The geography of speciation in butterflies (Lepidoptera,Papilionoidea): an empirical test of theoretical models

Despite the fundamental role of speciation in formation of biodiversity, the genetic and ecological mechanisms related to this process, as well as the geography of speciation are still poorly known. In our research we have used methods of molecular phylogenetics and phylogeography to reconstruct the stages of speciation in two model groups of butterflies. Phylogeographic analysis showed that speciation in Agrodiaetus blues started in allopatry. An additional age-range correlation test also revealed a pattern consistent with allopatric speciation. However, the formation of new wing colors, the characters most important for maintenance of pre-zygotic reproductive isolation, was shown to occur after transition from allopatry to sympatry. Analysis of karyotypes, mitochondrial and nuclear molecular markers in the Wood White butterfly Leptidea sinapis L. showed that clinal speciation may have occurred in this case; this process is theoretically possible but difficult to document.     

Reproductive isolation and patterns of genetic differentiation in a cryptic butterfly species complex

Molecular studies of natural populations are often designed to detect and categorize hidden layers of cryptic diversity, and an emerging pattern suggests that cryptic species are more common and more widely distributed than previously thought. However, these studies are often decoupled from ecological and behavioural studies of species divergence. Thus, the mechanisms by which the cryptic diversity is distributed and maintained across large spatial scales are often unknown. In 1988, it was discovered that the common Eurasian Wood White butterfly consisted of two species (Leptidea sinapis and Leptidea reali), and the pair became an emerging model for the study of speciation and chromosomal evolution. In 2011, the existence of a third cryptic species (Leptidea juvernica) was proposed. This unexpected discovery raises questions about the mechanisms preventing gene flow and about the potential existence of additional species hidden in the complex. Here, we compare patterns of genetic divergence across western Eurasia in an extensive data set of mitochondrial and nuclear DNA sequences with behavioural data on inter‐ and intraspecific reproductive isolation in courtship experiments. We show that three species exist in accordance with both the phylogenetic and biological species concepts and that additional hidden diversity is unlikely to occur in Europe. The Leptidea species are now the best studied cryptic complex of butterflies in Europe and a promising model system for understanding the formation of cryptic species and the roles of local processes, colonization patterns and heterospecific interactions for ecological and evolutionary divergence.     

Host plant preference and performance of the sibling species of butterflies <Emphasis Type="Italic">Leptidea sinapis</Emphasis> and <Emphasis Type="Italic">Leptidea reali</Emphasis>: a test of the trade-off hypothesis for food specialisation

A large proportion of phytophagous insect species are specialised on one or a few host plants, and female host plant preference is predicted to be tightly linked to high larval survival and performance on the preferred plant(s). Specialisation is likely favoured by selection under stable circumstances, since different host plant species are likely to differ in suitability—a pattern usually explained by the “trade-off hypothesis”, which posits that increased performance on a given plant comes at a cost of decreased performance on other plants. Host plant specialisation is also ascribed an important role in host shift speciation, where different incipient species specialise on different host plants. Hence, it is important to determine the role of host plants when studying species divergence and niche partitioning between closely related species, such as the butterfly species pair Leptidea sinapis and Leptidea reali. In Sweden, Leptidea sinapis is a habitat generalist, appearing in both forests and meadows, whereas Leptidea reali is specialised on meadows. Here, we study the female preference and larval survival and performance in terms of growth rate, pupal weight and development time on the seven most-utilised host plants. Both species showed similar host plant rank orders, and larvae survived and performed equally well on most plants with the exceptions of two rarely utilised forest plants. We therefore conclude that differences in preference or performance on plants from the two habitats do not drive, or maintain, niche separation, and we argue that the results of this study do not support the trade-off hypothesis for host plant specialisation, since the host plant generalist Leptidea sinapis survived and performed as well on the most preferred meadow host plant Lathyrus pratensis as did Leptidea reali although the generalist species also includes other plants in its host range. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Habitat requirements for the conservation of wood cricket (Nemobius sylvestris) (Orthoptera: Gryllidae) on the Isle of Wight, UK

In the UK, wood cricket (Nemobius sylvestris) is a ‘Species of Conservation Concern’, being restricted to only three areas in southern England. Little information is available on the specific habitat requirements of this species. In 2006, a field investigation within three woodlands on the Isle of Wight was undertaken to identify its habitat preferences. Factors positively influencing wood cricket presence within woodlands included the presence of a well-developed leaf litter layer, relatively low ground vegetation cover and height, low canopy cover and relatively short distances between individual populations. Regression models identified the degree of isolation and variables describing vegetation structure as the main predictors for wood cricket presence within woodland fragments. The results of this study indicate the preference of wood cricket for open wooded edges. Conservation efforts for this species should focus on continuation of regular management activities aimed at providing permanent open edge habitat within woodlands, to maintain viable populations.     

