Heliconius Erato Research Articles

Shared and divergent expression domains on mimeticHeliconiuswings

Heliconius butterfly wing patterns show repeated convergence between species and have adaptive value in mimicry and mate choice, offering an opportunity to connect adaptive changes in phenotype with their underlying genotypes. Here we study forewing ommochrome pigmentation in Heliconius melpomene. We clone two new ommochrome pathway genes for the Lepidoptera, karmoisin and kynurenine formamidase (kf ), and analyze the expression patterns of all known ommochrome genes across pupal wing development. In combination with published work, this generates the first comparative gene expression data for the co-mimics Heliconius erato and H. melpomene. In both species cinnabar expression correlates with the forewing band, but the expression pattern of vermillion differs significantly between the mimics. This demonstrates that both shared and divergent expression patterns are associated with mimetic phenotypes between Heliconius species. Two genes not studied in H. erato, scarlet and possibly kf, also show enhanced expression in the forewing band of H. melpomene, implying co-ordinated upregulation of several members of this biosynthetic pathway during pattern formation.

Construction and sequence sampling of deep-coverage, large-insert BAC libraries for three model lepidopteran species.

BackgroundManduca sexta, Heliothis virescens, and Heliconius erato represent three widely-used insect model species for genomic and fundamental studies in Lepidoptera. Large-insert BAC libraries of these insects are critical resources for many molecular studies, including physical mapping and genome sequencing, but not available to date.ResultsWe report the construction and characterization of six large-insert BAC libraries for the three species and sampling sequence analysis of the genomes. The six BAC libraries were constructed with two restriction enzymes, two libraries for each species, and each has an average clone insert size ranging from 152–175 kb. We estimated that the genome coverage of each library ranged from 6–9 ×, with the two combined libraries of each species being equivalent to 13.0–16.3 × haploid genomes. The genome coverage, quality and utility of the libraries were further confirmed by library screening using 6~8 putative single-copy probes. To provide a first glimpse into these genomes, we sequenced and analyzed the BAC ends of ~200 clones randomly selected from the libraries of each species. The data revealed that the genomes are AT-rich, contain relatively small fractions of repeat elements with a majority belonging to the category of low complexity repeats, and are more abundant in retro-elements than DNA transposons. Among the species, the H. erato genome is somewhat more abundant in repeat elements and simple repeats than those of M. sexta and H. virescens. The BLAST analysis of the BAC end sequences suggested that the evolution of the three genomes is widely varied, with the genome of H. virescens being the most conserved as a typical lepidopteran, whereas both genomes of H. erato and M. sexta appear to have evolved significantly, resulting in a higher level of species- or evolutionary lineage-specific sequences.ConclusionThe high-quality and large-insert BAC libraries of the insects, together with the identified BACs containing genes of interest, provide valuable information, resources and tools for comprehensive understanding and studies of the insect genomes and for addressing many fundamental questions in Lepidoptera. The sample of the genomic sequences provides the first insight into the constitution and evolution of the insect genomes.

On the reliability of a simple method for scoring phenotypes to estimate heritability: A case study with pupal color in Heliconius erato phyllis , Fabricius 1775 (Lepidoptera, Nymphalidae)

In this paper, two methods for assessing the degree of melanization of pupal exuviae from the butterfly Heliconius erato phyllis, Fabricius 1775 (Lepidoptera, Nymphalidae, Heliconiini) are compared. In the first method, which was qualitative, the exuviae were classified by scoring the degree of melanization, whereas in the second method, which was quantitative, the exuviae were classified by optical density followed by analysis with appropriate software. The heritability (h2) of the degree of melanization was estimated by regression and analysis of variance. The estimates of h 2 were similar with both methods, indicating that the qualitative method could be particularly suitable for field work. The low estimates obtained for heritability may have resulted from the small sample size (n = 7-18 broods, including the parents) or from the allocation-priority hypothesis in which pupal color would be a lower priority trait compared to morphological traits and adequate larval development.

