), an unusual feature for a spider population. Mating frequency could not be determined in the field, but laboratory trials indicated that encounters between the sexes usually resulted in immediate copulation. No guarding of females or serious fights between males were observed. Males were observed to steal prey from females and to prey on subadult females. High feeding rates and lack of aggressiveness in males are tactics that secure survival into the egg-laying period. However males also mated frequently early in the maturity season; this is seen as a tactic to maximize the total amount of sperm surviving to the time of fertilization. Thus, survival and multiple matings are the key factors for male success.

MATING BEHAVIOUR AND MALE MATE CHOICE IN TRIBOLIUM CASTANEUM (COLEOPTERA, TENEBRIONIDAE)

Mate choice by males has received less attention than female choice, despite the recognition that males can incur non-trivial reproductive costs through mating. In this study we investigate male mate choice in the red flour beetle, Tribolium castaneum. We determine whether male beetles have evolved sensitivity that enables them to discriminate between females and we then examine how males discriminate between females that present different reproductive potentials. Recently-emerged adult females are immature, and we investigate when egg-laying maturity develops, and whether male mating attempts with immature females provide reproductive pay-offs. We show that males are the sex most likely to initiate mating attempts (more than 85% of male-female contacts are male-initiated). Therefore, we test male mating initiations when presented with choices between: (i) immature and mature virgin females; (ii) mature virgin females and previously mated mature females; and (iii) mature females previously mated either with different male or with the test males. Last male sperm precedence exists in T. castaneum and females are polyandrous. Matings with immature females are therefore likely to generate lower fertilization successes than matings with mature females. Furthermore, males are likely to achieve higher total fertilization success when they mate if they precede a rival male's sperm than if they precede their own sperm. Accordingly, we find that: (i) males copulate more frequently with mature, than with immature females; (ii) males do differentiate between virgin and mated females; and (iii) males prefer females that have been inseminated by a different male to those previously inseminated by the test males themselves. The results demonstrate that male T. castaneum recognise female status, display mate choice, and prefer to inseminate females which provide greater reproductive potential.

Sperm Precedence and the Potential of the Nuptial Gift to Function as Paternal Investment in the Tettigoniid Steropleurus stali Bolivar (Orthoptera: Tettigoniidae: Epphippigerinae)

The spermatophores of the majority of species of tettigoniid consist of two parts: an ampulla which contains the sperm and a spermatophylax which the female eats following the end of copulation. One hypothesis for the function of the spermatophylax is that it is maintained by selection for paternal investment. In order for this to be possible, the donating male must fertilize a significant proportion of the eggs which stand to benefit from his spermatophylax nutrients. In order to determine whether or not this was the case, the pattern of sperm precedence was examined in the tettigoniid Steropleurus stali, using the sterile-male technique. The duration of the female's refractory period, the number of eggs laid in this period and the number of mature eggs stored by females were also recorded. The results indicated pronounced last-male sperm precedence (median P 2 = 0.95). Taking into account the large number of mature eggs typically stored by females of this species, together with the mean number of eggs laid in the female's refractory period, it appears that females are likely to have re-mated before laying eggs which could contain nutrients donated by her previous mate. This means that one male's spermatophylax nutrients are likely to be incorporated into eggs which will be fertilized by the sperm of males that subsequently mate with the female. The very large spermatophylax of S. stali is therefore unlikely to be maintained by selection for paternal investment.

Genetic linkage maps of the ground crickets Allonemobius fasciatus and Allonemobius socius using RAPD and allozyme markers

The ground crickets, Allonemobius fasciatus and A. socius are sister species and form a mosaic hybrid zone in the eastern United States. In this study, we developed RAPD markers and constructed genetic linkage maps for the two species, as a first step toward mapping quantitative trait loci that control conspecific sperm precedence. The A. fasciatus map consists of 25 markers mapped to 8 linkage groups, spanning 400.1 contiguous centiMorgans (cM). The average distance between two framework markers is 23.5 cM. The A. socius map consists of 20 markers mapped to 6 linkage groups, spanning 400.5 contiguous cM. The average distance between two framework markers is 30.4 cM. A sex chromosome was identified in the A. socius map. These are the first reported genetic linkage maps for Orthoptera.Key words: Allonemobius, allozyme, cricket, hybrid zone, linkage maps, RAPD.

