Mechanisms of reproductive isolation among sympatric broadcast-spawning corals of the Montastraea annularis species complex

Many coral species spawn simultaneously and have compatible gametes, leading to controversy over the nature of species boundaries and the frequency with which hybridization occurs. Three western Atlantic corals, Montastraea annularis, M. faveolata, and M. franksi, typify this controversy; they all spawn sympatrically on the same evenings after the fall full moons. Here we show, in both Panama and the Bahamas for multiple years, how a variety of mechanisms may act in concert to reproductively isolate all three species. Field studies indicate that M. franksi spawns two hours earlier than the other two species, and the eggs released during this earlier period disperse an average of 500 m by the time the other species spawn. Field measures of fertilization indicate that peak fertilization occurs when spawning synchrony is high and that corals that spawn at the tails of the spawning distributions have greatly reduced fertilization success. Laboratory studies indicate that there is a gametic incompatibility between M. faveolata and the other two species. There are regional differences in the gametic compatibility of M. franksi and M. annularis. In Panama, the two species are completely compatible, whereas in the Bahamas, M. franksi sperm can fertilize M. annularis eggs but the reciprocal cross often fails. Gamete age influences patterns of fertilization, such that very young eggs seem resistant to fertilization and old sperm lose viability after two hours. In sum, the combination of temporal differences in spawning, sperm aging, gamete dispersal and dilution, and gametic incompatibility act in various combinations among the three species, making it unlikely that hybrid fertilization would occur.     

Weak prezygotic isolating mechanisms in threatened Caribbean Acropora corals

The Caribbean corals, Acropora palmata and A. cervicornis, recently have undergone drastic declines primarily as a result of disease. Previous molecular studies have demonstrated that these species form a hybrid (A. prolifera) that varies in abundance throughout the range of the parental distribution. There is variable unidirectional introgression across loci and sites of A. palmata genes flowing into A. cervicornis. Here we examine the efficacy of prezygotic reproductive isolating mechanisms within these corals including spawning times and choice and no-choice fertilization crosses. We show that these species have subtly different mean but overlapping spawning times, suggesting that temporal isolation is likely not an effective barrier to hybridization. We found species-specific differences in gametic incompatibilities. Acropora palmata eggs were relatively resistant to hybridization, especially when conspecific sperm are available to outcompete heterospecific sperm. Acropora cervicornis eggs demonstrated no evidence for gametic incompatibility and no evidence of reduced viability after aging four hours. This asymmetry in compatibility matches previous genetic data on unidirectional introgression.     

Quantitative analysis of gametic incompatibility between closely related species of neotropical sea urchins

Species of the sea urchin genus Echinometra found on the two coasts of Panamá are recently diverged and only partially isolated by incomplete barriers to interspecific fertilization. This study confirms previous work that revealed incompatibility between the eggs of the Atlantic E. lucunter and the sperm of the other two neotropical species, whereas eggs of its sympatric congener E. viridis and allopatric E. vanbrunti are largely compatible with heterospecific sperm. Here we quantify fertilization using a range of sperm dilutions. We demonstrate a much stronger block to cross-species fertilization of E. lucunter eggs than was previously shown at fixed sperm concentrations, and mild incompatibility of the other two species' eggs where previous crosses between species were not distinguishable from within-species controls. Additionally, we present evidence for intraspecific variation in egg receptivity towards heterospecific sperm. Our findings here again discount the "reinforcement model" as a viable explanation for the pattern of prezygotic isolation. Gamete incompatibility in these Echinometra has appeared recently-within the last 1.5 million years-but is weaker in sympatry than in allopatry. Accidents of history may help explain why incompatibility of eggs emerged in one species and not in others. Compensatory sexual selection on sperm in this species could follow, and promote divergence of proteins mediating sperm-egg recognition.     