A hybrid zone provides evidence for incipient ecological speciation inHeliconiusbutterflies

In Heliconius butterflies, it has been proposed that speciation occurs through a combination of divergence in ecological habitat preferences and mimetic colour patterns. Here we test this hypothesis by investigating a parapatric form of the widespread species Heliconius erato. Mendelian (colour patterns) and molecular genetic data permit us to address hypotheses about introgression and genetic differentiation between different populations. Combined analysis of colour pattern, microsatellite loci and mitochondrial DNA showed that Heliconius erato venus and Heliconius erato chestertonii form a bimodal hybrid zone implying partial reproductive isolation. In a sample of 121 individuals collected in sympatry, 25% were hybrids representing a significant deficit of heterozygotes compared to the Hardy-Weinberg expectation. Seven microsatellite loci, analysed for a subset of these individuals, showed marked differentiation between the parental taxa, and unambiguously identified two genotypic clusters concordant with our phenotypic classification of individuals. Mitochondrial DNA analysis showed H. erato venus as a monophyletic group well differentiated from H. erato chestertonii, implying a lack of historical introgression between the populations. Heliconius erato chestertonii is therefore an incipient species that maintains its integrity despite high levels of hybridization. Moreover, H. erato chestertonii is found at higher altitudes than other races of H. erato and has a distinct colour pattern and mimetic relationship. Hence, there are now two examples of parapatric incipient species related to H. erato, H. himera and H. erato chestertonii, both of which are associated with higher altitudes, more arid habitats and distinct mimetic relationships. This implies that parapatric habitat adaptation is a likely cause of speciation in this group.

Borboletas (Lepidoptera: Hesperioidea e Papilionoidea) visitantes florais no Jardim Botânico da Universidade Federal de Santa Maria, Santa Maria, RS, Brasil

Butterfl ies (Lepidoptera: Papilionoidea and Hesperioidea) visiting fl owers in the Botanical Garden of the Federal University of Santa Maria, Santa Maria, RS, Brazil. Urban environments, such as parks and gardens, may offer many alimentary resources, besides shelter and favorable conditions, for butterfl y survival. This study aimed to make an inventory of butterfl ies visiting fl owers in the Botanical Garden ofthe Federal University of Santa Maria (UFSM). From March 2006 to March 2007, the fl oral visitors were observed weekly for 2h. After 108 hours’ observations, 1114 visits by 39 butterfl y species, associated with 43 plant species (21 families), were confi rmed. Among the butterfl ies, Nymphalidae had the highest richness of species (S = 18), followed by Hesperiidae (S = 8), Pieridae (S = 7), Papilionidae (S = 4) and Lycaenidae (S = 2). The pierid Phoebis philea philea was the most frequent species (187 visits), followed by hesperiids Urbanus proteus proteus (100), U. teleus (73) and the nymphalid Heliconius erato phyllis (71). Lantana camara (Verbenaceae), Eupatorium laevigatum (Asteraceae), Russelia equisetiformis (Scrophulariaceae) and Stachytarpheta cayennensis (Verbenaceae) were the most visited plants. The Botanical Garden of UFSM is an example of an urban park that seems to provide fl oral resources for the feeding of many butterfl y species, beingalso a potential refuge for species from forest areas nearby.

Spectrum of Cyanide Toxicity and Allocation in Heliconius erato and Passiflora Host Plants

Spectrum of Cyanide Toxicity and Allocation in Heliconius erato and Passiflora Host Plants

Borboletas (Lepidoptera: Hesperioidea e Papilionoidea) do campus da Universidade Federal de Santa Maria, Santa Maria, Rio Grande do Sul

