Dispersal-related life-history trade-offs in a butterfly metapopulation

1. Recent studies on butterflies have documented apparent evolutionary changes in dispersal rate in response to climate change and habitat change. These studies often assume a trade-off between dispersal rate (or flight capacity) and reproduction, which is the rule in wing-dimorphic species but might not occur equally in wing-monomorphic species such as butterflies. 2. To investigate the relationship between dispersal rate and fecundity in the Glanville fritillary butterfly Melitaea cinxia we recorded lifetime individual movements, matings, ovipositions, and maximal life span in a large (32 x 26 m) population cage in the field. Experimental material was obtained from 20 newly established and 20 old local populations within a large metapopulation in the Aland Islands in Finland. 3. Females of the Glanville fritillary from newly established populations are known to be more dispersive in the field, and in the cage they showed significantly greater mobility, mated earlier, and laid more egg clutches than females from old populations. The dispersive females from new populations exhibited no reduced lifetime fecundity in the cage, but they had a shorter maximal life span than old-population females. 4. These results challenge the dispersal-fecundity trade-off for nonmigratory butterflies but instead suggest a physiological trade-off between high metabolic performance and reduced maximal life span. High metabolic performance may explain high rates of dispersal and oviposition in early life. 5. In fragmented landscapes, an ecological trade-off exists between being more dispersive and hence spending more time in the landscape matrix vs. having more time for reproduction in the habitat. We estimate with a dispersal model parameterized for the Glanville fritillary that the lifetime egg production is 4% smaller on average in the more dispersive butterflies in a representative landscape, with much variation depending on landscape structure in the neighbourhood of the natal patch, from--26 to 45% in the landscape analysed in this paper.     

Transcriptome Analysis Reveals Signature of Adaptation to Landscape Fragmentation

We characterize allelic and gene expression variation between populations of the Glanville fritillary butterfly (Melitaea cinxia) from two fragmented and two continuous landscapes in northern Europe. The populations exhibit significant differences in their life history traits, e.g. butterflies from fragmented landscapes have higher flight metabolic rate and dispersal rate in the field, and higher larval growth rate, than butterflies from continuous landscapes. In fragmented landscapes, local populations are small and have a high risk of local extinction, and hence the long-term persistence at the landscape level is based on frequent re-colonization of vacant habitat patches, which is predicted to select for increased dispersal rate. Using RNA-seq data and a common garden experiment, we found that a large number of genes (1,841) were differentially expressed between the landscape types. Hexamerin genes, the expression of which has previously been shown to have high heritability and which correlate strongly with larval development time in the Glanville fritillary, had higher expression in fragmented than continuous landscapes. Genes that were more highly expressed in butterflies from newly-established than old local populations within a fragmented landscape were also more highly expressed, at the landscape level, in fragmented than continuous landscapes. This result suggests that recurrent extinctions and re-colonizations in fragmented landscapes select a for specific expression profile. Genes that were significantly up-regulated following an experimental flight treatment had higher basal expression in fragmented landscapes, indicating that these butterflies are genetically primed for frequent flight. Active flight causes oxidative stress, but butterflies from fragmented landscapes were more tolerant of hypoxia. We conclude that differences in gene expression between the landscape types reflect genomic adaptations to landscape fragmentation.     

Thermal biology of flight in a butterfly: genotype,flight metabolism,and environmental conditions

Knowledge of the effects of thermal conditions on animal movement and dispersal is necessary for a mechanistic understanding of the consequences of climate change and habitat fragmentation. In particular, the flight of ectothermic insects such as small butterflies is greatly influenced by ambient temperature. Here, variation in body temperature during flight is investigated in an ecological model species, the Glanville fritillary butterfly (Melitaea cinxia). Attention is paid on the effects of flight metabolism, genotypes at candidate loci, and environmental conditions. Measurements were made under a natural range of conditions using infrared thermal imaging. Heating of flight muscles by flight metabolism has been presumed to be negligible in small butterflies. However, the results demonstrate that Glanville fritillary males with high flight metabolic rate maintain elevated body temperature better during flight than males with a low rate of flight metabolism. This effect is likely to have a significant influence on the dispersal performance and fitness of butterflies and demonstrates the possible importance of intraspecific physiological variation on dispersal in other similar ectothermic insects. The results also suggest that individuals having an advantage in low ambient temperatures can be susceptible to overheating at high temperatures. Further, tolerance of high temperatures may be important for flight performance, as indicated by an association of heat‐shock protein (Hsp70) genotype with flight metabolic rate and body temperature at takeoff. The dynamics of body temperature at flight and factors affecting it also differed significantly between female and male butterflies, indicating that thermal dynamics are governed by different mechanisms in the two sexes. This study contributes to knowledge about factors affecting intraspecific variation in dispersal‐related thermal performance in butterflies and other insects. Such information is needed for predictive models of the evolution of dispersal in the face of habitat fragmentation and climate change.     

