Paul E. Eady

Female genotype affects male success in sperm competition

The central question addressed by most studies of sperm competition is: ‘what determines which males sperm are used at fertilization?’ Empirical and theoretical studies that address this question have traditionally focused on adaptations which enhance male fertilization success while treating the female as a receptacle in which sperm competition is played out. Here we provide evidence which suggests that female genotype strongly influences the outcome of sperm competition. When the sperm of two males are in competition the proportion of offspring fathered by the second male to mate (P2) was found to be highly repeatable only if the male pair were mated to three different, but genetically similar females (full–sisters to each other; unrelated to either of the males). In contrast, if a male pair were mated to three females that were unrelated then P2 was either non–repeatable or marginally repeatable. We also show that male success in sperm competition is determined, to a large extent, by gamete and/or male–female compatibility. This conclusion is derived from the observation that P2 was repeatable among full–sisters mated to different, yet genetically similar male pairings, whilst P2 was non–repeatable among full–sisters mated to different, genetically distinct male pairings.

Why do male Callosobruchus maculatus beetles inseminate so many sperm

Male Callosobruchus maculatus F. (Coleoptera: Bruchidae) inseminate more sperm than females can effectively store in their spermathecae. This study examines the adaptive significance of “excess” sperm transfer by measuring components of male and female reproductive success in response to manipulating the number of sperm inseminated. The number of sperm transferred during copulation was reduced from 56,000 ±4,462 to 8,700±1,194 by sequentially mating males to virgin females. Reducing the number of sperm inseminated by the first male to mate had no effect on the extent of sperm precedence, but reducing the number of sperm inseminated by the second male resulted in a significant reduction in the extent of sperm precedence. When large numbers of sperm are inseminated the remating refractory period of females is increased. These results indicate that males transferring large numbers of sperm during copulation have a two-fold advantage at fertilization; they are more effective at preempting previously stored sperm and they are likely to father more offspring by delaying the time of female remating. The transfer of “excess” sperm does not appear to serve as nonpromiscuous male mating effort; the number of eggs laid, their fertility and the subsequent survival of zygotes were unaffected by manipulating the number of sperm inseminated. The underlying mechanisms of sperm precedence were also examined. Simple models of sperm displacement failed to accurately predict the patterns of sperm precedence observed in this species. However, the results do not provide conclusive evidence against the models but rather serve to highlight our limited understanding of the movement of sperm within the females reproductive tract.

GENETIC ARCHITECTURE OF METABOLIC RATE: ENVIRONMENT SPECIFIC EPISTASIS BETWEEN MITOCHONDRIAL AND NUCLEAR GENES IN AN INSECT

The extent to which mitochondrial DNA (mtDNA) variation is involved in adaptive evolutionary change is currently being reevaluated. In particular, emerging evidence suggests that mtDNA genes coevolve with the nuclear genes with which they interact to form the energy producing enzyme complexes in the mitochondria. This suggests that intergenomic epistasis between mitochondrial and nuclear genes may affect whole‐organism metabolic phenotypes. Here, we use crossed combinations of mitochondrial and nuclear lineages of the seed beetle Callosobruchus maculatus and assay metabolic rate under two different temperature regimes. Metabolic rate was affected by an interaction between the mitochondrial and nuclear lineages and the temperature regime. Sequence data suggests that mitochondrial genetic variation has a role in determining the outcome of this interaction. Our genetic dissection of metabolic rate reveals a high level of complexity, encompassing genetic interactions over two genomes, and genotype × genotype × environment interactions. The evolutionary implications of these results are twofold. First, because metabolic rate is at the root of life histories, our results provide insights into the complexity of life‐history evolution in general, and thermal adaptation in particular. Second, our results suggest a mechanism that could contribute to the maintenance of nonneutral mtDNA polymorphism.