Este trabalho teve por objetivo investigar a composição, riqueza e diversidade das borboletas encontradas na área do campus Camobi da Universidade Federal de Santa Maria - UFSM, Santa Maria, Rio Grande do Sul. As amostragens foram realizadas bimestralmente de setembro de 2005 a setembro de 2006, com esforço amostral padronizado em cerca de sete horas/rede entomológica/ocasião. No total de 113 horas de amostragem, foram registrados 872 indivíduos, distribuídos em 89 espécies. Do total de indivíduos, 575 (65,9%) pertencem à família Nymphalidae, 174 (19,9%) a Hesperiidae, 88 (10,1%) Pieridae, 21 (2,4%) Lycaenidae, 10 (1,2%) Papilionidae e quatro (0,5%) Riodinidae. Do total de espécies, 40 (44,9%) pertencem à família Nymphalidae, 25 (28,1%) à Hesperiidae, 11 (12,4%) Pieridae, oito (8,9%) Lycaenidae, três (3,4%) Papilionidae e duas (2,3%) Riodinidae. As cinco espécies mais abundantes do campus Camobi foram Pyrgus orcus (Stoll, 1780) (N = 78) (Hesperiidae) e os Nymphalidae Hermeuptychia hermes (Fabricius, 1775) (N = 70), Anartia amathea roeselia (Eschscholtz, 1821) (N = 55), Vanessa braziliensis (Moore, 1883) (N = 51) e Heliconius erato phyllis (Fabricius, 1775) (N = 51). Destas, apenas H. hermes e V. braziliensis, consideradas comuns em campos ou ambientes abertos, não são características de ambientes perturbados. Estudos anteriores registraram mais 12 espécies de borboletas, elevando a riqueza local de espécies para 101. Apesar de estar sujeito a forte ação antrópica, o campus Camobi ainda é capaz de abrigar uma fauna expressiva de borboletas.

Interspecific sexual attraction because of convergence in warning colouration: is there a conflict between natural and sexual selection in mimetic species?

When species converge in their colour patterns because of mimicry, and those patterns are also used in mate recognition, there is a probability of conflicting selection pressures. Closely related species that mimic one another are particularly likely to face such confusion because of similarities in their courtship behaviour and ecology. We conducted experiments in greenhouse conditions to study interspecific attraction between two mimetic butterfly species, Heliconius erato and Heliconius melpomene. Both species spent considerable time approaching and courting females of the co-mimic species. Experiments using wing models demonstrated the importance of colour pattern in this interspecific attraction. Although males of H. melpomene were attracted to their co-mimics as much as to their own females, H. erato males were more efficient at distinguishing conspecifics, possibly using wing odours. Although preliminary, these results suggest that the use of additional cues may have evolved in H. erato to reduce the cost of convergence in visual signals with H. melpomene. Overall, our results showed that there might be a cost of mimetic convergence because of a reduction in the efficiency of species recognition. Such cost may contribute to explain the apparently stable diversity in Müllerian mimetic patterns in many tropical butterfly assemblages.

Races of Heliconius erato (Nymphalidae: Heliconiinae) Found on Different Sides of the Andes Show Wing Size Differences

Heliconius erato (Linnaeus, 1758) is a brightly colored, variably patterned butterfly that is distributed from Mexico to Central and South America. It is adapted to altitudes that range from sea level to 1,600 m. and is frequently found flying in open pastures in disturbed forest and secondary growth (DeVries 1987). Its geographical races have different coloration patterns that range from orange and black, to red, yellow, and black, and to iridescent blue and pink (this pattern of coloration is shared with the co-mimic H. melpomene , (Sheppard et al. 1985). There are approximately 30 parapatric races that mate randomly in narrow hybridization zones (Emsley 1964; Mallet 1986). Brower (1994) grouped H. erato races into 2 main clades. One clade includes races that are located on the eastern side of the Andes, while the second clade is found on the western side of the Andes. Here I report the presence of statistically significant differences in wing size between individuals representing the 2 clades. I show that individuals of H. erato from the eastern side of the Andes are larger than the races from the western side. Ecological studies were performed to understand whether these size differences are due to environmental, genetic, or maternal factors (HayRoe 1996, 2004) The Lepidoptera collection of the McGuire Center for Lepidoptera and Biodiversity, FLMNH, was used to measure the size (forewing length) of different races of H. erato . The number (n) of individuals measured is shown in Table 1. Wing length was measured from the base of the wing to the apex. A Vernier caliper (Spi 2000) graduated to 0.1 mm was used to make the measurements. Four geographical races, H. e. colombina, H. e. hydara , H. e. cyrbia, and H. e. petiverana, represent the H. erato population from the western side of the Andes. These races were compared with H. e. phyllis , H. e. favorinus, and H. e. magnifica found on the eastern side of the Andes. Systat program V 10 and ANOVA were used to analyze the results and a Tukey test was used for multiple comparisons when ANOVA gave significant F values. Measurements taken from the museum specimens support the hypothesis that geographical races from the eastern side of the Andes tend to be larger than the geographical races from the western side of the Andes (Table 1). Geographical factors alone had a large effect on size ( F = 254; df = 1, 179; P < 0.001), but the races of erato found on the western side of the Andes were also significantly different in size from each other ( F = 14; df = 5, 179; P < 0.001) (Table 1). The races from the eastern region are relatively constant with respect to their body size. In contrast, the races from the western side of the Andes show more variability in sizes. In the eastern region, stabilizing selection probably acts against extreme phenotypes and favors the more common variant. This mode of selection may reduce variation and maintains the larger sizes. The races of the western side of the Andes may be exposed to greater ecological pressures, such as competitive interactions, as discussed by Benson et al. (1976). According to Brower (1994) the vicariant separation of the races could have occurred during the Cenozoic era, specifically the Pleistocene, during the formation of large mountains (including the Andes) around the world.