The effect of female mating history on sperm precedence in the two-spot ladybird, Adalia bipunctata (Coleoptera, Coccinellidae)

Effects of two different mating regimes on sperm precedence in the two-spot ladybird, Adalia bipunctata, were studied using the polymorphic gene for melanism as a marker for paternity. Virgin nonmelanic females (homozygous recessive) were mated to nonmelanic male(s) and then, after laying fertilized eggs, were mated to a melanic male of known genotype. The results after the two successive single matings showed a highly variable degree of paternity of the second male. Initial multiple mating with nonmelanic males did not alter the pattern of paternity after the subsequent single mating with a melanic male, but it had two other effects: (1) the female showed an increase in rejection behavior, and (2) a longer copulation was required for high success of the melanic male. Additional observations in which families were reared from beetles collected in copula in the field demonstrated that sperm competition also occurs under natural conditions. The outcome of the competition was variable with frequent sperm mixing. Key words: copulation duration, mating history, rejection, sperm competition, sperm precedence. [Behav Ecol 9:559-565 (1998)]

Mating behaviour in the cellar spider,Pholcus phalangioides, indicates sperm mixing

In insects and spiders, the pattern of sperm priority is often attributed to the shape of the spermathecae and should entail marked consequences for mating behaviour. Since last-male priority is assumed to occur in haplogyne spiders such as the cellar spider, females of this species are predicted to be more attractive to males shortly before, than shortly after, egg laying and males may guard females after copulation until oviposition. To test these predictions, I individually marked spiders of a natural population and recorded their position and the distance between potential mating partners twice a day over 100 days. The distance between female and male was taken as a measure of the female’s attractiveness. The behaviour of cellar spider males was not in accordance with the predictions; females were visited throughout the observation period with no significant increase in attractiveness before egg laying and there was no evidence for mate guarding. However, female attractiveness was correlated with female size, which was correlated with the number of eggs laid. Behaviour and genital morphology suggest sperm mixing occurs in this species. This is discussed in the light of conflicting data on sperm priority.

INFERENCE OF SPERM COMPETITION FROM BROODS OF FIELD-CAUGHT DROSOPHILA.

In field studies of multiple mating and sperm competition there typically is no experimental control over the number of times that a female mates, the interval between matings, or the genetic identity of multiple fathers contributing to a brood. Irrespective of this complexity, high-resolution molecular markers can be used to assign paternity with considerable confidence. This study employed two highly heterozygous microsatellite loci to assess multiple paternity and sperm displacement in a sample of broods taken from a natural population of Drosophila melanogaster. The large number of alleles present at each of the loci makes it difficult to derive explicit maximum-likelihood estimates for multiple paternity and sperm displacement from brood samples. Monte Carlo simulations were used to estimate maximum-likelihood parameters for the distribution of female remating frequency and the proportion of offspring sired by the second or subsequent mating males. Estimates were made based on genotypes scored at two distinct marker loci because they were found to give statistically homogeneous results. Fitting a Poisson distribution of number of matings, the mean number of males mated by a female was 1.82. The sperm displacement parameter estimated from doubly mated females were 0.79 and 0.86 for the two loci (0.83 for the joint estimate). The overall probability that a multiply mated female will be misclassified as singly mated was only 0.006, which indicates that microsatellites can provide excellent resolution for identifying multiple mating. In addition, microsatellites can be used to generate relatively precise estimates of sperm precedence in brood-structured samples from a natural population.

