On the evolutionary consequences of increasing litter size with multiple paternity in wild boar (Sus scrofa scrofa)

Understanding how some species may be able to evolve quickly enough to deal with anthropogenic pressure is of prime interest in evolutionary biology, conservation, and management. Wild boar (Sus scrofa scrofa) populations keep growing all over Europe despite increasing hunting pressure. In wild boar populations subject to male‐selective harvesting, the initially described polygynous mating system may switch to a promiscuous/polyandrous one. Such a change in the mating system, where potentially more males sire a litter at one reproductive event, may be associated with the retention of high genetic diversity and an increase of litter size. We tested these hypotheses by estimating the number of sires per litter based on a six‐year long monitoring of a wild boar population subject to particularly high harvesting pressure. Our results show a high and stable genetic diversity and high rates of multiple paternity compared to other populations, thus depicting a promiscuous/polyandrous mating system in this population. We also show that litter size is positively linked to the number of sires, suggesting that multiple paternity increases fecundity. We finally discuss that multiple paternity may be one of the factors allowing rapid evolution of this population by maintaining both genetic and phenotypic diversity.     

Unequal contribution of sexes in the origin of dog breeds

Dogs (Canis familiaris) were domesticated from the gray wolf (Canis lupus) at least 14,000 years ago, and there is evidence of dogs with phenotypes similar to those in modern breeds 4000 years ago. However, recent genetic analyses have suggested that modern dog breeds have a much more recent origin, probably <200 years ago. To study the origin of contemporaneous breeds we combined the analysis of paternally inherited Y chromosome markers with maternally inherited mitochondrial DNA and biparentally inherited autosomal microsatellite markers in both domestic dogs and their wild ancestor, the gray wolf. Our results show a sex bias in the origin of breeds, with fewer males than females contributing genetically, which clearly differs from the breeding patterns in wild gray wolf populations where both sexes have similar contributions. Furthermore, a comparison of mitochondrial DNA and Y chromosome diversity in dog groups recognized by the World Canine Organization, as well as in groups defined by the breeds' genetic composition, shows that paternal lineages are more differentiated among groups than maternal lineages. This demonstrates a lower exchange of males than of females between breeds belonging to different groups, which illustrates how breed founders may have been chosen.     

Why do females mate multiply? A review of the genetic benefits

The aim of this review is to consider the potential benefits that females may gain from mating more than once in a single reproductive cycle. The relationship between non-genetic and genetic benefits is briefly explored. We suggest that multiple mating for purely non-genetic benefits is unlikely as it invariably leads to the possibility of genetic benefits as well. We begin by briefly reviewing the main models for genetic benefits to mate choice, and the supporting evidence that choice can increase offspring performance and the sexual attractiveness of sons. We then explain how multiple mating can elevate offspring fitness by increasing the number of potential sires that compete, when this occurs in conjunction with mechanisms of paternity biasing that function in copula or post-copulation. We begin by identifying cases where females use pre-copulatory cues to identify mates prior to remating. In the simplest case, females remate because they identify a superior mate and 'trade up' genetically. The main evidence for this process comes from extra-pair copulation in birds. Second, we note other cases where pre-copulatory cues may be less reliable and females mate with several males to promote post-copulatory mechanisms that bias paternity. Although a distinction is drawn between sperm competition and cryptic female choice, we point out that the genetic benefits to polyandry in terms of producing more viable or sexually attractive offspring do not depend on the exact mechanism that leads to biased paternity. Post-copulatory mechanisms of paternity biasing may: (1) reduce genetic incompatibility between male and female genetic contributions to offspring; (2) increase offspring viability if there is a positive correlation between traits favoured post-copulation and those that improve performance under natural selection; (3) increase the ability of sons to gain paternity when they mate with polyandrous females. A third possibility is that genetic diversity among offspring is directly favoured. This can be due to bet-hedging (due to mate assessment errors or temporal fluctuations in the environment), beneficial interactions between less related siblings or the opportunity to preferentially fertilise eggs with sperm of a specific genotype drawn from a range of stored sperm depending on prevailing environmental conditions. We use case studies from the social insects to provide some concrete examples of the role of genetic diversity among progeny in elevating fitness. We conclude that post-copulatory mechanisms provide a more reliable way of selecting a genetically compatible mate than pre-copulatory mate choice. Some of the best evidence for cryptic female choice by sperm selection is due to selection of more compatible sperm. Two future areas of research seem likely to be profitable. First, more experimental evidence is needed demonstrating that multiple mating increases offspring fitness via genetic gains. Second, the role of multiple mating in promoting assortative fertilization and increasing reproductive isolation between populations may help us to understand sympatric speciation.     

