Lygaeus Simulans Research Articles

Mating failure shapes the patterns of sperm precedence in an insect

Estimates of last male sperm precedence (P2) are often used to infer mechanisms of sperm competition, a form of post-copulatory sexual selection. However, high levels of mating failure (i.e. copulations resulting in no offspring) in a population can lead to misinterpretations of sperm competition mechanisms. Through simulations, García-González (2004) illustrated how mating failure could cause bimodal distributions of paternity with peaks at P2 = 0 and 1, under a random sperm mixing mechanism. Here, we demonstrate this effect empirically with the seed bug Lygaeus simulans, a species known to exhibit high levels of mating failure (40–60%), using a morphological marker to estimate paternity. Contrary to previous findings in a sister species, we did not find strong evidence for last male sperm precedence. There was a tendency towards last male precedence (P2 = 0.58) but within the expected range for random sperm mixing. Instead, P2 was highly variable, with a bimodal distribution, as predicted by García-González (2004). After taking mating failure into account, the strongest driver of paternity outcome was copulation duration. Furthermore, we found evidence that mating failure could partly be a female-associated trait. Some doubly-mated females were more likely to produce no offspring or produce offspring from two different sires than expected by chance. Therefore, some females are more prone to experience mating failure than others, a result that mirrors an earlier result in male L. simulans. Our results confirm that mating failure needs to be considered when interrogating mechanisms of post-copulatory sexual selection.Significance statementMating failure arises when animals fail to produce offspring across their lifetime. This may be due to a failure to find a mate or a failure to produce offspring after one or more apparently successful matings. Sperm competition is when ejaculates of rival males compete to fertilize a female’s eggs. Estimates of second male paternity (P2) are often used to infer mechanisms of sperm competition (i.e. which male “wins” and how). However, García-González (2004) suggested that high levels of mating failure can skew paternity (i.e. give spuriously high/low levels of P2) and lead to misinterpretations of these mechanisms. We carried out sperm competition experiments on Lygaeus simulans seed bugs using a morphological marker to estimate paternity. We show empirically that mating failure does skew patterns of paternity, causing a bimodal distribution of P2. Therefore, by disrupting patterns of sperm competition, mating failure influences both the action of post-copulatory sexual selection and also our understanding of the mechanisms of sperm competition.

The fitness effects of a pale mutant in the aposematic seed bug Lygaeus simulans indicate pleiotropy between warning coloration and life history.

Conspicuous warning colors that signal chemical or other defenses are common in the natural world. For instance, such aposematic warning patterns of red‐and‐black or yellow‐and‐black are common among insect taxa, particularly in the order Hemiptera, often forming the basis of Batesian and/or Müllerian mimicry rings. In addition, it has been repeatedly noted that color polymorphisms or mutants that influence pigmentation can show pleiotropy with other behavioral, physiological, or life‐history traits. Here, we describe a pale mutant of the seed bug Lygaeus simulans that appeared in our laboratory population in 2012, which differs in color to the wild‐type bugs. Through multigenerational experimental crosses between wild‐type and pale mutant L. simulans, we first show that the pale phenotype segregates as a single Mendelian locus, with the pale allele being recessive to the wild type. Next, we show (a) that there is a large heterozygous advantage in terms of fecundity, (b) that pale females suffer reduced longevity, and (c) that pale males have increased body length compared to wild‐type homozygotes. Our data therefore suggest that the color locus is pleiotropic with a number of life‐history traits, opening the door for a more complete genetic analysis of aposematic coloration in this species. In addition, this phenotype will be useful as a visible genetic marker, providing a tool for investigating sperm competition and other post‐copulatory drivers of sexual selection in this species.

Can females choose to avoid mating failure in the seed bug Lygaeus simulans?

