Pervasive genetic associations between traits causing reproductive isolation in Heliconius butterflies

Ecological speciation proceeds through the accumulation of divergent traits that contribute to reproductive isolation, but in the face of gene flow traits that characterize incipient species may become disassociated through recombination. Heliconius butterflies are well known for bright mimetic warning patterns that are also used in mate recognition and cause both pre- and post-mating isolation between divergent taxa. Sympatric sister taxa representing the final stages of speciation, such as Heliconius cydno and Heliconius melpomene, also differ in ecology and hybrid fertility. We examine mate preference and sterility among offspring of crosses between these species and demonstrate the clustering of Mendelian colour pattern loci and behavioural loci that contribute to reproductive isolation. In particular, male preference for red patterns is associated with the locus responsible for the red forewing band. Two further colour pattern loci are associated, respectively, with female mating outcome and hybrid sterility. This genetic architecture in which ‘speciation genes’ are clustered in the genome can facilitate two controversial models of speciation, namely divergence in the face of gene flow and hybrid speciation.     

Behavioural,ecological and genetic evidence confirm the occurrence of host‐associated differentiation in goldenrod gall‐midges

Host‐associated differentiation (HAD) is considered a step towards ecological speciation and an important mechanism promoting diversification in phytophagous insects. Although the number of documented cases of HAD is increasing, these still represent only a small fraction of species and feeding guilds among phytophagous insects, and most reports are based on a single type of evidence. Here we employ a comprehensive approach to present behavioural, morphological, ecological and genetic evidence for the occurrence of HAD in the gall midge Dasineura folliculi (Diptera: Cecidomyiidae) on two sympatric species of goldenrods (Solidago rugosa and S. gigantea). Controlled experiments revealed assortative mating and strong oviposition fidelity for the natal‐host species. Analysis of mitochondrial DNA showed an amount of genetic divergence between the two host‐associated populations compatible with cryptic species rather than host races. Lower levels of within‐host genetic divergence, gall development and natural‐enemy attack in the S. gigantea population suggest this is the derived host.     

Geographic contrasts between pre‐ and postzygotic barriers are consistent with reinforcement in Heliconius butterflies

Identifying the traits causing reproductive isolation and the order in which they evolve is fundamental to understanding speciation. Here, we quantify prezygotic and intrinsic postzygotic isolation among allopatric, parapatric, and sympatric populations of the butterflies Heliconius elevatus and Heliconius pardalinus. Sympatric populations from the Amazon (H. elevatus and H. p. butleri) exhibit strong prezygotic isolation and rarely mate in captivity; however, hybrids are fertile. Allopatric populations from the Amazon (H. p. butleri) and Andes (H. p. sergestus) mate freely when brought together in captivity, but the female F1 hybrids are sterile. Parapatric populations (H. elevatus and H. p. sergestus) exhibit both assortative mating and sterility of female F1s. Assortative mating in sympatric populations is consistent with reinforcement in the face of gene flow, where the driving force, selection against hybrids, is due to disruption of mimicry and other ecological traits rather than hybrid sterility. In contrast, the lack of assortative mating and hybrid sterility observed in allopatric populations suggests that geographic isolation enables the evolution of intrinsic postzygotic reproductive isolation. Our results show how the types of reproductive barriers that evolve between species may depend on geography.     

Testing host‐associated differentiation in a quasi‐endophage and a parthenogen on native trees

Host‐associated differentiation (HAD) is the formation of genetically divergent host‐associated sub‐populations. Evidence of HAD has been reported for multiple insect herbivores to date, but published studies testing more than one herbivore for any given host‐plant species pair is limited to herbivores on goldenrods. This limits the number of pair‐wise comparisons that can be made about insect life‐history traits that might facilitate or inhibit host‐race development in general. Two traits previously proposed to facilitate HAD include endophagy and parthenogenesis. We tested for HAD in two herbivores, a quasi‐endophagous caterpillar and a parthenogenetic aphid, feeding on two closely related species of hickories. We found that the quasi‐endophage is panmictic, whereas the parthenogen exhibits HAD on their sympatric host plants, pecan and water hickory, at a geographic mesoscale. This is an important first step in the characterization of HAD in multiple insect herbivores using North American hickories, a host‐plant system with many shared parthenogens.     

