Premating Isolation Research Articles

Heterospecific mating and species recognition in the planktonic marine copepods Temora stylifera and T. longicornis

Many planktonic copepods use diffusible pheromone or hydromechanical signals to remotely detect the presence of potential mates. To determine whether these mating signals also play a role in species recognition and mate choice, we observed and video recorded (3D) mate-finding and pursuit behaviors in heterospecific and conspecific mating crosses in a pair of congeneric, partially sympatric species (Temora stylifera and T. longicornis) in the laboratory. The species appear to have asymmetrical pre-mating isolation, with T. longicornis males readily pursuing T. stylifera females to mate contact and capture, but with little mate-finding activity observed in the reverse cross. Males of T. longicornis pursuing heterospecific females executed a number of behaviors known to facilitate successful pheromone trail following and mate capture in conspecific mating, including accelerated swimming in a ‘signal-scanning’ mode to recover a lost pheromone trail, reversal of the tracking direction in cases when the male initiated tracking in the incorrect direction, and accelerated swimming speeds when in the presence of a pheromone signal but prior to locating the trail. Detailed analyses of mate-tracking behavior in T. longicornis male × T. stylifera female crosses gave no indication that males were aware they were pursuing heterospecific females prior to mate contact, indicating that diffusible pheromone and hydromechanical signals are not used, either singly or in combination, for species recognition in this mating pair. Heterospecific mating attempts among sympatric, congeneric copepods may commonly proceed to mate capture, and incur fitness costs to either or both mating partners.

DIVERGENT SELECTION, NOT LIFE-HISTORY PLASTICITY VIA FOOD LIMITATION, DRIVES MORPHOLOGICAL DIVERGENCE BETWEEN PREDATOR REGIMES INGAMBUSIA HUBBSI

We recently tested the hypothesis of ecological speciation in a post-Pleistocene radiation of Bahamas mosquitofish (Gambusia hubbsi) inhabiting inland blue holes (vertical, water-filled caves) on Andros Island, the Bahamas (Langerhans et al. 2007). Combining morphological, molecular, and behavioral data, our results suggested that enhanced premating isolation evolved as a byproduct of differences in body shape between predator regimes, due to assortative mating for body shape. Our results strongly supported the hypothesis of ecological speciation, and parallel results were observed across species within the genus, suggesting a past history of such ecological speciation within the Gambusia lineage. Downhower et al. (in press) question the morphological results described for G. hubbsi in our paper, contending that prior work on life-history variation in G. hubbsi (primarily Downhower et al. 2000, 2002) invalidates our conclusions. Here we show that this contention is based on a mischaracterization of our study, inaccurate quotation of our paper, and misinterpretation of the relevance of their life-history data for morphological results observed in our study. Downhower et al. (in press) suggest—without providing any direct evidence—that differences in body shape between predator regimes in G. hubbsi reflect correlative effects of phenotypic plasticity in life-history traits in response to variation in food availability. To make such a conclusion, we would have to set aside the following facts: (1) morphological differences were observed in adult males, who do not exhibit the life-history traits in question (e.g., number and size of offspring), (2) results matched well-supported a priori predictions of body shape variation based on divergent selection on locomotor performance, (3) results matched empirical results observed in other fish presumably experiencing similar forms of divergent selection between predator regimes, (4) the observed morphological differences are known to influence swimming performance in Gambusia fish and consequently influence endurance and survival with predators, (5) consistent results were observed for males both within and between Gambusia species, (6) any differences in food availability among blue holes are unknown, and (7) available evidence to date indicates that differences in body shape partly reflect genetic divergence, and not solely phenotypic plasticity. The criticisms in Downhower et al. (in press) can be summarized as three arguments: (1) incorrect attribution of evidence for divergent selection between predator regimes in G. hubbsi to several previous