Functional genomics of life history variation in a butterfly metapopulation

In fragmented landscapes, small populations frequently go extinct and new ones are established with poorly understood consequences for genetic diversity and evolution of life history traits. Here, we apply functional genomic tools to an ecological model system, the well-studied metapopulation of the Glanville fritillary butterfly. We investigate how dispersal and colonization select upon existing genetic variation affecting life history traits by comparing common-garden reared 2-day adult females from new populations with those from established older populations. New-population females had higher expression of abdomen genes involved in egg provisioning and thorax genes involved in the maintenance of flight muscle proteins. Physiological studies confirmed that new-population butterflies have accelerated egg maturation, apparently regulated by higher juvenile hormone titer and angiotensin converting enzyme mRNA, as well as enhanced flight metabolism. Gene expression varied between allelic forms of two metabolic genes (Pgi and Sdhd), which themselves were associated with differences in flight metabolic rate, population age and population growth rate. These results identify likely molecular mechanisms underpinning life history variation that is maintained by extinction-colonization dynamics in metapopulations.     

Life history of the Glanville fritillary butterfly in fragmented versus continuous landscapes

Habitat loss and fragmentation threaten the long‐term viability of innumerable species of plants and animals. At the same time, habitat fragmentation may impose strong natural selection and lead to evolution of life histories with possible consequences for demographic dynamics. The Baltic populations of the Glanville fritillary butterfly (Melitaea cinxia) inhabit regions with highly fragmented habitat (networks of small dry meadows) as well as regions with extensive continuous habitat (calcareous alvar grasslands). Here, we report the results of common garden studies on butterflies originating from two highly fragmented landscapes (FL) in Finland and Sweden and from two continuous landscapes (CL) in Sweden and Estonia, conducted in a large outdoor cage (32 by 26 m) and in the laboratory. We investigated a comprehensive set of 51 life‐history traits, including measures of larval growth and development, flight performance, and adult reproductive behavior. Seventeen of the 51 traits showed a significant difference between fragmented versus CL. Most notably, the growth rate of postdiapause larvae and several measures of flight capacity, including flight metabolic rate, were higher in butterflies from fragmented than CL. Females from CL had shorter intervals between consecutive egg clutches and somewhat higher life‐time egg production, but shorter longevity, than females from FL. These results are likely to reflect the constant opportunities for oviposition in females living in continuous habitats, while the more dispersive females from FL allocate more resources to dispersal capacity at the cost of egg maturation rate. This study supports theoretical predictions about small population sizes and high rate of population turnover in fragmented habitats selecting for increased rate of dispersal, but the results also indicate that many other life‐history traits apart from dispersal are affected by the degree of habitat fragmentation.     

Flight metabolic rate has contrasting effects on dispersal in the two sexes of the Glanville fritillary butterfly

Evolution of dispersal is affected by context-specific costs and benefits. One example is sex-biased dispersal in mammals and birds. While many such patterns have been described, the underlying mechanisms are poorly understood. Here, we study genetic and phenotypic traits that affect butterfly flight capacity and examine how these traits are related to dispersal in male and female Glanville fritillary butterflies (Melitaea cinxia). We performed two mark–recapture experiments to examine the associations of individuals’ peak flight metabolic rate (MR_peak) and Pgi genotype with their dispersal in the field. In a third experiment, we studied tethered flight in the laboratory. MR_peak was negatively correlated with dispersal distance in males but the trend was positive in females, and the interaction between MR_peak and sex was significant for long-distance dispersal. A similar but nonsignificant trend was found in relation to molecular variation at Pgi, which encodes a glycolytic enzyme: the genotype associated with high MR_peak tended to be less dispersive in males but more dispersive in females. The same pattern was repeated in the tethered flight experiment: the relationship between MR_peak and flight duration was positive in females but negative in males. These results suggest that females with high flight capacity are superior in among-population dispersal, which facilitates the spatial spreading of their reproductive effort. In contrast, males with high flight capacity may express territorial behaviour, and thereby increase the number of matings, whereas inferior males may be forced to disperse. Thus, flight capacity has opposite associations with dispersal rate in the two sexes.     