CRYPTIC CHOICE OF CONSPECIFIC SPERM CONTROLLED BY THE IMPACT OF OVARIAN FLUID ON SPERM SWIMMING BEHAVIOR

Despite evidence that variation in male–female reproductive compatibility exists in many fertilization systems, identifying mechanisms of cryptic female choice at the gamete level has been a challenge. Here, under risks of genetic incompatibility through hybridization, we show how salmon and trout eggs promote fertilization by conspecific sperm. Using in vitro fertilization experiments that replicate the gametic microenvironment, we find complete interfertility between both species. However, if either species’ ova were presented with equivalent numbers of both sperm types, conspecific sperm gained fertilization precedence. Surprisingly, the species’ identity of the eggs did not explain this cryptic female choice, which instead was primarily controlled by conspecific ovarian fluid, a semiviscous, protein‐rich solution that bathes the eggs and is released at spawning. Video analyses revealed that ovarian fluid doubled sperm motile life span and straightened swimming trajectory, behaviors allowing chemoattraction up a concentration gradient. To confirm chemoattraction, cell migration tests through membranes containing pores that approximated to the egg micropyle showed that conspecific ovarian fluid attracted many more spermatozoa through the membrane, compared with heterospecific fluid or water. These combined findings together identify how cryptic female choice can evolve at the gamete level and promote reproductive isolation, mediated by a specific chemoattractive influence of ovarian fluid on sperm swimming behavior.     

Males with high genetic similarity to females sire more offspring in sperm competition in Peron's tree frog Litoria peronii

Recent work has confirmed that genetic compatibility among mates can be an important determinant of siring success in sperm competition experiments and in free-ranging populations. Most of this work points towards mate choice of less related mates. However, there may also be the potential for mate choice for intermediate or even genetically similar mates to prevent outbreeding depression or hybridization with closely related taxa. We studied relatedness effects on post-copulatory gametic choice and/or sperm competition in an external fertilizer, Peron's tree frog (Litoria peronii), since external fertilizers offer exceptional control in order to test gametic interaction effects on probability of paternity and zygote viability. Sperm competition experiments were done blindly with respect to genetic relatedness among males and females. Thereafter, paternity of offspring was assigned using eight microsatellite loci. Three hybridization trials between L. peronii and a closely related sympatric species Litoria tyleri were also carried out. In the sperm competition trials, males that are more genetically similar to the female achieved higher siring success compared with less genetically similar males. The hybridization trials confirmed that the two species can interbreed and we suggest that the risk of hybridization may contribute to selection benefits for genetically more similar males at fertilization. To our knowledge, this study is the first to show evidence for post-copulatory selection of sperm from genetically more similar individuals within a natural population.     

Differential fertilization success between two populations of eastern oyster, Crassostrea virginica

Identification of mechanisms promoting prezygotic reproductive isolation and their prevalence are key goals in evolutionary biology because of their potential role in speciation. In marine broadcast-spawning species, molecular interactions between gamete surface proteins are more important than mating behavior for determining reproductive compatibility. Evidence for differential fertilization capacity has been reported from experiments utilizing competing sperm from two males sampled within populations and between species, but to our knowledge conspecific populations that might have diverged in allopatry have never been tested on the basis of sperm competition. In the present study, the gametic compatibility and embryo survivorship from matings between two allopatric populations of Crassostrea virginica, the eastern oyster, on either side of a genetic step cline were investigated. Fertilization success, embryo survival, and paternity data all indicated an absence of strong reproductive barriers between the two oyster populations, implicating other mechanisms for maintenance of the cline step. Sperm from northern male oysters showed a tendency to produce more larvae than expected when competing with sperm from southern male oysters. Although the northern male advantage was not strong, the trend implies that long-distance dispersal across the step cline might more successfully result in north-to-south gene flow than the reverse, providing a mechanistic hypothesis explaining the asymmetric cline shape.     

Ocean acidification alters sperm responses to egg-derived chemicals in a broadcast spawning mussel

The continued emissions of anthropogenic carbon dioxide are causing progressive ocean acidification (OA). While deleterious effects of OA on biological systems are well documented in the growth of calcifying organisms, lesser studied impacts of OA include potential effects on gamete interactions that determine fertilization, which are likely to influence the many marine species that spawn gametes externally. Here, we explore the effects of OA on the signalling mechanisms that enable sperm to track egg-derived chemicals (sperm chemotaxis). We focus on the mussel Mytilus galloprovincialis, where sperm chemotaxis enables eggs to bias fertilization in favour of genetically compatible males. Using an experimental design based on the North Carolina II factorial breeding design, we test whether the experimental manipulation of seawater pH (comparing ambient conditions to predicted end-of-century scenarios) alters patterns of differential sperm chemotaxis. While we find no evidence that male–female gametic compatibility is impacted by OA, we do find that individual males exhibit consistent variation in how their sperm perform in lowered pH levels. This finding of individual variability in the capacity of ejaculates to respond to chemoattractants under acidified conditions suggests that climate change will exert considerable pressure on male genotypes that can withstand an increasingly hostile fertilization environment.     