It is becoming increasingly clear that copulation does not necessarily always lead to offspring production in many organisms, despite fertilization success presumably being under both strong natural and sexual selection. In the seed bug Lygaeus simulans, between 40% and 60% of copulations fail to produce offspring, with this ‘mating failure’ representing a significantly repeatable male-associated trait. Mating has been demonstrated to be costly in this species and, as such, we might expect females to minimize the chance of mating failure by displaying a preference for males with higher insemination success where possible. After assaying males for mating failure, we asked whether females preferred males with a history of successful inseminations versus unsuccessful inseminations in pairwise mate choice trials. Contrary to our expectations, females showed no preference for more successful over less successful males. Moreover, females showed no preference for larger males in the choice trials, even though larger males were significantly more likely to successfully inseminate females in the initial assay. This apparent lack of female precopulatory choice suggests that postcopulatory choice mechanisms may be key to mating failure in this species. However, this does not necessarily explain why females pay the cost of mating with males they will then reject via postcopulatory processes. More generally, our results suggest that mating failure may play a largely underappreciated role in mating systems evolution, influencing both the cost of choosiness, and the costs and benefits of polyandry.

Variation in pre- and post-copulatory sexual selection on male genital size in two species of lygaeid bug.

Sexual selection has been shown to be the driving force behind the evolution of the sometimes extreme and elaborate genitalia of many species. Sexual selection may arise before and/or after mating, or vary according to other factors such as the social environment. However, bouts of selection are typically considered in isolation. We measured the strength and pattern of selection acting on the length of the male intromittent organ (or processus) in two closely related species of lygaeid seed bug: Lygaeus equestris and Lygaeus simulans. In both species, we measured both pre- and post-copulatory selection. For L. equestris, we also varied the experimental choice design used in mating trials. We found contrasting pre- and post-copulatory selection on processus length in L. equestris. Furthermore, significant pre-copulatory selection was only seen in mating trials in which two males were present. This selection likely arises indirectly due to selection on a correlated trait, as the processus does not interact with the female prior to copulation. In contrast, we were unable to detect significant pre- or post-copulatory selection on processus length in L. simulans. However, a formal meta-analysis of previous estimates of post-copulatory selection on processus length in L. simulans suggests that there is significant stabilising selection across studies, but the strength of selection varies between experiments. Our results emphasise that the strength and direction of sexual selection on genital traits may be multifaceted and can vary across studies, social contexts and different stages of reproduction.Significance statementAnimal genitalia vary greatly in size and complexity across species, and selection acting on genital size and shape can be complex. In this study, we show that the length of the penis in two species of seed bug is subject to complex patterns of selection, varying depending on the social context and whether selection is measured before or after mating. In one of the species, we show unexpectedly that penis length is correlated with male mating success, despite the fact that the penis does not interact with the female prior to mating. Our results highlight the fact that genitalia may be subject to both direct and indirect selection at different stages of mating and that to fully understand the evolution of such traits we should combine estimates of selection arising from these multiple episodes.Electronic supplementary materialThe online version of this article (doi:10.1007/s00265-016-2082-6) contains supplementary material, which is available to authorized users.

Natural breakage of the very long intromittent organ of the seed bug Lygaeus simulans (Heteroptera: Lygaeidae)

The male intromittent organ of the seed bug Lygaeus simulans ends in a long, sclerotized structure which is used to transfer sperm during mating. Observations suggest that this structure becomes brittle and is liable to breakage after being artificially exposed to the air for an extended period of time. In this study we investigate the frequency of intromittent organ breakage in L. simulans. We first examined the intromittent organ of a sample of males that mated once, and found that breakage was rare. We hypothesised that breakages are likely to be more frequent if a male is able to mate multiple times, and so we next paired males with a female for 21 days in order to provide the opportunity for multiple mating. Almost a quarter (22.5%) of these males exhibited signs of genital breakage. The point of breakage varied: for six males only the tip of the structure (around 6% of its length) was missing, whereas for three males over 50% of the structure was missing. However we were unable to locate any fragments of male genitalia in the reproductive tracts of any females that came into contact with these males. This suggests that breakages do not necessarily occur during mating itself, but instead probably occur as the intromittent organ is being retracted into the genital capsule following mating. In this species breakage may not significantly reduce male reproductive fitness as sperm transfer may still be possible.

