The costs of colour: plasticity of melanin pigmentation in an outbreaking polymorphic forest moth

Outside the context of industrial melanism, little is known about the physiological and ecological importance of genetic melanic polymorphisms in moths. Melanin pigments are synthesized from amino acid precursors and should therefore be costly to produce in nitrogen‐limited insects. A genetic melanic polymorphism is present in adult Malacosoma disstria Hübner (Lepidoptera: Lasiocampidae), a widespread forest moth with outbreaking population dynamics. We test the hypotheses that melanin‐based colouration is physiologically costly in M. disstria, that expression of melanin‐based colouration is a plastic trait which varies with population density and nutrition, and that the genetically based melanic phenotype is disadvantaged under nutritionally poor conditions. Two experiments were used to test these hypotheses. A field study compared pigmentation and phenotypic frequencies in moths collected from high‐ and low‐density populations. A laboratory experiment investigated the effects of larval nitrogen availability on adult pigmentation and phenotypic frequencies. High population density and nitrogen limitation reduced pigmentation and size of all moths, but phenotypic frequencies were not affected in either experiment. The effects of diet on both pigmentation and size were stronger for melanic moths than for typical moths. Our results show that adult melanism in M. disstria is physiologically costly, that colour expression is plastic despite its genetic component, and that the melanic phenotype may be disadvantaged under poor conditions but favoured under good conditions. We suggest that temporal variation in selection and trait plasticity help maintain polymorphism stability.     

A proposed selective mechanism based on metal chelation in industrial melanic moths

Industrial melanism, a phenomenon observed in some moths and especially in the case of the peppered moth (Biston betularia), has received much attention as an example of Darwinian evolution in action. The rapid rise in the proportion of the darker melanic form of the adult moth coincided with the advent of atmospheric pollution resulting from industrialization, and was ascribed to the improved camouflage of the melanotic insects against a background blackened by soot, which conferred a selective advantage in the avoidance of predation by birds. The topic of the increase in melanization during the initial period of industrial expansion and the reversal of the process after the introduction of the Clean Air Act has received much attention. Although there is sound experimental evidence to support selective avian predation as a major mechanism to account for the changes in the relative frequency of melanics, it is not clear that this is the only selective factor involved in industrial melanism. It is possible that other processes may have made a contribution to the preponderance of melanic variants. In the present study, the hypothesis is advanced that melanization may have conferred a selective advantage by protecting the insects from the toxic effects of metals by virtue of the strong metal chelating action of melanin. © 2013 The Linnean Society of London, Biological Journal of the Linnean Society, 2013, 109 , 298–301.     

Recent history of melanism in American peppered moths

Industrial melanism in peppered moths has been studied most intensively in Britain. The first melanic phenotype (effectively solid black) was recorded near Manchester in 1848. By 1895 about 98% of the specimens near Manchester were melanic, and this once rare phenotype had spread across regions of the country blackened by industrial soot. In rural, unpolluted regions, well away from industrial centers, the pale phenotype (peppered with white and black scales) remained the predominant form. During the latter half of the 20th century, following legislation designed to improve air quality, melanics began to decline in frequency and are now rare where once they had been common. Similar evolutionary changes have occurred elsewhere, but records from outside Britain are fragmentary. We have extended previous surveys of American peppered moth populations and present a composite picture of the recent decline in melanism in northern industrial states-Michigan and Pennsylvania-where melanic phenotypes decreased from more than 90% in 1959 to 6% by 2001. We contrast these changes to the near absence of melanism in a southern state-Virginia-during that same period. As in Britain, the decline in melanism in American peppered moths followed clean air legislation.     