Mechanisms of reinforcement in natural and simulated polymorphic populations

Reinforcement speciation is the process whereby selection against hybrids drives the evolution of enhanced pre-mating reproductive isolation. Work has focused on divergent mating preferences (assortative mating) but pre-mating isolation can also arise via various migration modification behaviours, such as divergent habitat preferences. The relative importance of these two different mechanisms of reinforcement remains unclear. A recent theoretical model (Yukilevich–True model) found that relative fixation probabilities between these mechanisms can vary. Additionally, natural populations of Timema cristinae walking-sticks exhibit variation (polymorphism) in both mechanisms, generating questions about the patterns expected for allele frequencies prior to fixation, during the early stages of the speciation process. In the present study, we report: (1) new analyses examining the correlation between fixation probabilities for assortative mating and migration modification in the Yukilevich–True model; (2) novel simulations examining allele frequencies in polymorphic populations; and (3) empirical patterns of reinforcement in T. cristinae in the context of theoretical predictions. Simulations of both types yielded congruent results, revealing that the outcome of reinforcement was dependent on the strength of selection. Under weak selection, reinforcement by either mechanism is unlikely. Under intermediate selection, the conditions favoring the rise and fixation of one mechanism favored the rise and fixation of the other. However, assortative mating evolved somewhat more readily than migration modification. Populations of T. cristinae, which experience such intermediate selection, supported these predictions. Under strong selection, the evolution of migration modification generally interfered with the evolution of assortative mating by decreasing migration between populations, thereby reducing selection for assortative mating. Congruence of the results for allele frequencies versus fixation probabilities suggests that similar patterns of reinforcement are expected during different stages of the speciation process. © 2008 The Linnean Society of London, Biological Journal of the Linnean Society, 2008, 95, 305–319.

Courtship plasticity reveals the evolution of dialects in allopatric fish populations

AbstractTraditionally, reproductive isolation between disjoint populations has been thought to emerge as a result of the accumulation of different mutations, genetic drift, or through the effects of natural and sexual selection^1^. Alternatively, the ability of an organism to express different phenotypes depending on the environment (i.e. phenotypic plasticity) could produce reproductive isolation^2,3^. Sexually selected traits are expected to be phenotypically plastic and can result in modifications of the species recognition system and thus originate new species^4^. Here we show that the population-characteristic male courtship behaviour of a fish (Girardinichthys multiradiatus) is modified in the presence of females from other populations, that this is due to the males responding to subtle cues from females, and that they fail to emulate the female's population-characteristic behaviour. We conclude that plasticity has led to the creation of local dialects in the courtship pattern that hampers communication between heterogametic individuals and promotes pre-mating isolation.

Reproductive Interference between the Common Ground-hopper Tetrix undulata and the Slender Ground-hopper Tetrix subulata (Orthoptera, Tetrigidae)

The coexistence of closely related species is often hampered by resource competition or reproductive interference (interspecific sexual interactions). Species utilising similar signal channels might face substantial problems when they co-occur. It has, therefore, been suggested that reinforcement might drive signal evolution in narrow suture zones of secondary contact. However, species with large overlapping ranges are usually not believed to interact sexually. The Slender Ground-hopper, Tetrix subulata, and the Common Ground-hopper, Tetrix undulata (Orthoptera: Tetrigidae) are sister species, which occur sympatrically in large parts of western and Central Europe, but rarely share the same habitat. It has been hypothesized that reproductive interference might account for their missing coexistence. Here, we test experimentally whether these two species interact sexually. Our results suggest an incomplete premating isolation of these ground-hoppers, as we recorded heterospecific courtship, mating attempts and mating. The number of conspecific copulations and mating attempts of T. subulata decreased substantially in the presence of T. undulata, whereas the latter species was not negatively affected. Males of both species preferred to approach females of T. undulata, whereas females of both species did not discriminate against heterospecific males. Further studies on the reproductive success are needed to clarify whether reproductive interference might influence habitat partitioning between these species.

Acoustic divergence in the communication of cryptic species of nocturnal primates (Microcebus ssp.)

BackgroundA central question in evolutionary biology is how cryptic species maintain species cohesiveness in an area of sympatry. The coexistence of sympatrically living cryptic species requires the evolution of species-specific signalling and recognition systems. In nocturnal, dispersed living species, specific vocalisations have been suggested to act as an ideal premating isolation mechanism. We studied the structure and perception of male advertisement calls of three nocturnal, dispersed living mouse lemur species, the grey mouse lemur (Microcebus murinus), the golden brown mouse lemur (M. ravelobensis) and the Goodman's mouse lemur (M. lehilahytsara). The first two species occur sympatrically, the latter lives allopatrically to them.ResultsA multi-parameter sound analysis revealed prominent differences in the frequency contour and in the duration of advertisement calls. To test whether mouse lemurs respond specifically to calls of the different species, we conducted a playback experiment with M. murinus from the field using advertisement calls and alarm whistle calls of all three species. Individuals responded significantly stronger to conspecific than to heterospecific advertisement calls but there were no differences in response behaviour towards statistically similar whistle calls of the three species. Furthermore, sympatric calls evoked weaker interest than allopatric advertisement calls.ConclusionOur results provide the first evidence for a specific relevance of social calls for speciation in cryptic primates. They furthermore support that specific differences in signalling and recognition systems represent an efficient premating isolation mechanism contributing to species cohesiveness in sympatrically living species.

