Light Morph Research Articles

Pleiotropic effects of melanin pigmentation: haemoparasite infection intensity but not telomere length is associated with plumage morph in black sparrowhawks.

There is increasing recognition of the potential pleiotropic effects of melanin pigmentation, particularly on immunity, with reports of variation in haemoparasite infection intensity and immune responses between the morphs of colour-polymorphic bird species. In a population of the black sparrowhawk (Accipiter melanoleucus) in western South Africa, light morphs have a higher haemoparasite infection intensity, but no physiological effects of this are apparent. Here, we investigate the possible effects of haemoparasite infection on telomere length in this species and explore whether relative telomere length is associated with either plumage morph or sex. Using quantitative polymerase chain reaction analysis, we confirmed that dark morphs had a lower haemoparasite infection intensity than light morphs. However, we found no differences in telomere length associated with either the haemoparasite infection status or morph in adults, although males have longer telomeres than females. While differences in haemoparasite intensity between morphs are consistent with pleiotropic effects of melanin pigmentation in the black sparrowhawk, we found no evidence that telomere length was associated with haemoparasite infection. Further work is needed to investigate the implications of possible pleiotropic effects of plumage morph and their potential role in the maintenance of colour polymorphism in this species.

Встречи крупных хищных птиц в Новосибирской области, Россия

Встречи крупных хищных птиц в Новосибирской области, Россия

Observing Evolution: Peppered Moths and the Discovery of Parallel Melanism

Observing Evolution: Peppered Moths and the Discovery of Parallel Melanism

Adaptive changes in energy reserves and effects of body melanization on thermal tolerance in Drosophila simulans

Seasonally polyphenic types have been documented in many Drosophilids, which differ significantly during thermal stress. Although Drosophila simulans is a sibling species to Drosophila melanogaster, both thrive in the temperate and tropical climates, but various climatic factors are expected to impact their distribution and abundance. As a result, D. simulans may use phenotypic plasticity to adapt to colder and drier circumstances in temperate zones, although such studies are less known. In the present study, our main aim was to find a link between adaptive plasticity and thermal tolerance in D. simulans. We characterized two morphs in D. simulans flies based on the abdominal melanization collected from the same locality and season, as this trait is highly associated with the larval developmental conditions. Our results suggested that flies reared from dark and light morph showed significant differences in the basal level of proline, carbohydrates (trehalose, glycogen), and lipids (cuticular lipids and total body lipids) within simulated seasons and morph lineages in D. simulans flies. We further showed that D. simulans reared from dark morph are better adapted to cold conditions, whereas light flies are more adapted to warm conditions. The flies, both from light and dark morph lineages, when reared at 15 °C, showed an increase in the level of total body lipids after acclimation at 0 °C but a decrease in the level of proline and carbohydrates (trehalose, glycogen). Heat acclimation increases glycogen levels in the flies from light morph lineage while decreases trehalose and proline.

Thermal Costs and Benefits of Replicated Color Evolution in the White Sands Desert Lizard Community.

AbstractTraits often contribute to multiple functions, complicating our understanding of the selective pressures that influence trait evolution. In the Chihuahuan Desert, predation is thought to be the primary driver of cryptic light coloration in three White Sands lizard species relative to the darker coloration of populations on adjacent dark soils. However, coloration also influences radiation absorption and thus animal body temperatures. We combined comparative physiological experiments and biophysical models to test for thermal consequences of evolving different color morphs in White Sands across the three species. While light and dark morphs have not evolved different physiological heat limits within species, differences in radiation absorption between morphs lead to body temperature differences that impact relative overheating risk and activity patterns. Moreover, for all three species, an idealized morph that matches the White Sands substrate would have considerably less activity time, by approximately 1 month, than existing light morphs. Overall, there are both benefits and costs to greater substrate matching, the balance of which may prevent the evolution of optimal crypsis. Our work highlights the importance of color in dictating thermal performance and the complexity inherent in understanding the evolution of coloration.