Niche separation in space and time between two sympatric sister species—a case of ecological pleiotropy

We investigate the niche separation in space and time between the Palearctic sister species Leptidea sinapis and L. reali (Lepidoptera, Pieridae) in central Sweden. Using field sampling, we show that L. reali is a habitat specialist confined to meadows, whereas L. sinapis is a habitat generalist also inhabiting forests. This difference in habitat utilization was corroborated by experimental release of laboratory-reared L. sinapis and L. reali in two adjacent forest and meadow habitats during their natural flight period; virtually all recaptured L. reali that were released in the forest were later caught in the meadow, whereas L. sinapis shifted equally often from meadow to forest as in the opposite direction. In the field, both species fly in May–June, but L. reali appears on average a week earlier in spring and has a substantial second generation in July, whereas L. sinapis is practically univoltine. When overwintered pupae were incubated under identical conditions in the laboratory, females did, however, not differ in phenology, and L. sinapis males actually emerged earlier than L. reali males. When larvae were reared at 23°C on the host plant Lotus corniculatus at a range of daylengths, both species produced a substantial proportion of directly developing individuals at an 18.5 h daylength or longer. When reared at 23°C and a 22 h daylength, L. reali showed an overall higher propensity to develop directly than L. sinapis on plant species originating from both the meadow and the forest habitat. Both Leptidea species showed a lower propensity to enter direct development on forest associated plants than on meadow associated plants. Hence, we suggest that the difference in phenology and voltinism between L. sinapis and L. reali is largely the result of environmentally implemented ecological pleiotropic effects caused by the between-species difference in habitat preference.     

Generation-dependent female choice: behavioral polyphenism in a bivoltine butterfly

Climatic and biotic circumstances vary as seasons shift, anddifferent cohorts of multivoltine species are likely subjectedto different selection regimes. The bivoltine butterfly Leptideareali (Réal's wood white; Lepidoptera: Pieridae) appearsduring May and June in central Sweden and has a partial secondgeneration in late July. We manipulated both generations toappear simultaneously and performed laboratory mating experimentsthat showed the presence of a behavioral polyphenism in matingpropensity, which is induced during the developmental stages.Females of the summer generation expressed higher mating propensitiesthan spring generation females. Spring females showed an increasein mating propensity with increasing age, whereas summer femalesaccepted most matings already when they were only 1 or 2 daysold. It is likely that larval time constraints, a lower abundanceof males and a lower risk of accepting a male of their univoltinesister species Leptidea sinapis (wood white), have relaxed selectionon mate discrimination among summer generation females. A majorchallenge for future research is to further investigate thedevelopmental pathways causing the polyphenism and the adaptiveimplications of cohort-dependent behaviors.     

Separation possibilities and genital measurement variations in two cryptic species of European pierid butterflies,Leptidea juvernica Williams, 1946 and L. sinapis (Linnaeus, 1758)

Of the three recently separated cryptic butterfly species of the Leptidea sinapis complex, the two species L. sinapis and L. juvernica occur sympatrically and syntopically in central Europe. As the separation of these species requires genital or genetic characters, their correct identification is a fundamental problem to be solved prior to any biological and distributional studies. In the present study a morphometric approach was applied to test for separation possibilities and to examine genital measurement variations based on large population samples of the two species (347 females and 636 males). Butterflies were collected at 456 localities distributed across Poland. Specimens of both sexes could be separated using either the shape or the length of the antrum bursae (females), phallus and saccus (males). Intraspecific seasonal differences in these measurements may affect the discrimination of males only. Genital characters were significantly larger in butterflies of the spring brood than in those of the second brood. In females, antrum bursae length ranges were separated by an interval, allowing for the exact identification of each specimen. In males, no such intervals were found between phallus and saccus length ranges, which slightly overlapped. Discriminant analysis resulted in 100% of males correctly classified. The present study confirmed the validity of interspecific differences in the shape of phallus and saccus for species identification without measurements. The length of these two characters discriminates the two species when separation is limited to specimens from the same generations. As an alternative for discriminant analysis, either phallus length and vinculum width or phallus length and the ratio of the ventral edge of genital capsule length and saccus length (VEL/SL ratio) is recommended for the practical separation of the two species. The differences in the shape and size of the copulatory organs of the two species seem to indicate reproductive isolation due to mechanical incompatibility of their genitals.     