Highly conserved gene order and numerous novel repetitive elements in genomic regions linked to wing pattern variation in Heliconius butterflies

BackgroundWith over 20 parapatric races differing in their warningly colored wing patterns, the butterfly Heliconius erato provides a fascinating example of an adaptive radiation. Together with matching races of its co-mimic Heliconius melpomene, H. erato also represents a textbook case of Müllerian mimicry, a phenomenon where common warning signals are shared amongst noxious organisms. It is of great interest to identify the specific genes that control the mimetic wing patterns of H. erato and H. melpomene. To this end we have undertaken comparative mapping and targeted genomic sequencing in both species. This paper reports on a comparative analysis of genomic sequences linked to color pattern mimicry genes in Heliconius.ResultsScoring AFLP polymorphisms in H. erato broods allowed us to survey loci at approximately 362 kb intervals across the genome. With this strategy we were able to identify markers tightly linked to two color pattern genes: D and Cr, which were then used to screen H. erato BAC libraries in order to identify clones for sequencing. Gene density across 600 kb of BAC sequences appeared relatively low, although the number of predicted open reading frames was typical for an insect. We focused analyses on the D- and Cr-linked H. erato BAC sequences and on the Yb-linked H. melpomene BAC sequence. A comparative analysis between homologous regions of H. erato (Cr-linked BAC) and H. melpomene (Yb-linked BAC) revealed high levels of sequence conservation and microsynteny between the two species. We found that repeated elements constitute 26% and 20% of BAC sequences from H. erato and H. melpomene respectively. The majority of these repetitive sequences appear to be novel, as they showed no significant similarity to any other available insect sequences. We also observed signs of fine scale conservation of gene order between Heliconius and the moth Bombyx mori, suggesting that lepidopteran genome architecture may be conserved over very long evolutionary time scales.ConclusionHere we have demonstrated the tractability of progressing from a genetic linkage map to genomic sequence data in Heliconius butterflies. We have also shown that fine-scale gene order is highly conserved between distantly related Heliconius species, and also between Heliconius and B. mori. Together, these findings suggest that genome structure in macrolepidoptera might be very conserved, and show that mapping and positional cloning efforts in different lepidopteran species can be reciprocally informative.

An Antiaphrodisiac in Heliconius melpomene Butterflies

Gilbert (1976) suggested that male-contributed odors of mated females of Heliconius erato could enforce monogamy. We investigated the pheromone system of a relative, Heliconius melpomene, using chemical analysis, behavioral experiments, and feeding experiments with labeled biosynthetic pheromone precursors. The abdominal scent glands of males contained a complex odor bouquet, consisting of the volatile compound (E)-beta-ocimene together with some trace components and a less volatile matrix made up predominately of esters of common C16- and C18-fatty acids with the alcohols ethanol, 2-propanol, 1-butanol, isobutanol, 1-hexanol, and (Z)-3-hexenol. This bouquet is formed during the first days after eclosion, and transferred during copulation to the females. Virgin female scent glands do not contain these compounds. The transfer of ocimene and the esters was shown by analysis of butterflies of both sexes before and after copulation. Additional proof was obtained by males fed with labeled D-13C6- glucose. They produced 13C-labeled ocimene and transferred it to females during copulation. Behavioral tests with ocimene applied to unmated females showed its repellency to males. The esters did not show such activity, but they moderated the evaporation rate of ocimene. Our investigation showed that beta-ocimene is an antiaphrodisiac pheromone of H. melpomene.