The impact of mating systems and dispersal on fine‐scale genetic structure at maternally,paternally and biparentally inherited markers

For decades, studies have focused on how dispersal and mating systems influence genetic structure across populations or social groups. However, we still lack a thorough understanding of how these processes and their interaction shape spatial genetic patterns over a finer scale (tens—hundreds of metres). Using uniparentally inherited markers may help answer these questions, yet their potential has not been fully explored. Here, we use individual‐level simulations to investigate the effects of dispersal and mating system on fine‐scale genetic structure at autosomal, mitochondrial and Y chromosome markers. Using genetic spatial autocorrelation analysis, we found that dispersal was the major driver of fine‐scale genetic structure across maternally, paternally and biparentally inherited markers. However, when dispersal was restricted (mean distance = 100 m), variation in mating behaviour created strong differences in the comparative level of structure detected at maternally and paternally inherited markers. Promiscuity reduced spatial genetic structure at Y chromosome loci (relative to monogamy), whereas structure increased under polygyny. In contrast, mitochondrial and autosomal markers were robust to differences in the specific mating system, although genetic structure increased across all markers when reproductive success was skewed towards fewer individuals. Comparing males and females at Y chromosome vs. mitochondrial markers, respectively, revealed that some mating systems can generate similar patterns to those expected under sex‐biased dispersal. This demonstrates the need for caution when inferring ecological and behavioural processes from genetic results. Comparing patterns between the sexes, across a range of marker types, may help us tease apart the processes shaping fine‐scale genetic structure.     

Paternity assessment in free-ranging wild boar (Sus scrofa) – Are littermates full-sibs?

Multiple paternity within litters occurs in various groups of mammals exhibiting different mating systems. Using seven genetic markers (i.e., microsatellites), we investigated the paternity of littermates in free-ranging wild boar (Sus scrofa) in a Mediterranean habitat. Using the software CERVUS 2.0 we estimated the probability of detecting multiple paternity across all loci (D), the probability of paternity (W) and a statistic Δ that allows the assignment of paternity to the most likely male with strict and relaxed levels of confidence. Multiple paternity was inferred for one of the nine analysed litters at the 80% confidence level. This suggests that a single male may control the access to receptive adult females and it shows that multiple paternity is not very common in the studied free-ranging wild boar population. Despite the possible occurrence of sperm competition and/or female cryptic choice, mate guarding seems to play a significant role in sexual selection. To better understand the wild boar's mating strategies further studies analysing the reproductive success of both sexes and under different environmental conditions should be conducted.     