Selective bird predation on the peppered moth: the last experiment of Michael Majerus

Colour variation in the peppered moth Biston betularia was long accepted to be under strong natural selection. Melanics were believed to be fitter than pale morphs because of lower predation at daytime resting sites on dark, sooty bark. Melanics became common during the industrial revolution, but since 1970 there has been a rapid reversal, assumed to have been caused by predators selecting against melanics resting on today's less sooty bark. Recently, these classical explanations of melanism were attacked, and there has been general scepticism about birds as selective agents. Experiments and observations were accordingly carried out by Michael Majerus to address perceived weaknesses of earlier work. Unfortunately, he did not live to publish the results, which are analysed and presented here by the authors. Majerus released 4864 moths in his six-year experiment, the largest ever attempted for any similar study. There was strong differential bird predation against melanic peppered moths. Daily selection against melanics (s ≈ 0.1) was sufficient in magnitude and direction to explain the recent rapid decline of melanism in post-industrial Britain. These data provide the most direct evidence yet to implicate camouflage and bird predation as the overriding explanation for the rise and fall of melanism in moths.     

The Genetic,Morphological, and Physiological Characterization of a Dark Larval Cuticle Mutation in the Butterfly,Bicyclus anynana

Studies on insect melanism have greatly contributed to our understanding of natural selection and the ultimate factors influencing the evolution of darkly pigmented phenotypes. Research on several species of melanic lepidopteran larvae have found that low levels of circulating juvenile hormone (JH) titers are associated with a melanic phenotype, suggesting that genetic changes in the JH biosynthetic pathway give rise to increased deposition of melanin granules in the cuticle in this group. But does melanism arise through different molecular mechanisms in different species? The present study reports on a Bicyclus anynana (Lepidoptera: Nymphalidae) dark larvae single locus mutation, in which larvae exhibit a darker cuticle relative to wild type. Unlike other lepidopteran melanic larvae mutations, this one is autosomal recessive and does not appear to involve a deficiency in JH titers. Unlike JH deficiency mutants, dark larvae mutants display similar growth rates and sexual behaviors as wild type, and topical application of a JH analogue failed to rescue the wild type cuticular coloration. Finally, transmission electron microscopy showed that sclerotization or deposition of diffuse melanin, rather than deposition of melanin granules, produces the dark coloration found in the cuticle of this species. We conclude that different molecular mechanisms underlie larval melanism in different species of Lepidoptera.     

Melanin‐Based Coloration Covaries with Hiding and Exploratory Behavior in Male Spanish Terrapins

Melanin‐based coloration reflects mostly social status and dominance in males of several species. However, the relation of melanism with other suites of behavioral traits has received less attention. Here, we examined whether the melanic coloration of the shell of male Spanish terrapins Mauremys leprosa was related to several behavioral patterns. To test this, we simulated predatory attacks of different risk levels and measured the time that the turtles spent hiding entirely in their own shells (i.e. appearance times). We also measured the activity level of the turtles in a novel‐environment test. The results showed that melanin‐based coloration was related to antipredatory behavior. Male terrapins with a greater extent of black coloration in their shells demonstrated shorter appearance times after a predator attack. However, this pattern was significant only when turtles were subjected to a high risk. In addition, darker turtles had longer latency times in a novel environment. The results of the present study support the hypothesis that melanin‐based coloration may be associated with some behavioral functions. Our study provides a good framework for future studies concerning the effect of melanin‐coloration on multiple behavioral traits that could be applied to further research using other animal models.     

Greater cuticular melanism is not associated with greater immunogenic response in adults of the polymorphic mountain stone weta, Hemideina maori

Abstract.  1. Greater immune function is associated with the high-density melanic phase of polyphenic insects, appearing to compensate for density-dependent increases in susceptibility to parasites and/or pathogens. Other types of discrete variation in cuticular colour occur in insects (which may or may not be associated with melanin pigmentation), but whether this variation is predictive of immune ability has not been investigated.
2. In the mountain stone weta Hemideina maori , a black morph and yellow banded morph occur. These morphs are not seasonally polyphenic and have discrete haplotype genetic markers. Black individuals are typically found at lower local densities than yellow individuals, contrary to relations between cuticular melanism and density seen in polyphenic insects.
3. Yellow males and females had greater melanotic encapsulation responses upon immune challenge than did black males and females, but these differences were not associated with differences in temperature selection between morphs. Morph differences in melanotic encapsulation responses were somewhat related to differences between morphs in haemocyte concentrations.
4. These results indicate that a common form of immune expression is not heightened with dark coloration in the mountain stone weta. Thus, earlier findings of greater immunity associated with darker cuticles in phase polyphenic insects cannot be extended to insects with other forms of discrete colour variation. These findings will help in elucidating causes and consequences of such colour polymorphism, which is widespread in several insect orders.     

