Oviposition specificity in single vs. cluster egg-laying butterflies: a discrimination phase in Colias eurytheme?

Summary Oviposition preferences and rates of the pierid butterfly Colias eurytheme were examined in laboratory tests. Oviposition rates with only a non-preferred host plant present were not lower than rates observed when both preferred and non-preferred hosts were present. There was no correlation between strength of preference in simultaneous choice tests and the extent to which oviposition rate declined when the preferred host was absent. The results are consistent with the hypothesis that butterflies which lay their eggs singly will accept non-preferred hosts for oviposition more readily than cluster egg-layers.     

Butterfly Response to Microclimatic Conditions Following Ponderosa Pine Restoration

Efforts to restore ponderosa pine ecosystems to open, park‐like conditions that predominated prior to European‐American settlement result in altered stand structure and increased landscape heterogeneity, potentially altering habitat suitability for invertebrates and other forest organisms. We examined the responses of two butterfly species, Colias eurytheme and Neophasia menapia, to microclimatic changes at structural edges created by experimental restoration treatments in northern Arizona. We monitored microclimate, including air temperature, light intensity, and vapor pressure deficit (VPD), on several mornings during butterfly releases. We placed adult butterflies at east‐ and west‐facing edges approximately one half‐hour before dawn to determine their behavioral response to microclimatic differences between east‐ and west‐facing edges. After sunrise, all three microclimatic variables were higher at east‐facing edges, and the difference in microclimate between the two edge orientations increased through early morning. For both species, butterflies placed at east‐facing edges flew earlier than butterflies at west‐facing edges. Colias eurytheme, an open‐habitat species, tended to move toward the treated forest during initial flight, while movements of Neophasia menapia, a forest‐dwelling species, did not differ from random flight. Our results indicate that butterflies respond to microclimatic factors associated with restoration treatments, while responses to structural changes in habitat vary among species, based on habitat and food plant preferences. These changes in forest structure and microclimate may affect the distribution of many mobile invertebrates in forested landscapes undergoing restoration treatments.     

An AFLP-based interspecific linkage map of sympatric, hybridizing Colias butterflies

Colias eurytheme and C. philodice are sister species with broad sympatry in North America. They hybridize frequently and likely share a significant portion of their genomes through introgression. Both taxa have been ecologically well characterized and exploited to address a broad spectrum of evolutionary issues. Using AFLP markers, we constructed the first linkage map of Colias butterflies. The map is composed of 452 markers spanning 2541.7 cM distributed over 51 linkage groups (40 major groups and 11 small groups with 2-4 markers). Statistical tests indicate that these AFLP markers tend to cluster over the map, with the coefficient of variation of interval sizes being 1.236 (95% C.I. is 1.234-1.240). This nonrandom marker distribution can account for the nonequivalence between the number of linkage groups and the actual haploid chromosome number (N = 31). This study presents the initial step for further marker-assisted research on Colias butterflies, including QTL and introgression analyses. Further investigation of the genomes will help us understand better the roles of introgression and natural selection in the evolution of hybridizing species and devise more appropriate strategies to control these pests.     

PREDICTING HOST RANGE EVOLUTION: COLONIZATION OF CORONILLA VARIA BY COLIAS PHILODICE (LEPIDOPTERA: PIERIDAE)

Extensive sympatry is currently arising between the common sulfur butterfly, Colias philodice Latreille (Lepidoptera: Pieridae) and a potential leguminous host plant, Coronilla varia (L.). In laboratory trials, larval surviviorship and growth were higher on the primary host, Medicago sativa (L.), than on the nonhost C. varia. However, because females reared from C. varia were on average more fecund than females reared from M. sativa, fitness on C. varia (approximately as survivorship times fecundity) was commensurate with fitness on M. sativa. Thus, it is predicted that selection would favor oviposition on C. varia, if such behavior were to arise. In addition, significant among-family variation exists for several measures of larval performance on both C. varia and M. sativa, indicating that C. philodice can potentially respond to selection for increased performance on each species. Moreover, larval performance was significantly positively correlated across these species, suggesting that selection for increased performance on each species will facilitate, not constrain, evolution of increased performance on the other. It is concluded that behavioral rather than physiological barriers currently account for the absence of C. philodice from C. varia and that, if such barriers are overcome, C. philodice will expand its host plant range to include C. varia.     