Mitochondrial genetic structure reflects the geographical variation of elytral polymorphism frequency in Cheilomenes sexmaculata (Coleoptera: Coccinellidae)

• Many ladybird have colour polymorphism with geographical variation. • C. sexmaculata has colour polymorphism including dark and light morph types. • The mtCOI gene haplotypes of this species were integrated into two haplotype groups. • The higher frequencies haplotypes in lower latitudes tended to be light morph types. • The haplotypes in higher latitudes more frequently exhibited dark morph types. Many ladybird species are known to have an elytral colour polymorphism, which indicates geographical variation. The ladybird beetle Cheilomenes sexmaculata (Fabricius) exhibits elytral colour polymorphism and has expanded its distribution from 33°N to 36°N in Japan over 100 years since 1900. The mitochondrial COI gene haplotypes were integrated into two haplotype groups, with one group existing at higher frequencies in lower latitudes, the other group appearing at higher frequencies in higher latitudes. In addition, the dark morph types of this species increase with latitude, whereas the light types appear at higher relative frequencies in lower latitudes. In the present study, we first determined the morph types of individuals and examined the mitochondrial DNA COI gene. Second, we investigated the relationship between the genetic population structure based on the mitochondrial DNA COI gene and the morph types’ geographical variation. Results indicated that the mtCOI genetic structure was associated with the morph types by latitude; specifically, the haplotype group existing at higher frequencies in lower latitudes tended to be light morph types. In contrast, the haplotype dominant in higher latitudes more frequently exhibited dark morph types, indicating that dark morph types in the higher latitude genetic group may have led the distributional expansion toward higher latitudes since 1900 rather than the lower latitude light morph types.

Does melanin-based plumage coloration reflect health status of free-living birds in urban environments?

BackgroundEcological functions and processes in urban ecosystems are governed by various human activities. City-adapted and city-exploiting animal species are expected to present certain specific behavioral and physiological traits in comparison to city-avoiders or conspecific individual frequenting less urbanized or rural environments. A trait of high importance, the plumage color polymorphism has been selected as the main study model and was correlated with different morphological and physiological parameters to highlight its importance in determining the possible health status of urban Feral Pigeons (Columba livia) in North African urban habitats.MethodsDifferent body morphometrics, hematological and hemoparasitic parameters were quantified on free-living Feral Pigeons in urban environments of northern Algeria. Moreover, plumage melanin-based coloration (MBC) was measured and the data collected at the individual scale was correlated with the previous parameters using linear and non-linear modeling approaches.ResultsPlumage MBC scores of the sampled Feral Pigeons ranged between 0.3% and 74.8%. Among the 12 morphological traits measured, body weight, tail length and total length were deemed to be positively correlated with MBC. Darker morphs appeared to have more hemoparasites compared to lighter pigeons. Quite the same observation goes with the immunity but with non-linear trends. The number of monocytes and granulocytes increased with the increase in MBC levels in lighter morphs, while pigeons with high MBC scores exhibited negative relationships between MBC levels and the number of white blood cells.ConclusionsDespite the existence of a number of studies demonstrating phenotypic directional selection, further studies are undoubtedly necessary to understand in detail the underlying mechanisms in species life-history strategies between differently colored individuals. Findings of this correlative study open exciting perspectives revealing that MBC can be considered a good indicator of and health status and adaptation strategies to changes in urban environments.

Comparative Study of the Phenotypic Structure and Photoperiodic Responses of Female Asian Ladybirds Harmonia axyridis (Pallas) (Coleoptera, Coccinellidae) from Moscow, Belgorod, and Sochi

Insect invasions are often accompanied by changes in the phenotypic structure of the populations and in the photoperiodic responses that regulate seasonal cycles. The Asian ladybird Harmonia axyridis has been recently discovered in Krasnodar Territory, from whence it is now gradually colonizing Central Russia. Our comparative study of individuals collected in 2020 in Sochi, Belgorod Province, and Moscow Province shows that interpopulation differences in their phenotypic structure are nonsignificant. The proportion of the light morph succinea is 77–83%; the proportion of the most common dark morph, spectabilis, is 13–20%. Laboratory experiments show that females from all the studied populations exhibit only a weak photoperiodic response. Comparative analysis reveals that females from the more northerly Moscow and Belgorod populations have a somewhat greater tendency to diapause and slightly delay reproductive maturation.

Developmental and adult acclimation impact cold and drought survival of invasive tropical Drosophila kikkawai.

