Wing Melanization Research Articles

Resource-based trade-offs and the adaptive significance of seasonal plasticity in butterfly wing melanism.

Phenotypic plasticity is the ability of an organism to alter its phenotype in response to environmental cues. This can be adaptive if the cues are reliable predictors of impending conditions and the alterations enhance the organism's ability to capitalize on those conditions. However, since traits do not exist in isolation but as part of larger interdependent systems of traits (phenotypic integration), trade-offs between correlated plastic traits can make phenotypic plasticity non- or maladaptive. We examine this problem in the seasonally plastic wing melanism of a pierid (Order Lepidoptera, Family Pieridae) butterfly, Pieris rapae L. Several wing pattern traits are more melanized in colder than in warmer seasons, resulting in effective thermoregulation through solar absorption. However, other wing pattern traits, the spots, are less melanized during colder seasons than in warmer seasons. Although spot plasticity may be adaptive, reduced melanism of these spots could also be explained by resource-based trade-offs. Theory predicts that traits involved in resource-based trade-offs will be positively correlated when variation among individuals in resource acquisition is greater than variation among individuals in resource allocation strategies, and negatively correlated when variation in allocation is greater than variation in acquisition. Using data from both field studies and laboratory studies that manipulate dietary tyrosine, a melanin precursor, we show that when allocation to thermoregulatory melanism (ventral hindwing, and basal dorsal fore- and hindwing "shading") varies substantially this trait is negatively correlated with spot melanism. However, when there is less variation in allocation to thermoregulatory melanism we find these traits to be positively correlated; these findings are consistent with the resource-based trade-off hypothesis, which may provide a non- or maladaptive hypothesis to explain spot plasticity. We also show that increased dietary tyrosine results in increased spot melanism under some conditions, supporting the more general idea that melanism may involve resource-based costs.

Ornamentation diversified faster than eco-morphology across Nearctic dragonflies

AbstractEco-morphology and ornamentation are two phenotypic dimensions along which co-existing species often diverge, yet theory makes contrasting predictions about how these phenotypes diversify relative to each other. Some theory predicts that intense reproductive demands cause more pronounced divergence in ornamentation than in eco-morphology. Other theory predicts that preferences for condition-dependent ornamentation in species encountering divergent ecological conditions will facilitate rapid divergence in eco-morphology but not ornamentation. We evaluated these conflicting predictions in Nearctic Libelluloidea dragonflies by testing if the diversification of a condition-dependent ornament, male wing melanization, was slower and less pronounced between species than the diversification of two key eco-morphological traits, body size and relative wing size. We found that male wing melanization evolved much faster than either body size or relative wing size. Furthermore, in contrast to the patterns for either eco-morphological trait, the best-supported models of diversification in male wing melanization indicate that the majority of divergence arose between the most closely related species. These results reveal that the primary axis of divergence between closely related Libelluloidea dragonflies is ornamentation rather than eco-morphology. Our study therefore suggests that evolutionary responses to disparate reproductive demands may be fundamental to the persistence and co-existence of closely related species.

Comparing the roles of climate, predation and phylogeography in driving wing colour variation in Ranchman’s tiger moth (Arctia virginalis)

Abstract In Lepidoptera, as an explanation for darker phenotypes occurring in colder areas, wing melanism has been proposed to increase solar thermal gain. Alternatively, trade-offs with aposematic signalling and ultraviolet protection have been proposed as explanations for variation in melanism. To investigate the roles of temperature, humidity, solar radiation and predation in driving melanism in the Ranchman’s tiger moth (Arctia virginalis), we characterized wing melanism in 23 populations across the range. We also conducted predation experiments using artificial moths and carried out genetic analyses to examine population structure and to test whether wing coloration was hereditary. We found that wing melanism was positively associated with mean temperature during the flight season, which was the best predictor of melanism rates. Wing melanism also exhibited a negative association with humidity and a weak positive association with insolation. We also found two loci weakly associated with wing melanism and showed that melanism is likely to be highly hereditary but not closely associated with population differentiation. Our results contrast with previous findings that melanism is associated with colder conditions and higher predation risk and suggest that humidity and protection against ultraviolet radiation are potential drivers of variation in wing melanism that have been overlooked.