Multiple gene genealogies reveal asymmetrical hybridization and introgression among strongylocentrotid sea urchins

The evolution of incompatibilities between eggs and sperm is thought to play important roles in establishing and maintaining reproductive isolation among species of broadcast-spawning marine invertebrates. However, the effectiveness of gametic isolation in initiating the speciation process and/or in limiting the introgression of genes among species at later stages of divergence remains largely unknown. In the present study, we collected DNA sequence data from five loci in four species of Strongylocentrotus sea urchins ( S. droebachiensis , S. pallidus , S. purpuratus , and S. franciscanus ) to test whether the susceptibility of S. droebachiensis eggs to fertilization by heterospecific sperm results in gene flow between species. Despite the potential for introgression, a small but statistically significant signal of introgression was observed only between the youngest pair of sister taxa ( S. pallidus and S. droebachiensis ) that was strongly asymmetrical (from the former into the latter). No significant gene flow was observed for either S. purpuratus or S. franciscanus despite the ability of their sperm to readily fertilize the eggs of S. droebachiensis . Our results demonstrate that asymmetrical gamete compatibilities in strongylocentrotids can give rise to asymmetrical patterns of introgression but suggest that gamete traits alone cannot be responsible for maintaining species integrities. The genetic boundaries between strongylocentrotid urchin species in the northeast Pacific appear to be related to postzygotic isolating mechanisms that scale with divergence times and not intrinsic gametic incompatibilities per se .     

Gamete compatibility between marine and estuarine Acanthopagrus spp. (Sparidae) and their hybrids

On Australia's south‐east coast, hybridization between estuary‐restricted black bream Acanthopagrus butcheri Munro and its migratory coastal congener yellowfin bream Acanthopagrus australis (Günther) has led to estuarine populations largely composed of hybrids that are most genetically similar to A. butcheri. The fertilization success achieved when ova of estuary‐caught A. butcheri were fertilized with the cryogenically preserved sperm of either ocean‐caught A. australis or estuary‐caught A. butcheri‐like was compared. The experimental crosses, which by chance included both pure parental and hybrid bream, revealed no evidence that gametic incompatibility provides a barrier to fertilization among both pure species and their hybrids.     

几种同域分布姜花属植物的种间杂交亲和性

种间配子不亲和以及种间杂交种子活力低是众多的物种间杂交隔离机制中的两个方面。通过对云南澜沧地区分布的4种（型）姜花属植物——圆瓣姜花（Hedychium forrestii）、草果药(Hspicatum)、两类型滇姜花（Hyunnanense）间进行野外杂交试验,比较杂交结实率、每果种子数以及杂交种子的萌发参数等指标来分析4种（型）姜花之间的杂交亲和性和杂交后代表现,发现4种（型）姜花属植物配子间都具有不同程度的亲和性,但种间杂交种子的萌发适合度比同种授粉获得的种子低。结果表明,这4种（型）姜花属植物间配子亲和性不是有效的种间隔离机制,种间杂交种子活力低是一种不完全的隔离机制,它们在同域分布的地区,仍然有形成杂交后代的可能性。     

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.     

Sperm chromatin remodelling and Wolbachia-induced cytoplasmic incompatibility in Drosophila.

