Dorsal Hindwing Research Articles

A new species of silverline butterfly, Cigaritis Donzel, 1847 (Lepidoptera: Lycaenidae), from the Western Ghats biodiversity hotspot, India.

A new species, Cigaritis conjuncta sp. nov. (Lepidoptera: Lycaenidae), is described based on five male and four female specimens from Honey Valley, Kodagu District, Karnataka, which is part of the Western Ghats biodiversity hotspot, India. Cigaritis conjuncta sp. nov. is distinguished from all other congeners in India and Sri Lanka based on the following combination of diagnostic characters in the male: (a) dorsal forewing outer half, costal margin and nearly upper half of the cell black without orange markings, the remaining wing shining deep blue, (b) dorsal hindwing tornus pale orange-red with two black spots, (c) ventral forewing bands at end of discal cell conjoined, and (e) ventral hindwing subbasal and discal bands composed of spots that are not separated but conjoined to form broad bands that have irregular outlines, leaving only a narrow background colour in between. Female is similar on the ventral side, but entirely dark brown above. Male and female genitalia are dissected and figured for two paratypes of each sex, and natural history notes on the species are provided.

Descriptions of four new species of Capys from East and West Africa with notes on adult morphology and biogeography (Lepidoptera: Lycaenidae: Theclinae)

Four new species of the hairstreak butterfly genus Capys Hewitson, [1865] (Lepidoptera: Lycaenidae: Theclinae: Deudorigini) are described: C. arba Sáfián et Fric, sp. n. (type locality: Ethiopia, Dorze Lodge), C. moroto Sáfián et Collins, sp. n. (type locality: Uganda, Mount Moroto), C. robertsi Collins et Sáfián, sp. n. (type locality: Kenya, Mount Kenya, moorland above Marania Bredt), and C. smithi Takano et Sáfián, sp. n. (type locality: Ivory Coast, Comoé National Park). Morphological features such as male genitalia, dorsal hindwing surface, androconia and labial palps, previously used in the separation of Capys species, are reviewed. Biogeography of the newly described species is also discussed using evidence from molecular analysis. With 74 figures.

Multiple Loci Control Eyespot Number Variation on the Hindwings of Bicyclus anynana Butterflies.

The underlying genetic changes that regulate the appearance and disappearance of repeated traits, or serial homologs, remain poorly understood. One hypothesis is that variation in genomic regions flanking master regulatory genes, also known as input-output genes, controls variation in trait number, making the locus of evolution almost predictable. Another hypothesis implicates genetic variation in up- or downstream loci of master control genes. Here, we use the butterfly Bicyclus anynana, a species that exhibits natural variation in eyespot number on the dorsal hindwing, to test these two hypotheses. We first estimated the heritability of dorsal hindwing eyespot number by breeding multiple butterfly families differing in eyespot number and regressing eyespot numbers of offspring on midparent values. We then estimated the number and identity of independent genetic loci contributing to eyespot number variation by performing a genome-wide association study with restriction site-associated DNA sequencing from multiple individuals varying in number of eyespots sampled across a freely breeding laboratory population. We found that dorsal hindwing eyespot number has a moderately high heritability of ∼0.50 and is characterized by a polygenic architecture. Previously identified genomic regions involved in eyespot development, and novel ones, display high association with dorsal hindwing eyespot number, suggesting that homolog number variation is likely determined by regulatory changes at multiple loci that build the trait, and not by variation at single master regulators or input-output genes.

Real-Time In Vivo Imaging of the Developing Pupal Wing Tissues in the Pale Grass Blue Butterfly Zizeeria maha: Establishing the Lycaenid System for Multiscale Bioimaging.