Sperm precedence and sperm storage in multiply mated red flour beetles

In insects, the last male to mate with a female often gains access to a disproportionate number of subsequent fertilizations. This study examined last-male sperm precedence patterns in doubly and triply mated Tribolium castaneum females. Sperm storage processes were investigated by measuring the quantity of sperm stored within the female spermatheca following single, double, and triple matings. Both doubly mated and triply mated females exhibited high last-male sperm precedence for progeny produced during the first 48 h following the last mating, with females in both groups exhibiting parallel declines in sperm precedence 1 and 2 weeks later. The number of sperm stored by females increased by 33% between singly mated and doubly mated females, indicating that the spermatheca is filled to only two-thirds capacity following insemination by the first male. Based on the proportion of stored sperm from first and second matings, we tested predictions about sperm precedence values based on models of random sperm mixing. High initial last-male sperm precedence strongly supports stratification of last-male sperm. By 1–2 weeks after double matings, sperm precedence declined to levels indistinguishable from values expected under random mixing. These results provide insight into mechanisms of sperm storage and utilization in this species.

A female nervous system is necessary for normal sperm storage in Drosophila melanogaster: a masculinized nervous system is as good as none

A male Drosophila melanogaster deposits many more sperm in a female's bursa copulatrix than are stored in her ventral receptacle or paired spermathecae soon after copula has ended. The remaining sperm are expelled by the female. These observations suggest a sexual conflict over the processes involved in sperm storage. We used genetically manipulated flies to study the role of the central nervous system in sperm storage. Flies with female bodies but masculinized nervous systems, or isolated female abdomens, stored significantly fewer sperm than did control females. Furthermore, compared with control flies, there were relatively more sperm in the ventral receptacle and relatively fewer in the spermathecae. These results suggest that the female nervous input counteracts the male's attempts to force sperm into the ventral receptacle during copula and promotes active transport of sperm to the spermathecae during and after copula. The female is clearly a very active partner in influencing processes involved in sperm competition, especially as only stored sperm can be used later to fertilize eggs. To our knowledge, this is the first study to show directly the involvement of the female nervous system in sperm storage.

Snail sperm production characteristics vary with sperm competition risk

Sperm competition is widespread and influences both male investment in spermatogenic tissue and ejaculate characteristics. Sperm competition models assume trade-offs between sperm size and number, although such trade-offs may be difficult to detect. This study examines the effects of sperm competition risk on the sperm production characteristics of the freshwater snail Viviparus ater. In this prosobranch, females mate frequently and store sperm, generating sperm competition. Males produce two sperm morphs, fertile eupyrene sperm and non-fertilizing oligopyrene sperm. Non-fertilizing sperm may play a role in sperm competition and therefore, like fertilizing sperm, the number produced could vary relative to sperm competition risk. In addition, trade-offs between sperm number and sperm size may be expected. We manipulated the sex ratio of sexually mature snails and found the presence of rivals affected the ratio of oligopyrene/eupyrene sperm males produced. In experimental and natural populations, the number of oligopyrene sperm produced, but not the number of eupyrene sperm, was significantly higher when the sex ratio was male biased. Testis mass did not vary between experimental treatments. We also found a negative relationship between the number and size of oligopyrene sperm produced, which is consistent with evolutionary models of sperm competition, and is, to our knowledge, the first intraspecific demonstration of a trade-off between these traits.

Female genotypes affect sperm displacement in Drosophila.

Differential success of sperm is likely to be an important component of fitness. Extensive variation among male genotypes in competitive success of sperm in multiply mated females has been documented for Drosophila melanogaster. However, virtually all previous studies considered the female to be a passive vessel. Nevertheless, under certain conditions female fitness could be determined by her role in mediating use of sperm from multiple males. Here we ask whether females differ among genotypes in their tendency to exhibit last-male precedence. Competition of sperm from two tester male genotypes (bwD and B3-09, a third-chromosome isogenic line from Beltsville, MD) was quantified by doubly mating female lines that had been rendered homozygous for X, second, or third chromosomes isolated from natural populations. The composite sperm displacement parameter, P2', was highly heterogeneous among lines, whether or not viability effects were compensated, implying the presence of polymorphic genes affecting access of sperm to eggs. Genetic variation of this type is completely neutral in the absence of pleiotropy or interaction between variation in the two sexes.