Spectrum of Cyanide Toxicity and Allocation in Heliconius erato and Passiflora Host Plants

The larvae of three races of Heliconius erato were fed various species of Passiflora containing varying levels of cyanoglucosides. The mortality rate of larvae and pupae rose when larvae were fed species of Passiflora capable of releasing larger quantities of cyanide. When larvae were fed species of Passiflora with these properties, the resulting adult butterflies also released higher levels of cyanide. This may serve as a defense mechanism. The compounds responsible for the release of cyanide were not evenly distributed throughout the adult butterfly's body. The thorax contained the highest concentration of cyanogenic substances, followed by the head, wings, and abdomen. The younger tissues of Passiflora plants had higher levels of cyanide-releasing compounds than stems and mature leaves. Cyanogenic glycoside distribution within the plants is consistent with optimal allocation theory. The levels of cyanide-releasing substances in plants varied depending on the season.

Natural hybridization in heliconiine butterflies: the species boundary as a continuum.

BackgroundTo understand speciation and the maintenance of taxa as separate entities, we need information about natural hybridization and gene flow among species.ResultsInterspecific hybrids occur regularly in Heliconius and Eueides (Lepidoptera: Nymphalidae) in the wild: 26–29% of the species of Heliconiina are involved, depending on species concept employed. Hybridization is, however, rare on a per-individual basis. For one well-studied case of species hybridizing in parapatric contact (Heliconius erato and H. himera), phenotypically detectable hybrids form around 10% of the population, but for species in sympatry hybrids usually form less than 0.05% of individuals. There is a roughly exponential decline with genetic distance in the numbers of natural hybrids in collections, both between and within species, suggesting a simple "exponential failure law" of compatibility as found in some prokaryotes.ConclusionHybridization between species of Heliconius appears to be a natural phenomenon; there is no evidence that it has been enhanced by recent human habitat disturbance. In some well-studied cases, backcrossing occurs in the field and fertile backcrosses have been verified in insectaries, which indicates that introgression is likely, and recent molecular work shows that alleles at some but not all loci are exchanged between pairs of sympatric, hybridizing species. Molecular clock dating suggests that gene exchange may continue for more than 3 million years after speciation. In addition, one species, H. heurippa, appears to have formed as a result of hybrid speciation. Introgression may often contribute to adaptive evolution as well as sometimes to speciation itself, via hybrid speciation. Geographic races and species that coexist in sympatry therefore form part of a continuum in terms of hybridization rates or probability of gene flow. This finding concurs with the view that processes leading to speciation are continuous, rather than sudden, and that they are the same as those operating within species, rather than requiring special punctuated effects or complete allopatry. Although not qualitatively distinct from geographic races, nor "real" in terms of phylogenetic species concepts or the biological species concept, hybridizing species of Heliconius are stably distinct in sympatry, and remain useful groups for predicting morphological, ecological, behavioural and genetic characteristics.

Hydrogen cyanide release during feeding of generalist and specialist lepidopteran larvae on a cyanogenic plant, Passiflora capsularis

Abstract The hydrogen cyanide‐based interaction of a strongly cyanogenic plant, Passiflora capsularis, and larvae of two insect herbivores, a generalist (Spodoptera frugiperda) and a specialist (Heliconius erato), is examined in terms of the combined kinetics of the feeding process and the simultaneous hydrogen cyanide (HCN) liberation, as compared with the natural kinetics of hydrogen cyanide evolution by plant‐leaf tissue. There are marked differences in acceptance of P. capsularis by third‐instar larvae of specialist and generalist species. The former, H. erato, display a parsimonious ingestion rate of 0.74 ± 0.15 mg (fresh weight) min−1 comprising 18% active feeding time, whereas S. frugiperda larvae show a more erratic and restrained feeding involving 4% of the time at 0.45 ± 0.14 mg min−1. These S. frugiperda larvae ingest 124.4 ± 8.3 mg (fw) of the non‐cyanogeneic Spinacia oleracea leaves in 24 h compared with only 74.7 ± 20.1 mg of P. capsularis in the same period. The total hydrogen cyanide released naturally from wild specimens of P. capsularis plants is in the range 326–3901 μg g−1. Hydrogen cyanide evolution from macerated P. capsularis leaves takes place along a hyperbolic function with time and initial velocities of cyanide evolution are a linear function of total hydrogen cyanide. When feeding on P. capsularis leaves, H. erato releases only a minor fraction relative to total hydrogen cyanide (0.09%) and to the anticipated cyanide from the initial velocity (7%). By contrast, S. frugiperda evokes 5.8‐fold more than the anticipated hydrogen cyanide release from the plant. The findings are interpreted as diverging strategies by generalist and specialist insects in the utilization of hydrogen cyanide in cyanogenic plants.