FUNCTIONAL INCOMPATIBILITY BETWEEN THE FERTILIZATION SYSTEMS OF TWO ALLOPATRIC POPULATIONS OF CALLOSOBRUCHUS MACULATUS (COLEOPTERA: BRUCHIDAE)

Abstract Recent studies indicate that postcopulatory sexual selection may represent an important component of the speciation process by initiating reproductive isolation via the evolutionary divergence of fertilization systems. Using two geographically isolated populations of the polyandrous beetle Callosobruchus maculatus, we investigated divergence in fertilization systems by determining the extent of postcopulatory functional incompatibility. Through reciprocal, cross‐population matings we were able to separately estimate the effects of male and female population origin and their interaction on the extent of last‐male sperm precedence, female receptivity to further copulation and female oviposition. Our results indicate partial incompatibility between the fertilization systems of the two populations at all three functional levels. Males derived from the same population as females outcompete rival, allopatric males with respect to sperm preemption, sperm protection, and ability to stimulate female oviposition. This pattern is reciprocated in both populations indicating that postcopulatory, prezygotic events represent important mechanisms by which between‐population gene flow is reduced. We suggest the partial gametic isolation observed is a by‐product of the coevolution of male and female fertilization systems by a process of cryptic female choice. Our results are consistent with a mechanism akin to conventional mate choice models although they do not allow us to reject antagonistic sexual coevolution as the mechanism of cryptic female choice.

COPULATING WITH MULTIPLE MATES ENHANCES FEMALE FECUNDITY BUT NOT EGG-TO-ADULT SURVIVAL IN THE BRUCHID BEETLE CALLOSOBRUCHUS MACULATUS

Postcopulatory sexual selection theory has come a long way since the evolutionary implications of sperm competition were first spelled out by Parker (1970). However, one of the most enduring questions remains: why do females copulate with multiple males? Here we show that females copulating with multiple males lay more eggs than those copulating repeatedly with the same male. We also show egg‐to‐adult survival to be more variable when females copulate multiply with different males and less variable when they copulate multiply with the same male. This supports the notion that egg‐to‐adult survival may depend on the genetic compatibility of males and females. However, pre‐adult survival was highest when females copulated repeatedly with the same male rather than with different males. Thus, it would appear that polyandry in this species does not function to reduce the risk of embryo failure resulting from fertilization by genetically incompatible sperm.

Copulation, genital damage and early death in Callosobruchus maculatus

Antagonistic sexual coevolution stems from the notion that male and female interests over reproduction are in conflict. Such conflicts appear to be particularly obvious when male genital armature inflicts damage to the female reproductive tract resulting in reduced female longevity. However, studies of mating frequency, genital damage and female longevity are difficult to interpret because females not only sustain more genital damage, but also receive more seminal fluid when they engage in multiple copulations. Here, we attempt to disentangle the effects of genital damage and seminal fluid transfer on female longevity in the beetle Callosobruchus maculatus (Coleoptera: Bruchidae). Males copulating for the sixth time in succession inflicted greater levels of genital damage, but transferred smaller ejaculates in comparison with virgin males. The number of copulations performed by males was negatively related to female fecundity and positively related to female longevity, suggesting a trade-off between fecundity and longevity. However, inclusion of fecundity as a covariate revealed sperm and/or seminal fluid transfer to have a negative impact on female longevity above that caused by the fecundity–longevity trade-off. The consequences of multiple copulations on female longevity were examined. Females that mated twice laid more eggs and died sooner than those that mated once. However, incorporation of fecundity as a covariate into our statistical model removed the effect of female mating frequency on female longevity, indicating that double-mated females suffer greater mortality owing to the trade-off between fecundity and longevity. Males of this species are known to transfer very large ejaculates (up to 8% of their body weight), which may represent a significant nutritional benefit to females. However, the receipt of large ejaculates appears to carry costs. Thus, the interpretation of multiple mating experiments on female longevity and associated functional explanations of polyandry in this species are likely to be complex.