Genetic basis of stage-specific melanism: a putative role for a cysteine sulfinic acid decarboxylase in insect pigmentation

Melanism, the overall darkening of the body, is a widespread form of animal adaptation to particular environments, and includes bookcase examples of evolution by natural selection, such as industrial melanism in the peppered moth. The major components of the melanin biosynthesis pathway have been characterized in model insects, but little is known about the genetic basis of life-stage specific melanism such as cases described in some lepidopteran species. Here, we investigate two melanic mutations of Bicyclus anynana butterflies, called Chocolate and melanine, that exclusively affect pigmentation of the larval and adult stages, respectively. Our analysis of Mendelian segregation patterns reveals that the larval and adult melanic phenotypes are due to alleles at different, independently segregating loci. Our linkage mapping analysis excludes the pigmentation candidate gene black as the melanine locus, and implicates a gene encoding a putative pyridoxal phosphate-dependant cysteine sulfinic acid decarboxylase as the Chocolate locus. We show variation in coding sequence and in expression levels for this candidate larval melanism locus. This is the first study that suggests a biological function for this gene in insects. Our findings open up exciting opportunities to study the role of this locus in the evolution of adaptive variation in pigmentation, and the uncoupling of regulation of pigment biosynthesis across developmental stages with different ecologies and pressures on body coloration.     

Competitive trade-off strategies in ArcticDaphnia linked to melanism and UV-B stress

 Different high-Arctic, freshwater localities at Ny-A˚lesund, Svalbard (79°N) were examined for their UV-absorbing properties, depth and presence of melanic or non-melanic morphs of the planktonic crustacean Daphnia pulex. Light regimes in two localities with each of these morphs were measured by using underwater spectroradiometer. Most localities have low absorbance of short-waved light, but no clearcut relationship between UV transparency and occurrence of melanic morphs was detected. Yet, in the laboratory, the melanic morph showed far lower growth rates, thus being competitively inferior to the non-melanic. Conversely, the melanic morph was more resistant to UV light, suggesting a trade-off between the metabolic tax paid for the melanin synthesis and its UV-protecting abilities, or a staggered growth capacity possibly owing to polyploidy. Frequency of melanic or non-melanic clones could thus be directly linked to ambient UV-B stress and serve as an indicator thereof, but the apparently extensive need for UV protection under the Arctic light regimes is still puzzling, and the role of melanism and polyploidy in these organisms cannot be considered finally settled. Received: 29 September 1995/Accepted: 29 January 1996     

Fitness of insularia morphs of the peppered moth Biston betularia

In recent years the industrial melanic carbonaria morph in the moth Biston betularia (L.) has decreased rapidly in frequency in Britain as air pollution has decreased. The intermediate melanic insularia has shown a variable response. We have estimated the fitness of insularia, compared with the other two morphs, for several data sets. As a rule its fitness lies between that of carbonaria and typical, but nearer to typical and sometimes very close to it. The intermediate position is expected if fitness relates directly to phenotype. The results suggest that insularia may continue polymorphic while carbonaria is likely to disappear. The past high frequency of insularia in South Wales may have been due to an initial increase in insularia frequency before carbonaria reached the region. Differences in dynamics of frequency change in insularia and carbonaria are evidence against induction, which has sometimes been invoked to explain the spread of melanism in this species.  © 2004 The Linnean Society of London, Biological Journal of the Linnean Society , 2004, 82 , 359–366.     