Spatial and genetic structure of host‐associated differentiation in the parasitoid Copidosoma gelechiae

Host‐associated differentiation (HAD) appears to be an important driver of diversification in the hyperdiverse phytophagous and parasitoid insects. The gallmaking moth Gnorimoschema gallaesolidaginis has undergone HAD on two sympatric goldenrods (Solidago), and HAD has also been documented in its parasitoid Copidosoma gelechiae, with the intriguing suggestion that differentiation has proceeded independently in multiple populations. We tested this suggestion with analysis of Amplified Fragment Length Polymorphism (AFLP) markers for C. gelechiae collections from the midwestern and northeastern United States and eastern Canada. AFLP data were consistent with the existence of HAD, with between‐host F_ST significant before Bonferroni correction in two of seven sympatric populations. amova analysis strongly rejected a model of HAD with a single historical origin, and thus supported the repeated‐HAD hypothesis. Copidosoma gelechiae shows significant host‐associated divergence at a number of allozyme loci ( Stireman et al., 2006 ), but only weak evidence via AFLPs for genome‐wide differentiation, suggesting that this species is at a very early stage of HAD.     

Testing the host‐finding ability of a monophagous caterpillar in the field

1. Relatively few studies of the host‐finding ability of specialised, phytophagous insects involve direct observations of individual insects moving among intact hosts and non‐hosts. Information from such studies can inform the design of restoration programmes for species of conservation concern. 2. The movement of caterpillars of the threatened Oregon silverspot butterfly, Speyeria zerene hippolyta (Edwards) (Lepidoptera: Nymphalidae) was studied in the field in cleared arenas with 10 cm radii. Caterpillars were placed in the centre, surrounded by three individuals of their host, Viola adunca, and three different non‐host individuals, separated by bare ground. In a second experiment, second instars were placed between a host and a non‐host, 3–6 cm away. Caterpillars were observed to determine if they walked to their host more often than expected by chance. 3. Caterpillars walked to vegetation significantly more often than expected by chance. They did not, however, reach their hosts more often than expected, based on plant availability. 4. It is concluded that S. z. hippolyta caterpillars can distinguish vegetation from bare ground from 10 cm away. There is no evidence that they can distinguish their host plant from other herbaceous species at distances of 3 cm.     

Intraspecific difference among herbivore lineages and their host‐plant specialization drive the strength of trophic cascades

Trophic cascades – the indirect effect of predators on non‐adjacent lower trophic levels – are important drivers of the structure and dynamics of ecological communities. However, the influence of intraspecific trait variation on the strength of trophic cascade remains largely unexplored, which limits our understanding of the mechanisms underlying ecological networks. Here we experimentally investigated how intraspecific difference among herbivore lineages specialized on different host plants influences trophic cascade strength in a terrestrial tri‐trophic system. We found that the occurrence and strength of the trophic cascade are strongly influenced by herbivores’ lineage and host‐plant specialization but are not associated with density‐dependent effects mediated by the growth rate of herbivore populations. Our findings stress the importance of intraspecific heterogeneities and evolutionary specialization as drivers of trophic cascade strength and underline that intraspecific variation should not be overlooked to decipher the joint influence of evolutionary and ecological factors on the functioning of multi‐trophic interactions.     

Effects of quantitative and qualitative differences in volatiles from host‐ and non‐host‐infested maize on the attraction of the larval parasitoid Cotesia kariyai