ABSTRACTNarrow distribution patterns of tropical Drosophila species are limited by lower resistance to cold or drought. In the invasive tropical Drosophila kikkawai, we tested whether developmental and adult acclimations at cooler temperatures could enhance its stress resistance level. Adult acclimation of winter collected body color morphs revealed a significant increase in the level of cold resistance. For light morph, its abundance during winter is not consistent with thermal-melanism hypothesis. However, higher cold acclimation capacity, as well as storage of energy metabolites could support its winter survival. In the wild-caught light and intermediate morphs, there is a lack of trade-off between cold and heat resistance but not in the case of dark morph. Developmental plasticity (15°C) resulted in the fivefold increase of cold survival at 0°C; and a twofold increase in desiccation resistance but a modest reduction (∼28–35%) in heat resistance as compared to morph strains reared at 25°C. Drought acclimation changes were significantly higher as compared with cold or heat pretreatment. We observed a trade-off between basal resistance and acclimation capacity for cold, heat, or drought resistance. For homeostatic energy balance, adult acclimation responses (cold versus drought; heat versus drought) caused compensatory plastic changes in the levels of proline or trehalose (shared patterns) but different patterns for total body lipids. In contrast, rapid cold or heat hardening-induced changes in energy metabolites were different as compared to acclimation. The ability of D. kikkawai to significantly increase stress tolerance through plasticity is likely to support its invasion potential.

Clinal variation in the polymorphic Black Sparrowhawk Accipiter melanoleucus is unrelated to infection by the blood parasite Haemoproteus nisi

Blood parasites can impact host fitness and can, thus, act as selective agents in their host’s evolution. The melanocortin system linked to colouration of vertebrates may infer higher parasite resistance via pleiotropic properties of the genes involved. Black Sparrowhawks (Accipiter melanoleucus) are colour polymorphic (dark and light morph adults) and distributed clinally in South Africa, with more dark morphs in the Southwest where the breeding season coincides with the rainy season. There, dark morphs have lower infections of haemosporidian parasites Haemoproteus nisi, suggesting a higher parasitic resistance. Thus, it is hypothesized that the morph distribution may be an adaptive response to coping with varying parasite prevalence associated with rainfall levels, which may regulate the parasites’ invertebrate vectors. This hypothesis assumes a fitness cost of high parasite burden, which could be specifically important during the energy-demanding breeding season. To explore this, we (1) quantified Black Sparrowhawk H. nisi infections across South Africa, and explored (2) breeding performance and apparent annual survival of adults, and (3) nestling body condition in Cape Town in relation to infection levels. In contrast to the predictions, we found that parasite prevalence did not vary across the country, we found no fitness costs of higher infection levels for breeding birds, and also nestling body condition was independent of infection levels. Thus, we found no support for our hypothesis. Black Sparrowhawk polymorphism is unlikely to be driven by an adaptive function linked to dealing with higher infections with H. nisi in regions with wetter breeding seasons.

Alternative reproductive tactics are associated with sperm performance in invasive round goby from two different salinity environments.

During male–male competition, evolution can favor alternative reproductive tactics. This often results in a dominant morph that holds a resource, such as a nest for egg laying, which competes with a smaller sneaker morph that reproduces by stealing fertilizations. The salinity environment can influence male growth rates, for example, via osmoregulatory costs, which in turn may influence the use of sneaker tactics for small males competing for mating opportunities. Salinity can also affect sperm directly; however, little is known of how salinity influences sneaker tactics through sperm performance. We sampled males of the invasive round goby (Neogobius melanostomus) from two environments, a freshwater river and a brackish estuary. This fish has two male morphs: nest‐holding dark males and non‐nest‐holding light males. We examined the role of water salinity of 0, 8, and 16 on sperm performance and found that for estuarine males, a salinity of 0 reduced sperm velocity compared to a salinity of 8 and 16. Riverine males had low velocity in all salinities. Sperm viability also decreased by over 30% in 0 salinity, compared to 8 and 16, for fish from both environments. Gobies produce ejaculate contents in specialized glands that could in theory shield sperm in an adverse environment. However, gland contents did not improve sperm performance in our tests. Body mass and age estimates indicate that riverine males invested more in somatic growth compared to estuarine males. Estuarine light morph males had a high enough gonadosomatic index to indicate sneaker tactics. We propose that when sperm performance is low, such as for the riverine males, sneaker tactics are ineffective and will be selected against or phenotypically suppressed. Instead, we interpret the increased investment in somatic growth found in riverine males as a life‐history decision that is advantageous when defending a nest in the next reproductive season.

