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Coloration is perhaps one of the most prominent adaptations for survival and reproduction of many taxa. Coloration is of particular importance for aposematic species, which rely on their coloring and patterning acting as a warning signal to deter predators. Most research has focused on the evolution of warning coloration by natural selection. However, little information is available for color mutants of aposematic species, particularly at the genomic level. Here, I compare the transcriptomes of albino mutant caterpillars of the aposematic wood tiger moth (Arctia plantaginis) to those of their full sibs having their distinctive orange‐black warning coloration. The results showed >290 differentially expressed genes genome‐wide. Genes involved in the immune system, structural constituents of cuticular, and immunity were mostly downregulated in the albino caterpillars. Surprisingly, higher expression was observed in core melanin genes from albino caterpillars, suggesting that melanin synthesis may be disrupted in terminal ends of the pathway during its final conversion. Taken together, these results suggest that caterpillar albinism may not be due to a depletion of melanin precursor genes. In contrast, the albino condition may result from the combination of faulty melanin conversion late in its synthesis and structural deficiencies in the cuticular preventing its deposition. The results are discussed in the context of how albinism may impact individuals of aposematic species in the wild. Albinism in warningly colored caterpillars.

The spatial distribution of neutral genetic diversity is mainly influenced by barriers to dispersal. The nature of such barriers varies according to the dispersal means and capabilities of the organisms concerned. Although these barriers are often obvious on land, in the ocean they can be more difficult to identify. Determining the relative influence of physical and biotic factors on genetic connectivity remains a major challenge for marine ecologists. Here, we compare gene flow patterns of 7 littoral fish species from 6 families with a range of early-life-history traits sampled at the same geographic locations across common environmental discontinuities in the form of oceanic fronts in the Western Mediterranean. We show that these fronts represent major barriers to gene flow and have a strong influence on the population genetic structure of some fish species. We also found no significant relation between the early-life-history traits most commonly investigated (egg type, pelagic larval duration, and inshore-offshore spawning) and gene flow patterns, suggesting that other life-history factors should deserve attention. The fronts analyzed and the underlying physical mechanisms are not site-specific but common among the oceans, suggesting the generality of our findings.

Colour is often used as an aposematic warning signal, with predator learning expected to lead to a single colour pattern within a population. However, there are many puzzling cases where aposematic signals are also polymorphic. The wood tiger moth, Arctia plantaginis , displays bright hindwing colours associated with unpalatability, and males have discrete colour morphs which vary in frequency between localities. In Finland, both white and yellow morphs can be found, and these colour morphs also differ in behavioural and life-history traits. Here, we show that male colour is linked to an extra copy of a yellow family gene that is only present in the white morphs. This white-specific duplication, which we name valkea, is highly upregulated during wing development. CRISPR targeting valkea resulted in editing of both valkea and its paralog, yellow-e, and led to the production of yellow wings. We also characterise the pigments responsible for yellow, white, and black colouration, showing that yellow is partly produced by pheomelanins, while black is dopamine-derived eumelanin. Our results add to a growing number of studies on the genetic architecture of complex and seemingly paradoxical polymorphisms, and the role of gene duplications and structural variation in adaptive evolution.

Antibiotics have long been used in the raising of animals for agricultural, industrial or laboratory use. The use of subtherapeutic doses in diets of terrestrial and aquatic animals to promote growth is common and highly debated. Despite their vast application in animal husbandry, knowledge about the mechanisms behind growth promotion is minimal, particularly at the molecular level. Evidence from evolutionary research shows that immunocompetence is resource-limited, and hence expected to trade off with other resource-demanding processes, such as growth. Here, we ask if accelerated growth caused by antibiotics can be explained by genome-wide trade-offs between growth and costly immunocompetence. We explored this idea by injecting broad-spectrum antibiotics into wood tiger moth (Arctia plantaginis) larvae during development. We follow several life-history traits and analyse gene expression (RNA-seq) and bacterial (r16S) profiles. Moths treated with antibiotics show a substantial depletion of bacterial taxa, faster growth rate, a significant downregulation of genes involved in immunity and significant upregulation of growth-related genes. These results suggest that the presence of antibiotics may aid in up-keeping the immune system. Hence, by reducing the resource load of this costly process, bodily resources may be reallocated to other key processes such as growth.

We report the assembly and annotation of the complete mitochondrial genome of the warningly-coloured wood tiger moth (Arctia plantaginis) and investigate its phylogenetic position within Arctiinae. The A.plantaginis mitogenome is 15,479 bp long with 13 protein-coding genes, 22 transfer RNAs, 2 ribosomal RNA genes, and an A + T-rich region (D-loop). The phylogenetic analyses based on 13 protein-coding genes showed A.plantaginis clustering within a clade of species with white wings and yellow or red bodies. This result can be useful in understanding the evolution of coloration in Arctiid moths.

