Glanville Fritillary Research Articles

Influence of Developmental Conditions on Immune Function and Dispersal-Related Traits in the Glanville Fritillary (Melitaea cinxia) Butterfly

Organisms in the wild are constantly faced with a wide range of environmental variability, such as fluctuation in food availability. Poor nutritional conditions influence life-histories via individual resource allocation patterns, and trade-offs between competing traits. In this study, we assessed the influence of food restriction during development on the energetically expensive traits flight metabolic rate (proxy of dispersal ability), encapsulation rate (proxy of immune defence), and lifespan using the Glanville fritillary butterfly, Melitaea cinxia, as a model organism. Additionally, we examined the direct costs of flight on individual immune function, and whether those costs increase under restricted environmental conditions. We found that nutritional restriction during development enhanced adult encapsulations rate, but reduced both resting and flight metabolic rates. However, at the individual level metabolic rates were not associated with encapsulation rate. Interestingly, individuals that were forced to fly prior to the immune assays had higher encapsulation rates than individuals that had not flown, suggesting that flying itself enhances immune response. Finally, in the control group encapsulation rate correlated positively with lifespan, whereas in the nutritional restriction group there was no relationship between these traits, suggesting that the association between encapsulation rate on adult lifespan was condition-dependent. Thus stressful events during both larval development (food limitation) and adulthood (forced flight) induce increased immune response in the adult butterflies, which may allow individuals to cope with stressful events later on in life.

Life history of theGlanville 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.

Lep-MAP: fast and accurate linkage map construction for large SNP datasets

Current high-throughput sequencing technologies allow cost-efficient genotyping of millions of single nucleotide polymorphisms (SNPs) for hundreds of samples. However, the tools that are currently available for constructing linkage maps are not well suited for large datasets. Linkage maps of large datasets would be helpful in de novo genome assembly by facilitating comprehensive genome validation and refinement by enabling chimeric scaffold detection, as well as in family-based linkage and association studies, quantitative trait locus mapping, analysis of genome synteny and other complex genomic data analyses. We describe a novel tool, called Lepidoptera-MAP (Lep-MAP), for constructing accurate linkage maps with ultradense genome-wide SNP data. Lep-MAP is fast and memory efficient and largely automated, requiring minimal user interaction. It uses simultaneously data on multiple outbred families and can increase linkage map accuracy by taking into account achiasmatic meiosis, a special feature of Lepidoptera and some other taxa with no recombination in one sex (no recombination in females in Lepidoptera). We demonstrate that Lep-MAP outperforms other methods on real and simulated data. We construct a genome-wide linkage map of the Glanville fritillary butterfly (Melitaea cinxia) with over 40 000 SNPs. The data were generated with a novel in-house SOLiD restriction site-associated DNA tag sequencing protocol, which is described in the online supplementary material. Java source code under GNU general public license with the compiled classes and the datasets are available from http://sourceforge.net/users/lep-map.

Long‐term metapopulation study of the Glanville fritillary butterfly (Melitaea cinxia): survey methods, data management, and long‐term population trends

Long-term observational studies conducted at large (regional) spatial scales contribute to better understanding of landscape effects on population and evolutionary dynamics, including the conditions that affect long-term viability of species, but large-scale studies are expensive and logistically challenging to keep running for a long time. Here, we describe the long-term metapopulation study of the Glanville fritillary butterfly (Melitaea cinxia) that has been conducted since 1991 in a large network of 4000 habitat patches (dry meadows) within a study area of 50 by 70 km in the Åland Islands in Finland. We explain how the landscape structure has been described, including definition, delimitation, and mapping of the habitat patches; methods of field survey, including the logistics, cost, and reliability of the survey; and data management using the EarthCape biodiversity platform. We describe the long-term metapopulation dynamics of the Glanville fritillary based on the survey. There has been no long-term change in the overall size of the metapopulation, but the level of spatial synchrony and hence the amplitude of fluctuations in year-to-year metapopulation dynamics have increased over the years, possibly due to increasing frequency of exceptional weather conditions. We discuss the added value of large-scale and long-term population studies, but also emphasize the need to integrate more targeted experimental studies in the context of long-term observational studies. For instance, in the case of the Glanville fritillary project, the long-term study has produced an opportunity to sample individuals for experiments from local populations with a known demographic history. These studies have demonstrated striking differences in dispersal rate and other life-history traits of individuals from newly established local populations (the offspring of colonizers) versus individuals from old, established local populations. The long-term observational study has stimulated the development of metapopulation models and provided an opportunity to test model predictions. This combination of empirical studies and modeling has facilitated the study of key phenomena in spatial dynamics, such as extinction threshold and extinction debt.