Contrasting genomic and phenotypic outcomes of hybridization between pairs of mimetic butterfly taxa across a suture zone

Hybrid zones, whereby divergent lineages come into contact and eventually hybridize, can provide insights on the mechanisms involved in population differentiation and reproductive isolation, and ultimately speciation. Suture zones offer the opportunity to compare these processes across multiple species. In this paper we use reduced‐complexity genomic data to compare the genetic and phenotypic structure and hybridization patterns of two mimetic butterfly species, Ithomia salapia and Oleria onega (Nymphalidae: Ithomiini), each consisting of a pair of lineages differentiated for their wing colour pattern and that come into contact in the Andean foothills of Peru. Despite similarities in their life history, we highlight major differences, both at the genomic and phenotypic level, between the two species. These differences include the presence of hybrids, variations in wing phenotype, and genomic patterns of introgression and differentiation. In I. salapia, the two lineages appear to hybridize only rarely, whereas in O. onega the hybrids are not only more common, but also genetically and phenotypically more variable. We also detected loci statistically associated with wing colour pattern variation, but in both species these loci were not over‐represented among the candidate barrier loci, suggesting that traits other than wing colour pattern may be important for reproductive isolation. Our results contrast with the genomic patterns observed between hybridizing lineages in the mimetic Heliconius butterflies, and call for a broader investigation into the genomics of speciation in Ithomiini ‐ the largest radiation of mimetic butterflies.     

The evolution of novel host use is unlikely to be constrained by trade‐offs or a lack of genetic variation

The genetic and ecological factors that shape the evolution of animal diets remain poorly understood. For herbivorous insects, the expectation has been that trade‐offs exist, such that adaptation to one host plant reduces performance on other potential hosts. We investigated the genetic architecture of alternative host use by rearing individual Lycaeides melissa butterflies from two wild populations in a crossed design on two hosts (one native and one introduced) and analysing the genetic basis of differences in performance using genomic approaches. Survival during the experiment was highest when butterfly larvae were reared on their natal host plant, consistent with local adaptation. However, cross‐host correlations in performance among families (within populations) were not different from zero. We found that L. melissa populations possess genetic variation for larval performance and variation in performance had a polygenic basis. We documented very few genetic variants with trade‐offs that would inherently constrain diet breadth by preventing the optimization of performance across hosts. Instead, most genetic variants that affected performance on one host had little to no effect on the other host. In total, these results suggest that genetic trade‐offs are not the primary cause of dietary specialization in L. melissa butterflies.     

Drivers of hybridization in a 66-generation record of Colias butterflies

Hybridization significantly affects the ecology and evolution of numerous plant and animal lineages. Most studies have focused on endogenous drivers of hybridization and neglected variation in exogenous factors, such as seasonal weather patterns. In this study, we take advantage of a unique dataset consisting of records of hybridization between the butterflies Colias eurytheme and C. eriphyle (Pieridae) for 66 generations (22 years) to investigate the importance of seasonal weather on the production and survival of hybrid offspring. Important seasonal weather variables for each parental species and hybrid offspring were determined using model averaging, and these weather variables, along with butterfly abundances, were analyzed using path analysis. The most important drivers of hybridization were the abundance of C. eriphyle, summer minimum temperature, and spring maximum temperature. In contrast, the abundance of C. eurytheme and weather variables prior to the current flight season were relatively unimportant for variation in hybrid abundance. Parental abundances were mostly driven by weather variables prior to the flight season possibly because these variables affect host plant quality. Our results suggest that exogenous, climatic factors can influence hybridization in natural systems, and that these factors can act both directly on hybrid abundance and indirectly through the population dynamics of parental species.     

Epizootiology of virus diseases in three lepidopterous insect species of alfalfa

Summary The incidence of virus infections in three lepidopterous insect species was studied from 1965 to 1968 in alfalfa fields in California. The insects were the alfalfa caterpillar,Colias eurytheme; the beet armyworm,Spodoptera exigua; and the alfalfa looper,Autographa californica. InC. eurytheme, the major virus was a nuclear polyhedrosis virus (NPV); inS. exigua, a granulosis virus (GV) and an NPV; inA. californica, a GV. Virus epizootics did not develop in very high densities ofC. eurytheme. Virus epizootics occurred in low host densities of the three insect species, especially in populations ofA. californica. The virus acted as a density-dependent factor in the regulation of the populations ofS. exigua andA. californica. Temperature, humidity and rainfall had no marked effect on the incidence of virus infections.     