Reproductive isolation between sympatric and allopatric Brazilian populations of Lutzomyia longipalpis s.l. (Diptera: Psychodidae)

Lutzomyia longipalpis s.l., the main vector of Leishmania chagasi in Latin America, is a species complex although the exact number of siblings is yet unknown. In Brazil, the siblings differ in male copulatory courtship songs and pheromones that most certainly act as pre-zygotic reproductive barriers. Here we analysed the reproductive isolation between three allopatric and two sympatric populations of Lu. longipalpis s.l. from Brazil. The results indicate a strong copulatory and pre-mating isolation between the three allopatric populations. In addition, the results also indicate a stronger pre-mating isolation between the two sympatric siblings than between the three allopatric ones, suggesting a role for reinforcement in the speciation of the Lu. longipalpis s.l. complex.

Existence of a pattern of reproductive character displacement inHomobasidiomycotabut not inAscomycota

Generally, stronger reproductive isolation is expected between sympatric than between allopatric sibling species. Such reproductive character displacement should predominantly affect premating reproductive isolation and can be due to several mechanisms, including population extinction, fusion of insufficiently isolated incipient species and reinforcement of reproductive isolation in response to low hybrid fitness. Experimental data on several taxa have confirmed these theoretical expectations on reproductive character displacement, but they are restricted to animals and a few plants. Using results reported in the literature on crossing experiments in fungi, we compared the degree and the nature of reproductive isolation between allopatric and sympatric species pairs. In accordance with theoretical expectations, we found a pattern of enhanced premating isolation among sympatric sibling species in Homobasidiomycota. By contrast, we did not find evidence for reproductive character displacement in Ascomycota at similar genetic distances. Both allopatric and sympatric species of Ascomycota had similarly low levels of reproductive isolation, being mostly post-zygotic. This suggests that some phylogeny-dependent life-history trait may strongly influence the evolution of reproductive isolation between closely related species. A significant correlation was found between degree of reproductive isolation and genetic divergence among allopatric species of Homobasidiomycota, but not among sympatric ones or among Ascomycota species.

Male Ant-mimicking Salticid Spiders Discriminate Between Retreat Silks of Sympatric Females: Implications for Pre-mating Reproductive Isolation

Mate recognition is essential to reproductive success especially under sympatry where encounters between the sexes of different species is likely. We examined the response of male ant-mimicking salticid spiders of the genus Myrmarachne and of five different color forms to retreat and dragline silks of sympatric females of six color forms to determine whether silk-based cues could be used as pre-mating isolation mechanisms and aid species determination. We found evidence for polymorphism within one species Myrmarachne plataleoides, a well-known mimic of the weaver ant Oecophylla smaragdina. Male color morphs of this species showed cross-interest in female silks of other color morphs of this species, but also exhibited greatest preference for the silk of their own color morph, providing evidence for assortative mating and the possibility of silk-based cues as mechanisms of incipient speciation via disruptive selection. Males of two other color forms exhibited unambiguous preference for the silk of their own color forms, suggesting that these two forms are distinct species. The silk-based findings were confirmed by no-choice mating experiments and opportunistic observations of natural matings.