Cotesia kariyai Watanabe (Hymenoptera: Braconidae) is a specialist larval parasitoid of Mythimna separata Walker (Lepidoptera: Noctuidae). Cotesia kariyai wasps use herbivore‐induced plant volatiles (HIPVs) to locate hosts. However, complex natural habitats are full of volatiles released by both herbivorous host‐ and non‐host‐infested plants at various levels of intensity. Therefore, the presence of non‐hosts may affect parasitoid decisions while foraging. Here, the host‐finding efficiency of naive C. kariyai from HIPVs influenced by host‐ and non‐host‐infested maize [Zea mays L. (Poaceae)] plants was investigated with a four‐arm olfactometer. Ostrinia furnacalis Guenée (Lepidoptera: Crambidae) was selected as a non‐host species. One unit (1 U) of host‐ or non‐host‐infested plant was prepared by infesting a potted plant with five host or seven non‐host larvae. In two‐choice bioassays, host‐infested plants fed upon by different numbers of larvae, and various units of host‐ and non‐host‐infested plants (infestation units; 1 U, 2 U, and 3 U) were arranged to examine the effects of differences in volatile quantity and quality on the olfactory responses of C. kariyai with the assumption that volatile quantity and quality changes with differences in numbers of insects and plants. Cotesia kariyai was found to perceive quantitative differences in volatiles from host‐infested plants, preferring larger quantities of volatiles from larger numbers of larvae or plants. Also, the parasitoids discriminated between healthy plants, host‐infested plants, and non‐host‐infested plants by recognising volatiles released from those plants. Cotesia kariyai showed a reduced preference for host‐induced volatiles, when larger numbers of non‐host‐infested plants were present. Therefore, quantitative and qualitative differences in volatiles from host‐ and non‐host‐infested plants appear to affect the decision of C. kariyai during host‐habitat searching in multiple tritrophic systems.     

A meta‐analysis of dispersal in butterflies

Dispersal has recently gained much attention because of its crucial role in the conservation and evolution of species facing major environmental changes such as habitat loss and fragmentation, climate change, and their interactions. Butterflies have long been recognized as ideal model systems for the study of dispersal and a huge amount of data on their ability to disperse has been collected under various conditions. However, no single ‘best’ method seems to exist leading to the co‐occurrence of various approaches to study butterfly mobility, and therefore a high heterogeneity among data on dispersal across this group. Accordingly, we here reviewed the knowledge accumulated on dispersal and mobility in butterflies, to detect general patterns. This meta‐analysis specifically addressed two questions. Firstly, do the various methods provide a congruent picture of how dispersal ability is distributed across species? Secondly, is dispersal species‐specific? Five sources of data were analysed: multisite mark‐recapture experiments, genetic studies, experimental assessments, expert opinions, and transect surveys. We accounted for potential biases due to variation in genetic markers, sample sizes, spatial scales or the level of habitat fragmentation. We showed that the various dispersal estimates generally converged, and that the relative dispersal ability of species could reliably be predicted from their relative vagrancy (records of butterflies outside their normal habitat). Expert opinions gave much less reliable estimates of realized dispersal but instead reflected migration propensity of butterflies. Within‐species comparisons showed that genetic estimates were relatively invariable, while other dispersal estimates were highly variable. This latter point questions dispersal as a species‐specific, invariant trait.     

Species mtDNA genetic diversity explained by infrapopulation size in a host‐symbiont system

Understanding what shapes variation in genetic diversity among species remains a major challenge in evolutionary ecology, and it has been seldom studied in parasites and other host‐symbiont systems. Here, we studied mtDNA variation in a host‐symbiont non‐model system: 418 individual feather mites from 17 feather mite species living on 17 different passerine bird species. We explored how a surrogate of census size, the median infrapopulation size (i.e., the median number of individual parasites per infected host individual), explains mtDNA genetic diversity. Feather mite species genetic diversity was positively correlated with mean infrapopulation size, explaining 34% of the variation. As expected from the biology of feather mites, we found bottleneck signatures for most of the species studied but, in particular, three species presented extremely low mtDNA diversity values given their infrapopulation size. Their star‐like haplotype networks (in contrast with more reticulated networks for the other species) suggested that their low genetic diversity was the consequence of severe bottlenecks or selective sweeps. Our study shows for the first time that mtDNA diversity can be explained by infrapopulation sizes, and suggests that departures from this relationship could be informative of underlying ecological and evolutionary processes.     