Heterospecific courtship,minority effects and niche separation between cryptic butterfly species

Species interacting in varied ecological conditions often evolve in different directions in different local populations. The butterflies of the cryptic Leptidea complex are sympatrically distributed in different combinations across their Eurasian range. Interestingly, the same species is a habitat generalist in some regions and a habitat specialist in others, where a sibling species has the habitat generalist role. Previous studies suggest that this geographically variable niche divergence is generated by local processes in different contact zones. By varying the absolute and relative densities of Leptidea sinapis and Leptidea juvernica in large outdoor cages, we show that female mating success is unaffected by conspecific density, but strongly negatively affected by the density of the other species. Whereas 80% of the females mated when a conspecific couple was alone in a cage, less than 10% mated when the single couple shared the cage with five pairs of the other species. The heterospecific courtships can thus affect the population fitness, and for the species in the local minority, the suitability of a habitat is likely to depend on the presence or absence of the locally interacting species. If the local relative abundance of the different species depends on the colonization order, priority effects might determine the ecological roles of interacting species in this system.     

Oviposition preferences of a threatened butterfly Leptidea morsei (Lepidoptera: Pieridae) at the western border of its range

The oviposition preferences of Leptidea morsei at the western border of its range (SE Slovenia) were studied with the aim of integrating this information into species habitat management recommendations. The characteristics of egg-laying habitat were examined at the landscape, patch, plant and leaf levels. Because sightings of adults in the field were infrequent, intensive searching for previously laid eggs on Lathyrus niger plants was chosen as the survey method. The main oviposition habitats of L. morsei in Slovenia were found to be the forest edges and light stands of thermophilous oak, oak-hornbeam and mesic beech forests (landscape level). The egg-laying sites were L. niger plants growing individually or in small stands, and oriented mostly towards W, SW, and S. The oviposition plants were between 33 and 120 cm high, with those growing in half to full shade preferred. Eggs were deposited singly, mainly on the underside of leaflets of terminal leaves, located mostly on the top quarter of plants irrespective of their height and growth form. The most important factors affecting egg-placement are sun exposure and the size of L. niger stands at ovipositing sites (patch level), leaf exposure on the plant in vertical and horizontal directions (plant level), leaf age and the leaf parts where the risk of leaf and egg desiccation is lower (leaf level). Management strategies for the main egg-laying habitats of the species are proposed.     

Demographics and spatial ecology in a population of cryptic wood white butterfly <Emphasis Type="Italic">Leptidea juvernica</Emphasis> in Northern Ireland

The cryptic wood white Leptidea juvernica is one of a newly-discovered species complex comprising three morphologically similar species in Europe. In the British Isles, L. juvernica is absent from Great Britain, but is widespread in Ireland, where it has experienced recent declines; it is classed as a priority species in Northern Ireland. Using a mark–recapture approach, this study aims to elucidate the population and spatial ecology of L. juvernica based on a population resident on a small, suburban site and to propose conservation measures. The results demonstrated that populations of L. juvernica, even on small sites, can reach high numbers during the peak flight season. Unusually for European Pieridae, there was only weak evidence of protandry in this population, possibly reflecting weather conditions prior to the flight season. The spatial distribution and abundance of L. juvernica was associated with the distribution of its larval host-plants, as well as maintaining close proximity to south-facing habitat edges for shelter. Males had a closer association to sheltered habitat edges, whilst females were found more commonly on open ground with a shorter sward which was the preferred egg-laying habitat. Long vegetation in sheltered areas was important for roosting during periods of non-activity. These results inform conservation measures which will benefit L. juvernica; appropriate measures focus on habitat management providing a mosaic of open, semi-natural grassland interspersed with tall vegetation, scrub, and trees.     