Induced defenses of Veronica spicata: Variability in herbivore-induced volatile organic compounds

Plants release volatile organic compounds (VOCs) that have many eco-physiological functions. Induction of plant VOCs is known to occur upon herbivory. Herbivore-induced VOCs are involved in the attraction of predators and parasitoids, a phenomenon known as an indirect defense of plants. We measured the VOC profiles of the wild species Veronica spicata with and without larval feeding and oviposition by the specialist butterfly Melitaea cinxia. V. spicata showed great plasticity when deploying indirect defences. The induction of several ubiquitous terpenoids and green leaf volatiles (GLVs) was associated with larval feeding, whereas the increase of two ketones, 6-methyl-5-hepten-2-one and t-geranylacetone and the suppression of GLVs were associated with oviposition by the butterfly.

Molecular cloning and expression in vitro of a carboxylesterase gene from the Glanville fritillary butterfly (Melitaea cinxia)

Carboxylesterase (EC 3.1.1.1) is a member of the carboxyl/cholinesterase (CCE) superfamily, which is widely distributed in animals, plants and microorganisms. This enzyme has been known to be associated with insecticide resistance and detoxification. Although CCEs have been extensively studied in insects, including lepidopterans, the research on butterflies, a major subgroup in Lepidoptera, is still poor. In the present study, we cloned a CCE gene (McCCE1) from the Glanville fritillary butterfly (Melitaea cinxia, Lepidoptera: Nymphalidae). The full-length cDNA encoding McCCE1 was 1786bp, containing a 1641bp open reading frame encoding 546 amino acids, a 38bp 5′-untranslated region (5′-UTR), and a 107bp 3′-UTR with a poly(A) tail. The functionally conserved amino acids in McCCE1 shared the 55% identity with the cytoplasmic esterase CCE017a in Helicoverpa armigera (Lepidoptera: Noctuidae), which has been associated with detoxification. Assays in vitro showed that the recombinant McCCE1 could hydrolyze α- and β-naphthyl acetate. Thus, the present study adds to the body of knowledge concerning the detoxification of pesticides by lepidopterans.

The effect of multi‐species host density on superparasitism and sex ratio in a gregarious parasitoid

Abstract The ecological factors that influence key life‐history traits such as brood size and sex ratio are enormously important to the survival and population dynamics of insect species. The effects of host density on life‐history characteristics of a parasitoid, Pteromalus apum, were examined in a field experiment conducted in the Åland Islands, Finland. This gregarious parasitoid preys on two co‐occurring butterfly species, Melitaea cinxia and Melitaea athalia. The abundances of both butterfly species contribute to host density for the parasitoid. The goals of the study were to: (i) test sex allocation theory in a field setting by evaluating how host density and species affect parasitoid brood size and sex ratio; and (ii) understand how parasitoid foraging behaviour and co‐occurrence of multiple host species relate to life‐history traits of the parasitoid. Local host density and species were experimentally manipulated and natural parasitism was then allowed to occur. Larger brood sizes were found at low host density due to higher rates of superparasitism. Further, parasitoid brood size and sex ratio (proportion male progeny) were positively related, with a much stronger effect at low host density. These results illustrate that host density affects life‐history and population‐level traits of parasitoids. The fact that low host density was associated with high superparasitism (larger brood size) supports predictions based on the ‘apparent commensal’ indirect interaction between the host species. This work highlights the value of research that integrates interactions between species with the study of foraging behaviour and life‐history traits.