Dark wing pigmentation as a mechanism for improved flight efficiency in the Larinae

There are many hypotheses explaining the diversity of colours and patterns found in nature, but they are often difficult to examine empirically. Recent studies show the dark upperside of gliding birds’ wings could reduce drag by decreasing the density of surrounding air. It may therefore be expected that species with darker wings have less efficient morphology than their paler counterparts. I conducted an analysis of the Larinae (gulls), which exhibit extreme variation in wing (mantle and wingtip) melanization, to test whether wing loading is a predictor of wing darkness. I found that, for each standard deviation increase in wing loading, mantle darkness is predicted to increase by 1.2 shades on the Kodak grey scale. Wing loading is also positively related to the proportion of black on wingtips. Furthermore, heavier species have lower aspect ratio wings, suggesting that dark wings have evolved to improve the trade-off between maneuverability and long distance flight.

Natural Selection on Adults Has Trait-Dependent Consequences for Juvenile Evolution in Dragonflies.

AbstractAlthough natural selection often fluctuates across ontogeny, it remains unclear what conditions enable selection in one life-cycle stage to shape evolution in others. Organisms that undergo metamorphosis are useful for addressing this topic because their highly specialized life-cycle stages cannot always evolve independently despite their dramatic life-history transition. Using a comparative study of dragonflies, we examined three conditions that are hypothesized to allow selection in one stage to affect evolution in others. First, we tested whether lineages with less dramatic metamorphosis (e.g., hemimetabolous insects) lack the capacity for stage-specific evolution. Rejecting this hypothesis, we found that larval body shape evolves independently from selection on adult shape. Next, we evaluated whether stage-specific evolution is limited for homologous and/or coadapted structures. Indeed, we found that selection for larger wings is associated with the evolution of coadapted larval sheaths that store developing wing tissue. Finally, we assessed whether stage-specific evolution is restricted for traits linked to a single biochemical pathway. Supporting this hypothesis, we found that species with more wing melanization in the adult stage have evolved weaker melanin immune defenses in the larval stage. Thus, our results collectively show that natural selection in one stage imposes trait-dependent constraints on evolution in others.

Beyond thermal melanism: association of wing melanization with fitness and flight behaviour in a butterfly

Cold developmental conditions can greatly affect adult life history of ectotherms in seasonal habitats. Such effects are mostly negative, but sometimes adaptive. Here, we tested how cold conditions experienced during pupal development affect adult wing melanization of an insect ectotherm, the Glanville fritillary butterfly, Melitaea cinxia. We also assessed how in turn previous cold exposure and increased melanization can shape adult behaviour and fitness, by monitoring individuals in a seminatural set-up. We found that, despite pupal cold exposure inducing more melanization, wing melanization was not linked to adult thermoregulation preceding flight, under the conditions tested. Conversely, wing-vibrating behaviour had a major role in producing heat preceding flight. Moreover, more melanized individuals were more mobile across the experimental set-up. This may be caused by a direct impact of melanization on flight ability or a more indirect impact of coloration on behaviours such as mate search strategies and/or eagerness to disperse to more suitable mating habitats. We also found that more melanized individuals of both sexes had reduced mating success and produced fewer offspring, which suggests a clear fitness cost of melanization. Whether the reduced mating success is dictated by impaired mate search behaviour, reduced physical condition leading to a lower dominance status or weakened visual signalling remains unknown. In conclusion, while there was no clear role of melanization in providing a thermal advantage under our seminatural conditions, we found a fitness cost of being more melanized, which potentially impacted adult space use behaviour.

