Stickleback Species Pair Research Articles

Courtship behavior, nesting microhabitat, and assortative mating in sympatric stickleback species pairs.

The maintenance of reproductive isolation in the face of gene flow is a particularly contentious topic, but differences in reproductive behavior may provide the key to explaining this phenomenon. However, we do not yet fully understand how behavior contributes to maintaining species boundaries. How important are behavioral differences during reproduction? To what extent does assortative mating maintain reproductive isolation in recently diverged populations and how important are “magic traits”? Assortative mating can arise as a by‐product of accumulated differences between divergent populations as well as an adaptive response to contact between those populations, but this is often overlooked. Here we address these questions using recently described species pairs of three‐spined stickleback (Gasterosteus aculeatus), from two separate locations and a phenotypically intermediate allopatric population on the island of North Uist, Scottish Western Isles. We identified stark differences in the preferred nesting substrate and courtship behavior of species pair males. We showed that all males selectively court females of their own ecotype and all females prefer males of the same ecotype, regardless of whether they are from species pairs or allopatric populations. We also showed that mate choice does not appear to be driven by body size differences (a potential “magic trait”). By explicitly comparing the strength of these mating preferences between species pairs and single‐ecotype locations, we were able to show that present levels of assortative mating due to direct mate choice are likely a by‐product of other adaptations between ecotypes, and not subject to obvious selection in species pairs. Our results suggest that ecological divergence in mating characteristics, particularly nesting microhabitat may be more important than direct mate choice in maintaining reproductive isolation in stickleback species pairs.

The ecological stage changes benefits of mate choice and drives preference divergence.

Preference divergence is thought to contribute to reproductive isolation. Ecology can alter the way selection acts on female preferences, making them most likely to diverge when ecological conditions vary among populations. We present a novel mechanism for ecologically dependent sexual selection, termed 'the ecological stage' to highlight its ecological dependence. Our hypothesized mechanism emphasizes that males and females interact over mating in a specific ecological context, and different ecological conditions change the costs and benefits of mating interactions, selecting for different preferences in distinct environments and different male traits, especially when traits are condition dependent. We test key predictions of this mechanism in a sympatric three-spine stickleback species pair. We used a maternal half-sib split-clutch design for both species, mating females to attractive and unattractive males and raising progeny on alternate diets that mimic the specialized diets of the species in nature. We estimated the benefits of mate choice for an indicator trait (male nuptial colour) by measuring many fitness components across the lifetimes of both sons and daughters from these crosses. We analysed fitness data using a combination of aster and mixed models. We found that many benefits of mating with high-colour males depended on both species and diet. These results support the ecological stage hypothesis for sticklebacks. Finally, we discuss the potential role of this mechanism for other taxa and highlight its ability to enhance reproductive isolation as speciation proceeds, thus facilitating the evolution of strong reproductive isolation. This article is part of the theme issue 'Towards the completion of speciation: the evolution of reproductive isolation beyond the first barriers'.

Genetics of adaptation: Experimental test of a biotic mechanism driving divergence in traits and genes.

The genes underlying adaptations are becoming known, yet the causes of selection on genes—a key step in the study of the genetics of adaptation—remains uncertain. We address this issue experimentally in a threespine stickleback species pair showing exaggerated divergence in bony defensive armor in association with competition‐driven character displacement. We used semi‐natural ponds to test the role of a native predator in causing divergent evolution of armor and two known underlying genes. Predator presence/absence altered selection on dorsal spines and allele frequencies at the Msx2a gene across a generation. Evolutionary trajectories of alleles at a second gene, Pitx1, and the pelvic spine trait it controls, were more variable. Our experiment demonstrates how manipulation of putative selective agents helps to identify causes of evolutionary divergence at key genes, rule out phenotypic plasticity as a sole determinant of phenotypic differences, and eliminate reliance on fitness surrogates. Divergence of predation regimes in sympatric stickleback is associated with coevolution in response to resource competition, implying a cascade of biotic interactions driving species divergence. We suggest that as divergence proceeds, an increasing number of biotic interactions generate divergent selection, causing more evolution in turn. In this way, biotic adaptation perpetuates species divergence through time during adaptive radiation in an expanding number of traits and genes.