Host‐driven divergence in the parasitic plant Orobanche minor Sm. (Orobanchaceae)

Many parasitic angiosperms have a broad host range and are therefore considered to be host generalists. Orobanche minor is a nonphotosynthetic root parasite that attacks a range of hosts from taxonomically disparate families. In the present study, we show that O. minor sensu lato may comprise distinct, genetically divergent races isolated by the different ecologies of their hosts. Using a three‐pronged approach, we tested the hypothesis that intraspecific taxa O. minor var. minor and O. minor ssp. maritima parasitizing either clover (Trifolium pratense) or sea carrot (Daucus carota ssp. gummifer), respectively, are in allopatric isolation. Morphometric analysis revealed evidence of divergence but this was insufficient to define discrete, host‐specific taxa. Intersimple sequence repeat (ISSR) marker‐based data provided stronger evidence of divergence, suggesting that populations were isolated from gene flow. Phylogenetic analysis, using sequence‐characterized amplified region (SCAR) markers derived from ISSR loci, provided strong evidence for divergence by clearly differentiating sea carrot‐specific clades and mixed‐host clades. Low levels of intrapopulation SCAR marker sequence variation and floral morphology suggest that populations on different hosts are probably selfing and inbreeding. Morphologically cryptic Orobanche taxa may therefore be isolated from gene flow by host ecology. Together, these data suggest that host specificity may be an important driver of allopatric speciation in parasitic plants.     

Nutrition as a facilitator of host‐race formation: the shift of a stem‐boring beetle to a gall host

1. The importance of host‐race formation to herbivorous insect diversity depends on the likelihood that successful populations can be established on a new plant host. A previously unexplored ecological aid to success on a novel host is better nutritional quality. The role of nutrition was examined in the shift of the stem‐boring beetle Mordellistena convicta to fly‐induced galls on goldenrod and the establishment there of a genetically distinct gall host race. 2. First, larvae of the host race inhabiting stems of Solidago gigantea were transplanted into stems and galls of greenhouse‐grown S. gigantea plants. At the end of larval development, the mean mass of larvae transplanted to galls was significantly greater than the mass of larvae transplanted to stems, indicating a likely nutritional benefit during the shift. This advantage was slightly but significantly diminished when the gall‐inducing fly feeding at the centre of the gall died early in the season. Additionally, there was a suggestion of a trade‐off in the increased mortality of smaller beetle larvae transplanted into galls. 3. In a companion experiment, S. gigantea gall‐race beetle larvae were likewise transplanted to S. gigantea stems and galls. Besides the expected greater mass in galls, the larvae also exhibited adaptations to the gall nutritional environment: larger inherent size, altered tunnelling behaviour, and no diminution of mass pursuant to gall‐inducer mortality. 4. In a third line of inquiry, chemical analyses of field‐collected S. gigantea plants revealed higher levels of mineral elements important to insect nutrition in galls as compared with stems.     

Effects of the maternal and pre‐adult host plant on adult performance and preference in the pea aphid,Acyrthosiphon pisum

Abstract 1. The taxon known as the pea aphid, Acyrthosiphon pisum, is composed of a series of host plant associated populations and is widely used as a model system to explore ecological speciation and the evolution of specialisation. It is thus important to know how maternal and pre‐adult experience influences host plant utilisation in this species. 2. The relative importance of the maternal and pre‐adult host plant for adult fecundity and host preference was investigated using three aphid clones collected from Lathyrus pratensis and maintained on Lathyrus or Vicia faba. 3. No significant effects of the maternal host plant on offspring fecundity were detected. 4. The host plant on which the aphid grew up influenced adult fecundity, although in a complex way that depended on both the adult host plant species and when after transfer to the test plant fecundity was assessed. 5. All three clones preferred to colonise Lathyrus over Vicia, and this preference was stronger for aphids raised on Lathyrus. 6. The significance of the results for studies of the evolution of specialisation and speciation that employ A. pisum is discussed.     