To systematically analyze biological changes with spatiotemporal dynamics, it is important to establish a system that is amenable for real-time in vivo imaging at various size levels. Herein, we focused on the developing pupal wing tissues in the pale grass blue butterfly, Zizeeria maha, as a system of choice for a systematic multiscale approach in vivo in real time. We showed that the entire pupal wing could be monitored throughout development using a high-resolution bright-field time-lapse imaging system under the forewing-lift configuration; we recorded detailed dynamics of the dorsal and ventral epithelia that behaved independently for peripheral adjustment. We also monitored changes in the dorsal hindwing at the compartmental level and directly observed evaginating scale buds. We also employed a confocal laser microscopy system with multiple fluorescent dyes for three-dimensional observations at the tissue and cellular levels. We discovered extensive cellular clusters that may be functionally important as a unit of cellular communication and differentiation. We also identified epithelial discal and marginal dents that may function during development. Together, this lycaenid forewing system established a foundation to study the differentiation process of epithelial cells and can be used to study biophysically challenging mechanisms such as the determination of color patterns and scale nanoarchitecture at the multiscale levels.

Fine nanostructural variation in the wing pattern of a moth Chiasmia eleonora Cramer (1780)

Butterflies and moths possess diverse patterns on their wings. Butterflies employ miscellaneous colour in the wings whereas moths use a combination of dull colours like white, grey, brown and black for the patterning of their wings. The exception is some of the toxic diurnal moths which possess bright wing colouration. Moths possess an obscure pattern in the dorsal part of the wings which may be a line, zigzag or swirl. Such patterns help in camouflage during resting period. Thus, the dorsal wing pattern of the moth is used for both intra- as well as inter-specific signal communication. Chiasmia eleonora is a nocturnal moth of greyish black colouration. The dorsal hindwing possesses yellow and black colour patches. A whitecoloured oblique line crosses both left and right fore- and hindwings to form a V-shaped pattern across the dorsal wing. This V-shaped pattern possesses a UV signal. Closer to the body, the colour appears darker, which fades towards the margin. The fine nanostructural variation is observed throughout the wings. This study elucidates the wing pattern of the geometrid moth C. eleonora using high-resolution microscopy techniques that has not been described in previous studies.

Classical lepidopteran wing scale colouration in the giant butterfly-moth Paysandisia archon.

The palm borer moth Paysandisia archon (Castniidae; giant butterfly-moths) has brown dorsal forewings and strikingly orange-coloured dorsal hindwings with white spots surrounded by black margins. Here, we have studied the structure and pigments of the wing scales in the various coloured wing areas, applying light and electron microscopy and (micro)spectrophotometry, and we analysed the spatial reflection properties with imaging scatterometry. The scales in the white spots are unpigmented, those in the black and brown wing areas contain various amounts of melanin, and the orange wing scales contain a blue-absorbing ommochrome pigment. In all scale types, the upper lamina acts as a diffuser and the lower lamina as a thin film interference reflector, with thickness of about 200 nm. Scale stacking plays an important role in creating the strong visual signals: the colour of the white eyespots is created by stacks of unpigmented blue scales, while the orange wing colour is strongly intensified by stacking the orange scales.

Giving eyespots a shiner: Pharmacologic manipulation of the Io moth wing pattern.

Our knowledge of wing pattern formation in Lepidoptera has advanced significantly in recent years due to the careful examination of several groups of butterflies. The eyespot is a prominent feature of Lepidoptera wing pattern, especially in the family Saturniidae. The present study examined how sulfated polysaccharides affected the wing pattern formation of the Io moth, Automeris io (Saturniidae). Prepupae and pupae of this species were subjected to injections of heparin and cold shock. While the cold shock had little to no effect on wing pattern, the aberrations resulting from heparin injections were moderate to profound and depended on the dose and the stage at which injection was made. The changes consisted of expansion of the black ring around the dorsal hindwing eyespots and distortion of discal spots on both dorsal and ventral sides of forewings, suggesting a possible link between genetic controls of these elements. Several different types of scales form the normal color pattern of Automeris io, and heparin-induced changes correspond to changes in shape of scales. The resulting aberrations are dubbed 'Black Eye' and 'Comet Eye.' Other known aberrations of Automeris io eyespots are summarized, illustrated, and named.