Population structure of pierid butterflies IV. Genetic and physiological investment in offspring by male Colias

Summary Population structure encompasses all the rules by which a population's gametes come together, including genetic and physiological investment in offspring. We document female use of nutrients donated by males at mating, and complete sperm precedence, in Colias eurytheme Boisduval. The effect of these phenomena on the population structure of this species is discussed.     

Development of ultraviolet-reflecting butterfly scales: How to make an interference filter

Light and electron microscope studies of development of the ultraviolet-reflecting scales of male Colias eurytheme butterflies show that basic developmental processes are similar to those of other scales. The ridges form between bundles of microfilaments and as they form they buckle to produce the lamellae seen in the adult scales. There is evidence that the buckling may be purely in response to mechanical stress and that some of the bundles of microfilaments may produce such stresses.     

Individual variation in oviposition preference in the butterfly,Colias eurytheme

Summary We examined intra-population variation in oviposition preference in the pierid butterfly, Colias eurytheme. Females' preferences were tested in the laboratory, using two-way choice tests between the potential hosts, alfalfa and vetch. There were consistent differences in oviposition preference among females within a population. Larval and adult experience had little or no effect on females' preference. These results suggest that the intra-population variation in oviposition preference is genetically based, but the results of experiments designed to estimate the heritability of oviposition preference were not conclusive. We suggest that intra-population variation in host selection characters may play a key role in shifts to new host plants.     

Facultative monophagy as a consequence of prior feeding experience: behavioral and physiological specialization in Colias philodice larvae

Summary Although newly-emerged Colias philodice readily accept Medicago sativa, Melilotus alba, and Coronilla varia, fifth instar larvae reared on any single plant species display a highly significant induced feeding preference for their rearing host. Forced host-switching reveals that fifth instar relative growth rate (RGR) on M. sativa and M. alba is significantly reduced by prior feeding on either alternative host. Moreover, regardless of rearing diet, switching to a novel host during the fifth instar results in reduced RGR, relative consumption rate (RCR), efficiency of conversion of digested food (ECD), and pupal weight. These results support the hypothesis that induction of feeding preference is an adaptive response that predisposes larvae to feed on the plant species they are most capable of utilizing for growth.     

Does host outcrossing disrupt compatibility with heritable symbionts?

Vertically transmitted microbes are common in macro‐organisms and can enhance host defense against environmental stress. Because vertical transmission couples host and symbiont lineages, symbionts may become specialized to host species or genotypes. Specialization and contrasting reproductive modes of symbiotic partners could create incompatibilities between inherited symbionts and novel host genotypes when hosts outcross or hybridize. Such incompatibilities could manifest as failed colonization or poor symbiont growth in host offspring that are genetically dissimilar from their maternal host. Moreover, outcrossing between host species could influence both host and symbiont reproductive performance. We tested these hypotheses by manipulating outcrossing between populations and species of two grasses, Elymus virginicus and E. canadensis, that host vertically transmitted fungal endophytes (genus Epichloё). In both greenhouse and field settings, we found that host–symbiont compatibility was robust to variation in host genetic background, spanning within‐population, between‐population and between‐species crosses. Symbiont transmission into the F₁ generation was generally high and weakly affected by host outcrossing. Furthermore, endophytes grew equally well in planta regardless of host genetic background and transmitted at high frequencies into the F₂ generation. However, outcrossing, especially inter‐specific hybridization, reduced reproductive fitness of the host, and thereby the symbiont. Our results challenge the hypothesis that host genetic recombination, which typically exceeds that of symbionts, is a disruptive force in heritable symbioses. Instead, symbionts may be sufficiently generalized to tolerate ecologically realistic variation in host outcrossing.     