Male potential reproductive rate influences mate choice in a bushcricket

Male bushcrickets,Requena verticalis, provide their mates with a proteinaceous nuptial gift, which functions both as paternal investment in offspring and to ensure sperm transfer. When nutrients are limited, males have a lower potential reproductive rate than females, so the operational sex ratio becomes female biased. Males are then expected to discriminate more in their choice of mate for two reasons: (1) the relatively higher female potential reproductive rate should reduce the costs of rejecting a female; (2) multiple mating by females should increase variance in female mate quality, because of first male sperm precedence. Males are known to discriminate against old females, which are more likely to have mated previously. Our objective in this study was to partition experimentally the influences of male potential reproductive rate (manipulated by diet) and variance in female quality (estimated by age) and to assess their relative influence on the level of mate choice of male bushcrickets. The potential reproductive rate of males had a great impact on their choosiness, whereas variance in female age had no effect. These results support recent theoretical models that predict costs, rather than benefits, will primarily influence the level of mate choice.

Sperm competition in birds.

Sperm competition in birds occurs when a female is inseminated by more than one male during a single breeding cycle. Despite most birds being socially monogamous, sperm competition is widespread and results in frequent extra-pair paternity. Sperm competition is a fundamental part of sexual selection since it results in differential reproductive success among males. Male adaptations to sperm competition include relatively large testes, large sperm stores and long spermatozoa, mate guarding and frequent pair copulations. Females show no obvious morphological adaptations to sperm competition but, by controlling whether copulations are successful, they probably determine its frequency and extent. Despite this, the evolutionary benefits females acquire from extra-pair fertilizations are poorly understood. Experiments in which females are inseminated with equal numbers of spermatozoa from two males usually show last male sperm precedence. Understanding the mechanism of sperm competition requires understanding of why the last male to inseminate a female fertilizes a disproportionate number of eggs. The data from sperm competition studies on the domestic fowl, turkeys and zebra finches are consistent only with a passive sperm loss model of sperm competition. The mechanism is as follows: after insemination, spermatozoa enter the sperm storage tubules located in the oviduct, from which they are lost at a constant rate over days or weeks. All else being equal, the interval between two inseminations determines the probability of fertilization: the second of two inseminations fertilizes most eggs simply because, by the time fertilization occurs, fewer of these spermatozoa have been lost. Other factors also affect the outcome of sperm competition: the timing of insemination relative to oviposition, the differential fertilizing capacity of males and differences in the numbers of spermatozoa inseminated; as a consequence, last male sperm precedence is not automatic. On the basis of the mechanism of sperm competition, the optimal strategy for both males and females to maximize their likelihood of extra-pair fertilization is to copulate with an extra-pair partner as close as possible to the onset of oviposition.

Sperm competition in birds

Sperm competition in birds occurs when a female is inseminated by more than one male during a single breeding cycle. Despite most birds being socially monogamous, sperm competition is widespread and results in frequent extra-pair paternity. Sperm competition is a fundamental part of sexual selection since it results in differential reproductive success among males. Male adaptations to sperm competition include relatively large testes, large sperm stores and long spermatozoa, mate guarding and frequent pair copulations. Females show no obvious morphological adaptations to sperm competition but, by controlling whether copulations are successful, they probably determine its frequency and extent. Despite this, the evolutionary benefits females acquire from extra-pair fertilizations are poorly understood. Experiments in which females are inseminated with equal numbers of spermatozoa from two males usually show last male sperm precedence. Understanding the mechanism of sperm competition requires understanding of why the last male to inseminate a female fertilizes a disproportionate number of eggs. The data from sperm competition studies on the domestic fowl, turkeys and zebra finches are consistent only with a passive sperm loss model of sperm competition. The mechanism is as follows: after insemination, spermatozoa enter the sperm storage tubules located in the oviduct, from which they are lost at a constant rate over days or weeks. All else being equal, the interval between two inseminations determines the probability of fertilization: the second of two inseminations fertilizes most eggs simply because, by the time fertilization occurs, fewer of these spermatozoa have been lost. Other factors also affect the outcome of sperm competition: the timing of insemination relative to oviposition, the differential fertilizing capacity of males and differences in the numbers of spermatozoa inseminated; as a consequence, last male sperm precedence is not automatic. On the basis of the mechanism of sperm competition, the optimal strategy for both males and females to maximize their likelihood of extra-pair fertilization is to copulate with an extra-pair partner as close as possible to the onset of oviposition.