Male flat lizards prefer females with novel scents

In lizards, pheromonal cues are important for mate recognition and are expected to diverge during the speciation process. We tested for divergence in pheromonal mate and species recognition between male flat lizards (Platysaurus broadleyi) belonging to the same population, a different population, and a sister species. Males were given a choice between two refuges, an odourless control and one of the following: a conspecific female from the same population (sympatric), a conspecific female from a distant population (allopatric), and a female from their sister species (heterospecific), P. capensis. Males chose refuges treated with the scent of a heterospecific over an odourless control refuge but showed no preference for refuges that were scented with female conspecific (allopatric or sympatric) cues over the control. These results were consistent in the breeding and non-breeding seasons. In a second experiment males were offered a choice of female scents between sympatric and allopatric conspecifics; an allopatric conspecific and a heterospecific; and a sympatric conspecific and a heterospecific. Males showed a trend of preferring heterospecific over conspecific refuges and allopatric over sympatric conspecific-scented refuges, but these results were not significant at alpha <0.05. Contrary to our original expectations, these experiments do not provide any evidence for a pheromonal pre-mating isolation mechanism within this species complex. However, our results suggest a preference for novel female scents by males, consistent with selection for genetic diversity.

Male flat lizards prefer females with novel scents

In lizards, pheromonal cues are important for mate recognition and are expected to diverge during the speciation process. We tested for divergence in pheromonal mate and species recognition between male flat lizards (Platysaurus broadleyi) belonging to the same population, a different population, and a sister species. Males were given a choice between two refuges, an odourless control and one of the following: a conspecific female from the same population (sympatric), a conspecific female from a distant population (allopatric), and a female from their sister species (heterospecific), P. capensis. Males chose refuges treated with the scent of a heterospecific over an odourless control refuge but showed no preference for refuges that were scented with female conspecific (allopatric or sympatric) cues over the control. These results were consistent in the breeding and non-breeding seasons. In a second experiment males were offered a choice of female scents between sympatric and allopatric conspecifics; an allopatric conspecific and a heterospecific; and a sympatric conspecific and a heterospecific. Males showed a trend of preferring heterospecific over conspecific refuges and allopatric over sympatric conspecific-scented refuges, but these results were not significant at alpha <0.05. Contrary to our original expectations, these experiments do not provide any evidence for a pheromonal pre-mating isolation mechanism within this species complex. However, our results suggest a preference for novel female scents by males, consistent with selection for genetic diversity.

RAPID EVOLUTION OF CUTICULAR HYDROCARBONS IN A SPECIES RADIATION OF ACOUSTICALLY DIVERSE HAWAIIAN CRICKETS (GRYLLIDAE: TRIGONIDIINAE: LAUPALA)

Understanding the origin and maintenance of barriers to gene exchange is a central goal of speciation research. Hawaiian swordtail crickets (genus Laupala) represent one of the most rapidly speciating animal groups yet identified. Extensive acoustic diversity, strong premating isolation, and female preference for conspecific acoustic signals in laboratory phonotaxis trials have strongly supported divergence in mate recognition as the driving force behind the explosive speciation seen in this system. However, recent work has shown that female preference for conspecific male calling song does not extend to mate choice at close range among these crickets, leading to the hypothesis that additional sexual signals are involved in mate recognition and premating isolation. Here we examine patterns of variation in cuticular lipids among several species of Laupala from Maui and the Big Island of Hawaii. Results demonstrate (1) a rapid and dramatic evolution of cuticular lipid composition among species in this genus, (2) significant differences among males and females in cuticular lipid composition, and (3) a significant reduction in the complexity of cuticular lipid profiles in species from the Big Island of Hawaii as compared to two outgroup species from Maui. These results suggest that behavioral barriers to gene exchange in Laupala may be composed of multiple mate recognition signals, a pattern common in other cricket species.

The Role of Cuticular Hydrocarbons in Mating and Conspecific Recognition in the Closely Related Longicorn Beetles Pidonia grallatrix and P. takechii

The role of cuticular chemicals in mating behavior and their chemical components were studied in two sympatric flower-visiting longicorn beetles, Pidonia grallatrix and P. takechii. Mating experiments revealed that female cuticular chemicals elicit male mating behavior and that males can discriminate between conspecific and heterospecific females on the basis of contact chemicals. GC-MS analyses of whole-body extracts in the two species and both sexes determined that extracts contained a series of hydrocarbons including n-alkanes, n-alkenes, and methylalkanes. The relative abundance of some hydrocarbons differed between species and sexes, and canonical discriminant analysis showed discrimination of species and sex could be made unambiguously with several compounds. These results imply that the difference in cuticular hydrocarbons facilitates the premating isolation of sympatric Pidonia species.