Effects of ambient and preceding temperatures and metabolic genes on flight metabolism in the Glanville fritillary butterfly

Flight is essential for foraging, mate searching and dispersal in many insects, but flight metabolism in ectotherms is strongly constrained by temperature. Thermal conditions vary greatly in natural populations and may hence restrict fitness-related activities. Working on the Glanville fritillary butterfly (Melitaea cinxia), we studied the effects of temperature experienced during the first 2 days of adult life on flight metabolism, genetic associations between flight metabolic rate and variation in candidate metabolic genes, and genotype–temperature interactions. The maximal flight performance was reduced by 17% by 2 days of low ambient temperature (15 °C) prior to the flight trial, mimicking conditions that butterflies commonly encounter in nature. A SNP in phosphoglucose isomerase (Pgi) had a significant association on flight metabolic rate in males and a SNP in triosephosphate isomerase (Tpi) was significantly associated with flight metabolic rate in females. In the Pgi SNP, AC heterozygotes had higher flight metabolic rate than AA homozygotes following low preceding temperature, but the trend was reversed following high preceding temperature, consistent with previous results on genotype–temperature interaction for this SNP. We suggest that these results on 2-day old butterflies reflect thermal effect on the maturation of flight muscles. These results highlight the consequences of variation in thermal conditions on the time scale of days, and they contribute to a better understanding of the complex dynamics of flight metabolism and flight-related activities under conditions that are relevant for natural populations living under variable thermal conditions.     

Dispersal in the Glanville fritillary butterfly in fragmented versus continuous landscapes: comparison between three methods

1. Habitat fragmentation may lead to natural selection on dispersal rate and other life‐history traits. Both theoretical analyses and empirical studies suggest that habitat fragmentation may select either for increased or decreased dispersal depending on the traits of the species and the characteristics of the landscape. 2. Dispersal and movement rates in Glanville fritillary butterflies (Melitaea cinxia) originating from a continuous landscape in China and from a highly fragmented landscape in Finland were compared using three different methods. 3. The methods included replicated mark‐release‐recapture (MRR) experiments conducted in the natural environments in China and Finland, tracking with harmonic radar of captive‐reared but free‐flying butterflies in a common environment in the field, and replicated common garden experiments in a large outdoor population cage. 4. The results were largely consistent, showing that butterflies from the more continuous landscape in China had a lower movement rate than butterflies originating from the fragmented landscape in Finland. Butterflies originating from newly‐established populations in Finland moved significantly longer distances than butterflies originating from old populations in Finland or from China, demonstrating significant intra‐specific variation in dispersal rate in Finland. These results are consistent with model predictions for the Glanville fritillary. 5. The tracking experiment revealed a result that would have been impossible to obtain with MRR experiments: movement rate was influenced by a significant interaction between population origin (China vs. Finland) and ambient air temperature.     

On the ecology of the poplar admiral (<Emphasis Type="Italic">Limenitis populi</Emphasis>, Lepidoptera,Nymphalidae) in Eastern Fennoscandia

The ecology of the poplar admiral was studied in the periphery of its range for over 19 years (1990–2008), including seasonal phenomena, spatial distribution, and dynamics of abundance and dispersal of the species. The spatial organization of population of the species was studied in one of the local areas. The flying period of butterflies in Eastern Fennoscandia is divided into three phases: the observed flight of males, latent activity, and observed flight of females. The males emerge more than 10 days earlier than the females. The difference is likely to be one of the reasons for strong reduction in the butterfly abundance. The asynchrony of dynamics in different areas was explained by the influence of local factors. In the years of high abundance, the proportion of occupied areas increased. Analysis of the dispersal and abundance curves showed a trend for colonization of vacant areas. Within the occupied area, the butterflies are irregularly distributed. The presence of areas with constantly moist soils and aspen trees in the forest stands are the main factors responsible for butterfly aggregations. Observations of the marked individuals showed that the maximum distance covered by a butterfly was 4.8 km. The butterfly moved freely across all the study area, and no single accumulation of butterflies isolated from the others was recorded. All the movements are shown to be local. The distance of these movements, according to the exponential model, reached 6–7 km; the grouping itself is classified as a subpopulation, i.e., a structural unit of a large spatially differentiated population. The high mobility of the poplar admiral ensures migration of individuals between the populations and colonization of vacant habitats.     