两种共存网蛱蝶的卵块特征及其雌性成虫对产卵地的选择

对同一网络斑块中金堇蛱蝶(Euphydryas aurinia)和大网蛱蝶(Meiltaea phoebe)雌性成虫的产卵地进行了调查,以确定影响它们斑块质量的因素.结果表明,2种网蛱蝶雌性成虫均选择体积大的寄主植物、周围植被高度低、开放的区域产卵,它们的卵块主要分布在温暖向阳的坡面.金堇蛱蝶的卵块距离农田边缘为3.55±0.33 m (n=246),且集中分布在＜3 m区域内;大网蛱蝶卵块距离农田边缘为7.34±1.53 m (n=25),但在＜3 m区域内的数量少.研究表明,由于这2种网蛱蝶雌性成虫对产卵地要求的特异性,评价它们斑块质量以及对其制定保护管理措施时均应有所不同.     

Development,survival, and phenotypic plasticity in anthropogenic landscapes: trade-offs between offspring quantity and quality in the nettle-feeding peacock butterfly

Habitats selected for development may have important fitness consequences. This is relevant within the framework of niche shifts in human-dominated landscapes. Currently, the peacock butterfly (Aglais io) occurs ubiquitously, covering many habitat types, whereas its distribution used to be much more restricted. Indeed, its host plant (stinging nettle Urtica dioica) was limited to natural forest gaps on relatively nitrogen-rich soil, but due to land use changes and eutrophication, host plants are now quasi-omnipresent in Western Europe. In order to assess the impact of specific anthropogenic habitat types on host plant quality and environmental conditions for phenotypic trait values, an experiment was conducted in woodlands, field margins, and urban gardens. Larval development was studied in field enclosures, and adult traits were analyzed to test predicted effects of warmer and more nitrogen-rich conditions in field margins compared to woodlands and urban gardens. Survival to the adult stage was highest in woodlands and lowest in field margins, and whilst development time did not differ amongst habitat types, butterflies that developed in field margins were larger and had higher lipid content and wing loadings than conspecifics from woodlands and urban gardens. Nettles in field margins provided warmer microclimates. However, and contrary to predictions, the nitrogen level within host plant leaves was highest in woodlands. Hence, anthropogenic landscapes may pose a conflict for choosing what is ultimately the best breeding habitat, as survival was highest in woodlands (followed by urban gardens), but adults with highest fitness predictions were produced in field margins (and secondarily urban gardens).     

Evidence-based National Recovery Plan for <Emphasis Type="Italic">Leptidea sinapis</Emphasis> (wood white butterfly) in Southern Britain

We present a key part of the evidence base used to develop a National Recovery Plan for Leptidea sinapis a declining and threatened species in Britain. Knowledge of the current sites has been updated along with the macro- and micro-habitat resource requirements of the species and its autecology to set the basis for a conservation strategy for 2010 and beyond. The three main findings are that: L. sinapis uses a greater diversity of biotopes within and between sites than was previously thought; population dynamics are complex with 2–3 year peaks and troughs of annual abundance indices and two generations per year throughout its range; and dispersal ability is also greater than previously stated. This evidence has been used to develop an Action Plan for 2010 and beyond. The importance of landscape-scale management and effective collaboration between scientists, land managers and volunteers is emphasised.     

Untangling the effects of fire, grazing, and land-use legacies on grassland butterfly communities

Many grassland ecosystems are disturbance-dependent, having evolved under the pressures of fire and grazing. Restoring these disturbances can be controversial, particularly when valued resources are thought to be disturbance-sensitive. We tested the effects of fire and grazing on butterfly species richness and population density in an economically productive grassland landscape of the central U.S. Three management treatments were applied: (1) patch-burn graze—rotational burning of three spatially distinct patches within a pasture, and moderately-stocked cattle grazing (N?=?5); (2) graze-and-burn—burning entire pasture every 3?years, and moderately-stocked cattle grazing (N?=?4); and (3) burn-only—burning entire pasture every 3?years, but no cattle grazing (N?=?4). Butterfly abundance was sampled using line transect distance sampling in 2008 and 2009, with six 100-m transects per pasture. Butterfly species richness did not respond to management treatment, but was positively associated with pre-treatment proportion of native plant cover. Population density of two prairie specialists (Cercyonis pegala and Speyeria idalia) and one habitat generalist (Danaus plexippus) was highest in the burn-only treatment, whereas density of one habitat generalist (Cupido comyntas) was highest in the patch-burn graze treatment. Treatment application affected habitat structural characteristics including vegetation height and cover of bare ground. Historic land uses have reduced native plant cover and permitted exotic plant invasion; for some butterfly species, these legacies had a greater influence than management treatments on butterfly density. Conservation of native insect communities in altered grasslands might require native plant restoration in addition to restoration of disturbance processes.     