Plastic larval development in a butterfly has complex environmental and genetic causes and consequences for population dynamics

1. In insects, the length of larval development time typically influences adult body size and individual fitness, and hence development time can be expected to respond in an adaptive manner to variation in environmental conditions. In the wild, larval growth may be influenced by individual condition, which can be affected by population-level parameters such as population density and abundance and quality of resources. 2. We sampled larvae of the Glanville fritillary butterfly (Melitaea cinxia) from 514 local populations across a large metapopulation before the winter diapause and reared the larvae in common garden conditions after diapause. Here, we report that small post-diapause larvae prolonged their development via an extra larval instar, apparently to compensate for their 'bad start' after diapause. The number of instars was additionally a plastic response to environmental conditions, as the frequency of the extra instar increased under cooler thermal conditions. 3. The benefit of the extra instar is clear, as it allows individuals to develop into larger adults, but the cost is delayed adult eclosion, which is likely to select against the extra instar especially in males, in which early eclosion is critical for mating success. In support of this, the frequency of the extra instar was significantly lower in males (7%) than in females (42%). 4. Polymorphisms in three genes, serpin-1, vitellin-degrading protease precursor and phosphoglucose isomerase, which are known to influence development in insects, were associated with the occurrence of the extra instar. 5. At the level of local populations, the frequency of the extra instar was higher in newly established populations than that in old local ones, possibly reflecting maternal effects, as new populations are often established by females with heavy investment in dispersal. The frequency of the extra instar in turn correlated with the change in population size over 1 year and the risk of local extinction in the natural metapopulation of the Glanville fritillary. 6. Our results highlight the importance of the physiological condition of individuals in shaping subsequent life-history events and even population dynamics.

Evolutionary Ecological Genomics

The recognition that evolution can happen on ecological timescales (Hairston et al. 2005; Pelletier et al. 2009; Ellner et al. 2011; Becks et al. 2012) has prompted the integration of ecology and evolution, while easier access to high-throughput sequencing technologies has increased the number of genetic nonmodel species entering the ‘omics’ era (e.g. Turner et al. 2010; Colbourne et al. 2011; Jones et al. 2012). We are now in a position to identify the genetic basis of adaptation and the mechanisms of adaptive responses in the wild. It nonetheless remains a challenge to go beyond descriptive measures of patterns of genetic variation and to identify the evolutionary processes driving species adaptation and evolution. This special issue represents a broad cross-section of research into evolutionary adaptation at the genetic level. The approaches used vary from classic QTL studies to RAD sequencing and RNAseq. Given the rapid advance of sequencing technology, we fully expect that ‘genomic’ as defined here will be merely ‘genetic’ in a few years, but we nonetheless hope that the results and methods described in this special issue will serve as a blueprint for future work in this field.

GENETIC VARIATION IN HIF SIGNALING UNDERLIES QUANTITATIVE VARIATION IN PHYSIOLOGICAL AND LIFE-HISTORY TRAITS WITHIN LOWLAND BUTTERFLY POPULATIONS

Oxygen conductance to the tissues determines aerobic metabolic performance in most eukaryotes but has cost/benefit tradeoffs. Here we examine in lowland populations of a butterfly a genetic polymorphism affecting oxygen conductance via the hypoxia-inducible factor (HIF) pathway, which senses intracellular oxygen and controls the development of oxygen delivery networks. Genetically distinct clades of Glanville fritillary (Melitaea cinxia) across a continental scale maintain, at intermediate frequencies, alleles in a metabolic enzyme (succinate dehydrogenase, SDH) that regulates HIF-1α. One Sdhd allele was associated with reduced SDH activity rate, twofold greater cross-sectional area of tracheoles in flight muscle, and better flight performance. Butterflies with less tracheal development had greater post-flight hypoxia signaling, swollen & disrupted mitochondria, and accelerated aging of flight metabolic performance. Allelic associations with metabolic and aging phenotypes were replicated in samples from different clades. Experimentally elevated succinate in pupae increased the abundance of HIF-1α and expression of genes responsive to HIF activation, including tracheal morphogenesis genes. These results indicate that the hypoxia inducible pathway, even in lowland populations, can be an important axis for genetic variation underlying intraspecific differences in oxygen delivery, physiological performance, and life history.

Variation in a Host-Parasitoid Interaction across Independent Populations.