Mechanisms of premating isolation between Helicoverpa armigera (Hübner) and Helicoverpa assulta (Guenée) (Lepidoptera: Noctuidae)

Helicoverpa armigera and Helicoverpa assulta are sympatric sibling species, and in the laboratory they can interbreed and produce viable offspring. To assess the contributions of temporal barriers and sexual barriers to premating isolation, we investigated both the temporal rhythms of calling behavior and pheromone titers of H. armigera and H. assulta females and the behavioral responses of males to conspecific and heterospecific calling females in a wind tunnel. Both H. armigera and H. assulta females called throughout the scotophase, and there was more calling during the second half of the scotophase than during the first half. Maximal pheromone titer and maximal calling activity in H. armigera synchronously occurred at the sixth hour into the scotophase, whereas, in H. assulta, the maximal pheromone titer occurred 2 h before the peak of calling. Pheromone blend ratios of the two species were opposite and, within each species, changes in the ratio within the scotophase and at different ages were relatively small. Males of both H. armigera and H. assulta responded strongly to their conspecific calling females in the wind tunnel and completed the whole courtship sequence. In contrast, they did not land and had no copulation attempts in response to heterospecific calling females. These results show that the two species do not have obvious temporal differences in calling behavior and pheromone production, and the specificity of sex pheromone blend emitted by females plays a key role in their premating isolation. In addition, we summarized the potential isolation mechanisms of H. armigera and H. assulta.

Disassociation between weak sexual isolation and genetic divergence in a hermaphroditic land snail and implications about chirality

Examination of the association between reproductive isolation and genetic divergence in a variety of organisms is essential for elucidating the mechanisms causing speciation. However, such studies are lacking for hermaphrodites. We measured premating (sexual) isolation in species pairs of the hermaphroditic land snail Albinaria and we compared it with their genetic divergence. We did not find substantial sexual isolation barriers between the species studied. The absence of strong sexual isolation between species implies its minor effect in the evolution of this genus, because distributional, population and life-history characteristics of Albinaria make mate-choice possibly redundant. Furthermore, we found disassociation between genetic divergence and sexual isolation, suggesting that they do not form necessarily a cause-effect duet. However, Albinaria voithii, the only dextral Albinaria species, shows strong sexual isolation against the other sinistral species. We discuss whether change in coiling either has triggered instantaneous speciation, or is an example of character displacement.

Divergent preferences of Rhagoletis pomonella host races for olfactory and visual fruit cues

AbstractThe apple and hawthorn‐infesting host races of Rhagoletis pomonella Walsh (Diptera: Tephritidae) represent a model for incipient sympatric speciation. Recent studies have shown that apple and hawthorn flies differ in their responses to host‐fruit odor. Each host race preferentially orients to the volatile chemicals emanating from the surface of its natal fruit and is antagonized by non‐natal fruit volatiles. Fruit odor discrimination is evolutionarily important due to its contributory role in host‐specific mating, generating partial premating reproductive isolation between the races in sympatry. Visual cues are also involved in R. pomonella host searching, yet the extent to which they are used to discriminate between hosts has not been fully resolved, nor has their interaction with olfactory cues. Here, we report results from a series of two‐way choice experiments conducted in the field on resident apple and hawthorn flies at a site near Fennville, MI, USA. Four key findings emerged from the study: (1) both olfactory and (2) visual cues increased host discrimination of apple and hawthorn flies for their respective natal fruit, (3) the presence of a single appropriate volatile blend on a natal or non‐natal visual model generally increased the ability of flies to discriminate for their natal fruit, and (4) the presence of both natal and non‐natal olfactory cues on fruit models did not enhance host discrimination above levels observed for visual models alone. We discuss the implications of our results for interpreting the findings of two‐way host choice experiments and for sympatric speciation in phytophagous insects.

Sympatric, Simultaneous, and Random Mating Between Markless Trout (Iwame; Oncorhynchus iwame) and Red-spotted Masu Salmon (Amago; Oncorhynchus masou ishikawae)

The markless trout (iwame, Oncorhynchus iwame) has long been suspected not to be an independent species, but rather a markless mutant form of the red-spotted masu salmon (amago, O. masou ishikawae). Nevertheless, no field study has examined this issue. Here we report a field study on the reproductive ecology of iwame and amago coexisting in the upper stream of the Inabe River. We found that iwame and amago mate sympatrically, simultaneously, and randomly, and observed five cases of heterogeneous oviposition between the two. Our results suggest that no pre-mating reproductive isolation exists between iwame and amago. Because previous studies have shown the absence of post-mating isolation, we propose that iwame and amago are the same species. Iwame should be considered as a markless form of the red-spotted masu salmon, and O. iwame as a junior synonym of O. masou ishikawae.