A candidate locus for variation in dispersal rate in a butterfly metapopulation

Frequent extinctions of local populations in metapopulations create opportunities for migrant females to establish new populations. In a metapopulation of the Glanville fritillary butterfly (Melitaea cinxia), more mobile individuals are more likely to establish new populations, especially in habitat patches that are poorly connected to existing populations. Here we show that flight metabolic rate and the frequency of a specific allele of the metabolic enzyme phosphoglucose isomerase (pgi) were both highest in newly established, isolated populations. Furthermore, genotypes with this pgi allele had elevated flight metabolic rates. These results suggest that genetic variation in pgi or a closely linked locus has a direct effect on flight metabolism, dispersal rate, and thereby on metapopulation dynamics in this species. These results also contribute to an emerging understanding of the mechanisms by which population turnover in heterogeneous landscapes may maintain genetic and phenotypic variation across populations.     

Heritability of flight and resting metabolic rates in the Glanville fritillary butterfly

Dispersal capacity is a key life‐history trait especially in species inhabiting fragmented landscapes. Evolutionary models predict that, given sufficient heritable variation, dispersal rate responds to natural selection imposed by habitat loss and fragmentation. Here, we estimate phenotypic variance components and heritability of flight and resting metabolic rates (RMRs) in an ecological model species, the Glanville fritillary butterfly, in which flight metabolic rate (FMR) is known to correlate strongly with dispersal rate. We modelled a two‐generation pedigree with the animal model to distinguish additive genetic variance from maternal and common environmental effects. The results show that FMR is significantly heritable, with additive genetic variance accounting for about 40% of total phenotypic variance; thus, FMR has the potential to respond to selection on dispersal capacity. Maternal influences on flight metabolism were negligible. Heritability of flight metabolism was context dependent, as in stressful thermal conditions, environmentally induced variation dominated over additive genetic effects. There was no heritability in RMR, which was instead strongly influenced by maternal effects. This study contributes to a mechanistic understanding of the evolution of dispersal‐related traits, a pressing question in view of the challenges posed to many species by changing climate and fragmentation of natural habitats.     

Variation in migration propensity among individuals maintained by landscape structure

Metapopulation dynamics lead to predictable patterns of habitat occupancy, population density and trophic structure in relation to landscape features such as habitat patch size and isolation. Comparable patterns may occur in behavioural, physiological and life‐history traits but remain little studied. In the Glanville fritillary butterfly, females in newly established populations were more mobile than females in old populations. Among females from new populations, mobility decreased with increasing connectivity (decreasing isolation), but in females from old populations mobility increased with connectivity. The [ATP]/[ADP] ratio of flight muscles following controlled activity showed the same pattern as mobility in relation to population age and connectivity, suggesting that physiological differences in flight metabolic performance contribute to the observed variation in mobility. We demonstrate with an evolutionary metapopulation model parameterised for the Glanville fritillary that increasing spatial variation in landscape structure increases variance in mobility among individuals in a metapopulation, supporting the general notion that complex landscape structure maintains life‐history variation.     

Molecular-level variation affects population growth in a butterfly metapopulation

The dynamics of natural populations are thought to be dominated by demographic and environmental processes with little influence of intraspecific genetic variation and natural selection, apart from inbreeding depression possibly reducing population growth in small populations. Here we analyse hundreds of well-characterised local populations in a large metapopulation of the Glanville fritillary butterfly (Melitaea cinxia), which persists in a balance between stochastic local extinctions and recolonisations in a network of 4,000 discrete habitat patches. We show that the allelic composition of the glycolytic enzyme phosphoglucose isomerase (Pgi) has a significant effect on the growth of local populations, consistent with previously reported effects of allelic variation on flight metabolic performance and fecundity in the Glanville fritillary and Colias butterflies. The strength and the sign of the molecular effect on population growth are sensitive to the ecological context (the area and spatial connectivity of the habitat patches), which affects genotype-specific gene flow and the influence of migration on the dynamics of local populations. The biological significance of the results for Pgi is underscored by lack of any association between population growth and allelic variation at six other loci typed in the same material. In demonstrating, to our knowledge for the first time, that molecular variation in a candidate gene affects population growth, this study challenges the perception that differential performance of individual genotypes, leading to differential fitness, is irrelevant to population dynamics. These results also demonstrate that the spatial configuration of habitat and spatial dynamics of populations contribute to maintenance of Pgi polymorphism in this species.     