Antagonistic relationships between parasitoids and their insect hosts involve multiple traits and are shaped by their ecological and evolutionary context. The parasitoid wasp Cotesia melitaearum and its host butterfly Melitaea cinxia occur in several locations around the Baltic sea, with differences in landscape structure, population sizes and the histories of the populations. We compared the virulence of the parasitoid and the susceptibility of the host from five populations in a reciprocal transplant-style experiment using the progeny of five independent host and parasitoid individuals from each population. The host populations showed significant differences in the rate of encapsulation and parasitoid development rate. The parasitoid populations differed in brood size, development rate, pupal size and adult longevity. Some trait differences depended on specific host-parasitoid combinations, but neither species performed systematically better or worse in experiments involving local versus non-local populations of the other species. Furthermore, individuals from host populations with the most recent common ancestry did not perform alike, and there was no negative effect due to a history of inbreeding in the parasitoid. The complex pattern of variation in the traits related to the vulnerability of the host and the ability of the parasitoid to exploit the host may reflect multiple functions of the traits that would hinder simple local adaptation.

High genetic load in an old isolated butterfly population

We investigated inbreeding depression and genetic load in a small (N(e) ∼ 100) population of the Glanville fritillary butterfly (Melitaea cinxia), which has been completely isolated on a small island [Pikku Tytärsaari (PT)] in the Baltic Sea for at least 75 y. As a reference, we studied conspecific populations from the well-studied metapopulation in the Åland Islands (ÅL), 400 km away. A large population in Saaremaa, Estonia, was used as a reference for estimating genetic diversity and N(e). We investigated 58 traits related to behavior, development, morphology, reproductive performance, and metabolism. The PT population exhibited high genetic load (L = 1 - W(PT)/W(ÅL)) in a range of fitness-related traits including adult weight (L = 0.12), flight metabolic rate (L = 0.53), egg viability (L = 0.37), and lifetime production of eggs in an outdoor population cage (L = 0.70). These results imply extensive fixation of deleterious recessive mutations, supported by greatly reduced diversity in microsatellite markers and immediate recovery (heterosis) of egg viability and flight metabolic rate in crosses with other populations. There was no significant inbreeding depression in most traits due to one generation of full-sib mating. Resting metabolic rate was significantly elevated in PT males, which may be related to their short lifespan (L = 0.25). The demographic history and the effective size of the PT population place it in the part of the parameter space in which models predict mutation accumulation. This population exemplifies the increasingly common situation in fragmented landscapes, in which small and completely isolated populations are vulnerable to extinction due to high genetic load.

Direct and trans-generational responses to food deprivation during development in the Glanville fritillary butterfly

Life history characteristics and resulting fitness consequences manifest not only in an individual experiencing environmental conditions but also in its offspring via trans-generational effects. We conducted a set of experiments to assess the direct and trans-generational effects of food deprivation in the Glanville fritillary butterfly Melitaea cinxia. Food availability was manipulated during the final stages of larval development and performance was assessed during two generations. Direct responses to food deprivation were relatively minor. Food-deprived individuals compensated, via increased development time, to reach a similar mass as adults from the control group. Delayed costs of compensatory growth were observed, as food-deprived individuals had either reduced fecundity or lifespan depending on the type of feeding treatment they had experienced (intermittent vs. continuous). Female food deprivation did not directly affect her offspring's developmental trajectory, but the way the offspring coped with food deprivation. Offspring of mothers from control or intermittent starvation treatments reached the size of those in the control group via increased development time when being starved. In contrast, offspring of mothers that had experienced 2 days of continuous food deprivation grew even larger than control animals, when deprived of food themselves. Offspring of food-deprived Glanville fritillary initially showed poor immune response to parasitism, but not later on in development.

Impact of grazing management on hibernating caterpillars of the butterfly Melitaea cinxia in calcareous grasslands

Semi-natural grasslands are increasingly grazed by large herbivores for nature conservation purposes. For many insects such grazing is essential for the conservation of their habitat, but at the same time, populations decrease at high grazing intensity. We hypothesised that grazing management may cause increased butterfly mortality, especially for life-stages with low mobility, such as hibernating caterpillars. To test this, we measured the effect of sheep grazing on overwinter larval survival. We used the Glanville fritillary (Melitaea cinxia), which has gregarious caterpillars hibernating in silk nests, as a model species. Caterpillar nests were monitored throughout the hibernating period in calcareous grassland reserves with low and high intensity sheep grazing and in an ungrazed control treatment. After grazing, 64 % of the nests at the high intensity grazing treatment were damaged or missing, compared to 8 and 12 % at the ungrazed and low intensity grazing treatment, respectively. Nest volume and caterpillar survival were 50 % lower at the high intensity grazing treatment compared to both ungrazed and low intensity grazing treatments. Nest damage and increased mortality were mainly caused by incidental ingestion of the caterpillars by the sheep. It is likely that grazing similarly affects other invertebrates, depending on their location within the vegetation and their ability to actively avoid herbivores. This implies that the impact of grazing strongly depends on the timing of this management in relation to the phenology of the species. A greater focus on immature and inactive life-stages in conservation policy in general and particularly in action plans for endangered species is required to effectively preserve invertebrate diversity.