Rethinking the thermal melanism hypothesis: rearing temperature and coloration in pygmy grasshoppers

Selection for efficient conversion of solar radiation to body heat has favored the evolution of dark coloration in many ectotherms. The thermal melanism hypothesis posits that dark coloration is beneficial under conditions of low ambient temperatures because it results in faster heating rates and higher body temperatures. Fast heating rates, however, may come at a cost of overheating unless compensated for by thermal physiology or behaviour. Pygmy grasshopper (Orthoptera, Tetrigidae) populations that inhabit fire-ravaged areas characterized by blackened backgrounds and hot surface temperatures due to high absorbance of solar radiation show an increased frequency of black phenotypes. I raised the progeny of wild-captured Tetrix undulata in cold and hot temperatures and used data on color patterns and survival in a greenhouse to examine whether a cold thermal environment triggered the development of melanic coloration or differently affected survival of melanic versus non-melanic individuals. My results indicate that melanism was not influenced by rearing temperature but by genes or epigenetic maternal effects. Temperature also did not affect survival. However, melanic individuals produced by melanic mothers survived longer than melanic individuals produced by non- melanic mothers, whereas non-melanic individuals produced by non-black mothers survived longer than melanic individuals produced by non-black mothers. This suggests a mismatch between color and physiology in offspring belonging to a different color morph than their mother. Future investigations into the evolution of melanism should consider conflicting selection pressures on thermal capacity and camouflage as well as the influence of correlated responses to selection on traits associated with coloration.     

Effects of polymorphic melanism and larval diet on life history traits of Malacosoma disstria moths

In this study we investigated the presence and possible genetic basis of polymorphic melanism in the forest tent caterpillar (Malacosoma disstria) moth. Adult moths were classified into pattern-based phenotypes and wing darkness was measured to quantify the degree of melanization. We found that two distinct phenotypes, melanic and simple, are present in these moths. Although the full melanic phenotype is sex-limited to males, it is partially expressed in females. We also provide support for the theory that the melanic allele is autosomal and dominant. The effects of larval diet quality on the survival, development and wing melanization of each phenotype were studied by rearing larvae on the foliage of either a primary or secondary host. Diet quality did not differentially affect the two phenotypes; however, melanic males were found to be smaller than simple males regardless of larval diet. Such inherent developmental differences between the two phenotypes could have important consequences for the frequencies of the two morphs.     

Melanin‐based colour polymorphism responding to climate change

Climate warming leads to a decrease in biodiversity. Organisms can deal with the new prevailing environmental conditions by one of two main routes, namely evolving new genetic adaptations or through phenotypic plasticity to modify behaviour and physiology. Melanin‐based colouration has important functions in animals including a role in camouflage and thermoregulation, protection against UV‐radiation and pathogens and, furthermore, genes involved in melanogenesis can pleiotropically regulate behaviour and physiology. In this article, I review the current evidence that differently coloured individuals are differentially sensitive to climate change. Predicting which of dark or pale colour variants (or morphs) will be more penalized by climate change will depend on the adaptive function of melanism in each species as well as how the degree of colouration covaries with behaviour and physiology. For instance, because climate change leads to a rise in temperature and UV‐radiation and dark colouration plays a role in UV‐protection, dark individuals may be less affected from global warming, if this phenomenon implies more solar radiation particularly in habitats of pale individuals. In contrast, as desertification increases, pale colouration may expand in those regions, whereas dark colourations may expand in regions where humidity is predicted to increase. Dark colouration may be also indirectly selected by climate warming because genes involved in the production of melanin pigments confer resistance to a number of stressful factors including those associated with climate warming. Furthermore, darker melanic individuals are commonly more aggressive than paler conspecifics, and hence they may better cope with competitive interactions due to invading species that expand their range in northern latitudes and at higher altitudes. To conclude, melanin may be a major component involved in adaptation to climate warming, and hence in animal populations melanin‐based colouration is likely to change as an evolutionary or plastic response to climate warming.     