INFLUENCE OF ANTIBIOTIC TREATMENT AND WOLBACHIA CURING ON SEXUAL ISOLATION AMONG DROSOPHILA MELANOGASTER CAGE POPULATIONS

Speciation depends on the establishment of reproductive isolation between populations of the same species. Whether assortative mating evolves as a by-product of adaptation is a major question relevant to the origin of species by reproductive isolation. The long-term selection populations used here were originally established 30 years ago from a single cage population (originating from a maternal one) and subsequently subjected to divergent selection for tolerance of toxins in food (heavy metals versus ethanol) to investigate this question. Those populations now differ in sexual isolation and Wolbachia infection status. Wolbachia are common and widespread bacteria infecting arthropods and nematodes. Attention has recently focused on their potential role in insect speciation, due to post-mating sperm-egg incompatibilities induced by the bacteria. In this paper we examine the potential effect of Wolbachia on the level of sexual isolation. By antibiotic curing, we show that removal of Wolbachia decreases levels of mate discrimination (sexual isolation index) between populations by about 50%. Backcrossing experiments confirm that this effect is due to infection status rather than to genetic changes in the populations resulting from antibiotic treatment. Antibiotic treatment has no effect on mate discrimination level between uninfected populations. Our findings suggest that the presence of Wolbachia (or another undetected bacterial associate) act as an additive factor contributing to the level of pre-mating isolation between these Drosophila melanogaster populations. Given the ubiquity of bacterial associates of insects, such effects could be relevant to some speciation events.

INFLUENCE OF ANTIBIOTIC TREATMENT AND WOLBACHIA CURING ON SEXUAL ISOLATION AMONG DROSOPHILA MELANOGASTER CAGE POPULATIONS

Speciation depends on the establishment of reproductive isolation between populations of the same species. Whether assortative mating evolves as a by-product of adaptation is a major question relevant to the origin of species by reproductive isolation. The long-term selection populations used here were originally established 30 years ago from a single cage population (originating from a maternal one) and subsequently subjected to divergent selection for tolerance of toxins in food (heavy metals versus ethanol) to investigate this question. Those populations now differ in sexual isolation and Wolbachia infection status. Wolbachia are common and widespread bacteria infecting arthropods and nematodes. Attention has recently focused on their potential role in insect speciation, due to post-mating sperm-egg incompatibilities induced by the bacteria. In this paper we examine the potential effect of Wolbachia on the level of sexual isolation. By antibiotic curing, we show that removal of Wolbachia decreases levels of mate discrimination (sexual isolation index) between populations by about 50%. Backcrossing experiments confirm that this effect is due to infection status rather than to genetic changes in the populations resulting from antibiotic treatment. Antibiotic treatment has no effect on mate discrimination level between uninfected populations. Our findings suggest that the presence of Wolbachia (or another undetected bacterial associate) act as an additive factor contributing to the level of pre-mating isolation between these Drosophila melanogaster populations. Given the ubiquity of bacterial associates of insects, such effects could be relevant to some speciation events.

Is premating isolation in Drosophila overestimated due to uncontrolled factors?

Sexual isolation in Drosophila is typically measured by multiple-choice mating tests. While many environmental variables during such tests are controlled by the researcher, there are some factors that are usually uncontrolled. We demonstrate, using Drosophila melanogaster and D. pseudoobscura flies, that the temperature of rearing, pre-adult density, and level of consanguinity, can all produce differences in mating propensity between genetically equivalent flies. These differences in mating propensity, in turn, can give rise to statistically significant results in multiple-choice mating tests, leading to positive isolation values and the artifactual inference of sexual isolation between populations. This fact agrees with a nonrandom excess of significant positive tests found in a review of the literature of Drosophila intraspecific mating choice. An overestimate of true cases of sexual isolation in Drosophila in the literature can, therefore, not be ruled out.