Ultrastructural variation tune wing coloration of a moth Asota caricae Fabricius, 1775.

Butterflies and moths develop highly ordered coloration in their wing for signal transmission. We have investigated the ultrastructural arrangement of wing coloration of a moth Asota caricae, applying light, optical polarized, and scanning electron microscopy, and spectrophotometry. The forewing of the moth is brown in color with a white spot at the center. The hindwing is golden yellow in color with many black patches in it. The ventral part of the forewing and dorsal hindwing share the similar color pattern. The ventral part of the hindwing has dull coloration in comparison to the dorsal one although the pattern remains same. The spectrometry analysis reveals various patterns of absorbance and reflectance spectra for various colors. The peak observed for various colors remain same although the intensity of peak changes. Bright colors possess highly ordered structures whereas irregular structures are found in dull colored scales. The color variation observed due to dorsal and ventral part of the wing is due to the minute difference observed in terms of ultrastructural arrangement revealed by scanning electron microscope. The color pattern of A. caricae is due to variation of microstructures present within the scale.

Giving eyespots a shiner: Pharmacologic manipulation of the Io moth wing pattern

Our knowledge of wing pattern formation in Lepidoptera has advanced significantly in recent years due to the careful examination of several groups of butterflies. The eyespot is a prominent feature of Lepidoptera wing pattern, especially in the family Saturniidae. The present study examined how sulfated polysaccharides affected the wing pattern formation of the Io moth, Automeris io (Saturniidae). Prepupae and pupae of this species were subjected to injections of heparin and cold shock. While the cold shock had little to no effect on wing pattern, the aberrations resulting from heparin injections were moderate to profound and depended on the dose and the stage at which injection was made. The changes consisted of expansion of the black ring around the dorsal hindwing eyespots and distortion of discal spots on both dorsal and ventral sides of forewings, suggesting a possible link between genetic controls of these elements. Several different types of scales form the normal color pattern of Automeris io, and heparin-induced changes correspond to changes in shape of scales. The resulting aberrations are dubbed ‘Black Eye’ and ‘Comet Eye.’ Other known aberrations of Automeris io eyespots are summarized, illustrated, and named.

Indicators of recent mating success in the pipevine swallowtail butterfly (Battus philenor) and their relationship to male phenotype

A key determinant of the intensity of sexual selection acting on a trait is how variation in that trait is related to variance in reproductive success of individuals. This connection compels efforts to assess lifetime mating number and how it varies among individuals in a population. In the Lepidoptera, female mating success can be assessed relatively easily by counting by the number of spermatophores in the female’s copulatory sac but male mating success in the field can often only be documented by observing copulations. Here we report a method for identifying whether or not males have recently mated that relies on the effect of mating on the appearance of the male’s reproductive tract in the pipevine swallowtail, Battus philenor. In this species laboratory experiments reveal that during mating, components of a male’s reproductive tract become shorter, decrease in mass, and change in appearance, irrespective of male age. These changes persist for at least two days after mating. After documenting these indicators of recent mating, we examined the reproductive tract of 68 field-caught males and found that twelve (17.6%) showed strong evidence of having mated recently. We found that older males were more likely to have recently mated. In addition, the color of the dorsal hindwing, a feature that females use in mate choice, was significantly greener in males, that according to our criteria, had recently-mated than in males that had not.

Colour pattern homology and evolution in Vanessa butterflies (Nymphalidae: Nymphalini): eyespot characters.