Temperature treatments during larval development reveal extensive heritable and plastic variation in gene expression and life history traits

Little is known about variation in gene expression that affects life history traits in wild populations of outcrossing species. Here, we analyse heritability of larval development traits and associated variation in gene expression in the Glanville fritillary butterfly (Melitaea cinxia) across three ecologically relevant temperatures. We studied the development of final-instar larvae, which is greatly affected by temperature, and during which stage larvae build up most of the resources for adult life. Larval development time and weight gain varied significantly among families sampled from hundreds of local populations, indicating substantial heritable variation segregating in the large metapopulation. Global gene expression analysis using common garden-reared F2 families revealed that 42% of the >8000 genes surveyed exhibited significant variation among families, 39% of the genes showed significant variation between the temperature treatments, and 18% showed a significant genotype-by-environment interaction. Genes with large family and temperature effects included larval serum protein and cuticle-binding protein genes, and the expression of these genes was closely correlated with the rate of larval development. Significant expression variation in these same categories of genes has previously been reported among adult butterflies originating from newly established versus old local populations, supporting the notion of a life history syndrome put forward based on ecological studies and involving larval development and adult dispersal capacity. These findings suggest that metapopulation dynamics in heterogeneous environments maintain heritable gene expression variation that affects the regulation of life history traits.

Diet Quality Can Play a Critical Role in Defense Efficacy against Parasitoids and Pathogens in the Glanville Fritillary (Melitaea cinxia)

Numerous herbivorous insect species sequester noxious chemicals from host plants that effectively defend against predators, and against parasitoids and pathogens. Sequestration of these chemicals may be expensive and involve a trade off with other fitness traits. Here, we tested this hypothesis. We reared Glanville fritillary butterfly (Melitaea cinxia L.) larvae on plant diets containing low- and high-levels of iridoid glycosides (IGs) (mainly aucubin and catalpol) and tested: 1) whether IGs affect the herbivore's defense against parasitoids (measured as encapsulation rate) and bacterial pathogens (measured as herbivore survival); 2) whether parasitoid and bacterial defenses interact; and 3) whether sequestration of the plant's defense chemicals incurs any life history costs. Encapsulation rates were stronger when there were higher percentages of catalpol in the diet. Implanted individuals had greater amounts of IGs in their bodies as adults. This suggests that parasitized individuals may sequester more IGs, increase their feeding rate after parasitism, or that there is a trade off between detoxification efficiency and encapsulation rate. Larval survival after bacterial infection was influenced by diet, but probably not by diet IG content, as changes in survival did not correlate linearly with the levels of IGs in the diet. However, M. cinxia larvae with good encapsulation abilities were better defended against bacteria. We did not find any life history costs of diet IG concentration for larvae. These results suggest that the sequestering of plant defense chemicals can help herbivorous insects to defend against parasitoids.

A long life in the fast lane: positive association between peak metabolic rate and lifespan in a butterfly

High peak metabolic rates may provide a performance advantage, but it may also entail a physiological cost. A long-held assumption is that high mass-specific energy expenditure is associated with short lifespan. To examine the relationship between energy expenditure and lifespan we asked two questions. First, do individuals have a consistent rate of metabolism throughout their life? Second, is metabolic rate correlated with lifespan? We analysed the repeatability of measurements of resting (RMR) and peak flight metabolic rate (MR(peak)) throughout the life of the Glanville fritillary butterfly (Melitaea cinxia). Measurements of MR(peak) showed significant repeatability. Senescence occurred only shortly before death. RMR showed a U-shaped relationship with age and very low repeatability. Intraspecific association between metabolic rates and lifespan was tested under three conditions: in the laboratory, under field conditions and in a laboratory experiment with repeated flight treatments. There was a significant correlation between MR(peak) and lifespan in all three experiments, but the correlation was positive, not negative. RMR was not correlated with lifespan. Both MR(peak) and lifespan may reflect physiological condition and therefore be positively correlated. Individuals with a large resource pool may be able to invest in mechanisms that slow down ageing. Individuals with high metabolic capacity may also possess adaptations against ageing. Molecular polymorphism in the gene phosphoglucose isomerase (Pgi) was significantly associated with both MR(peak) and lifespan, and may have coevolved with defence mechanisms against senescence. Generalisations such as 'live fast, die young' may be too simple to explain the complex processes affecting ageing and lifespan.