Population genetics of two chromatic morphs of the Chagas disease vector Rhodnius pallescens Barber, 1932 in Panamá

Rhodnius pallescens is the principal vector of Chagas disease in Panama. Recently a dark chromatic morph has been discovered in the highlands of Veraguas Province. Limited genetic studies have been conducted with regards to the population structure and dispersal potential of Triatominae vectors, particularly in R. pallescens. Next generation sequencing methods such as RADseq and complete mitochondrial DNA (mtDNA) genome sequencing have great potential for examining vector biology across space and time. Here we utilize a RADseq method (3RAD), along with complete mtDNA sequencing, to examine the population structure of the two chromatic morpho types of R. pallescens in Panama. We sequenced 105 R. pallescens samples from five localities in Panama. We generated a 2216 SNP dataset and 6 complete mtDNA genomes. RADseq showed significant differentiation among the five localities (FCT = 0.695; P = .004), but most of this was between localities with the dark vs. light chromatic morphs (Veraguas vs. Panama Oeste). The mtDNA genomes showed a 97–98% similarity between dark and light chromatic morphs across all genes and a 502 bp insert in light morphs. Thus, both the RADseq and mtDNA data showed highly differentiated clades with essentially no gene flow between the dark and light chromatic morphs from Veraguas and central Panama respectively. We discuss the growing evidence showing clear distinctions between these two morpho types with the possibility that these are separate species, an area of research that requires further investigation. Finally, we discuss the cost-effectiveness of 3RAD which is a third of the cost compared to other RADseq methods used recently in Chagas disease vector research.

Pigeons in the sun: Thermal constraints of eumelanic plumage in the rock pigeon (Columba livia).

In wild vertebrates, several species exhibit eumelanic color polymorphism with the coexistence of dark and light morphs. The maintenance of such polymorphism suggests the existence of a selective balance between the morphs and a large body of literature has reported the costs and benefits of darker plumage coloration in birds. Among them, it has been suggested that melanin and dark plumage could entail high energetic costs especially under hot and sunny climates. However, to my knowledge, the thermal constraints of sun exposure have rarely been studied in polymorphic species. Here, we tested the impact of eumelanic plumage coloration on plumage and body temperatures, and evaporative cooling behavior in the polymorphic rock pigeon (Columbia livia). We experimentally exposed light and dark pigeons to direct sun radiation for 1h while a few birds were maintained in the shade as controls. We found that sun exposure was associated with increased plumage temperature, and this effect was greater for darker pigeons. In addition, we found that sun exposure was also associated with higher cloacal temperature but for dark pigeons only. Finally, light and dark pigeons were more likely to show cooling evaporative behavior when exposed to sun and as their cloacal temperature increases. Altogether, these results suggest that darker pigeons may have a lower ability to cope with heat and solar radiations and that dark plumage can be associated with thermal costs in this polymorphic species.

Evidence of non-random mating in a colour polymorphic raptor, the Booted Eagle

Sexual selection and non-random mating are considered, among others, determinant mechanisms for the maintenance of genetic colour polymorphism in some bird species. We analyse the mechanisms, which, in parallel with Mendelian inheritance, may be acting in the maintenance and evolution of the morph ratio in a two-morph raptor species, using observational data of successful breeding individuals and their offspring from long-term studies conducted in three Spanish populations. Our results showed that the dark offspring produced in breeding events involving mixed-morph adult pairs far exceeds the expected value under the Hardy–Weinberg equilibrium, especially in the case of pairs formed by a light male and a dark female. In addition, the low number of dark eaglets born from pairs formed by light individuals (indistinctly homozygous or heterozygous) indicates that the number of breeding events of heterozygous (both the male and female) light morph pairs, was much lower than expected. As the plausible existence of a transmission ratio distortion phenomenon in heterozygous light morph males does not, alone, explain the disproportionate number of dark eaglets observed, our results suggest that one or two selective mating phenomena may be occurring in this polymorphic system. The first one could be a disassortative mating process whereby heterozygous light males preferentially mate with dark females, based on the imprint of the colour morph of their mother. The second phenomenon would only affect light morph individuals, which would preferentially mate with heterozygous individuals of the opposite sex, selected according to secondary sexual characteristics or behavioural traits that are unknown at the moment.

Pollen colour morphs take different paths to fitness.