Ocelli are serially repeated colour patterns on the wings of many butterflies. Eyespots are elaborate ocelli that function in predator avoidance and deterrence as well as in mate choice. A phylogenetic approach was used to study ocelli and eyespot evolution in Vanessa butterflies, a genus exhibiting diverse phenotypes among these serial homologs. Forty-four morphological characters based on eyespot number, arrangement, shape and the number of elements in each eyespot were defined and scored. Ocelli from eight wing cells on the dorsal and ventral surfaces of the forewing and hindwing were evaluated. The evolution of these characters was traced over a phylogeny of Vanessa based on 7750 DNA base pairs from 10 genes. Our reconstruction predicts that the ancestral Vanessa had 5 serially arranged ocelli on all four wing surfaces. The ancestral state on the dorsal forewing and ventral hindwing was ocelli arranged in two heterogeneous groups. On the dorsal hindwing, the ancestral state was either homogenous or ocelli arranged in two heterogeneous groups. On the ventral forewing, we determined that the ancestral state was organized into three heterogeneous groups. In Vanessa, almost all ocelli are individuated and capable of independent evolution relative to other colour patterns except for the ocelli in cells -1 and 0 on the dorsal and ventral forewings, which appear to be constrained to evolve in parallel. The genus Vanessa is a good model system for the study of serial homology and the interaction of selective forces with developmental architecture to produce diversity in butterfly colour patterns.

A new species of Cogia from Oaxaca, Mexico (Lepidoptera: Hesperiidae: Eudaminae).

A new species of Cogia A. Butler, 1870, is described from two localities ranging from 1470 to 2000 m elevation in the Sierra Madre del Sur of Oaxaca, Mexico; it occurs in cloud forest habitats and appears to be endemic to Mexico. Cogia buena, n. sp., is closely related to C. mala Evans, 1953 and C. aventinus (Godman & Salvin, 1894); these three species are the only known Cogia taxa whose males lack a hair tuft on the dorsal hindwing, and all have similar genitalia.

Color pattern evolution in Vanessa butterflies (Nymphalidae: Nymphalini): non-eyespot characters.

A phylogenetic approach was used to study color pattern evolution in Vanessa butterflies. Twenty-four color pattern elements from the Nymphalid ground plan were identified on the dorsal and ventral surfaces of the fore- and hind wings. Eyespot characters were excluded and will be examined elsewhere. The evolution of each character was traced over a Bayesian phylogeny of Vanessa reconstructed from 7750 DNA base pairs from 10 genes. Generally, the correspondence between character states on the same surface of the two wings is stronger on the ventral side compared to the dorsal side. The evolution of character states on both sides of a wing correspond with each other in most extant species, but the correspondence between dorsal and ventral character states is much stronger in the forewing than in the hindwing. The dorsal hindwing of many species of Vanessa is covered with an extended Basal Symmetry System and the Discalis I pattern element is highly variable between species, making this wing surface dissimilar to the other wing surfaces. The Basal Symmetry System and Discalis I may contribute to behavioral thermoregulation in Vanessa. Overall, interspecific directional character state evolution of non-eyespot color patterns is relatively rare in Vanessa, with a majority of color pattern elements showing non-variable, non-directional, or ambiguous character state evolution. The ease with which the development of color patterns can be modified, including character state reversals, has likely made important contributions to the production of color pattern diversity in Vanessa and other butterfly groups.

The colouration toolkit of the Pipevine Swallowtail butterfly, Battus philenor: thin films, papiliochromes, and melanin

The ventral hindwings of Pipevine Swallowtail butterflies, Battus philenor, display a colourful pattern, created by variously coloured wing scales. Reflectance and transmittance measurements of single scales indicate that the cream and orange scales contain papiliochrome pigments, while brown, black and blue scales contain melanin. Microspectrophotometry and scatterometry of both sides of the wing scales show that the lower lamina acts as a thin film, with reflection properties dependent on the scale's pigmentation. Notably in the orange scales, the reflectance spectrum of the lower lamina is tuned to the pigment's absorbance spectrum. The dorsal hindwings of the male (but not the female) B. philenor are blue-green iridescent. At oblique illumination, the light reflected by the male's dorsal hindwings can be highly polarised, which may have a function in intersexual signalling.