The wave speed of intergradation zone in two-species lattice Müllerian mimicry model

A spatially explicit model is studied to analyse the movement of coupled clines in two-species Müllerian mimicry system as exemplified by the comimicking helicoiine butterflies in Central-South America Heliconius erato and Heliconius melpomene. In this system, a pair of comimicking wing patterns of two species (mimicry ring) is found in a geographical region but another pair of wing patterns is found in a different geographical region. The distribution of mimicry rings thus forms a spatial mosaic in a large geographical scale, and the mechanism responsible for their stable maintenance has been a long-standing question in evolutionary biology. We here examine the speed of the movement of boundaries that divide the regions inhabited by different mimetic morphs in each comimicking species, by assuming coupled two-state stochastic cellular automatons where the flipping rate of the site occupied by a mimetic morph depends on the local density of the same morph and of the comimicking morph in the other species. The speed of cline movement shows a complex dependence on the coupling parameter between mimetic species—greater coupling of comimicking morphs between species slows down the cline movement only when the reduction in predation rate exhibits diminishing return to the increase of local mimetic morph density. The analytical predictions are confirmed by the results of Monte Carlo simulations. The speed of advance is quite different from that predicted from the conventional reaction-diffusion model, indicating that demographic stochasticity plays a critical role in determining the speed of cline movement. We also examine if the spatial heterogeneity in migration rate can stably maintain clines.

Localization of Müllerian Mimicry Genes on a Dense Linkage Map ofHeliconius erato

We report a dense genetic linkage map of Heliconius erato, a neotropical butterfly that has undergone a remarkable adaptive radiation in warningly colored mimetic wing patterns. Our study exploited natural variation segregating in a cross between H. erato etylus and H. himera to localize wing color pattern loci on a dense linkage map containing amplified fragment length polymorphisms (AFLP), microsatellites, and single-copy nuclear loci. We unambiguously identified all 20 autosomal linkage groups and the sex chromosome (Z). The map spanned a total of 1430 Haldane cM and linkage groups varied in size from 26.3 to 97.8 cM. The average distance between markers was 5.1 cM. Within this framework, we localized two major color pattern loci to narrow regions of the genome. The first gene, D, responsible for red/orange elements, had a most likely placement in a 6.7-cM region flanked by two AFLP markers on the end of a large 87.5-cM linkage group. The second locus, Sd, affects the melanic pattern on the forewing and was found within a 6.3-cM interval between flanking AFLP loci. This study complements recent linkage analysis of H. erato's comimic, H. melpomene, and forms the basis for marker-assisted physical mapping and for studies into the comparative genetic architecture of wing-pattern mimicry in Heliconius.

Effect of nitrogen on Passiflora suberosa L. (Passifloraceae) and consequences for larval performance and oviposition in Heliconius erato phyllis (Fabricius) (Lepidoptera: Nymphalidae)

The present study examines the influence of nitrogen on growth rates, morphological and nutritional characteristics of Passiflora suberosa L., and the corresponding consequences for Heliconius erato phyllis (Fabricius) growth and oviposition. The treatments consisted of different nitrogen doses (0, 150 and 300 mg L(-1)) applied to the soil. Newly emerged larvae were individually reared in the laboratory (25+/-1 degree C) on branches of P. suberosa obtained from plants grown under the three nitrogen levels, and field collected females were submitted to oviposition choice tests. P. suberosa growth rates, leaf area and internode length increased significantly when grown with nitrogen enriched soil. P. suberosa presented significantly greater leaf toughness when grown without nitrogen addition. Leaves of this species also presented lower water content and toughness. Nitrogen addition increased the concentration of nitrogen and potassium on plant leaf tissues. The greater nitrogen concentration on plants did not affect H. erato phyllis larval survival, but led to a reduction in larval development time and increased adult size. Females significantly preferred to lay eggs in plants cultivated in nitrogen enriched soil. This choice was positively correlated with larval performance, which could be a possible explanation for the maintenance of that behaviour, as well as the choice of larger branches as reported in previous studies.