The genomic landscape at a late stage of stickleback speciation: High genomic divergence interspersed by small localized regions of introgression.

Speciation is a continuous process and analysis of species pairs at different stages of divergence provides insight into how it unfolds. Previous genomic studies on young species pairs have revealed peaks of divergence and heterogeneous genomic differentiation. Yet less known is how localised peaks of differentiation progress to genome-wide divergence during the later stages of speciation in the presence of persistent gene flow. Spanning the speciation continuum, stickleback species pairs are ideal for investigating how genomic divergence builds up during speciation. However, attention has largely focused on young postglacial species pairs, with little knowledge of the genomic signatures of divergence and introgression in older stickleback systems. The Japanese stickleback species pair, composed of the Pacific Ocean three-spined stickleback (Gasterosteus aculeatus) and the Japan Sea stickleback (G. nipponicus), which co-occur in the Japanese islands, is at a late stage of speciation. Divergence likely started well before the end of the last glacial period and crosses between Japan Sea females and Pacific Ocean males result in hybrid male sterility. Here we use coalescent analyses and Approximate Bayesian Computation to show that the two species split approximately 0.68–1 million years ago but that they have continued to exchange genes at a low rate throughout divergence. Population genomic data revealed that, despite gene flow, a high level of genomic differentiation is maintained across the majority of the genome. However, we identified multiple, small regions of introgression, occurring mainly in areas of low recombination rate. Our results demonstrate that a high level of genome-wide divergence can establish in the face of persistent introgression and that gene flow can be localized to small genomic regions at the later stages of speciation with gene flow.

Genomic landscape of early ecological speciation initiated by selection on nuptial colour.

Ecological speciation is the evolution of reproductive isolation as a consequence of direct divergent natural selection or ecologically mediated divergent sexual selection. While the genomic signature of the former has been extensively studied in recent years, only few examples exist for genomic differentiation where environment-dependent sexual selection has played an important role. Here, we describe a very young (~90years old) population of threespine sticklebacks exhibiting phenotypic and genomic differentiation between two habitats within the same pond. We show that differentiation among habitats is limited to male throat colour and nest type, traits known to be subject to sexual selection. Divergence in these traits mirrors divergence in much older benthic and limnetic stickleback species pairs from North American west coast lakes, which also occur in sympatry but are strongly reproductively isolated from each other. We demonstrate that in our population, differences in throat colour and breeding have been stable over a decade, but in contrast to North American benthic and limnetic stickleback species, these mating trait differences are not accompanied by divergence in morphology related to feeding, predator defence or swimming performance. Using genomewide SNP data, we find multiple genomic islands with moderate differentiation spread across several chromosomes, whereas the rest of the genome is undifferentiated. The islands contain potential candidate genes involved in visual perception of colour. Our results suggest that phenotypic and multichromosome genomic divergence of these morphs was driven by environment-dependent sexual selection, demonstrating incipient speciation after only a few decades of divergence in sympatry.

Predator experience overrides learned aversion to heterospecifics in stickleback species pairs.

Predation risk can alter female mating decisions because the costs of mate searching and selecting attractive mates increase when predators are present. In response to predators, females have been found to plastically adjust mate preference within species, but little is known about how predators alter sexual isolation and hybridization among species. We tested the effects of predator exposure on sexual isolation between benthic and limnetic threespine sticklebacks (Gasterosteus spp.). Female discrimination against heterospecific mates was measured before and after females experienced a simulated attack by a trout predator or a control exposure to a harmless object. In the absence of predators, females showed increased aversion to heterospecifics over time. We found that predator exposure made females less discriminating and precluded this learned aversion to heterospecifics. Benthic and limnetic males differ in coloration, and predator exposure also affected sexual isolation by weakening female preferences for colourful males. Predator effects on sexual selection were also tested but predators had few effects on female choosiness among conspecific mates. Our results suggest that predation risk may disrupt the cognitive processes associated with mate choice and lead to fluctuations in the strength of sexual isolation between species.