Wolbachia pipientis is an obligate bacterial endosymbiont, which has successfully invaded approximately 20% of all insect species by manipulating their normal developmental patterns. Wolbachia-induced phenotypes include parthenogenesis, male killing, and, most notably, cytoplasmic incompatibility. In the future these phenotypes might be useful in controlling or modifying insect populations but this will depend on our understanding of the basic molecular processes underlying insect fertilization and development. Wolbachia-infected Drosophila simulans express high levels of cytoplasmic incompatibility in which the sperm nucleus is modified and does not form a normal male pronucleus when fertilizing eggs from uninfected females. The sperm modification is somehow rescued in eggs infected with the same strain of Wolbachia. Thus, D. simulans has become an excellent model organism for investigating the manner in which endosymbionts can alter reproductive programs in insect hosts. This paper reviews the current knowledge of Drosophila early development and particularly sperm function. Developmental mutations in Drosophila that are known to affect sperm function will also be discussed.incompatibility.     

几种同域分布姜花属植物的种间杂交亲和性及杂交后代种子活力

种间配子不亲和以及种间杂交种子活力低是众多的物种间杂交隔离机制中的两个方面。通过对云南澜沧地区分布的4种（型）姜花属植物——圆瓣姜花（Hedychium forrestii）、草果药（H.spicatum）、两类型滇姜花（H.yunnanense）间进行野外杂交试验,比较杂交结实率、每果种子数以及杂交种子的萌发参数等指标来分析4种（型）姜花之间的杂交亲和性和杂交后代表现,发现4种（型）姜花属植物配子间都具有不同程度的亲和性,但种间杂交种子的萌发适合度比同种授粉获得的种子低。结果表明,这4种（型）姜花属植物间配子亲和性不是有效的种间隔离机制,种间杂交种子活力低是一种不完全的隔离机制,它们在同域分布的地区,仍然有形成杂交后代的可能性。     

RAPID EVOLUTION OF ASYMMETRIC REPRODUCTIVE INCOMPATIBILITIES IN STALK‐EYED FLIES

The steps by which isolated populations acquire reproductive incompatibilities remain poorly understood. One potentially important process is postcopulatory sexual selection because it can generate divergence between populations in traits that influence fertilization success after copulation. Here we present a comprehensive analysis of this form of reproductive isolation by conducting reciprocal crosses between variably diverged populations of stalk‐eyed flies (Teleopsis dalmanni). First, we measure seven types of reproductive incompatibility between copulation and fertilization. We then compare fertilization success to hatching success to quantify hybrid inviability. Finally, we determine if sperm competition acts to reinforce or counteract any incompatibilities. We find evidence for multiple incompatibilities in most crosses, including failure to store sperm after mating, failure of sperm to reach the site of fertilization, failure of sperm to fertilize eggs, and failure of embryos to develop. Local sperm have precedence over foreign sperm, but this effect is due mainly to differences in sperm transfer and reduced hatching success. Crosses between recently diverged populations are asymmetrical with regard to the degree and type of incompatibility. Because sexual conflict in these flies is low, postcopulatory sexual selection, rather than antagonistic coevolution, likely causes incompatibilities due to mismatches between male and female reproductive traits.     

GAMETIC INCOMPATIBILITY BETWEEN SPECIES OF THE SEA URCHIN ECHINOMETRA ON THE TWO SIDES OF THE ISTHMUS OF PANAMA

The Pliocene rise of the Central American Isthmus has resulted in numerous “geminate pairs,” i.e., closely related species, one on each coast. Such species pairs can provide information on the evolution of isolating mechanisms in allopatry and on the relationship between genetic divergence and reproductive isolation in populations separated at a known time. The sea urchin genus Echinometra has one species, E. vanbrunti, in the eastern Pacific, and two, E. lucunter and E. viridis, in the Caribbean. E. viridis is morphologically distinct from the other two species, leading to the conclusion that E. lucunter and E. vanbrunti constitute a geminate pair. Allozyme data, on the other hand, place the speciation event of the two currently sympatric species after the rise of the Isthmus. We report fertilization experiments between the gametes of the three species performed to determine degree of reproductive isolation. Crosses between E. viridis and E. vanbrunti produce rates of fertilization almost equal to those manifested in homogamic crosses. Sperm of E. lucunter can fertilize eggs of the other two species, but few of its eggs permit fertilization by heterospecific sperm. Contrary to the predictions of the “speciation by reinforcement” hypothesis, degree of incompatibility between the allopatric E. lucunter and E. vanbrunti is higher than between the sympatric E. lucunter and E. viridis. Despite the incomplete and unidirectional nature of their gametic isolation, E. lucunter and E. viridis maintain their genetic integrities. Consideration of the likely phylogenetic relationships between the three species suggests that incompatibility of E. lucunter eggs with heterospecific sperm has evolved in the last 3.5 million years, after the rise of the Isthmus. There is no correlation between genetic divergence and strength of reproductive isolation, either within Echinometra, or among the geminate species pairs of echinoids that have been studied to date. Because recognition between echinoid gametes depends on the chemical compositions of a sperm protein and an egg glycoprotein, the appearance of gametic isolation would require only the fixation of a few mutations in each population on either side of a geographic barrier and could be independent of any other kind of genetic divergence. Thus, in animals with external fertilization, speciation need not be accompanied by major genomic reorganization.     