Benefits of polyandry: Molecular evidence from field‐caught dung beetles

When females mate with multiple males, they set the stage for postcopulatory sexual selection via sperm competition and/or cryptic female choice. Surprisingly little is known about the rates of multiple mating by females in the wild, despite the importance of this information in understanding the potential for postcopulatory sexual selection to drive the evolution of reproductive behaviour, morphology and physiology. Dung beetles in the genus Onthophagus have become a laboratory model for studying pre‐ and postcopulatory sexual selection, yet we still lack information about the reproductive behaviour of female dung beetles in natural populations. Here, we develop microsatellite markers for Onthophagus taurus and use them to genotype the offspring of wild‐caught females and to estimate natural rates of multiple mating and patterns of sperm utilization. We found that O. taurus females are highly polyandrous: 88% of females produced clutches sired by at least two males, and 5% produced clutches with as many as five sires. Several females (23%) produced clutches with significant paternity skew, indicating the potential for strong postcopulatory sexual selection in natural populations. There were also strong positive correlations between the number of offspring produced and both number of fathers and paternity skew, which suggests that females benefit from mating polyandrously by inciting postcopulatory mechanisms that bias paternity towards males that can sire more viable offspring. This study evaluates the fitness consequences of polyandry for an insect in the wild and provides strong evidence that female dung beetles benefit from multiple mating under natural conditions.     

Estimation of genetically effective breeding numbers using a rejection algorithm approach

Polygynous mating results in nonrandom sampling of the adult male gamete pool in each generation, thereby increasing the rate of genetic drift. In principle, genetic paternity analysis can be used to infer the effective number of breeding males (Nebm). However, this requires genetic data from an exhaustive sample of candidate males. Here we describe a new approach to estimate Nebm using a rejection algorithm in association with three statistics: Euclidean distance between the frequency distributions of maternally and paternally inherited alleles, average number of paternally inherited alleles and average gene diversity of paternally inherited alleles. We quantify the relationship between these statistics and Nebm using an individual-based simulation model in which the male mating system varied continuously between random mating and extreme polygyny. We evaluate this method using genetic data from a natural population of highly polygynous fruit bats (Cynopterous sphinx). Using data in the form of mother-offspring genotypes, we demonstrate that estimates of Nebm are very similar to independent estimates based on a direct paternity analysis that included data on candidate males. Our method also permits an evaluation of uncertainty in estimates of Nebm and thus facilitates inferences about the mating system from genetic data. Finally, we investigate the sensitivity of our method to sample size, model assumptions, adult population size and the mating system. These analyses demonstrate that the rejection algorithm provides accurate estimates of Nebm across a broad range of demographic scenarios, except when the true Nebm is high.     

Female polyandry affects their sons’ reproductive success in the red flour beetle Tribolium castaneum

A potential benefit to females mating with multiple males is the increased probability that their sons will inherit traits enhancing their pre‐ or post‐mating ability to obtain fertilizations. We allowed red flour beetle (Tribolium castaneum) females to mate on three consecutive days either repeatedly to the same male or to three different males. This procedure was carried out in 20 replicate lines, 10 established with wild‐type, and 10 with the Chicago black morph, a partially dominant phenotypic marker. The paternity achieved by the sons of females from polyandrous vs. monandrous lines of contrasting morph was assessed in the F1, F2 and F3 generation by mating wild‐type stock females to two experimental males and assigning the progeny to either sire based on phenotype. The sons of polyandrous wild‐type females achieved significantly higher paternity when mating in the second male role than the sons of monandrous wild‐type females. By contrast, when mating in the first male role, males produced by females from polyandrous lines tended to have lower paternity than males from monandrous lines. Both effects were independent of the number of mates of the black competitor’s mother, and interacted significantly with the number of progeny laid by the female. These results provide the first evidence that manipulating the number of mates of a female can influence her sons’ mating success and suggest a potential trade‐off between offence and defence in this species.     

Multiple paternity and mating group size in the European earwig,Forficula auricularia

1. The patterns of multiple paternity among the progeny of females are key properties of genetic mating systems. Female multiple mating should evolve due to direct or indirect benefits, but it may also partly be driven by the encounter rate with different potential mates. 2. In this study this hypothesis was experimentally tested in the European earwig (Forficula auricularia L.) by establishing experimental mating groups that differed in the number of males and females (i.e. density). The number of sires and mean sibling relatedness in each clutch were estimated using microsatellite‐based paternity analysis. 3. As predicted, the mean number of sires per clutch was significantly increased, and sibling relatedness decreased, in the higher density treatment where more potential male mates were available. This change was less than proportional to the number of males in the mating groups, indicating that mechanisms limiting multiple paternity in large mating groups were involved. There were no significant relationships between female reproductive success or male siring success with morphology (body size, weight, and forceps size). 4. The present results show that multiple paternity in F. auricularia clutches is partly determined by the availability of male mates and suggest that this effect is modulated by mechanisms in males and/or females that limit multiple paternity.     