Costly sexual harassment in a beetle

Abstract The optimal number of mating partners for females rarely coincides with that for males, leading to sexual conflict over mating frequency. In the bruchid beetle Callosobruchus maculatus, the fitness consequences to females of engaging in multiple copulations are complex, with studies demonstrating both costs and benefits to multiple mating. However, females kept continuously with males have a lower lifetime egg production compared with females mated only once and then isolated from males. This reduction in fitness may be a result of damage caused by male genitalia, which bear spines that puncture the female’s reproductive tract, and/or toxic elements in the ejaculate. However, male harassment rather than costs of matings themselves could also explain the results. In the present study, the fitness costs of male harassment for female C. maculatus are estimated. The natural refractory period of females immediately after their first mating is used to separate the cost of harassment from the cost of mating. Male harassment results in females laying fewer eggs and this results in a tendency to produce fewer offspring. The results are discussed in the context of mate choice and sexual selection.

Sperm transfer and storage in relation to sperm competition in Callosobruchus maculatus

This paper examines the underlying mechanisms of sperm competition in the beetle Callosobruchus maculatus (F.) (Coleoptera: Bruchidae). Recently developed mathematical models of sperm competition are combined with an empirical investigation of the processes of sperm transfer and storage. During a single insemination virgin males transfer approximately 46000 sperm, 85% more sperm than females can effectively store in their spermathecae. Many of these sperm remain in the bursa copulatrix where they are apparently rapidly degraded and can therefore play no role in fertilization. The spermatheca (primary site of sperm storage) is filled by a single insemination and sperm are lost from this organ at a constant rate. This rate of sperm loss from the spermatheca is insufficient for sperm mixing (without displacement) or sperm stratification to account for the degree of last male sperm precedence measured as P2; the proportion of offspring fathered by the second male to mate reported for this species (P2 = 0.83, when two inseminations are separated by 24 h). Models of sperm displacement correctly predict high levels of sperm precedence although the precision of these predictions is limited because the proportion of sperm entering the spermatheca cannot be accurately determined. The results suggested that last male sperm precedence in C. maculatus the result of sperm displacement, although the exact mechanism of displacement (sperm-for-sperm or fluid displacement) remains unknown. Possible constraints imposed by female genital anatomy on sperm displacement are discussed.

Postcopulatory, prezygotic reproductive isolation

Prezygotic reproductive isolation has traditionally been studied from a precopulatory perspective even though postcopulatory events have been known to influence success at fertilization. Postcopulatory, prezygotic reproductive isolation (gametic isolation) has received relatively little attention mainly because the focus of research has been on the events that occur either earlier (pre-mating) or later (postzygotic) in the chain of events that lead to successful reproduction. However, recent evidence from an array of taxa from sea urchins to beetles reveals that postejaculatory, prezygotic events may be an important factor in reproductive isolation. Such a mechanism may be the driving force behind speciation in some taxa, such as free-spawning marine invertebrates, whilst it may complement pre-mating and postzygotic mechanisms in others. In this paper we draw an analogy between sperm–egg interactions and classical sexual selection theory and argue that gametic incompatibility may arise through cryptic female choice.

The effect of single, double, and triple matings on the lifetime fecundity of Callosobruchus analis and Callosobruchus maculatus (Coleoptera: Bruchidae)

This study examined the effect of single, double, and triple matings on female lifetime fecundity in two closely related species of beetle, Callosobruchus analis and C. maculatus. Multiple mating resulted in elevated lifetime fecundities in both species, although the pattern of elevation differed between the two species. When oviposition resource was plentiful the elevation was apparent after a second but not a third copulation for C. maculatus and after a second and after a third for C. analis. By altering the availability of oviposition sites to C. maculatus females, we were able to alter the pattern of fecundity elevation between matings. When oviposition sites were limited, fecundity increased after a third mating but not a second mating. We suggest that the pattern of fecundity elevation associated with multiple mating in these species is primarily the result of oviposition stimulants delivered by males, rather than a nutritional “donation” brought about by the metabolism of ejaculates. We also suggest that the intra- and interspecific differences in the relationship between mating frequency and fecundity are the result of differences in egg maturation rates.