Parasitoid wasps may well be the most species-rich animal group on Earth, and host–parasitoid interactions may thereby be one of the most common types of species interactions. Understanding the major mechanisms underlying diversification in parasitoids should be a high priority, not the least in order to predict consequences from high extinction rates currently observed. The two major hypotheses explaining host-associated diversification are the escape-and-radiate hypothesis and the oscillation hypothesis, where the former assumes that key innovations are major drivers of radiation bursts, whereas the latter rather assumes that diversification depends on processes acting on the standing genetic variation that influences host use. This paper reviews the recent literature on parasitoid speciation in light of these major hypotheses to identify potential key innovations and host use variability underlying diversification. The paper also calls upon recent theoretical advances from a similar system, plant–butterfly interactions, to provide shortcuts in the development of theories explaining the high diversity of parasitoid wasps.

Generalist plant-feeding insects are characterised by a broad host repertoire that can comprise several families or even different orders of plants. The genetic and physiological mechanisms underlying the use of such a wide host range are still not fully understood. Earlier studies indicate that the consumption of different host plants is associated with host-specific gene expression profiles. It remained, however, unclear if and how larvae can alter these profiles in the case of a changing host environment. Using the polyphagous comma butterfly (Polygonia c-album) we show that larvae can adjust their transcriptional profiles in response to a new host plant. The switch to some of the host plants, however, resulted in a larger transcriptional response and, thus, seems to be more challenging. At a physiological level, no correspondence for these patterns could be found in larval performance. This suggests that a high transcriptional but also phenotypic flexibility are essential for the use of a broad and diverse host range. We furthermore propose that host switch tests in the laboratory followed by transcriptomic investigations can be a valuable tool to examine not only plasticity in host use but also subtle and/or transient trade-offs in the evolution of host plant repertoires.

In this study, we investigated whether patterns of gene expression in larvae feeding on different plants can explain important aspects of the evolution of insect-plant associations, such as phylogenetic conservatism of host use and re-colonization of ancestral hosts that have been lost from the host repertoire. To this end, we performed a phylogenetically informed study comparing the transcriptomes of 4 nymphalid butterfly species in Polygonia and the closely related genus Nymphalis. Larvae were reared on Urtica dioica, Salix spp., and Ribes spp. Plant-specific gene expression was found to be similar across butterfly species, even in the case of host plants that are no longer used by two of the butterfly species. These results suggest that plant-specific transcriptomes can be robust over evolutionary time. We propose that adaptations to particular larval food plants can profitably be understood as an evolved set of modules of co-expressed genes, promoting conservatism in host use and facilitating re-colonization. Moreover, we speculate that the degree of overlap between plant-specific transcriptomes may correlate with the strength of trade-offs between plants as resources and hence to the probability of colonizing hosts and complete host shifts.

1. One of the main challenges faced by ecologists today is to understand and predict how species interactions will respond to the current environmental change. It is likely that these changes will have a stronger effect on phylogenetic lineages that depend on intimate and specialised ecological interactions, such as most herbivorous insects.

2. In this review, we highlight the aspects that we consider are fundamental for understanding how species interactions change over time. We start by reviewing terminology and conclude that commonly used terms have undesired connotations with regard to what we know about how hosts are acquired and lost over time.

3. Based on that, we suggest host repertoire as a better term to describe the use of multiple hosts than the host range or diet breadth, both of which fail to capture that host use is often non-contiguous and tend to emphasise the total number of hosts, while ignoring host identity.

4. Another important issue that we highlight is the differentiation between fundamental and realised host repertoires, where the latter is the set of hosts that are actually used in nature, whereas the first includes all hosts that can potentially be used by the species. We show that the characterisation of the fundamental host repertoire is key to understanding how insect-plant interactions will respond to the ecological opportunities instigated by environmental change.

The study of herbivorous insects underpins much of the theory that concerns the evolution of species interactions. In particular, Pieridae butterflies and their host plants have served as a model system for studying evolutionary arms races. To learn more about the coevolution of these two clades, we reconstructed ancestral ecological networks using stochastic mappings that were generated by a phylogenetic model of host-repertoire evolution. We then measured if, when, and how two ecologically important structural features of the ancestral networks (modularity and nestedness) evolved over time. Our study shows that as pierids gained new hosts and formed new modules, a subset of them retained or recolonised the ancestral host(s), preserving connectivity to the original modules. Together, host-range expansions and recolonisations promoted a phase transition in network structure. Our results demonstrate the power of combining network analysis with Bayesian inference of host-repertoire evolution to understand changes in complex species interactions over time.

Climate and land use change can alter the incidence and strength of biotic interactions, with important effects on the distribution, abundance and function of species. To assess the importance of these effects and their dynamics, studies quantifying how biotic interactions change in space and time are needed. We studied interactions between nettle-feeding butterflies and their shared natural enemies (parasitoids) locally and across 500 km latitudinal gradient in Sweden. We also examined the potential impact of the range-expansion of the butterfly Araschnia levana on resident butterflies via shared parasitoids, by studying how parasitism in resident butterflies covaries with the presence or absence of the newly-established species. We collected 6777 larvae of four nettle-feeding butterfly species (Aglais urticae, Aglais io, Ar. levana and Vanessa atalanta), over two years, at 19 sites distributed along the gradient. We documented the parasitoid complex for each butterfly species and measured their overlap, and analysed how parasitism rates were affected by butterfly species assemblage, variations in abundance, time, and the arrival of Ar. levana. Parasitoids caused high mortality, with substantial overlap in the complex of parasitoids associated with the four host butterflies. Levels of parasitism differed significantly among butterflies and were influenced by the local butterfly species assemblage. Our results also suggest that parasitism in resident butterflies is elevated at sites where Ar. levana has been established for a longer period. In our study system, variations in butterfly species assemblages were associated in a predictable way with substantial variations in rates of parasitism. This relationship is likely to affect the dynamics of the butterfly host species, and potentially cascade to the larger number of species with which they interact. These results highlight the importance of indirect interactions and their potential to reorganise ecological communities, especially in the context of shifts in species distributions in a warmer world.