Loss of sexual isolation in a hybridizing stickleback species pair

Abstract One approach to understand the importance of reproductive barriers to the speciation process is to study the breakdown of barriers between formerly distinct species. One reproductive barrier, sexual isolation, reduces gene flow between species through differences in mate preferences and mating signals and is likely important for species formation and maintenance. We measure sexual isolation in two limnetic-benthic threespine stickleback species pairs (Gasterosteus spp.). One species pair maintains strong reproductive isolation while the other species pair has recently collapsed into a hybrid swarm. We compare the strength of sexual isolation in the hybridizing pair to the currently isolated pair. We provide the first evidence that sexual isolation has been lost in the hybridizing pair and show furthermore that preferences females have for conspecific mates and the traits they use to distinguish conspecific and heterospecific males contribute to this loss. This work highlights the fragility of reproductive isolation between young species pairs and considers the role of sexual isolation in speciation.

Weak habitat isolation in a threespine stickleback (Gasterosteusspp.) species pair

Reproductive isolation is central to the study of speciation. Multiple isolating barriers may prevent species from hybridizing, although their individual strength and the interactions between them are rarely measured. We quantified habitat isolation in a recently diverged threespine stickleback species pair (Gasterosteus aculeatus complex) and controlled for any such interactions. Using enclosures in an outdoor pond, we confirm that males of the two species strongly prefer different nesting habitats: limnetic males build nests in open habitats, whereas benthic males nest under vegetation. However, forcing males to nest in their nonpreferred habitat did not reduce the probability of spawning by females. As a result, habitat isolation between the species is estimated to be weak. We compared the strength of habitat isolation estimated in the present study with estimates of other behavioural barriers using previously published data. We discovered that, although total mating isolation between the species is strong, the contributions of differences in body size and male nuptial colour are similarly individually weak. Instead, interactions with other, undetermined species-specific traits were responsible for most of the isolation resulting from differences in body size and, in benthics, colour. This is one of the first attempts to estimate individual isolating barriers at the same time as controlling for interactions. © 2013 The Linnean Society of London, Biological Journal of the Linnean Society, 2013, 110, 466‐476.

Functional basis of ecological divergence in sympatric stickleback

BackgroundThe evolution of ecological divergence in closely related species is a key component of adaptive radiation. However, in most examples of adaptive radiation the mechanistic basis of ecological divergence remains unclear. A classic example is seen in the young benthic and limnetic stickleback species pairs of British Columbia. In each pair the benthic species feeds on littoral macroinvertebrates whereas the limnetic feeds on pelagic zooplankton. Previous studies indicate that in both short-term feeding trials and long-term enclosure studies, benthics and limnetics exhibit enhanced performance on their own resource but fare more poorly on the other species’ resource. We examined the functional basis of ecological divergence in the stickleback species pair from Paxton Lake, BC, using biomechanical models of fish feeding applied to morphological traits. We examined the consequences of morphological differences using high speed video of feeding fish.ResultsBenthic stickleback possess morphological traits that predict high suction generation capacity, including greatly hypertrophied epaxial musculature. In contrast, limnetic stickleback possess traits thought to enhance capture of evasive planktonic prey, including greater jaw protrusion than benthics and greater displacement advantage in both the lower jaw-opening lever system and the opercular four-bar linkage. Kinematic data support the expectations from the morphological analysis that limnetic stickleback exhibit faster strikes and greater jaw protrusion than benthic fish, whereas benthics exert greater suction force on attached prey.ConclusionsWe reveal a previously unknown suite of complex morphological traits that affect rapid ecological divergence in sympatric stickleback. These results indicate that postglacial divergence in stickleback involves many functional systems and shows the value of investigating the functional consequences of phenotypic divergence in adaptive radiation.

Mapping the genomic architecture of ecological speciation in the wild: does linkage disequilibrium hold the key?

The hunt for the genes underlying ecological speciation has now closed in on a number of candidates, but making the link from genotype to phenotype continues to pose a significant challenge. This is partly because genetic studies in many systems remain impeded by long generation times or an inability to perform controlled crosses. Now, in this issue of Molecular Ecology, Malek etal. (2012) demonstrate the utility of a novel admixture mapping approach that can be used to identify genomic regions contributing to adaptive trait divergence between natural populations. Remarkably, they validate their approach by mapping traits associated with mate choice in a wild limnetic and benthic threespine stickleback (Gasterosteus aculeatus) species pair, finding several loci associated with male nuptial coloration and shape. While this study benefited from tried-and-true microsatellites in a well-characterized species with a detailed genetic map (and genome sequence), the field is quickly moving towards the use of next-generation sequencing, especially for nonmodel systems. The ability to characterize molecular polymorphisms for any system suggests that molecular ecologists working on virtually any species may benefit from applying Malek etal.'s approach, if naturally admixed populations are available.