Background Microbes play a role in their host's fundamental ecological, chemical, and physiological processes. Host life-history traits from defence to growth are therefore determined not only by the abiotic environment and genotype but also by microbiota composition. However, the relative importance and interactive effects of these factors may vary between organisms. Such connections remain particularly elusive in Lepidoptera, which have been argued to lack a permanent microbiome and have microbiota primarily determined by their diet and environment. We tested the microbiome specificity and its influence on life-history traits of two colour genotypes of the wood tiger moth ( Arctia plantaginis ) that differ in several traits, including growth. All individuals were grown in the laboratory for several generations with standardized conditions. We analyzed the bacterial community of the genotypes before and after a reciprocal frass (i.e., larval faeces) transplantation and followed growth rate, pupal mass, and the production of defensive secretion. Results After transplantation, the fast-growing genotype grew significantly slower compared to the controls, but the slow-growing genotype did not change its growth rate. The frass transplant also increased the volume of defensive secretions in the fast-growing genotype but did not affect pupal mass. Overall, the fast-growing genotype appeared more susceptible to the transplantation than the slow-growing genotype. Microbiome differences between the genotypes strongly suggest genotype-based selective filtering of bacteria from the diet and environment. A novel cluster of insect-associated Erysipelotrichaceae was exclusive to the fast-growing genotype, and specific Enterococcaceae were characteristic to the slow-growing genotype. These Enterococcaceae became more prevalent in the fast-growing genotype after the transplant, which suggests that a slower growth rate is potentially related to their presence. Conclusions We show that reciprocal frass transplantation can reverse some genotype-specific life-history traits in a lepidopteran host. The results indicate that genotype-specific selective filtering can fine-tune the bacterial community at specific life stages and tissues like the larval frass, even against a background of a highly variable community with stochastic assembly. Altogether, our findings suggest that the host's genotype can influence its susceptibility to being colonized by microbiota, impacting key life-history traits.

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'.

Aim: To investigate the phylogeography of the aposematic wood tiger moth (Parasemia plantaginis) across its Holarctic distribution and to explore how its genetic structure relates to geographical differences in hindwing warning coloration of males and females. Males have polymorphic hindwing coloration, while female hindwing coloration varies continuously, but no geographical analyses of coloration or genetic structure exist. Location: The Holarctic. Methods: We sequenced a fragment of the mitochondrial cytochrome c oxidase subunit I gene (COI) from 587 specimens. We also examined more current population structure by genotyping 569 specimens at 10 nuclear microsatellite loci. Species distribution modelling for present conditions and the Last Glacial Maximum (LGM) was performed to help understand genetic structure. Geographical patterns in hindwing warning coloration were described from 1428 specimens and compared to the genetic analyses. Results: We found only two instances of genetic divergence that coincided with distinct, yet imperfect, shifts in male hindwing coloration in the Caucasus region and Japan. A shift in female hindwing colour did not appear to be associated with genetic structure. A change from sexual monomorphism to sexual dimorphism was also observed. Mitogenetic (mtDNA) structure does not show the influence of glacial refugia during the LGM. Climate shifts following the LGM appear to have isolated the red Caucasus populations and other southerly populations. Populations at opposite ends of the moth's distribution showed high levels of differentiation in the microsatellite data analysis compared to the shallow mitogenetic structure, supporting a more recent divergence. Main conclusions: Parasemia plantaginis populations appeared to have been historically well connected, but current populations are much more differentiated. This raises the possibility that incipient speciation may be occurring in portions of the species' distribution. Some changes in colour align to genetic differences, but others do not, which suggests a role for selective and nonselection based influences on warning signal variation.

Intraspecific competition is a pervasive phenomenon with important ecological and evolutionary consequences, yet its effect in natural populations remains controversial. Although numerous studies suggest that in many cases populations across all organisms are limited by density-dependent processes, this conclusion often relies on correlative data. Here, using an experimental approach, we examined the effect of intraspecific competition on population regulation of the ant Aphaenogaster senilis . In this species females are philopatric while males disperse by flying over relatively long distances. All colonies were removed from 15 experimental plots, except for one focal colony in each plot, while 15 other plots remained unmanipulated. After the first reproductive season, nest density in the experimental plots returned to a level nonsignificantly different from that in the control plots, which was not expected if the populations were indeed regulated by density-independent phenomena. In both the control plots and the experimental plots colonies remained overdispersed throughout the experiment, suggesting colony mutual exclusion. Nests outside the plots rapidly extended their foraging span, but we did not detect any significant inward migration into the experimental plots. Experimental reduction in density did not significantly affect the focal colonies' biomass, measured just before the first reproductive season. However, the ratio of males to workers-pupae biomasses was smaller in experimental plots, suggesting that colonies there had redirected part of the resources normally allocated to male production to the production instead of new workers. Microsatellite analysis indicated that, after the reproductive season, many colonies in the experimental plots were headed by a young queen that was the mother of the brood but not of the old workers, indicating that reduction in colony density stimulated fission of the remaining colonies. Finally, at the end of the experiment, 14 months after experimental reduction in density, colonies that derived from fission were smaller in the experimental than in the control plots, suggesting that the former had undergone fission at a smaller size than in control plots, which presumably allowed them to colonize the emptied areas. We conclude that colonies adjust resource allocation and colony fission to the degree of intraspecific competition.

Polyandry is widespread among many animal taxa, yet the benefits for females are still debated. The two main hypotheses to explain its evolution are the direct benefits and the genetic benefits hypotheses, which are not mutually exclusive. We tested both in the wood tiger moth Arctia plantaginis (Arctiidae) by comparing fitness components in single and multiple mated females. We measured female longevity and number of eggs laid (i.e. direct benefits), as well as offspring hatching success and survival (i.e. genetic benefits). Fitness components did not differ between single and multiple mated females; therefore, there was no evidence to support either direct or genetic benefits hypotheses, or any costs. After paternity analyses, we obtained qualitatively similar results by comparing clutches effectively sired by one male with clutches sired by two males, regardless of the number of times a female mated. We further investigated the proximate mechanisms driving the outcome of paternity patterns. First male precedence, last male precedence, and mixed paternity were present in equal proportions, although there was a trend towards last male sperm precedence in later clutches. Interestingly, in polyandric females, the age of the second male positively affected the number of eggs laid and the number of surviving offspring, indicating an advantage for older males, possibly due to a higher parental investment. We suggest in light of recent theoretical work that the acceptance of more partners in female A. plantaginis may have evolved to ensure fertilization and avoid the risk of virgin death.