MATE PREFERENCE FOR A PHENOTYPICALLY PLASTIC TRAIT IS LEARNED, AND MAY FACILITATE PREFERENCE-PHENOTYPE MATCHING

Fixed, genetically determined, mate preferences for species whose adult phenotype varies with rearing environment may be maladaptive, as the phenotype that is most fit in the parental environment may be absent in the offspring environment. Mate preference in species with polyphenisms (environmentally dependent alternative phenotypes) should therefore either not focus on polyphenic traits, be polyphenic themselves, or learned each generation. Here, we test these alternative hypotheses by first describing a female-limited seasonal polyphenism in a sexually dimorphic trait in the butterfly Bicyclus anynana, dorsal hindwing spot number (DHSN), and then testing whether male and female mate preferences for this trait exist, and whether they are seasonally polyphenic, or learned. Neither naïve males nor naïve females in either seasonal form exhibited mating preferences for DHSN. However, males, but not females, noticed DHSN variation and learned mate preferences for DHSN. These results suggest that individuals may accommodate environmentally dependent variation in morphological traits via learned mate preferences in each generation, and that learned mate preference plasticity can be sexually dimorphic.

Male wing color properties predict the size of nuptial gifts given during mating in the Pipevine Swallowtail butterfly (Battus philenor)

In many animals, males bear bright ornamental color patches that may signal both the direct and indirect benefits that a female might accrue from mating with him. Here we test whether male coloration in the Pipevine Swallowtail butterfly, Battus philenor, predicts two potential direct benefits for females: brief copulation duration and the quantity of materials the male passes to the female during mating. In this species, males have a bright iridescent blue field on the dorsal hindwing surface, while females have little or no dorsal iridescence. Females preferentially mate with males who display a bright and highly chromatic blue field on their dorsal hindwing. In this study, we show that the chroma of the blue field on the male dorsal hindwing and male body size (forewing length) significantly predict the mass of material or spermatophore that a male forms within the female's copulatory sac during mating. We also found that spermatophore mass correlated negatively with copulation duration, but that color variables did not significantly predict this potential direct benefit. These results suggest that females may enhance the material benefits they receive during mating by mating with males based on the coloration of their dorsal hindwing.

Male-specific Iridescent Coloration in the Pipevine Swallowtail (Battus philenor) is Used in Mate Choice by Females but not Sexual Discrimination by Males

We investigated the potential roles in behavioral interactions of sexually dichromatic iridescent blue coloration found on the dorsal hindwing of male Pipevine Swallowtails (Battus philenor). Behavioral experiments in a large enclosure addressed whether male dorsal hindwing coloration mediated sexual recognition by males, female choice of mates, or both. Models presented to males in the enclosure produced responses that show that males discriminate females from other males using chemical but not visual cues. In contrast, ablation of male dorsal iridescence significantly reduced male mating success with virgin females in the enclosure. The results support the hypothesis that male-specific iridescent coloration in this species is a signal that is important for females in assessing either male species identity or quality in intersexual interactions.

No Evidence that Male Choice Contributes to the Maintenance of a Shared, Sex-Limited Trait in Mimetic and Non-mimetic Female Tiger Swallowtail Butterflies, Papilio glaucus