Adaptive plasticity in wing melanisation of a montane butterfly across a Himalayan elevational gradient

Traits that are significant to the thermal ecology of temperate or montane species are expected to prominently co‐vary with the thermal environment experienced by an organism. The Himalayan Pieris canidia butterfly exhibits considerable variation in wing melanisation. We investigated: (i) whether variation in wing melanisation and (ii) activity period of this montane butterfly was influenced by the seasonally and elevationally changing thermal landscape. We discovered that wing melanisation varied across elevation, seasons, sex, and wing surfaces, with the variation strongly structured in space and time: colder seasons and higher elevations produced more melanic individuals. Notably, melanisation did not vary uniformly across all wing surfaces: (i) melanisation of the ventral hindwing co‐varied much more prominently with elevation, but (ii) melanisation on all other surfaces varied with seasonal changes in the thermal environment. Observed wing surface‐specific patterns indicated thermoregulatory function for this variation in melanisation. Such wing surface‐specific responses to seasonal and elevational variation in temperature have rarely been reported in montane insects. Moreover, daily and seasonal thermal cycles were found to strongly influence activity periods of this species, suggesting the potential limits to wing melanisation plasticity. Overall, these results showed that the seasonal and elevational gradients in temperature influence the thermal phenotype as well as activity periods of this Himalayan butterfly. It will be critical to study the phenotypic evolution of such montane insects in response to the ongoing climate change, which is already showing significant signs in this iconic mountain range.

Genotype-environment interactions rule the response of a widespread butterfly to temperature variation.

Understanding how organisms adapt to complex environments is a central goal of evolutionary biology and ecology. This issue is of special interest in the current era of rapidly changing climatic conditions. Here, we investigate clinal variation and plastic responses in life history, morphology and physiology in the butterfly Pieris napi along a pan-European gradient by exposing butterflies raised in captivity to different temperatures. We found clinal variation in body size, growth rates and concomitant development time, wing aspect ratio, wing melanization and heat tolerance. Individuals from warmer environments were more heat-tolerant and had less melanised wings and a shorter development, but still they were larger than individuals from cooler environments. These findings suggest selection for rapid growth in the warmth and for wing melanization in the cold, and thus fine-tuned genetic adaptation to local climates. Irrespective of the origin of butterflies, the effects of higher developmental temperature were largely as expected, speeding up development; reducing body size, potential metabolic activity and wing melanization; while increasing heat tolerance. At least in part, these patterns likely reflect adaptive phenotypic plasticity. In summary, our study revealed pronounced plastic and genetic responses, which may indicate high adaptive capacities in our study organism. Whether this may help such species, though, to deal with current climate change needs further investigation, as clinal patterns have typically evolved over long periods.

Latitudinal and altitudinal variation in ecologically important traits in a widespread butterfly

AbstractUnderstanding how organisms adapt to complex environments lies at the very heart of evolutionary biology and ecology, and is of particular concern in the current era of anthropogenic global change. Variation in ecologically important traits associated with environmental gradients is considered to be strong evidence for adaptive responses. Here, we study phenotypic variation along a latitudinal and an altitudinal cline in 968 field-collected males of the widespread European butterfly Pieris napi. In contrast to our expectations, body size decreased with increasing latitude and altitude, suggesting that warmer rather than cooler conditions may be more beneficial for individual development in this species. Higher altitudes but not latitudes seemed to be associated with increased flight performance, suggesting stronger challenges for flight activity in high-altitude environments (e.g. due to strong wind). Moreover, wing melanization increased while yellow reflectance decreased towards colder environments in both clines. Thus, increased melanization under thermally challenging conditions seems to compromise investment into a sexually selected trait, resulting in a trade-off. Our study, although exclusively based on field-collected males, revealed indications of adaptive patterns along geographical clines. It documents the usefulness of field-collected specimens, and the strength of comparing latitudinal and altitudinal clines to identify traits being potentially under thermal selection.

Evaluating responses to temperature during pre-metamorphosis and carry-over effects at post-metamorphosis in the wood tiger moth (Arctia plantaginis).