Parasitic mites influence intra‐ and interpopulational variation in sperm length in a simultaneous hermaphrodite land snail (Gastropoda: Helicidae)

Sperm morphology can be highly variable among species, but less is known about patterns of population differentiation within species. Sperm morphology is under strong sexual selection, may evolve rapidly, and often co‐varies with other reproductive traits that differ between populations. We investigated variation in sperm morphology in the simultaneous hermaphrodite land snail Arianta arbustorum in relation to parasitic mite infection. Variation in total sperm length and sperm head length was assessed in 23 populations sampled across the distributional range of the species in Central and Northern Europe. We found a pronounced variation in total sperm length among the populations studied, with a difference of 11.0% of total sperm length between the shortest and longest population means. Differences among populations explained 62.9% of the variance in total sperm length, differences among individual snails within population 23.4% and differences within individual snail 13.7%. Mantel tests showed that interpopulation differences in total sperm length increased significantly with geographical distance between populations. A minimal adequate model revealed that parasitic infection had a positive effect and longitude a negative effect on total sperm length. Thus, independent of the population examined, mite‐infected individuals of A. arbustorum produced larger sperm than uninfected snails and total sperm length decreased from west to east. Sperm head length also varied among populations, but it was not influenced by any of the factors examined. In a subsample of 12 populations restricted to the mountains of Switzerland (elevational range 440–2485 m a.s.l.), total sperm length decreased with increasing elevation. Our results suggest that selection pressures acting among populations may differ from those acting within. Stabilizing selection might be a possible mechanism for producing the reduced variation observed in sperm length within a population. © 2014 The Linnean Society of London, Biological Journal of the Linnean Society, 2014, 113 , 1036–1046.     

Asymmetric conspecific sperm precedence in relation to spawning times in the Montastraea annularis species complex (Cnidaria: Scleractinia)

In broadcast spawners, prezygotic reproductive isolation depends on differences in the spatial and temporal patterns of gamete release and gametic incompatibility. Typically, gametic incompatibility is measured in no‐choice crosses, but conspecific sperm precedence (CSP) can prevent hybridization in gametes that are compatible in the absence of sperm competition. Broadcast spawning corals in the Montastraea annularis species complex spawn annually on the same few evenings. Montastraea franksi spawns an average of 110 min before M. annularis, with a minimum gap of approximately 40 min. Gametes are compatible in no‐choice heterospecific assays, but it is unknown whether eggs exhibit choice when in competition. Hybridization depends on either M. franksi eggs remaining unfertilized and in proximity to M. annularis when the latter species spawns or M. franksi sperm remaining in sufficient viable concentrations when M. annularis spawns. We found that the eggs of the early spawning M. franksi demonstrate strong CSP, whereas CSP appears to be lacking for M. annularis eggs. This study provides evidence of diverging gamete affinities between these recently separated species and suggests for the first time that selection may favour CSP in earlier spawning species when conspecific sperm is diluted and aged and is otherwise at a numeric and viability disadvantage with heterospecific sperm.     