Synergistic and antagonistic interaction between different branches of the immune system is related to melanin‐based coloration in nestling tawny owls

When exposed to parasites, hosts often mount energetically expensive immune responses, and this may alter resource allocation between competing life history traits including other components of the immune system. Here, we investigated whether a humoral immune challenge towards a vaccine reduces or enhances the cutaneous immune responses towards an injection of lipopolysaccharid (LPS, innate immunity) and phytohaemagglutinin (PHA, T‐cell immunity) in nestling tawny owls in interaction with the degree of plumage melanin‐based coloration. The humoral immune challenge enhanced the response to LPS similarly in differently coloured nestlings. In contrast, the same humoral immune challenge enhanced immune response to PHA in dark reddish melanic nestlings while reducing it in pale reddish melanic nestlings. Our results highlight that both antagonistic and synergistic interactions can take place among branches of immune system, and that the sign and magnitude of these interactions can vary with immune responses involved and the degree of melanin‐based coloration.     

Dorsal pattern polymorphism in female Iberian wall lizards: differences in morphology,dorsal coloration,immune response,and reproductive investment

Sex‐specific colour polymorphisms have been extensively documented in many different taxa. When polymorphism in colour pattern is restricted to females, the condition is known as female‐limited pattern polymorphism (FPP), which has been less commonly addressed in vertebrates. FPP is present in several lizard species, although most research on lizards has focused on carotenoid‐ and pteridine‐based coloration and not on melanin‐based polymorphisms. In the present study, we focus on Iberian wall lizards, Podarcis hispanicus, where two female melanin‐based dorsal patterns can be clearly distinguished: striped and reticulated‐blotched. We indirectly tested the hypothesis that selection acts differentially among P. hispanicus female morphs to create alternative morph‐specific phenotypic optima at different levels by investigating whether morphs differ in fitness proxies. We specifically examined whether the two female dorsal pattern morphs differed in adult morphology, dorsal coloration, immune response, reproductive investment, and growth. We did not find a relationship between melanin‐based coloration and hatchling growth and immune response, despite a correlation between these traits possibly being expected as a result of pleiotropy in the melanocortin system. However, our results show that female dorsal morphs in P. hispanicus differ in terms of adult morphology, dorsal coloration, and reproductive investment. Reticulated‐blotched P. hispanicus females had deeper heads and longer femora, less melanin, and more brownish coloration, and also had larger and heavier hatchlings than striped females.     

Darker female pigeons transmit more specific antibodies to their eggs than do paler ones

Melanin‐based coloration is widespread among vertebrates, yet the adaptive significance of such pigments remains elusive, particularly with regard to the link between melanin and immune‐mediated maternal effects. The aim of this study was to investigate whether melanin‐based coloration could signal the ability of mothers to mount a humoral response and to transfer maternal antibodies (Ab) to their young. We injected differently coloured (pale and dark) female feral pigeons (Columba livia) with Chlamydiae (a natural antigen) and Keyhole Limpet Haemocyanin (KLH, an artificial antigen), and found no significant difference in humoral response between differently coloured females. However, darker females transferred more Ab against Chlamydiae into their eggs than paler ones, despite similar circulating levels of Ab. In addition to this, melanin‐based coloration showed a high heritability value. This suggests that a genetically based coloured trait might be linked to the ability of females to transfer specific Ab against Chlamydiae (but not against KLH) to their offspring, independent of their ability to produce Ab. This suggests that transmission of maternal Ab is antigen dependent, and that melanin‐based coloration might signal female ability to transmit specific Ab against natural pathogens. © 2013 The Linnean Society of London     

Immune function, parasitization and extended phenotypes in colour polymorphic pygmy grasshoppers