The repeatability of mating failure in a polyandrous bug.

Mating failure, characterized by the lack of production of offspring following copulation, is relatively common across taxa yet is little understood. It is unclear whether mating failures are stochastic occurrences between incompatible mating partners or represent a persistent, meaningful phenotype on the part of one or other sex. Here we test this in the seed bug Lygaeus simulans, by sequentially mating families of males with randomly allocated unrelated females and calculating the repeatability of mating outcome for each individual male and family. Mating outcome is significantly repeatable within individual males but not across full‐sib brothers. We conclude that mating failure represents a consistent male‐associated phenotype with low heritability in this species, affected by as yet undetermined environmental influences on males.

Do the benefits of polyandry scale with outbreeding?

There have been many potential explanations put forward as to why polyandry often persists despite the multiple costs it can inflict on females. One such explanation is avoidance of costs associated with mating with genetically incompatible males. Genetic incompatibility can be thought of as a spectrum from individuals that are genetically too similar (inbreeding) to those that are too dissimilar (outbreeding or hybridization). Here we look for evidence that the level of outbreeding influences the benefits of polyandry in the seed bug Lygaeus equestris. Our system allows us to test for benefits of polyandry at levels of genetic similarity ranging from full siblings to heterospecifics, both in terms of egg production and hatching success. We found that while outbreeding level appeared to have no effect on fitness for intraspecific matings, and polyandry did not appear to result in any increase in fertility or fecundity, hybridization with a closely related species, Lygaeus simulans, carried considerable fitness costs. However, these costs could be rescued with a single mating to a conspecific. Thus, polyandry may be beneficial in populations that co-occur with closely related species and where there is reproductive interference. However, within-species genetic incompatibility is unlikely to be the driving force behind polyandry in this species. Furthermore, the mechanism underlying this rescue of fertility remains unclear as manipulation of male cuticular hydrocarbon profile, a possible mechanism by which females can assess male identity, had no effect on female offspring production.

Functional anatomy of the spermatheca and its duct in the seed bug Lygaeus simulans (Heteroptera: Lygaeidae)

Female genitalia of lygaeid bugs are characterized by a tube-shaped ductus receptaculi (spermathecal duct) connecting the bursa copulatrix with the highly coiled receptaculum seminis (spermatheca). In this study the morphology and functional anatomy of these structures in Lygaeus simulans were examined by light-, fluorescence- and electron microscopy. In addition, copulating animals were freeze fixed and their interconnected genital structures observed using light microscopy. The ductus receptaculi is separated from the receptaculum seminis by a complicated valve. The valve is nearly surrounded by the spermathecal muscle, which controls its opening. The ductus receptaculi leads into the proximal convoluted tube of the receptaculum seminis. Both the ductus receptaculi and the convoluted tube are composed of a single layer of epithelial cells lined by a thick electron dense apical cuticle. The distal part of the receptaculum seminis is a brownish, irregularly coiled, blind ending canal made of small epithelial cells covered with cuticle. Big glands are present in the epidermal layer. The cuticle of this distal part is much thinner and features concentric lamellae. The lumen of the receptaculum seminis cannot be expanded. For transfering sperm into the receptaculum the male aedeagus is equipped with a long, tube-like, sclerotized appendix (processus gonopori), which enlarges the ductus receptaculi considerably during copulation. For successful insemination the tip of the processus gonopori has to pass the valve. The convoluted tube, the valve and the surrounding spermathecal muscle may enable females to control insemination and egg fertilization.