Environmental determinants of threespine stickleback species pair evolution and persistence

Threespine stickleback ( Gasterosteus aculeatus ) species pairs are found in four watersheds in southwestern British Columbia, Canada, and are listed as Endangered under the federal Species at Risk Act. Their origin is thought to be through a double-invasion process followed by character displacement; however, this hypothesis does not consider whether speciation is dependent on unique environmental factors necessary to support two species with separate habitat and resource requirements, which may be essential both for their evolution and persistence. To test whether species pair lakes have unique attributes, we compared abiotic and biotic factors of species pair lakes to lakes with only a single population of stickleback. There were no clear environmental differences between species pair and non-species pair lakes, but species pairs were only present in lakes with low fish species diversity, suggesting that evolution and persistence of species pairs requires a simplified fish community. Our study suggests that colonization history rather than unique lake attributes (related to either physical habitat or trophic resources) facilitated the evolution of stickleback species pairs and that the fish assemblage in lakes may affect resource availability and speciation potential as strongly as the limnological attributes of the lakes themselves.

A benthic predatory fish does not cause selection on armour traits in three-spined stickleback Gasterosteus aculeatus (Gasterosteiformes: Gasterosteidae)

Predation can promote divergence between prey populations and contribute to ecological speciation. In theory, predators can also constrain prey population divergence. In coastal British Columbia, Canada, Gasterosteus aculeatus (three-spined stickleback) species pairs only occur in lakes with a single species of predatory fish: Oncorhynchus clarkii (the cutthroat trout). Similar lakes containing additional predatory fish species (Cottus asper, prickly sculpins; Oncorhynchus mykiss, rainbow trout) contain only single species of morphologically intermediate stickleback, suggesting that these predators prevent the coexistence of stickleback species pairs. We conducted a mesocosm experiment to investigate how prickly sculpins might constrain divergence, by quantifying their impact on survival and natural selection on antipredator (armour) traits in F2 stickleback from a cross between ecologically divergent populations. We tested three hypotheses: (1) sculpin predation on sticklebacks reduces survival in a way that could result in their exclusion from certain niches; (2) sculpins compete with stickleback; (3) sculpins respond to prey vulnerabilities in similar ways to cutthroat trout, tending to constrain rather than to enhance divergence. We found that sculpins significantly reduce stickleback survival, that their presence per se does not reduce growth in stickleback, and that predation did not result in selection on any of the armour traits measured, or on gill raker length, which is an important trophic trait. These results tend to refute hypotheses (2) and (3), while supporting hypothesis (1). © 2011 The Linnean Society of London, Biological Journal of the Linnean Society, 2011, 104, 877–885.

Modelling sympatric speciation by means of biologically plausible mechanistic processes as exemplified by threespine stickleback species pairs

We investigate the plausibility of sympatric speciation through a modelling study. We built up a series of models with increasing complexity while focussing on questioning the realism of model assumptions by checking them critically against a particular biological system, namely the sympatric benthic and limnetic species of threespine stickleback in British Columbia, Canada. These are morphologically adapted to their feeding habits: each performs better in its respective habitat than do hybrids with intermediate morphology. Ecological character displacement through disruptive selection and competition, and reinforcement through mating preferences may have caused their divergence. Our model assumptions include continuous morphological trait(s) instead of a dimorphic trait, and mating preferences based on the same trait(s) as selected for in food competition. Initially, morphology is intermediate. We apply disruptive selection against intermediates, frequency-dependent resource competition, and one of two alternative mating preference mechanisms. Firstly, preference is based on similarity where mating preference may result from “imprinting” on conspecifics encountered in their preferred foraging habitat. Here, speciation occurs easily—ecological hybrid inferiority is not necessary. Hybrid inferiority reinforces the stringency of assortative mating. Secondly, individual preferences exist for different trait values. Here, speciation occurs when linkage disequilibrium between trait and preference develops, and some hybrid inferiority is required. Finally, if the morphology subject to disruptive selection, frequency-dependent competition, and mate choice, is coded for by two loci, linkage disequilibrium between the two loci is required for speciation. Speciation and reinforcement of stringency of choosiness are possible in this case too, but rarely. Results demonstrate the contingency of speciation, with the same starting point not necessarily producing the same outcome. The study resulted in flagging issues where models often lack in biological realism and issues where more empirical studies could inform on whether assumptions are likely valid.