Insect metamorphosis is one of the most recognized processes delimiting transitions between phenotypes. It has been traditionally postulated as an adaptive process decoupling traits between life stages, allowing evolutionary independence of pre- and post-metamorphic phenotypes. However, the degree of autonomy between these life stages varies depending on the species and has not been studied in detail over multiple traits simultaneously. Here, we reared full-sib larvae of the warningly coloured wood tiger moth (Arctia plantaginis) in different temperatures and examined their responses for phenotypic (melanization change, number of moults), gene expression (RNA-seq and qPCR of candidate genes for melanization and flight performance) and life-histories traits (pupal weight, and larval and pupal ages). In the emerging adults, we examined their phenotypes (melanization and size) and compared them at three condition proxies: heat absorption (ability to engage flight), flight metabolism (ability to sustain flight) and overall flight performance. We found that some larval responses, as evidenced by gene expression and change in melanization, did not have an effect on the adult (i.e. size and wing melanization), whereas other adult traits such as heat absorption, body melanization and flight performance were found to be impacted by rearing temperature. Adults reared at high temperature showed higher resting metabolic rate, lower body melanization, faster heating rate, lower body temperature at take-off and inferior flight performance than cold-reared adults. Thus, our results did not unambiguously support the environment-matching hypothesis. Our results illustrate the importance of assessing multiple traits across life stages as these may only be partly decoupled by metamorphosis.This article is part of the theme issue ‘The evolution of complete metamorphosis'.

Sex differences in immunity in a natural population of bush-cricket (Orthoptera: Phaneropterinae)

Immune defense often differs between sexes, where females generally exhibit higher individual immunity than males. Such sexually dimorphic immune defense stems from differential investment into reproduction as females allocate more resources into mating. Contrary to prevailing predictions, some studies have found that females exhibit a weaker immune response than males. In this study, I examined both humoral (phenoloxidase activity and lytic activity) and cellular (encapsulation ability) immune parameters including cuticular melanization in a field population of a bush-cricket (Isophya reticulata) that is an endemic species in the northeastern part of Turkey. As expected, females showed a higher phenoloxidase activity than males, whereas there were no sex differences in lytic activity. Males expressed a higher encapsulation response than females. These results indicated that sex differences in immunity may be variable and females may not always exhibit higher immunity than males. Another interesting finding of this study was a positive relationship between stridulatory file length, phenoloxidase activity, and wing melanization. This finding demonstrated a connection between size differences in stridulatory apparatus that affect sexual signaling and immune response.

Using museum specimens to track morphological shifts through climate change.

Museum specimens offer a largely untapped resource for detecting morphological shifts in response to climate change. However, morphological shifts can be obscured by shifts in phenology or distribution or sampling biases. Additionally, interpreting phenotypic shifts requires distinguishing whether they result from plastic or genetic changes. Previous studies using collections have documented consistent historical size changes, but the limited studies of other morphological traits have often failed to support, or even test, hypotheses. We explore the potential of collections by investigating shifts in the functionally significant coloration of a montane butterfly, Colias meadii, over the past 60 years within three North American geographical regions. We find declines in ventral wing melanism, which correspond to reduced absorption of solar radiation and thus reduced risk of overheating, in two regions. However, contrary to expected responses to climate warming, we find melanism increases in the most thoroughly sampled region. Relationships among temperature, phenology and morphology vary across years and complicate the distinction between plastic and genetic responses. Differences in these relationships may account for the differing morphological shifts among regions. Our findings highlight the promise of using museum specimens to test mechanistic hypotheses for shifts in functional traits, which is essential for deciphering interacting responses to climate change.This article is part of the theme issue 'Biological collections for understanding biodiversity in the Anthropocene'.

Viral exposure effects on life-history, flight-related traits, and wing melanisation in the Glanville fritillary butterfly

Infections represent a constant threat for organisms and can lead to substantial fitness losses. Understanding how individuals, especially from natural populations, respond towards infections is thus of great importance. Little is known about immunity in the Glanville fritillary butterfly (Melitaea cinxia). As the larvae live gregariously in family groups, vertical and horizontal transmission of infections could have tremendous effects on individuals and consequently impact population dynamics in nature. We used the Alphabaculovirus type strain Autographa californica multiple nucleopolyhedrovirus (AcMNPV) and demonstrated that positive concentration-dependent baculovirus exposure leads to prolonged developmental time and decreased survival during larval and pupal development, with no sex specific differences. Viral exposure did not influence relative thorax mass or wing morphometric traits often related to flight ability, yet melanisation of the wings increased with viral exposure, potentially influencing disease resistance or flight capacity via thermal regulation. Further research is needed to explore effects under sub-optimal conditions, determine effects on fitness-related traits, and investigate a potential adaptive response of increased melanisation in the wings due to baculovirus exposure.