Population history and life history influence the migration rate of female Glanville fritillary butterflies

This study examines the causes of emigration from small fragments of suitable habitat in a species that has a distinct metapopulation structure, frequent turnover of local populations, and substantial migration among local populations and currently unoccupied habitat fragments. We conducted a field experiment in which 727 individuals of the Glanville fritillary butterfly ( Melitaea cinxia ) originating from four regions were marked and released simultaneously in a natural environment. In three of the four source regions, larvae for the experiment were collected from dozens of small local populations, some of which had been established in the previous summer (new populations), whereas the remaining populations were older. In two of the source regions, female butterflies prefer a host plant ( Veronica spicata ) that is not present in the release area, where there is only Plantago lanceolata , the preferred host plant of females from the other two source regions. We found that migration rate of males was unrelated to any of the factors studied in this experiment. In contrast, two factors influenced the migration rate of females. First, Veronica -preferring females had higher emigration rate than Plantago- preferring females from the Plantago -containing release patches, demonstrating that the individual perception of habitat quality significantly influences the migration rate of females. Second, females from newly-established populations were more dispersive than females from older populations, supporting the notion that metapopulation processes (recurrent colonizations) select for increased migration. The observed migration rate was not correlated with any body size measurements, and thus the observed differences in migration rate were apparently caused by differences in the behaviour of female butterflies rather than in their flight capacity.     

Takeoff temperatures in Melitaea cinxia butterflies from latitudinal and elevational range limits: a potential adaptation to solar irradiance

1. This study provides evidence that a heliophilic butterfly, the Glanville fritillary (Melitaea cinxia) has adapted differently to environmental variation across latitudes and elevations. 2. In cool air, basking M. cinxia orient themselves perpendicular to the sun's rays to gain heat and take off. During flight, solar heating is reduced because orientation perpendicular to the sun is no longer possible and convective cooling occurs. Consequently, M. cinxia have been shown to suffer net heat loss in flight, even in full sunshine. When flight duration is restricted in this way, the takeoff temperature becomes an important thermal adaptation. 3. Using a thermal imaging camera, takeoff temperatures were measured in experimental butterflies. Butterflies from the northern range limit in Finland took flight at slightly hotter temperatures than butterflies from the southern limit in Spain, and much hotter than butterflies from the elevational limit (1900–2300 m) in the French Alps. Butterflies from low‐elevation populations in southern France also took off much hotter than did the nearby Alpine population. 4. These results suggest that the influence of elevation is different from that of latitude in more respects than ambient temperature. Values of solar irradiance in the butterflies' flight season in each region show that insects from the coolest habitats, Finland and the Alps, experienced similar solar irradiance during basking, but that Finns experienced much lower irradiance in flight. This difference may have favored Finnish butterflies evolving higher takeoff temperatures than Alpine butterflies that also flew in cool air but benefited from more intense radiant energy after takeoff.     

Mobility and lifetime fecundity in new versus old populations of the Glanville fritillary butterfly

Life history theory often assumes a trade-off between dispersal and reproduction, and such a trade-off is commonly observed in wing-dimorphic insects. The results are less consistent for wing-monomorphic species, for which it is more difficult to assess dispersal capacity and rate. Three replicate experiments were carried out in consecutive years on the Glanville fritillary butterfly in a large outdoor population cage to study the relationship between lifetime egg production and mobility. The experimental material included females originating from newly-established and old populations, as previous studies have shown dispersal capacity to depend on population age. There was a consistent and significant interaction between mobility and population age, such that in newly-established populations mobile females had higher fecundity than less mobile females, while in old populations there was no such relationship. As selection favours individuals with the highest fecundity, selection pressure on mobility is likely to be different between the two population types, which may contribute to maintenance of variation in dispersal rate in the metapopulation as a whole. Several other female traits also affected lifetime fecundity, including lifespan, number of matings and date of eclosion, although these effects were not consistent across the years. These results highlight the importance of conducting experiments in more than one year before generalizing about patterns in life history variation.     