Colour phenotypes are often involved in communication and are thus under selection by species interactions. However, selection may also act on colour through correlated traits or alternative functions of biochemical pigments. Such forms of selection are instrumental in maintaining petal colour diversity in plants. Pollen colour also varies markedly, but the maintenance of this variation is little understood. In Campanula americana, pollen ranges from white to dark purple, with darker morphs garnering more pollinator visits and exhibiting elevated pollen performance under heat stress. Here, we generate an F2 population segregating for pollen colour and measure correlations with floral traits, pollen attributes and plant-level traits related to fitness. We determine the pigment biochemistry of colour variants and evaluate maternal and paternal fitness of light and dark morphs by crossing within and between morphs. Pollen colour was largely uncorrelated with floral traits (petal colour, size, nectar traits) suggesting it can evolve independently. Darker pollen grains were larger and had higher anthocyanin content (cyanidin and peonidin) which may explain why they outperform light pollen under heat stress. Overall, pollen-related fitness metrics were greater for dark pollen, and dark pollen sires generated seeds with higher germination potential. Conversely, light pollen plants produce 61% more flowers than dark, and 18% more seeds per fruit, suggesting a seed production advantage. Results indicate that light and dark morphs may achieve fitness through different means-dark morphs appear to have a pollen advantage whereas light morphs have an ovule advantage-helping to explain the maintenance of pollen colour variation.

Intergenerational fluctuations in colour morph frequencies may maintain elytral polymorphisms in the ladybird beetle Cheilomenes sexmaculata (Coleoptera: Coccinellidae)

AbstractPhenotypic polymorphisms are found in a wide array of taxa, and unravelling the mechanisms that maintain them is of great interest to evolutionary and ecological biologists. Temporal environmental heterogeneity may play a role in the maintenance of polymorphisms but is poorly understood. In the present study, we analysed trends in intergenerational elytral colour morph frequencies in relation to changes in fitness and life history traits (i.e. body size, mortality, fecundity, hatching rate and mate preference) in the ladybird beetle Cheilomenes sexmaculata (Coleoptera: Coccinellidae). A long-term field survey spanning nine years showed that the frequency of dark morphs increases over winter and then decreases in spring. Dark morphs may have an advantage in winter due to their higher tolerance of low temperatures compared with light morphs. Light-morph females were heavier in winter than dark-morph females. They also mated more frequently and had higher hatching rates, potentially causing an increase in light morphs in spring. These results suggest that fluctuations in morph frequencies resulting from the conflicting directions of selection pressures between overwintering and spring generations may help to maintain genetic polymorphism.

Response time of an avian prey to a simulated hawk attack is slower in darker conditions, but is independent of hawk colour morph.

To avoid predation, many species rely on vision to detect predators and initiate an escape response. The ability to detect predators may be lower in darker light conditions or with darker backgrounds. For birds, however, this has never been experimentally tested. We test the hypothesis that the response time of avian prey (feral pigeon Columbia livia f. domestica) to a simulated hawk attack (taxidermy mounted colour-polymorphic black sparrowhawk Accipiter melanoleucus) will differ depending on light levels or background colour. We predict that response will be slower under darker conditions, which would translate into higher predation risk. The speed of response of prey in relation to light level or background colour may also interact with the colour of the predator, and this idea underpins a key hypothesis proposed for the maintenance of different colour morphs in polymorphic raptors. We therefore test whether the speed of reaction is influenced by the morph of the hawk (dark/light) in combination with light conditions (dull/bright), or background colours (black/white). We predict slowest responses to morphs under conditions that less contrast with the plumage of the hawk (e.g. light morph under bright light or white background). In support of our first hypothesis, pigeons reacted slower under duller light and with a black background. However, we found no support for the second hypothesis, with response times observed between the hawk-morphs being irrespective of light levels or background colour. Our findings experimentally confirm that birds detect avian predators less efficiently under darker conditions. These conditions, for example, might occur during early mornings or in dense forests, which could lead to changes in anti-predator behaviours. However, our results provide no support that different morphs may be maintained in a population due to differential selective advantages linked to improved hunting efficiencies in different conditions due to crypsis.