The relationship between conspecific fertilization success and reproductive isolation among three congeneric sea urchins

Few data are available on the effectiveness of reproductive isolating mechanisms in externally fertilizing taxa. I investigated patterns of conspecific and heterospecific fertilization among three coexisting sea urchin species, Strongylocentrotus droebachiensis, S.franciscanus, and S. purpuratus. In the laboratory, both among and within species, eggs from individual females whose eggs are more easily fertilized by conspecific sperm are also most susceptible to heterospecific fertilization. At one extreme, S. droebachiensis requires an order of magnitude fewer conspecific sperm to fertilize eggs than do the other two species and shows very little distinction between conspecific and heterospecific sperm in no choice experiments. Strongylocentrotus franciscanus has an intermediate susceptibility to fertilization by heterospecific sperm. At the other extreme, S. purpuratus rarely cross-fertilizes. Field observations indicate that S. droebachiensis is often surrounded by heterospecific sea urchins. Genetic analysis of larvae produced during heterospecific spawning events indicate that hybrids are generally produced if male conspecifics are more than 1 m from a spawning female S. droebachiensis. Laboratory cultures indicate that these hybrids suffer high mortality relative to conspecific larvae. Comparisons of reproductive success of S. droebachiensis during single-species and multispecies spawning events indicate that the benefits of producing easily fertilized eggs under conditions of sperm limitation may outweigh the costs of losing some offspring to hybrid fertilization. Patterns of variability in heterospecific fertilization are considered in light of three hypotheses: phylogenetic relatedness, reinforcement selection, and sexual selection.     

Plant-like mating in an animal: sexual compatibility and allocation trade-offs in a simultaneous hermaphrodite with remote transfer of sperm

The importance of sexual compatibility between mates has only recently been realized in zoological research into sexual selection, yet its study has been central to botanical research for many decades. The reproductive characteristics of remote mating, an absence of precopulatory mate screening, internal fertilization and embryonic brooding are shared between passively pollinated plants and a phylogenetically diverse group of sessile aquatic invertebrates. Here, we further characterize the sexual compatibility system of one such invertebrate, the colonial ascidian Diplosoma listerianum. All 66 reciprocal pairings of 12 genetic individuals were carried out. Fecundities of crosses varied widely and suggested a continuous scale of sexual compatibility. Of the 11 animals from the same population c. 40% of crosses were completely incompatible with a further c. 20% having obvious partial compatibility (reduced fecundity). We are unaware of other studies documenting such high levels of sexual incompatibility in unrelated individuals. RAPD fingerprinting was used to estimate relatedness among the 12 individuals after a known pedigree was successfully reconstructed to validate the technique. In contrast to previous results, no correlation between genetic similarity and sexual compatibility was detected. The blocking of many genotypes of sperm is expected to severely modify realized paternity away from 'fair raffle' expectations and probably reduce levels of intra-brood genetic diversity in this obligatorily promiscuous mating system. One adaptive benefit may be to reduce the bombardment of the female reproductive system by outcrossed sperm with conflicting evolutionary interests, so as to maintain female control of somatic : gametic investment.     

Mating Compatibility between Geographic Populations of the Seed Beetle <Emphasis Type="Italic">Callosobruchus maculatus</Emphasis>

Allopatric populations sometimes display reproductive barriers in incipient form, and may thus reveal potential mechanisms of speciation. We conducted mating trials to estimate the degree of precopulatory (behavioral) and postcopulatory (gametic) prezygotic isolation between African and Asian populations of the seed beetle Callosobruchus maculatus. The populations differ in several fitness-related traits, and have been associated with different legume hosts. In single-pair trials, the probability of copulation within 10 min was lower when Asian females were paired with African males than it was in the other three pairing combinations. This pairing also provided evidence for gametic isolation, as Asian females mated once to African males laid fewer eggs than did females in the other treatments. Variation in fecundity could not be explained by the duration of copulation, which did not differ among pairing combinations. There was also no effect of pairing combination on the number of copulation attempts, latency to copulation, or rate of egg hatch. When females were simultaneously presented a male from each population, Asian males obtained a disproportionate share of the matings, and were more likely to disturb a pair already in copulo. Surprisingly, male size did not influence the probability of copulation in the single-male trials, the outcome of male–male competition, or the fecundity of once-mated females. Although the African and Asian populations showed some evidence of mating incompatibility, the absence of symmetrical isolating factors suggests minimal barriers to gene flow.