Recent advances in obtaining reduced representation libraries for next-generation sequencing permit phylogenomic analysis of species-rich, recently diverged taxa. In this study, we performed sequence capture with homemade PCR-generated probes to study diversification among closely related species in a large insect genus to examine the utility of this method. We reconstructed the phylogeny of Neptis Fabricius, a large and poorly studied nymphalid butterfly genus distributed throughout the Old World. We inferred relationships among 108 Neptis samples using 89 loci totaling up to 84 519 bp per specimen. Our taxon sample focused on Palearctic, Oriental and Australasian species, but included 8 African species and outgroups from 5 related genera. Maximum likelihood and Bayesian analyses yielded identical trees with full support for almost all nodes. We confirmed that Neptis is not monophyletic because Lasippa heliodore (Fabricius) and Phaedyma amphion (Linnaeus) are nested within the genus, and we redefine species groups for Neptis found outside of Africa. The statistical support of our results demonstrates that the probe set we employed is useful for inferring phylogenetic relationships among Neptis species and likely has great value for intrageneric phylogenetic reconstruction of Lepidoptera. Based on our results, we revise the following two taxa: Neptis heliodore comb. rev. and Neptis amphion comb. rev.

Intimate ecological interactions, such as those between parasites and their hosts, may persist over long time spans, coupling the evolutionary histories of the lineages involved. Most methods that reconstruct the coevolutionary history of such interactions make the simplifying assumption that parasites have a single host. Many methods also focus on congruence between host and parasite phylogenies, using cospeciation as the null model. However, there is an increasing body of evidence suggesting that the host ranges of parasites are more complex: that host ranges often include more than one host and evolve via gains and losses of hosts rather than through cospeciation alone. Here, we develop a Bayesian approach for inferring coevolutionary history based on a model accommodating these complexities. Specifically, a parasite is assumed to have a host repertoire, which includes both potential hosts and one or more actual hosts. Over time, potential hosts can be added or lost, and potential hosts can develop into actual hosts or vice versa. Thus, host colonization is modeled as a two-step process that may potentially be influenced by host relatedness. We first explore the statistical behavior of our model by simulating evolution of host-parasite interactions under a range of parameter values. We then use our approach, implemented in the program RevBayes, to infer the coevolutionary history between 34 Nymphalini butterfly species and 25 angiosperm families. Our analysis suggests that host relatedness among angiosperm families influences how easily Nymphalini lineages gain new hosts.

Many insects that live in temperate zones spend the cold season in a state of dormancy, referred to as diapause. As the insect must rely on resources that were gathered before entering diapause, keeping a low metabolic rate is of utmost importance. Organs that are metabolically expensive to maintain, such as the brain, can therefore become a liability to survival if they are too large. Insects that go through diapause as adults generally do so before entering the season of reproduction. This order of events introduces a conflict between maintaining low metabolism during dormancy and emerging afterward with highly developed sensory systems that improve fitness during the mating season. We investigated the timing of when investments into the olfactory system are made by measuring the volumes of primary and secondary olfactory neuropils in the brain as they fluctuate in size throughout the extended diapause life-period of adult Polygonia c-album butterflies. Relative volumes of both olfactory neuropils increase significantly during early adult development, indicating the importance of olfaction to this species, but still remain considerably smaller than those of nondiapausing conspecifics. However, despite butterflies being kept under the same conditions as before the dormancy, their olfactory neuropil volumes decreased significantly during the postdormancy period. The opposing directions of change in relative neuropil volumes before and after diapause dormancy indicate that the investment strategies governing structural plasticity during the two life stages could be functionally distinct. As butterflies were kept in stimulus-poor conditions, we find it likely that investments into these brain regions rely on experience-expectant processes before diapause and experience-dependent processes after diapause conditions are broken. As the shift in investment strategies coincides with a hard shift from premating season to mating season, we argue that these developmental characteristics could be adaptations that mitigate the trade-off between dormancy survival and reproductive fitness.

We show that females of 2 generalist butterflies improve their search efficacy after previous encounters of the same host in a way similar to search-image formation, especially if the butterfly-host relationship is historically old. Thus, by targeting a single host at a time, host search efficacy may be improved and constitute a selection pressure for specialization. This result can help explain the evolutionary trend toward host specialization in phytophagous insects that is not well understood. Abstract In phytophagous insects such as butterflies, there is an evolutionary trend toward specialization in host plant use. One contributing mechanism for this pattern may be found in female host search behavior. Since search attention is limited, generalist females searching for hosts for oviposition may potentially increase their search efficacy by aiming their attention on a single host species at a time, a behavior consistent with search image formation. Using laboratory reared and mated females of 2 species of generalist butterflies, the comma, Polygonia c-album, and the painted lady, Vanessa cardui (Lepidoptera: Nymphalidae), we investigated the probability of finding a specific target host (among nonhost distractors) immediately after being primed with an oviposition experience of the same host as compared with different host in indoor cages. We used species-specific host plants that varied with respect to growth form, historical age of the butterfly-host association, and relative preference ranking. We found improved search efficacy after previous encounters of the same host for some but not all host species. Positive priming effects were found only in hosts with which the butterfly has a historically old relationship and these hosts are sometimes also highly preferred. Our findings provides additional support for the importance of behavioral factors in shaping the host range of phytophagous insects, and show that butterflies can attune their search behavior to compensate for negative effects of divided attention between multiple hosts.