Intrasexual selection favours an immune-correlated colour ornament in a dragonfly.

Sexual signalling is predicted to shape the evolution of sex-specific ornamentation, and establishing the costs and benefits of ornamentation and the information that ornamentation provides to receivers is necessary to evaluating this adaptive function. Here, we assessed the adaptive function of a common colour ornament in insects, melanin wing ornamentation, using the dragonfly Pachydiplax longipennis. We hypothesized that greater ornamentation would improve territory-holding success by decreasing aggression that males receive from territorial rivals, but that more ornamented males may have shorter lifespans. Using mark-recapture field observations, we found that more ornamented males had greater territory-holding success and that viability selection did not act on wing melanization. We then compared the aggression of territorial rivals to decoy males before and after experimentally augmenting wing melanization, finding that males significantly reduced aggression following the manipulation. We next hypothesized that wing melanization would signal fighting ability to territorial rivals by reflecting condition via investment in the costly melanin synthesis pathway. We observed a positive relationship between ornamentation and the likelihood of winning territorial disputes, suggesting that wing melanization provides information about fighting ability to rivals. We also found a positive relationship between melanin-based immune defence and ornamentation, supporting a link between the signal and condition. We conclude that wing melanization is a condition-related signal of fighting ability and suggest that this may be a common mechanism promoting the evolution of melanin ornamentation.

Why Small Is Beautiful: Wing Colour Is Free from Thermoregulatory Constraint in the Small Lycaenid Butterfly, Polyommatus icarus.

We examined the roles of wing melanisation, weight, and basking posture in thermoregulation in Polyommatus Icarus, a phenotypically variable and protandrous member of the diverse Polyommatinae (Lycaenidae). Under controlled experimental conditions, approximating to marginal environmental conditions for activity in the field (= infrequent flight, long duration basking periods), warming rates are maximised with fully open wings and maximum body temperatures are dependent on weight. Variation in wing melanisation within and between sexes has no effect on warming rates; males and females which differ in melanisation had similar warming rates. Posture also affected cooling rates, consistent with cooling being dependent on convective heat loss. We hypothesise that for this small sized butterfly, melanisation has little or no effect on thermoregulation. This may be a factor contributing to the diversity of wing colours in the Polyommatinae. Because of the importance of size for thermoregulation in this small butterfly, requirements for attaining a suitable size to confer thermal stability in adults may also be a factor influencing larval feeding rates, development time and patterns of voltinism. Our findings indicate that commonly accepted views of the importance of melanisation, posture and size to thermoregulation, developed using medium and large sized butterflies, are not necessarily applicable to small sized butterflies.

Does Skipping a Meal Matter to a Butterfly's Appearance? Effects of Larval Food Stress on Wing Morphology and Color in Monarch Butterflies

In animals with complex life cycles, all resources needed to form adult tissues are procured at the larval stage. For butterflies, the proper development of wings involves synthesizing tissue during metamorphosis based on the raw materials obtained by larvae. Similarly, manufacture of pigment for wing scales also requires resources acquired by larvae. We conducted an experiment to test the effects of food deprivation in the larval stage on multiple measures of adult wing morphology and coloration of monarch butterflies (Danaus plexippus), a species in which long-distance migration makes flight efficiency critical. In a captive setting, we restricted food (milkweed) from late-stage larvae for either 24 hrs or 48 hrs, then after metamorphosis we used image analysis methods to measure forewing surface area and elongation (length/width), which are both important for migration. We also measured the brightness of orange pigment and the intensity of black on the wing. There were correlations between several wing features, including an unexpected association between wing elongation and melanism, which will require further study to fully understand. The clearest effect of food restriction was a reduction in adult wing size in the high stress group (by approximately 2%). Patterns observed for other wing traits were ambiguous: monarchs in the low stress group (but not the high) had less elongated and paler orange pigmentation. There was no effect on wing melanism. Although some patterns obtained in this study were unclear, our results concerning wing size have direct bearing on the monarch migration. We show that if milkweed is limited for monarch larvae, their wings become stunted, which could ultimately result in lower migration success.