Heritability of dispersal rate and other life history traits in the Glanville fritillary butterfly

Knowing the variances and heritabilities (h(2)) of life history traits in populations living under natural conditions is necessary for a mechanistic understanding of respective evolutionary processes. I estimated heritabilities of several life history traits, including dispersal rate, body mass, age at first reproduction, egg mass, clutch size and lifetime reproductive success, in the Glanville fritillary butterfly (Melitaea cinxia) using parent-offspring regression. Experiments were conducted under field conditions in a large population cage (32 x 26 m). Heritability estimates ranged from zero to almost one and several were significantly different from zero. Body size for both sexes, female age at first reproduction and egg weight were all moderately to highly heritable, whereas heritabilities were low or non-existent in clutch size and lifetime egg production. Heritability estimates for dispersal rate varied between the sexes, so that dispersal was heritable from mother to her female offspring only. This finding is consistent with previous results showing that the F1 female but not male offspring of females that naturally established new populations in the field are significantly more dispersive than butterflies in old populations.     

Fight or flight? – Flight increases immune gene expression but does not help to fight an infection

Flight represents a key trait in most insects, being energetically extremely demanding, yet often necessary for foraging and reproduction. Additionally, dispersal via flight is especially important for species living in fragmented landscapes. Even though, based on life‐history theory, a negative relationship may be expected between flight and immunity, a number of previous studies have indicated flight to induce an increased immune response. In this study, we assessed whether induced immunity (i.e. immune gene expression) in response to 15‐min forced flight treatment impacts individual survival of bacterial infection in the Glanville fritillary butterfly (Melitaea cinxia). We were able to confirm previous findings of flight‐induced immune gene expression, but still observed substantially stronger effects on both gene expression levels and life span due to bacterial infection compared to flight treatment. Even though gene expression levels of some immunity‐related genes were elevated due to flight, these individuals did not show increased survival of bacterial infection, indicating that flight‐induced immune activation does not completely protect them from the negative effects of bacterial infection. Finally, an interaction between flight and immune treatment indicated a potential trade‐off: flight treatment increased immune gene expression in naïve individuals only, whereas in infected individuals no increase in immune gene expression was induced by flight. Our results suggest that the up‐regulation of immune genes upon flight is based on a general stress response rather than reflecting an adaptive response to cope with potential infections during flight or in new habitats.     

Effects of Increased Flight on the Energetics and Life History of the Butterfly Speyeria mormonia

Movement uses resources that may otherwise be allocated to somatic maintenance or reproduction. How does increased energy expenditure affect resource allocation? Using the butterfly Speyeria mormonia, we tested whether experimentally increased flight affects fecundity, lifespan or flight capacity. We measured body mass (storage), resting metabolic rate and lifespan (repair and maintenance), flight metabolic rate (flight capacity), egg number and composition (reproduction), and food intake across the adult lifespan. The flight treatment did not affect body mass or lifespan. Food intake increased sufficiently to offset the increased energy expenditure. Total egg number did not change, but flown females had higher early-life fecundity and higher egg dry mass than control females. Egg dry mass decreased with age in both treatments. Egg protein, triglyceride or glycogen content did not change with flight or age, but some components tracked egg dry mass. Flight elevated resting metabolic rate, indicating increased maintenance costs. Flight metabolism decreased with age, with a steeper slope for flown females. This may reflect accelerated metabolic senescence from detrimental effects of flight. These effects of a drawdown of nutrients via flight contrast with studies restricting adult nutrient input. There, fecundity was reduced, but flight capacity and lifespan were unchanged. The current study showed that when food resources were abundant, wing-monomorphic butterflies living in a continuous meadow landscape resisted flight-induced stress, exhibiting no evidence of a flight-fecundity or flight-longevity trade-off. Instead, flight changed the dynamics of energy use and reproduction as butterflies adopted a faster lifestyle in early life. High investment in early reproduction may have positive fitness effects in the wild, as long as food is available. Our results help to predict the effect of stressful conditions on the life history of insects living in a changing world.