Indirect interaction between butterfly species mediated by a shared pupal parasitoid

AbstractIndirect interactions among species can greatly affect their abundances and the structure of the community they live in. Using a field experiment, we tested the hypothesis that congeneric butterfly species interact indirectly through a shared pupal parasitoid. We predicted that symmetrical apparent competition would lead to high parasitism of both species, and the effect would increase with regional butterfly abundance. Instead, parasitism of one host, Melitaea cinxia, was reduced in the presence of the second host, M. athalia. Parasitism of M. athalia did not differ whether or not M. cinxia was present. This pattern did not vary with regional butterfly abundance, though overall rate of parasitism did. Details of the experiment suggest that the apparent commensalism occurred because M. cinxia pupae are protected by silk tents whereas M. athalia are exposed, causing locally foraging parasitoids to favour the more accessible host where the two are present together. The local short‐term apparent commensalism favouring M. cinxia opposes the landscape scale trend, in which parasitism increases where butterfly density is high. The outcome of this study illustrates short‐term apparent commensalism, that host suitability can depend on relative accessibility, and that indirect interactions occurring at different scales may be in opposition.

Bayesian state-space modeling of metapopulation dynamics in the Glanville fritillary butterfly

The complexity of mathematical models of ecological dynamics varies greatly, and it is often difficult to judge what would be the optimal level of complexity in a particular case. Here we compare the parameter estimates, model fits, and predictive abilities of two models of metapopulation dynamics: a detailed individual-based model (IBM) and a population-based stochastic patch occupancy model (SPOM) derived from the IBM. The two models were fitted to a 17-year time series of data for the Glanville fritillary butterfly (Melitaea cinxia) inhabiting a network of 72 small meadows. The data consisted of biannual counts of larval groups (IBM) and the annual presence or absence of local populations (SPOM). The models were fitted using a Bayesian state-space approach with a hierarchical random effect structure to account for observational, demographic, and environmental stochasticities. The detection probability of larval groups (IBM) and the probability of false zeros of local populations (SPOM) in the observati...

Differential Performance of a Specialist and Two Generalist Herbivores and Their Parasitoids on Plantago lanceolata

The ability to cope with plant defense chemicals differs between specialist and generalist species. In this study, we examined the effects of the concentration of the two main iridoid glycosides (IGs) in Plantago lanceolata, aucubin and catalpol, on the performance of a specialist and two generalist herbivores and their respective endoparasitoids. Development of the specialist herbivore Melitaea cinxia was unaffected by the total leaf IG concentration in its host plant. By contrast, the generalist herbivores Spodoptera exigua and Chrysodeixis chalcites showed delayed larval and pupal development on plant genotypes with high leaf IG concentrations, respectively. This result is in line with the idea that specialist herbivores are better adapted to allelochemicals in host plants on which they are specialized. Melitaea cinxia experienced less post-diapause larval and pupal mortality on its local Finnish P. lanceolata than on Dutch genotypes. This could not be explained by differences in IG profiles, suggesting that M. cinxia has adapted in response to attributes of its local host plants other than to IG chemistry. Development of the specialist parasitoid Cotesia melitaearum was unaffected by IG variation in the diet of its host M. cinxia, a response that was concordant with that of its host. By contrast, the development time responses of the generalist parasitoids Hyposoter didymator and Cotesia marginiventris differed from those of their generalist hosts, S. exigua and C. chalcites. While their hosts developed slowly on high-IG genotypes, development time of H. didymator was unaffected. Cotesia marginiventris actually developed faster on hosts fed high-IG genotypes, although they then had short adult longevity. The faster development of C. marginiventris on hosts that ate high-IG genotypes is in line with the “immunocompromized host” hypothesis, emphasizing the potential negative effects of toxic allelochemicals on the host’s immune response.