How attractive is the girl next door? An assessment of spatial mate acquisition and paternity in the solitary Cape dune mole-rat, Bathyergus suillus

Behavioural observations of reproduction and mate choice in wild fossorial rodents are extremely limited and consequently indirect methods are typically used to infer mating strategies. We use a combination of morphological, reproductive, spatial, and genetic data to investigate the reproductive strategy of a solitary endemic species, the Cape dune mole-rat Bathyergus suillus. These data provide the first account on the population dynamics of this species. Marked sexual dimorphism was apparent with males being both significantly larger and heavier than females. Of all females sampled 36% had previously reproduced and 12% were pregnant at the time of capture. Post-partum sex ratio was found to be significantly skewed in favour of females. The paternity of fifteen litters (n?=?37) was calculated, with sires assigned to progeny using both categorical and full probability methods, and including a distance function. The maximum distance between progeny and a putative sire was determined as 2149 m with males moving between sub-populations. We suggest that above-ground movement should not be ignored in the consideration of mate acquisition behaviour of subterranean mammals. Estimated levels of multiple paternity were shown to be potentially as high as 26%, as determined using sibship and sire assignment methods. Such high levels of multiple paternity have not been found in other solitary mole-rat species. The data therefore suggest polyandry with no evidence as yet for polygyny.     

High levels of polyandry,but limited evidence for multiple paternity,in wild populations of the western rock lobster (Panulirus cygnus)

Polyandry, where multiple mating by females results in the temporal and spatial overlap of ejaculates from two or more males, is taxonomically widespread and occurs in varying frequencies within and among species. In decapods (crabs, lobsters, crayfish, and prawns), rates of polyandry are likely to be variable, but the extent to which patterns of multiple paternity reflect multiple mating, and thus are shaped by postmating processes that bias fertilization toward one or a subset of mated males, is unclear. Here, we use microsatellite markers to examine the frequency of multiple mating (the presence of spermatophores from two or more males) and patterns of paternity in wild populations of western rock lobster (Panulirus cygnus). Our data confirm that >45% of females had attached spermatophores arising from at least two males (i.e., confirming polyandry), but we found very limited evidence for multiple paternity; among 24 clutches sampled in this study, only two arose from fertilizations by two or more males. Single inferred paternal genotypes accounted for all remaining progeny genotypes in each clutch, including several instances when the mother had been shown to mate with two or more males. These findings highlight the need for further work to understand whether polyandry is adaptive and to uncover the mechanisms underlying postmating paternity biases in this system.     

High frequency of multiple paternity in the largest rookery of Mediterranean loggerhead sea turtles

Mating systems are a central component in the evolution of animal life histories and in conservation genetics. The patterns of male reproductive skew and of paternal shares in batches of offspring, for example, affect genetic effective population size. A prominent characteristic of mating systems of sea turtles seem to be a considerable intra- and interspecific variability in the degree of polyandry. Because of the difficulty of observing the mating behaviour of sea turtles directly in the open sea, genetic paternity analysis is particularly useful for gaining insights into this aspect of their reproductive behaviour. We investigated patterns of multiple paternity in clutches of loggerhead sea turtles in the largest Mediterranean rookery using four highly variable microsatellite loci. Furthermore, we tested for a relationship between the number of fathers detected in clutches and body size of females. More than one father was detected in the clutches of 14 out of 15 females, with two clutches revealing the contribution of at least five males. In more than half the cases, the contributions of different fathers to a clutch did not depart from equality. The number of detected fathers significantly increased with increasing female body size. This relationship indicates that males may prefer to mate with large, and therefore productive, females. Our results suggest that polyandry is likely to increase effective population size compared to a population in which females would mate with only one male; male reproductive contributions being equal.     