Insemination and fertilization in the seed bug Lygaeus simulans (Heteroptera: Lygaeidae)

In the laboratory, not all females of the seed bug Lygaeus simulans Deckert, 1985, produced fertilized eggs after copulation: 26.7% of the females were not inseminated and 5% were inseminated but did not lay fertilized eggs; only in 40% of the couples did copulation result in fertile eggs. The remaining 28.3% of couples refrained from mating. Duration of copulation was associated with insemination and fertilization: (i) fertile eggs were produced by only one couple that copulated for less than 60 min and all those that copulated for more than 360 min, (ii) probability of fertilization increased steadily with duration of copulation between 60 and 360 min, and (iii) duration of copulation was significantly different for couples that showed different insemination status.A possible morphological explanation for this rime dependency was revealed by examining the genitalia of 69 couples freeze-fixed in copula after different periods in copulation. Because of the intricate structure of the genitalia in L. simulans, a male takes a long time to manoeuver its intromittent organ into the narrow insemination duct of the female. Only if completely inserted is the tip of the intromittent organ close enough for successful ejaculation of sperm into the spermatheca. The freeze-fixing experiment revealed that it usually took the male more than 30 min to locate the entrance to the insemination duct and another 30 min for full penetration. This explains why copulations that lasted less than 60 min failed, since insemination began only after intromission was complete.The experiments, therefore, indicated that there is a relationship between the complex morphology of the genitalia and the low rates of insemination and fertilization in L. simulans.

Selection of male traits during and after copulation in the seedbug Lygaeus simulans (Heteroptera, Lygaeidae)

In order to evaluate selection of male morphological traits during copulation, a laboratory experiment was performed with the promiscuous seedbug Lygaeus simulans. Three male traits suspected as putative targets of selection were measured: weight, fluctuating asymmetry of a measure on the forewings, and length of a conspicuous genital structure, the processus gonopori. As fitness measures we considered total fecundity (number of fertilized eggs), insemination and fertilization success (established if a female laid fertilized eggs after copulation), and the interval between copulation and oviposition. Eighty-four males were allowed a single copulation with one virgin female each. Out of 67 copulations, 27 (40.2%) resulted in fertilized eggs and the oviposition latency ranged from 6 to 26 days. Regressions of male traits on the fitness measures showed significant phenotypical selection of two male traits: (1) males of average weight are more likely to achieve fertilization and (2) the oviposition latency was shorter for males with lower asymmetry. The copulation–oviposition interval may be especially important for male fertilization success because Lygaeus males perform copulatory mate guarding and the last male copulating with a female fertilizes most of the eggs. No selection of the genitalic trait was detected.

Selection of male traits during and after copulation in the seedbug Lygaeus simulans (Heteroptera, Lygaeidae)

Abstract In order to evaluate selection of male morphological traits during copulation, a laboratory experiment was performed with the promiscuous seedbug Lygaeus simulans . Three male traits suspected as putative targets of selection were measured: weight, fluctuating asymmetry of a measure on the forewings, and length of a conspicuous genital structure, the processus gonopori. As fitness measures we considered total fecundity (number of fertilized eggs), insemination and fertilization success (established if a female laid fertilized eggs after copulation), and the interval between copulation and oviposition. Eighty-four males were allowed a single copulation with one virgin female each. Out of 67 copulations, 27 (40.2%) resulted in fertilized eggs and the oviposition latency ranged from 6 to 26 days. Regressions of male traits on the fitness measures showed significant phenotypical selection of two male traits: (1) males of average weight are more likely to achieve fertilization and (2) the oviposition latency was shorter for males with lower asymmetry. The copulation–oviposition interval may be especially important for male fertilization success because Lygaeus males perform copulatory mate guarding and the last male copulating with a female fertilizes most of the eggs. No selection of the genitalic trait was detected.

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

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

Über Lygaeus simulans spec. nov. und L. equestris (Linnaeus, 1758), zwei nahe verwandte paläarktische Lygaeinae (Heteroptera, Lygaeidae)

On Lygaeus simulans spec. nov. and L. equestris (Linneaus, 1758), Two Closely Related Palearctic Lygaeinae (Heteroptera, Lygaeidae)The binomen Lygaeus equestris applies to two distinct but related sympatric species, both of palearctic distribution. L. simulans spec. nov. is described and the differences between the new species and L. equestris s. str. are discussed.