MULTITRAIT,HOST‐ASSOCIATED DIVERGENCE AMONG SETS OF BUTTERFLY POPULATIONS: IMPLICATIONS FOR REPRODUCTIVE ISOLATION AND ECOLOGICAL SPECIATION

When populations use different resources, they tend to diverge in traits that affect performance on those resources. The extent and complexity of divergence that is achieved will depend on gene flow, genetic constraints, and the character of divergent selection. We describe divergent host adaptation among Californian populations of the Melitaeine butterfly, Euphydryas editha. Divergence in seven traits created parallel phenotypic suites, each suite associated with the use of a different host species, either Collinsia torreyi or Pedicularis semibarbata. The suites involved alighting responses of adults (probably to visual stimuli), chemosensory responses to leaf surfaces, vertical positioning of adults and larvae (probably due to geotaxis), partitioning of reproductive effort among clutches, and larval performance. Remarkably, the divergent suites did not occur sympatrically, despite ubiquitous co‐occurrence of the hosts, and we know of only one site where any Collinsia species is used sympatrically with another host. In contrast, E. editha often uses two host genera sympatrically when neither of them is Collinsia. We suggest that adaptation to Collinsia is incompatible with adaptation to other hosts and may generate extrinsic postzygotic reproductive isolation among populations. Despite the apparent rarity of host‐shift‐associated speciation in Melitaeine butterflies, adoption of Collinsia as a host may lead to allopatric ecological speciation.     

Molecular evidence for hybridization in Colias (Lepidoptera: Pieridae): are Colias hybrids really hybrids?

Gene flow and hybridization among species dramatically affect our understanding of the species as a biological unit, species relationships, and species adaptations. In North American Colias eurytheme and Colias eriphyle, there has been historical debate over the extent of hybridization occurring and the identity of phenotypically intermediate individuals as genetic hybrids. This study assesses the population structure of these two species to measure the extent of hybridization and the genetic identity of phenotypic intermediates as hybrids. Amplified fragment length polymorphism (AFLP) marker analysis was performed on 378 specimens collected from northern California and Nevada. Population structure was inferred using a Bayesian/Markov chain Monte Carlo method, which probabilistically assigns individuals to genetic clusters. Three genetic clusters provided the best fit for the data. C. eurytheme individuals were primarily assigned to two closely related clusters, and C. eriphyle individuals were mostly assigned to a third, more distantly related cluster. There appeared to be significant hybridization between the two species. Individuals of intermediate phenotype (putative hybrids) were found to be genetically indistinguishable from C. eriphyle, indicating that previous work based on the assumption that these intermediate forms are hybrids may warrant reconsideration.     

Trade-offs in larval performance on normal and novel hosts

The evolution of host specialization in phytophagous insects is generally thought to involve genetic trade-offs that prevent individuals from maximizing fitness simultaneously on two or more hosts. Several hypotheses, however, have suggested that trade-offs may not be evident in experiments comparing larval performance on normal and novel hosts. Tests on survivorship, growth rate, and pupal mass among families of the swallowtail butterfly Papilio oregonius on its normal host and on a novel host provide support for these hypotheses, although they do not discriminate among them. Families differed in their relative performance on the hosts, but there was no evidence of a negative genetic correlation between hosts for any of the measures of performance. In addition, there were no correlations among the different measures, corroborating an earlier result suggesting that these different components of performance in the P. machaon species group are under at least partially separate genetic control. These results and similar results published for other insects have now produced a body of studies indicating that genetic trade-offs in individual components of larval performance may not be a major factor preventing shifts onto novel host plants. Trade-offs leading to the evolution of host specialization are more likely to involve coordination among the various components of performance together with ecological factors that allow higher fitness on one host than on others.     

Concordance of genetic and phenotypic characters across a sapsucker hybrid zone

Hybridization has presented a challenge for taxonomists and conservation biologists, since hybridizing forms could be stable evolutionary entities or ephemeral forms that are blending together. However, hybrid zones also provide a unique opportunity for evolutionary biologists who study the interaction between gene flow and reproductive isolation in speciation. Three forms of woodpeckers (sapsuckers; genus Sphyrapicus) in North America that are mostly geographically separated but hybridize with each other where they come into contact present a remarkable system for the study of hybridization. We provide the first comprehensive analysis of phenotypic and genetic variation across a hybrid zone between two of these forms, the red‐breasted Sphyrapicus ruber and yellow‐bellied S. varius sapsuckers. The objective was to infer whether selection maintains the differences between forms. Our analysis of eight morphometric and 20 plumage traits, and two molecular markers showed clear differences between the forms and roughly concordant clinal variation across a narrow hybrid zone. Thirty percent of sampled birds in the hybrid zone had mixed west/east genotypes at the genetic markers examined. The center of the genetic cline was located 20 km west of the crest of the Rocky Mountains. The width of the zone was 122 km, narrower than would be expected under neutral blending given reasonable estimates of the age of the zone and individual dispersal distances. Heterozygote deficit and cytonuclear disequilibrium at the centre of the hybrid zone suggested nonrandom mating or limited hybridization. Given these patterns and lack of evidence for habitat segregation we conclude that this hybrid zone is maintained by selection, most likely in the form of hybrid inferiority. This study provides an illustrative example of extensive hybridization between stable entities, providing additional evidence against the historical practice of treating hybridizing forms as members of the same species.     