The roles of geography and founder effects in promoting host‐associated differentiation in the generalist bogus yucca moth Prodoxus decipiens

There is ample evidence that host shifts in plant‐feeding insects have been instrumental in generating the enormous diversity of insects. Changes in host use can cause host‐associated differentiation (HAD) among populations that may lead to reproductive isolation and eventual speciation. The importance of geography in facilitating this process remains controversial. We examined the geographic context of HAD in the wide‐ranging generalist yucca moth Prodoxus decipiens. Previous work demonstrated HAD among sympatric moth populations feeding on two different Yucca species occurring on the barrier islands of North Carolina, USA. We assessed the genetic structure of P. decipiens across its entire geographic and host range to determine whether HAD is widespread in this generalist herbivore. Population genetic analyses of microsatellite and mtDNA sequence data across the entire range showed genetic structuring with respect to host use and geography. In particular, genetic differentiation was relatively strong between mainland populations and those on the barrier islands of North Carolina. Finer scale analyses, however, among sympatric populations using different host plant species only showed significant clustering based on host use for populations on the barrier islands. Mainland populations did not form population clusters based on host plant use. Reduced genetic diversity in the barrier island populations, especially on the derived host, suggests that founder effects may have been instrumental in facilitating HAD. In general, results suggest that the interplay of local adaptation, geography and demography can determine the tempo of HAD. We argue that future studies should include comprehensive surveys across a wide range of environmental and geographic conditions to elucidate the contribution of various processes to HAD.     

No host‐associated differentiation in the gall wasp Diplolepis rosae (Hymenoptera: Cynipidae) on three dog rose species

Differences in quality and quantity of secondary compounds, as well as in leaf traits of host plants, may influence the host choice of herbivores. Different host preferences could lead to host‐associated differentiation, the first step of sympatric speciation. In the present study, we investigated whether the rose gall wasp Diplolepis rosae L. (Hymenoptera: Cynipidae) shows genetic differentiation related to its host plants (Rosa canina L., Rosa corymbifera Borkh., and Rosa rubiginosa L.). These three host species radiated recently and subsequently expanded their range. Therefore, we expected a diversification within the closely‐associated phytophagous insects. The process of genetic differentiation should be intensified in D. rosae by its close relationship to the host plant, as well as by its parthenogenetic reproduction (infection rate by Wolbachia sp. of almost 100%). However, using 106 polymorphic amplified fragment length polymorphism markers, we found no genetic differentiation among the wasps from different host plants. The population structuring between geographical localities was also low, suggesting considerable gene flow between sites. In part, the low genetic differentiation between sites is explained by the wide distribution of host species and hybrids between host plants. Hybrids with intermediate traits may facilitate the gene flow between wasp populations exploiting different host species. © 2011 The Linnean Society of London, Biological Journal of the Linnean Society, 2011, 102 , 369–377.     

No evidence for Z‐chromosome rearrangements between the pied flycatcher and the collared flycatcher as judged by gene‐based comparative genetic maps

Revealing the genetic basis of reproductive isolation is fundamental for understanding the speciation process. Chromosome speciation models propose a role for chromosomal rearrangements in promoting the build up of reproductive isolation between diverging populations and empirical data from several animal and plant taxa support these models. The pied flycatcher and the collared flycatcher are two closely related species that probably evolved reproductive isolation during geographical separation in Pleistocene glaciation refugia. Despite the short divergence time and current hybridization, these two species demonstrate a high degree of intrinsic post‐zygotic isolation and previous studies have shown that traits involved in mate choice and hybrid viability map to the Z‐chromosome. Could rearrangements of the Z‐chromosome between the species explain their reproductive isolation? We developed high coverage Z‐chromosome linkage maps for both species, using gene‐based markers and large‐scale SNP genotyping. Best order maps contained 57–62 gene markers with an estimated average density of one every 1–1.5 Mb. We estimated the recombination rates in flycatcher Z‐chromosomes to 1.1–1.3 cM/Mb. A comparison of the maps of the two species revealed extensive co‐linearity with no strong evidence for chromosomal rearrangements. This study does therefore not provide support the idea that sex chromosome rearrangements have caused the relatively strong post‐zygotic reproductive isolation between these two Ficedula species.     