Habitat utilization by ovipositing females and larvae of the Marsh fritillary (Euphydryas aurinia) in a mosaic of meadows and croplands

The butterfly Euphydryas aurinia occurs as a classical metapopulation in Yanjiaping village, Heibei province, China. In the patch network under study there were 38 habitat patches. Most patches were uncultivated, consisting of meadows and fallows, while others had been cultivated, i.e., used as small croplands. In the cultivated patches, the habitat for the butterfly can be classified into two types: meadow type (MTH) and cropland type (CTH). In contrast, the uncultivated habitat patches consists only of MTH. We examined the habitat utilization of ovipositing females and larvae of E. aurinia to assess the effect of mixed land use on the butterfly’s occurrence. More egg clusters and pre-hibernating larval groups were distributed throughout CTH than MTH. This dependence on CTH may be due to the preference of egg-laying females for large-sized host plants, which were mainly concentrated in CTH. Compared to the MTH, the mortality rate in CTH was lower. Therefore, for both eggs and pre-hibernating larvae, the quality of CTH was higher than that of MTH. The conditions in MTH, on the other hand, were important for the development of post-hibernation larvae. A combination of extensive farming and animal husbandry has created a mosaic of meadows and croplands in the habitats studied here, which is typical of rural areas in some developing countries. This study implies that patchy land use typical of traditional agriculture facilitates the long-term persistence of E. aurinia.     

Fox-hunting in England and Wales: its contribution to the management of woodland and other habitats

Hunting foxes with hounds has been a countryside pursuit in Britain since the 17th Century, but its effect nationally on habitat management is little understood by the general public. A survey questionnaire was distributed to 163 mounted fox hunts of England and Wales to quantify their management practices in woodland and other habitat. Ninety-two hunts (56%), covering 75,514 km², returned details on woodland management motivated by the improvement of their sport. The management details were verified via on-site visits for a sample of 200 woodlands. Following verification, the area of woodlands containing the management was conservatively estimated at 24,053 (±2241) ha, comprising 5.9% of woodland area within the whole of the area hunted by the 92 hunts. Management techniques included: tree planting, coppicing, felling, ride and perimeter management. A case study in five hunt countries in southern England examined, through the use of botanical survey and butterfly counts, the consequences of the hunt management on woodland ground flora and butterflies. Managed areas had, within the last 5 years, been coppiced and rides had been cleared. Vegetation cover in managed and unmanaged sites averaged 86% and 64%, respectively, and managed areas held on average 4 more plant species and a higher plant diversity than unmanaged areas (Shannon index of diversity: 2.25 vs. 1.95). Both the average number of butterfly species (2.2 vs. 0.3) and individuals counted (4.6 vs. 0.3) were higher in the managed than unmanaged sites.     

Movement Patterns Differ between Sexes and Depend on Weather Conditions in the Butterfly <Emphasis Type="Italic">Lycaena tityrus</Emphasis>

Habitat loss and anthropogenic climate change are important threats to biodiversity conservation. Owing to the concomitantly deteriorating habitat quality, individuals are often forced to disperse to new habitats, rendering dispersal an ecologically important process. However, dispersal ability may differ within and among populations, and is further dependent on environmental conditions. We therefore studied sexual differences in and environmental effects on movement patterns in the sooty copper butterfly Lycaena tityrus. As predicted, males were more active and covered longer distances than females, presumably owing to mate location and territorial disputes. Males alighted more often on flowers than females, probably to fuel their high flight activity, while females showed a high affinity to host-plants for egg-laying. Our findings provide a striking example of sex-related differences in animal behavior, as revealed by the use of customary smartphones, which apparently can comprise suitable means to reveal biologically significant behavioural patterns. More problematic than the technical device used seems to be the challenge of following individual butterflies for long enough in the field, such that any extrapolations to dispersal seem difficult.