In animal species that have morphological polymorphisms maintained by unique or divergent selection pressures, understanding the preservation of shared traits is important for identifying the factors that are influencing overall evolutionary processes. In the Eastern tiger swallowtail butterfly, Papilio glaucus, females are dimorphic. One morph (‘dark-morph’) is mostly black and mimics the toxic pipevine swallowtail, Battus philenor. These females have large amounts of blue coloration on the dorsal hind wings that enhances their mimetic resemblance. Conversely, the alternate female type (‘yellow-morph’) is similar to males in coloration with the exception of extensive dorsal blue coloration, comparable to dark-morph females. Such coloration is almost completely absent in males. We examined dorsal blue coloration in P. glaucus to determine if mimetic resemblance in dark morphs is predominantly responsible for the maintenance of dorsal blue color in both female types, or whether mate recognition and/or sexual selection by males has a stronger influence on this trait. We measured the relative amount and variance of dorsal and ventral blue coloration in females of both color morphs, as well as males. We also compared these measurements to similar ones taken in the sister species, P. canadensis (which does not exhibit female dimorphism). Lastly, we investigated mate recognition and preferences of wild males. Our results suggest that mimetic resemblance may be more important than sexual selection for sustaining dorsal blue coloration in dark-morph females and that yellow-morphs could have elevated levels of blue due to currently unknown genetic associations. Although trait correlation between sexes is common, intrasexual trait correlation in a sex-limited, polymorphic species has not been frequently observed.

Iridescent hindwing patches in the Pipevine Swallowtail: differences in dorsal and ventral surfaces relate to signal function and context

Summary 1. Iridescent colour signals are directional but, like diffusely reflected colours, vary within and among species in ways that may be adaptations to specific types of receivers in specific light environments. 2. The hindwings of pipevine swallowtail butterflies exhibit brilliant blue and iridescent colour patches on the ventral surface in both sexes and on the dorsal wing surface in males. Evidence suggests that the ventral iridescent blue is a component of the warning coloration of this distasteful species, while the dorsal blue iridescent wing area is a sexual signal. Given differences in the function and ecological context of signal production, we analysed reflectance spectra from the iridescent blue areas of both field‐caught and laboratory‐reared animals to test several predictions about the iridescent colour patches on these wing surfaces. 3. The ventral blue patches in the warning coloration of males and females should be most visible early and late in the day, due to wing orientation relative to sun angle. We therefore predicted that these iridescent colour patches would be brighter and of longer wavelengths than the male dorsal blue patch displayed during midday courtships. The prediction about reflectance intensity was supported but the prediction about hue was not. 4. We predicted that the sexually selected dorsal hindwing iridescence of males would be more variable among individuals and condition dependent than the naturally selected ventral iridescent colour patches. To assess variation and condition dependence, laboratory‐reared and field‐captured individuals were compared. The prediction about variation was not supported, but only the dorsal wing surfaces showed evidence of being condition dependent. 5. We investigated whether development of dorsal and ventral blue iridescence was coupled by determining correlations in colour properties between the wing surfaces. Our finding of positive correlations indicated a potential developmental constraint in the evolution of colour differences between the two wing surfaces. 6. Results of this study suggest that some properties of iridescent coloration on the hindwing of the pipevine swallowtail (especially intensity) may have been fine‐tuned by evolution in response to prevailing ambient light conditions and viewing perspectives that differ among types of signal receivers.

Effect of larval‐rearing density on adult life‐history traits and developmental stability of the dorsal eyespot pattern in the speckled wood butterfly,Pararge aegeria

AbstractBehavioural stresses such as crowding are thought to incur a metabolic cost to insect larvae, and fluctuating asymmetry (FA) has been shown to be a possible indicator of this stress. A study of a Madeiran population of the butterflyPararge aegeriaL. (Lepidoptera: Satyrinae) shows that larval crowding affects larval development and growth, with female larvae being more adversely affected than males. It was predicted that if larval crowding increases FA, male and femaleP. aegeriaadults may show different responses to this stress. The FA of five different wing pattern elements on the dorsal hindwings of adult male and femaleP. aegeriathat had been reared at different larval densities was measured. Crowding inP. aegeriaresulted in a trait‐specific and sex‐specific increase in FA. Although a significant correlation between FA and longevity was not observed, there was a significant correlation with egg‐laying rate. Stressed females increased their egg‐laying rate. An increased egg‐laying rate may carry a further fitness cost through the offspring of stressed females, because high egg‐laying rates are associated with a decline in oviposition site quality.