Turnover of sex chromosomes and speciation in fishes.

Closely related species of fishes often have different sex chromosome systems. Such rapid turnover of sex chromosomes can occur by several mechanisms, including fusions between an existing sex chromosome and an autosome. These fusions can result in a multiple sex chromosome system, where a species has both an ancestral and a neo-sex chromosome. Although this type of multiple sex chromosome system has been found in many fishes, little is known about the mechanisms that select for the formation of neo-sex chromosomes, or the role of neo-sex chromosomes in phenotypic evolution and speciation. The identification of closely related, sympatric species pairs in which one species has a multiple sex chromosome system and the other has a simple sex chromosome system provides an opportunity to study sex chromosome turnover. Recently, we found that a population of threespine stickleback (Gasterosteus aculeatus) from Japan has an X1X2Y multiple sex chromosome system resulting from a fusion between the ancestral Y chromosome and an autosome, while a sympatric threespine stickleback population has a simple XY sex chromosome system. Furthermore, we demonstrated that the neo-X chromosome (X 2) plays an important role in phenotypic divergence and reproductive isolation between these sympatric stickleback species pairs. Here, we review multiple sex chromosome systems in fishes, as well as recent advances in our understanding of the evolutionary role of sex chromosome turnover in stickleback speciation.

Colour plasticity and background matching in a threespine stickleback species pair

Examining differences in colour plasticity between closely-related species in relation to the heterogeneity of background colours found in their respective habitats may offer important insight into how cryptic colour change evolves in natural populations. In the present study, we examined whether nonbreeding dorsal body coloration has diverged between sympatric species of stickleback along with changes in habitat-specific background colours. The small, limnetic species primarily occupies the pelagic zone and the large, benthic species inhabits the littoral zone. We placed benthic and limnetic sticklebacks against extremes of habitat background colours and measured their degree of background matching and colour plasticity. Benthics matched the littoral background colour more closely than did the limnetics, although there was no difference between species in their resemblance to the pelagic background colour. Benthics were able to resemble both background colours by exhibiting greater directional colour plasticity in their dorsal body coloration than limnetics, which may be an adaptive response to the greater spectral heterogeneity of the littoral zone. The present study highlights how habitat-specific spectral characteristics may shape cryptic coloration differences between stickleback species.

Parasites may contribute to ‘magic trait’ evolution in the adaptive radiation of three-spined sticklebacks, Gasterosteus aculeatus (Gasterosteiformes: Gasterosteidae)

In the past decade, there has been a new effort to understand the ecology that drives population divergence and speciation. It is well established in theory that speciation is most likely to occur when a trait that is under divergent natural selection in different populations is also used in mate choice. Such traits have been dubbed ‘magic traits’ (Gavrilets, 2004) and, although there appears to be good evidence that they exist, the ecological mechanisms that underlie their divergence are not well understood. Size at maturity in three-spined sticklebacks is an archetypal example of a magic trait. The present study documents for the first time that differences in body size at maturity in sympatric species pairs of lacustrine three-spined sticklebacks in British Columbia, Canada, are caused by differences in age at maturity. It is also shown that there are differences between the sympatric species in the patterns of infection with a virulent cestode, Schistocephalus solidus. Although the evidence is circumstantial, these differences in infection are consistent with the hypothesis that they have contributed to the observed divergence in age and size at maturity in these populations. © 2009 The Linnean Society of London, Biological Journal of the Linnean Society, 2009, 96, 425–433.