Fitness Costs of Thermal Reaction Norms for Wing Melanisation in the Large White Butterfly (Pieris brassicae)

The large white butterfly, Pieris brassicae, shows a seasonal polyphenism of wing melanisation, spring individuals being darker than summer individuals. This phenotypic plasticity is supposed to be an adaptive response for thermoregulation in natural populations. However, the variation in individuals’ response, the cause of this variation (genetic, non genetic but inheritable or environmental) and its relationship with fitness remain poorly known. We tested the relationships between thermal reaction norm of wing melanisation and adult lifespan as well as female fecundity. Butterflies were reared in cold (18°C), moderate (22°C), and hot (26°C) temperatures over three generations to investigate variation in adult pigmentation and the effects of maternal thermal environment on offspring reaction norms. We found a low heritability in wing melanisation (h2 = 0.18). Rearing families had contrasted thermal reaction norms. Adult lifespan of males and females from highly plastic families was shorter in individuals exposed to hot developmental temperature. Also, females from plastic families exhibited lower fecundity. We did not find any effect of maternal or grand-maternal developmental temperature on fitness. This study provides new evidence on the influence of phenotypic plasticity on life history-traits’ evolution, a crucial issue in the context of global change.

Sympatric sexual signal divergence among North American Calopteryx damselflies is correlated with increased intra- and interspecific male–male aggression

Divergence of sexual signals in sympatry can arise as a consequence of (1) interspecific competition for resources, (2) selection against maladaptive hybridization, or (3) as a result of selection to reduce the cost of interspecific aggression; termed agonistic character displacement (ACD). Calopterygid damselflies have emerged as a model system for studying the evolution of divergent sexual signals due to the repeated evolution of sympatric species pairs with fully and partially melanized wings. Damselfly wing patterns function during both courtship and territory defense. However, the relative contributions of natural and sexual selection to phenotypic divergence and enhanced isolation in sympatry remain unclear in many cases. Here, we investigated the hypothesis that interference competition, in the form of increased interspecific male–male aggression, drives the evolution of character displacement in sympatry between two species of North American damselflies, Calopteryx aequabilis and Calopteryx maculata, that show no evidence of ecological divergence or ongoing hybridization. In paired behavioral trials, we found that interspecific male aggression related to territory defense varied between site, species, and as a function of the relative abundance of con- vs. hetero-specific males. Specifically, we found that large-spotted C. aequabilis males received increased intra- and interspecific aggression but that aggression against large-spotted males declined during the middle of the flight season when both species were equally abundant. Based on these results, we suggest that ACD leads to enhanced species recognition, and may be a common outcome of the antagonism between interspecific male–male competition and the countervailing force of intraspecific sexual selection favoring increased wing melanization among territorial damselfly species.

Females show greater changes in wing colour with latitude than males in the green-veined white butterfly,Pieris napi(Lepidoptera: Pieridae)

In butterflies, wing colour may simultaneously be under sexual selection in the context of mating selection and natural selection in the context of thermoregulation. In the present study, we collected mated females of the green-veined white butterfly (Pieris napi) from locations spanning 960 km of latitude across Fennoscandia, and investigated sex-specific latitudinal wing colour variation in their offspring raised under identical conditions. We measured wing colour characteristics, including reflectance at wavelengths 300–700 nm and the degree of wing melanization. At all latitudes, females reflected more light in the short wavelengths ( 450 nm), and they were more melanized than males. However, female wing colour varied more with latitude than that of males. Among females, long wavelength reflectance decreased, whereas short wavelength reflectance and melanization increased, towards the north. By contrast, among males, latitudinal variation was found only in the ventral hindwing melanization. These results are consistent with the idea that the balance between natural and sexual selection acting on wing colour changes with latitude differently in males than females. The dark wing colour of females in the north may be a thermoregulatory adaptation, although males may be constrained from evolving the dark dorsal wing colour favoured by natural selection because of constant sexual selection across latitudes. © 2012 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, ••, ••–••.

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.