Pupal Melanization in Heliconius erato phyllis (Lepidoptera; Nymphalidae): Genetic and Environmental Effects

This paper deals with estimates of heritability and the effect of two different kinds of environment on the melanization of pupae in the butterfly Heliconius erato phyllis. The results for heritability showed greater differences when the offspring were regressed on male or female values for pupal colour. Pupal colour followed a system of discrete scores, from 2 for the light pupae to 5 for the darkest one. Estimate of the heritability (h2) was 0.44 when the average score of the siblings were regressed on the mid-parental value. When regressed on the male parent, h2 was larger (0.54), being very low when regressed on the female parent (0.09). Estimate by the analysis of variance was also 0.44 when both male and female sibs were included in the analysis (for male sibs, h2=0.43 and for female sibs, 0.47). To test for the effect of the environment, each brood was divided in three, one being the control, the second subjected to a black environment when reaching the fifth instar and the third to a white environment. Those in the black environment originated dark pupae that scored 5 or 4; for those in the white environment, there was no difference with the controls. Caterpillars when entering the prepupal stage were also subjected to the black treatment (early and late prepupal stage, respectively, for treatments called black A and black B). A significant effect was observed only when early prepupal stage was subjected to the black environment.

Rufous-tailed jacamars and aposematic butterflies: do older birds attack novel prey?

Although avian predators are thought to drive the evolution of warning-color mimicry in butterflies, few empirical studies directly address this assumption from the predator’s perspective. Heliconius butterflies are textbook examples of Mullerian mimicry, with perhaps the most remarkable example being the Heliconius erato and Heliconius melpomene mimicry complex. Rufous-tailed jacamars, Galbula ruficauda (Galbulidae), are well-known butterfly predators and provide an excellent study organism to investigate patterns of attack behavior in warning-colored butterflies. I investigated patterns of attack behavior by presenting three aposematic butterflies to wild-caught jacamars in a cage trial in Venezuela. I presented 80 jacamars with three Heliconius butterflies: an unaltered wing pattern (local morph) and two altered wing patterns (novel morphs). Twenty-one of 40 males and 8 of 40 females attacked a butterfly with a novel wing pattern. Of the morphological variables measured, tail length was the only significant predictor of attack behavior. Individuals with relatively longer tails attacked novel butterflies more frequently than shorter tailed individuals. Because tail length tended to increase between seasons, results suggest that older birds are more likely to attack novel aposematic prey than are young birds, contrary to the expectations that younger adult birds (i.e., more likely to be naive) would attack novel Heliconius more frequently than older birds. Overall results support the role of specialized avian predators, like jacamars, as important agents in the evolution of warning-color mimicry in butterflies and the need to investigate different age classes of birds in mimicry studies. Key words: aposematism, Galbula ruficauda, Heliconius, jacamar, Mullerian mimicry. [Behav Ecol]

Genomic tools and cDNA derived markers for butterflies

The Lepidoptera have long been used as examples in the study of evolution, but some questions remain difficult to resolve due to a lack of molecular genetic data. However, as technology improves, genomic tools are becoming increasingly available to tackle unanswered evolutionary questions. Here we have used expressed sequence tags (ESTs) to develop genetic markers for two Müllerian mimic species, Heliconius melpomene and Heliconius erato. In total 1363 ESTs were generated, representing 330 gene objects in H. melpomene and 431 in H. erato. User-friendly bioinformatic tools were used to construct a nonredundant database of these putative genes (available at http://www.heliconius.org), and annotate them with blast similarity searches, InterPro matches and Gene Ontology terms. This database will be continually updated with EST sequences for the Papilionideae as they become publicly available, providing a tool for gene finding in the butterflies. Alignments of the Heliconius sequences with putative homologues derived from Bombyx mori or other public data sets were used to identify conserved PCR priming sites, and develop 55 markers that can be amplified from genomic DNA in both H. erato and H. melpomene. These markers will be used for comparative linkage mapping in Heliconius and will have applications in other phylogenetic and genomic studies in the Lepidoptera.