Phenotypic divergence and reproductive isolation between sympatric forms of Japanese threespine sticklebacks

The threespine stickleback (Gasterosteus aculeatus) species complex is well suited for identifying the types of phenotypic divergence and isolating barriers that contribute to reproductive isolation at early stages of speciation. In the present study, we characterize the patterns of genetic and phenotypic divergence as well as the types of isolating barriers that are present between two sympatric pairs of threespine sticklebacks in Hokkaido, Japan. One sympatric pair consists of an anadromous and a resident freshwater form and shows divergence in body size between the forms, despite the lack of genetic differentiation between them. The second sympatric pair consists of two anadromous forms, which originated before the last glacial period and are currently reproductively isolated. These two anadromous forms have diverged in many morphological traits as well as in their reproductive behaviours. Both sexual isolation and hybrid male sterility contribute to reproductive isolation between the anadromous species pair. We discuss the shared and unique aspects of phenotypic divergence and reproductive isolation in the Japanese sympatric pairs compared with postglacial stickleback species pairs. Further studies of these divergent species pairs will provide a deeper understanding of the mechanisms of speciation in sticklebacks. © 2007 The Linnean Society of London, Biological Journal of the Linnean Society, 2007, 91, 671–685.

Condition‐dependent expression of red colour differs between stickleback species

Sexual isolation may arise when male mating traits and female preferences differ between species. Such divergence in mating traits is likely to occur when the strength or targets of sexual selection differ. Therefore, by comparing the traits under sexual selection in closely related species and the nature of preference for those traits, we can gain insight into when sexual selection contributes to sexual isolation and how it does so. Collecting these data is no easy undertaking. To simplify this comparison, I use the presence and extent of condition dependence in traits to determine whether directional sexual selection is acting on them. Condition dependence thus serves as a signature of sexual selection. I investigate differences in sexual selection on red nuptial colour in limnetic-benthic species pairs of three-spined sticklebacks. I evaluate condition dependence by comparing the strength of the relationship between colour and condition, and the magnitude of variance in red nuptial colour to other colour traits and to nonsexual traits. I find that limnetic males have strong condition-dependent expression of red nuptial colour whereas benthic males have at most weak condition-dependent expression. Ancestral anadromous males show no condition dependence. This suggests that colour is under strong directional sexual selection only in limnetics and that this is the derived state. Moreover, I find that the strength of female preference for red is related to the extent of condition dependence. The extent of condition dependence is also associated with the importance of colour differences to mate recognition. These results show that differences between these species in the action of sexual selection underlie their sexual isolation.

Editorial and Retrospective 2007

Editorial and Retrospective 2007

Speciation in reverse: morphological and genetic evidence of the collapse of a three‐spined stickleback (Gasterosteus aculeatus) species pair

Historically, six small lakes in southwestern British Columbia each contained a sympatric species pair of three-spined sticklebacks (Gasterosteus aculeatus). These pairs consisted of a 'benthic' and 'limnetic' species that had arisen postglacially and, in four of the lakes, independently. Sympatric sticklebacks are considered biological species because they are morphologically, ecologically and genetically distinct and because they are strongly reproductively isolated from one another. The restricted range of the species pairs places them at risk of extinction, and one of the pairs has gone extinct after the introduction of an exotic catfish. In another lake, Enos Lake, southeastern Vancouver Island, an earlier report suggested that its species pair is at risk from elevated levels of hybridization. We conducted a detailed morphological analysis, as well as genetic analysis of variation at five microsatellite loci for samples spanning a time frame of 1977 to 2002 to test the hypothesis that the pair in Enos Lake is collapsing into a hybrid swarm. Our morphological analysis showed a clear breakdown between benthics and limnetics. Bayesian model-based clustering indicated that two morphological clusters were evident in 1977 and 1988, which were replaced by 1997 by a single highly variable cluster. The most recent 2000 and 2002 samples confirm the breakdown. Microsatellite analysis corroborated the morphological results. Bayesian analyses of population structure in a sample collected in 1994 indicated two genetically distinct populations in Enos Lake, but only a single genetic population was evident in 1997, 2000, and 2002. In addition, genetic analyses of samples collected in 1997, 2000, and 2002 showed strong signals of 'hybrids'; they were genetically intermediate to parental genotypes. Our results support the idea that the Enos Lake species pair is collapsing into a hybrid swarm. Although the precise mechanism(s) responsible for elevated hybridization in the lake is unknown, the demise of the Enos Lake species pair follows the appearance of an exotic crayfish, Pascifasticus lenisculus, in the early 1990s.