Restoration of an endangered plant,Hygrophila pogonocalyx,leads to an adaptive host shift of the chocolate pansy (Junonia iphita iphita)

Anthropogenic introduction of a plant species may cause novel encounters between the plant and local herbivores, and initiate evolutionary changes in host plant usage by herbivores. Until recently the endemic aquatic plant Hygrophila pogonocalyx was endangered and had a restricted distribution in Taiwan. Massive restoration efforts since 1997 have led to an expansion of the plant's distribution and a novel encounter between it and an Asian butterfly, the chocolate pansy, Junonia iphita (Nymphalidae). This butterfly appears to have colonized H. pogonocalyx, switching from its original host, Strobilanthes penstemonoides var. formosana. In the present study, we aimed to investigate whether the utilization of H. pogonocalyx as a host plant has initiated a differentiation between butterflies using the novel and the original hosts. To this purpose we collected butterflies from patches of the two host plants which grow sympatrically. We tested oviposition preference for the two hosts and larval performance on them. Female adults exhibited distinct oviposition preference toward the host plant their mothers preferred. Offspring showed greater survivorship and pupal weight when fed on the host plant their mothers preferred. Male adults displayed territorial behaviors on the host plant that their mothers had preferred. Finally, the survival rate of offspring produced from cross-mating between individuals with different host plant preference was lower than that of non-hybrids. Taken together, we suggest that genetic differentiation has occurred between individuals preferring H. pogonocalyx versus S. penstemonoides as host plants via host shifting. This process was likely induced by the mass restoration of the formerly rare and endangered plant species.     

Hybridization between two parapatric ranid frog species in the northern Sierra Nevada,California, USA

Contact zones between species provide a unique opportunity to test whether taxa can hybridize or not. Cross‐breeding or hybridization between closely related taxa can promote gene flow (introgression) between species, adaptation, or even speciation. Though hybridization events may be short‐lived and difficult to detect in the field, genetic data can provide information about the level of introgression between closely related taxa. Hybridization can promote introgression between species, which may be an important evolutionary mechanism for either homogenization (reversing initial divergence between species) or reproductive isolation (potentially leading to speciation). Here, we used thousands of genetic markers from nuclear DNA to detect hybridization between two parapatric frog species (Rana boylii and Rana sierrae) in the Sierra Nevada of California. Based on principal components analysis, admixture, and analysis of heterozygosity at species diagnostic SNPs, we detected two F1 hybrid individuals in the Feather River basin, as well as a weak signal of introgression and gene flow between the frog species compared with frog populations from two other adjacent watersheds. This study provides the first documentation of hybridization and introgression between these two species, which are of conservation concern.     

A quantitative genetic analysis of leaf beetle larval performance on two natural hosts: including a mixed diet

Published quantitative genetic studies of larval performance on different host plants have always compared performance on one host species or genotype vs. performance on another species or genotype. The fact that some insects may feed on more than one plant species during their development has been neglected. We executed a quantitative genetic analysis of performance with larvae of the leaf beetle Oreinaelongata, raised on each of two sympatric host plants or on a mixture of them. Growth rate was higher for larvae feeding on Adenostylesalliariae, intermediate on the mixed diet and lowest on Cirsium spinosissimum. Development time was shortest on A. alliariae, intermediate on mixed diet and longest on C. spinosissimum. Survival was higher on the mixed diet than on both pure hosts. Genetic variation was present for all three performance traits but a genotype by host interaction was found only for growth rate. However, the reaction norms for growth rate are unlikely to evolve towards an optimal shape because of a lack of heritability of growth rate in each single environment. We found no negative genetic correlations for performance traits among hosts. Therefore, our results do not support a hypothesis predicting the existence of between‐host trade‐offs in performance when both hosts are sympatric with an insect population. We conclude that the evolution of host specialized genotypes is unlikely in the study population.