Colour plumage polymorphism in the Booted Eagle: inheritance pattern and temporal stability of the morph frequencies

AbstractColour polymorphism occurs in several animal taxa and in around 3.5% of bird species. This phenomenon is disproportionately high in raptors, affecting approximately 30% of species. However, there are only four research studies in wildAccipitridaespecies and two inFalconidaespecies on colour plumage polymorphism. Here, we used the data collected over 23 years (1995‐2017) for non‐tagged individuals of three Spanish Booted Eagle (Hieraaetus pennatus) populations in order to evaluate the stability of morph frequencies during the study period, also for each sex and between sexes, and also the inheritance pattern of colour plumage polymorphism. Our results showed that morph ratios appeared stable over time and between sexes. In one of the studied populations, we found a biased proportion between dark males and dark females, with very few dark males. Our results suggest that this species follows the typical Mendelian inheritance pattern, the light allele being dominant, for which we propose a system involving two loci with epistasis and two alleles per loci. This system would explain all the cases of colour morphs observed in our studied populations and also that described in the main raptor identification guides. Although our results showed no sex‐linked morph inheritance, the offspring produced in mixed pairs with light males and dark females appear significantly linked to the colour morph of the female with a ratio 1:1.49 light/dark, contrarily to the ratio of 4.35:1 predicted by the Hardy‐Weinberg equilibrium equation. This fact suggests a transmission ratio distortion (TRD) phenomenon in heterozygous light morph males. Further investigation using molecular genetic markers will be necessary to check the genotypes of breeding adults and their offspring to confirm the inheritance mechanisms operating in this species.

Ambient temperatures differently influence colour morphs of the leaf beetle Chrysomela lapponica: Roles of thermal melanism and developmental plasticity

We asked whether ambient temperatures can affect morph frequencies within a subarctic population of the polymorphic leaf beetle Chrysomela lapponica through thermal melanism and/or developmental plasticity. Body temperature increased faster in beetles of dark morph than in beetles of light morph under exposure to artificial irradiation. Dark males ran faster than light males in both field and laboratory experiments, and this difference decreased with increasing ambient air temperature, from significant at 10 °C to non-significant at 20 °C and 26 °C. On cold days (6–14 °C), significantly more dark males than light males were found on their host plants in copula (40.8% and 27.3% respectively); on warm days (15–22 °C) this difference disappeared. Light females produced twice as many eggs as dark females; this difference did not depend on the ambient temperature. The proportion of dark morphs in the progenies of pairs with one dark parent was twice as high as that in the progenies of pairs in which both parents were light, and this proportion was greater when larvae developed at low (10 and 15 °C) than at high (20 and 25 °C) temperatures. We conclude that low temperatures may increase the frequencies of dark morphs in C. lapponica populations due to both the mating advantages of dark males over light males and developmental plasticity. Variation in frequencies of low-fecund dark morphs in the population, caused by among-year differences in temperature together with density-dependent selection, may contribute to the evolutionary dynamics of the colour polymorphism and may influence abundance fluctuations in these leaf beetle populations.

Flickers of speciation: Sympatric colour morphs of the arc-eye hawkfish, Paracirrhites arcatus, reveal key elements of divergence with gene flow.

One of the primary challenges of evolutionary research is to identify ecological factors that favour reproductive isolation. Therefore, studying partially isolated taxa has the potential to provide novel insight into the mechanisms of evolutionary divergence. Our study utilizes an adaptive colour polymorphism in the arc-eye hawkfish (Paracirrhites arcatus) to explore the evolution of reproductive barriers in the absence of geographic isolation. Dark and light morphs are ecologically partitioned into basaltic and coral microhabitats a few metres apart. To test whether ecological barriers have reduced gene flow among dark and light phenotypes, we evaluated genetic variation at 30 microsatellite loci and a nuclear exon (Mc1r) associated with melanistic coloration. We report low, but significant microsatellite differentiation among colour morphs and stronger divergence in the coding region of Mc1r indicating signatures of selection. Critically, we observed greater genetic divergence between colour morphs on the same reefs than that between the same morphs in different geographic locations. We hypothesize that adaptation to the contrasting microhabitats is overriding gene flow and is responsible for the partial reproductive isolation observed between sympatric colour morphs. Combined with complementary studies of hawkfish ecology and behaviour, these genetic results indicate an ecological barrier to gene flow initiated by habitat selection and enhanced by assortative mating. Hence, the arc-eye hawkfish fulfil theoretical expectations for the earliest phase of speciation with gene flow.