Female multiple mating in wild and laboratory populations of the two-spot ladybird, Adalia bipunctata

Female mating rate is an important variable for understanding the role of females in the evolution of mating systems. Polyandry influences patterns of sexual selection and has implications for sexual conflict over mating, as well as for wider issues such as patterns of gene flow and levels of genetic diversity. Despite this, remarkably few studies of insects have provided detailed estimates of polyandry in the wild. Here we combine behavioural and molecular genetic data to assess female mating frequency in wild populations of the two-spot ladybird, Adalia bipunctata (Coleoptera: Coccinellidae). We also explore patterns of sperm use in a controlled laboratory environment to examine how sperm from multiple males is used over time by females, to link mating with fertilization. We confirm that females are highly polyandrous in the wild, both in terms of population mating rates (approximately 20% of the population found in copula at any given time) and the number of males siring offspring in a single clutch (three to four males, on average). These patterns are consistent across two study populations. Patterns of sperm use in the laboratory show that the number of mates does not exceed the number of fathers, suggesting that females have little postcopulatory influence on paternity. Instead, longer copulations result in higher paternity for males, probably due to the transfer of larger numbers of sperm in multiple spermatophores. Our results emphasize the importance of combining field and laboratory data to explore mating rates in the wild.     

Parentage assignment reveals multiple paternity in the critically-endangered Guatemalan beaded lizard (Heloderma charlesbogerti)

Within captive management programs for species of conservation concern, understanding the genetic mating system is of fundamental importance, given its role in generating and maintaining genetic diversity and promoting opportunities for sperm competition. If a goal of a conservation program is reintroduction, knowledge of the mating system may also inform prediction models aimed at understanding how genetic diversity may be spatially organized, thus informing decisions regarding where and which individuals should be released to maximize genetic diversity in the wild population. Within captive populations, such information may also influence how animals are maintained in order to promote natural behaviors. Here we investigate the genetic mating system of the Guatemalan beaded lizard, Heloderma charlesbogerti, a member of an entire clade lacking such information. A group of adult male and female H. charlesbogerti co-habited a large outdoor enclosure for five years during the species’ perceived breeding season. Through genomic parentage analysis, 50% of clutches comprising multiple offspring were found to result from multiple paternity, with up to three males siring offspring within single clutches. Both males and females were observed to produce offspring with multiple partners within a given year. As such, within this captive environment, where opportunities existed for mating with multiple partners, the genetic mating system was found to be highly polygamous, with multiple paternity common within clutches. These findings are novel for the family Helodermatidae, and the results have broader implications about how reproductive opportunities should be managed within captive conservation programs.

     

Genetic parentage assessment in the crayfish Orconectes placidus,a high-fecundity invertebrate with extended maternal brood care

Microsatellite data have recently been introduced in the context of genetic maternity and paternity assignments in high-fecundity fish species with single-parent-tended broods. Here we extend such analyses to an aquatic invertebrate, the crayfish Orconectes placidus, in which gravid females carry large numbers of offspring. Genetic parentage analyses of more than 900 progeny from 15 wild crayfish broods revealed that gravid females were invariably the exclusive dams of the offspring they tended (i.e. there was no allomaternal care), and that most of the females had mated with multiple (usually two) males who contributed sometimes highly skewed numbers of offspring to a brood. Within any multiply sired brood, the unhatched eggs (or the hatched juveniles) from different fathers were randomly distributed across the mother's brood space. All of these genetic findings are discussed in the light of observations on the mating behaviours and reproductive biology of crayfish.     