Ecological and evolutionary consequences of host–parasite interactions have attracted considerable attention from evolutionary biologists. Previous studies have suggested that immune responsiveness may be genetically or developmentally linked with colour pattern, such that the evolution of animal colour patterns may be influenced by correlated responses to selection for parasite resistance. We studied interactions between the endoparasitic fly Leiophora innoxia (Meigen) (Diptera: Tachinidae) and its colour polymorphic pygmy grasshopper host Tetrix undulata (Sow.) (Orthoptera: Tetrigidae) to test for morph‐specific differences in parasitization and immune defence, and host‐induced variation in parasite phenotypes. Our results revealed that c. 2 and 30% of adult grasshoppers collected from the same natural population in two subsequent years, respectively were parasitized. Parasite prevalence was independent of host sex and colour morph. Pupae were larger if the parasite had developed in a female than in a male host, possibly reflecting host resource value or a physical constraint on larval growth imposed by host body size. Pupal size was also associated with host colour morph, with individuals that had developed in dark morphs being smaller at pupation compared to those that developed in paler morphs. However, immune defence, measured as the encapsulation response to a novel antigen, did not differ among individuals belonging to alternative colour morphs or sexes. Darker morphs warm up more quickly and prefer higher body temperatures than paler ones. Encapsulation was not influenced by maintenance temperature (15 vs. 30 °C), however, suggesting that indirect effects of coloration on parasite resistance mediated via differential body temperature are unlikely. The dependence of parasite body size on host colour morph may thus reflect plasticity of growth and development of the larvae in response to differential host body temperature, rather than variable host immune defence. © 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 85 , 373–383.     

The molecular basis of the plumage colour polymorphism in the Tahiti reed‐warbler Acrocephalus caffer

The endemic Tahiti reed‐warbler Acrocephalus caffer occurs in two distinct morphs, a typical or ‘yellow’ morph and a melanic or ‘dark’ morph, which are found together in the valleys of the eastern and central parts of the island of Tahiti (Society Islands, French Polynesia). We investigated the molecular basis of the plumage colour polymorphism in this species using sequences of the melanocortin‐1 receptor (MC1R), a gene often found associated to melanism in birds. We found that the MC1R genotype was perfectly associated with plumage colour in the Tahiti reed‐warbler, with the same nonsynonymous substitution that showed a correlation with phenotype in the Caribbean bananaquit Coereba flaveola. An heterozygous reed‐warbler at this site presented a melanic phenotype, suggesting that the melanic allele is dominant. All other Polynesian reed‐warbler species, which do not have a melanic morph, shared the ‘yellow’ nucleotide at this position. These results suggested that the same mutation point was linked to a melanic polymorphism in two unrelated passerine birds.     

Ecological genetics of Norwegian Daphnia. II. Distribution of Daphnia longispina genotypes in relation to short-wave radiation and water colour

In the mountain range of South Norway, transparent and melanic members of the Daphnia longispina complex occur. Melanic populations inhabit clearwater lakes and ponds, while transparent populations are found in ponds with slightly humic water. Mixed populations have not been detected. The distribution patterns of the two morphs are related to ambient levels of short-wave radiation, and the light absorptive properties of the inhabited waters. The pigment present is probably melanin, which is deposited in the dorsally directed parts of the carapace, head shield and antennae. Allozyme studies indicate that these distinct morphs are only remotely related, the alpine transparent group being closer to lowland, likewise transparent, populations. A melanic population found at Svalbard is closely related to the melanic mainland populations. Clonal diversity in ponds and shallow lakes is very low, in contrast to populations of deeper lakes.Deceased.     

Melanic frequencies in three species of moths in post industrial Britain

Changing patterns of morph frequency in three moth species ( Biston betularia , Odontoptera bidentata and Apocheima pilosaria ) have been investigated using data from the Rothamsted Insect Survey. All three exhibited industrial melanism during the period of high atmospheric pollution in Britain. Three historical and habitat types are compared, the old industrial north of England, rural Scotland, Wales and South-West England, and a southern English intermediate region of high human population density but generally low industrialization. Between 1974 and 1999 the carbonaria morph of B. betularia declined in frequency in the industrial region and is nearly absent from rural areas. It is the form which most closely tracks atmospheric change. It is shown that the insularia forms of B. betularia and the melanic morphs in the other two species have decreased in the industrial region, commencing later than carbonaria , but have maintained their presence and possibly reached equilibrium elsewhere. They may be non-industrial polymorphisms. B. betularia is rarer than the other species and all three species are at lower densities in industrial than in non-industrial regions. © 2002 The Linnean Society of London , Biological Journal of the Linnean Society , 2002, 75 , 475–482.