Female preference and larval performance of sunflower moth,Homoeosoma electellum,on sunflower pre‐breeding lines

The preference–performance hypothesis for insect herbivores predicts that adult females should preferentially choose hosts on which their offspring perform better. We tested this hypothesis for the sunflower moth, Homoeosoma electellum (Hulst) (Lepidoptera: Pyralidae), using 16 sunflower (pre‐breeding) lines, derived from a number of wild species of Helianthus, including Helianthus annuus L., Helianthus deserticola Heiser, Helianthus paradoxus Heiser, Helianthus praecox Engelm. & Gray ssp. hirtus (Heiser) Heiser, Helianthus praecox Engelm. & Gray ssp. runyonii (Heiser) Heiser, Helianthus petiolaris Nutt., Helianthus resinosus Small, and Helianthus tuberosus L. (Asteraceae), that are suitable for introducing wild sunflower germplasm into commercial cultivars. Female moths showed a range of ovipositional preference measures to the various lines. Combined data for three Helianthus species represented by multiple lines showed significant differences in female preference with respect to the parental species. Larval performance, determined by proportion of infested neonate larvae reaching the pupal stage, or mean pupal weight, varied across the lines and, as for the female preference data, also showed significant differences among the three parental Helianthus species represented by multiple lines. These data suggest that the characteristics in the pre‐breeding lines influencing female sunflower moth preference and larval performance likely originate from the parental species and may be consistently transferred to the derived pre‐breeding lines. Of particular note with regard to potential plant resistance mechanisms, lines derived from H. tuberosus showed consistent low preference–performance measures. Female preference and larval performance (for both measures) were strongly correlated, indicating that females preferred plants and lines on which larvae performed better, in support of the preference–performance hypothesis.     

Reevaluating the conceptual framework for applied research on host‐plant resistance

Applied research on host‐plant resistance to arthropod pests has been guided over the past 60 years by a framework originally developed by Reginald Painter in his 1951 book, Insect Resistance in Crop Plants. Painter divided the “phenomena” of resistance into three “mechanisms,” nonpreference (later renamed antixenosis), antibiosis, and tolerance. The weaknesses of this framework are discussed. In particular, this trichotomous framework does not encompass all known mechanisms of resistance, and the antixenosis and antibiosis categories are ambiguous and inseparable in practice. These features have perhaps led to a simplistic approach to understanding arthropod resistance in crop plants. A dichotomous scheme is proposed as a replacement, with a major division between resistance (plant traits that limit injury to the plant) and tolerance (plant traits that reduce amount of yield loss per unit injury), and the resistance category subdivided into constitutive/inducible and direct/indirect subcategories. The most important benefits of adopting this dichotomous scheme are to more closely align the basic and applied literatures on plant resistance and to encourage a more mechanistic approach to studying plant resistance in crop plants. A more mechanistic approach will be needed to develop novel approaches for integrating plant resistance into pest management programs.     

Geographic and host‐mediated population genetic structure in a cestode parasite of the three‐spined stickleback

Comparative studies of genetic diversity and population structure can shed light on the ecological and evolutionary factors that influence host–parasite interactions. Here we examined whether geography, time and genetic variation in Alaskan three‐spined stickleback (Gasterosteus aculeatus Linneaus) hosts shape the population genetic structure of the diphyllobothridean cestode parasite Schistocephalus solidus (Müller, 1776). Host lineages and haplotypes were identified by sequencing the mitochondrial cytochrome b gene, and host population structure was assessed by Bayesian clustering analysis of allelic variation at 11 microsatellite loci. Parasite population structure was characterized according to allelic variation at eight microsatellite loci. Mantel tests and canonical redundancy analysis were conducted to evaluate the proportion of parasite genetic variation attributable to time and geography vs. host lineage, haplotype, and genotypic cluster. Host and parasite population structure were largely discordant across the study area, probably reflecting differences in gene flow, environmental influences external to the host, and genomic admixture among host lineages. We found that geography explained the greatest proportion of parasite genetic variation, but that variation also reflects time, host lineage, and host haplotype. Associations with host haplotypes suggest that one parasite genotypic cluster exhibits a narrower host range, predominantly infecting the most common host haplotypes, whereas the other parasite cluster infects all haplotypes equally, including rare haplotypes. Although experimental infection trials might prove otherwise, distributional differences in hosts preferentially infected by S. solidus could underlie the observed pattern of population structure.