Paternity following multiple mating in ladybird <Emphasis Type="Italic">Harmonia axyridis</Emphasis>

We investigated factors which may determine realised paternity following multiple mating in a promiscuous ladybird Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae), namely the mating order and ability of males of different morphs to fertilize females. In experiments using males of two colour morphs, we found that the first male had a reproductive advantage over the second male (sperm precedence P2 = 0.38). Males of the melanic morph spectabilis had a reproductive advantage over males of the semi-melanic morph axyridis (P2 = 0.35). Analysis of paternity using microsatellites applied to young larvae, which avoids possible differential mortality of progeny, confirmed the precedence of the first male and higher paternity rate of colour morphs that are more frequent in the wild. Hatchability of eggs increased from 65% after the first mating to 85% after the second mating delayed by five days. Average duration of mating was 157 min and did not differ between the first and second mating and between the colour morphs of males.     

Sexual dimorphism and sexual selection in a montane scincid lizard (Eulamprus leuraensis)

Sex‐based divergences in body sizes and/or shapes within a species imply that selective forces act differently on morphological features in males versus females. That prediction can be tested with data on the relationship between morphology and reproductive output in females, and between morphology and realized paternity (based on genetic assignment tests) in males. In a sample of 81 field‐collected adult Blue Mountains water skinks (Eulamprus leuraensis), males and females averaged similar overall body sizes (snout–vent lengths (SVLs)). Reproductive success (based on 105 progeny produced by the females) increased with SVL at similar rates in both sexes (as expected from the lack of sexual size dimorphism). Multiple paternity was common. Males had larger heads than females of the same body size, and (as predicted) reproductive success increased with relative head size in males but not in females. Males also had relatively longer limbs and shorter trunks than females, but we did not detect significant sex differences in selection on those traits. Reproductive success in both sexes was increased by relatively longer hind limbs. Our data clarify mating systems in this endangered species, and suggest that mating systems are diverse within the genus Eulamprus.     

Multiple paternity in Ixodes ricinus (Acari: Ixodidae), assessed by microsatellite markers

This investigation examines multiple paternity in Ixodes ricinus (Acari: Ixodidae). Previous studies have shown that multiple mating occurs in this tick, but this is the first evaluation of multiple paternity. Three family groups were examined by a panel of polymorphic microsatellite loci; all ticks were bred from wild-collected engorged females with a copulating male attached. For most larvae, the attached males could be excluded as possible sire, and in the 3 tested families, at least 2 of 3 females mated successfully with more than 1 male. This finding suggests that multiple paternity is a common reproductive strategy in I. ricinus, which may have consequences for the ticks' dispersal success by increasing the genetic diversity in broods from single females colonizing new sites.     

Strong male/male competition allows for nonchoosy females: high levels of polygynandry in a territorial frog with paternal care

Our knowledge about genetic mating systems and the underlying causes for and consequences of variation in reproductive success has substantially improved in recent years. When linked to longitudinal population studies, cross-generational pedigrees across wild populations can help answer a wide suite of questions in ecology and evolutionary biology. We used microsatellite markers and exhaustive sampling of two successive adult generations to obtain population-wide estimates of individual reproductive output of males and females in a natural population of the Neotropical frog Allobates femoralis (Aromobatidae), a pan-Amazonian species that features prolonged iteroparous breeding, male territoriality and male parental care. Parentage analysis revealed a polygynandrous mating system in which high proportions of males (35.5%) and females (56.0%) produced progeny that survived until adulthood. Despite contrasting reproductive strategies, successfully reproducing males and females had similar numbers of mating partners that sired the adult progeny (both sexes: median 2; range 1-6); the numbers of their offspring that reached adulthood were also similar (both sexes: median 2; range 1-8). Measures of reproductive skew indicate selection on males only for their opportunity to breed. Reproductive success was significantly higher in territorial than in nonterritorial males, but unrelated to territory size in males or to body size in both sexes. We hypothesize that female polyandry in this species has evolved because of enhanced offspring survival when paternal care is allocated to multiple partners.