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Larval food limitation in butterflies: effects on adult resource allocation and fitness.

Allocation of larval food resources affects adult morphology and fitness in holometabolous insects. Here we explore the effects on adult morphology and female fitness of larval semi-starvation in the butterfly Speyeria mormonia. Using a split-brood design, food intake was reduced by approximately half during the last half of the last larval instar. Body mass and forewing length of resulting adults were smaller than those of control animals. Feeding treatment significantly altered the allometric relationship between mass and wing length for females but not males, such that body mass increased more steeply with wing length in stressed insects as compared to control insects. This may result in changes in female flight performance and cost. With regard to adult life history traits, male feeding treatment or mating number had no effect on female fecundity or survival, in agreement with expectations for this species. Potential fecundity decreased with decreasing body mass and relative fat content, but there was no independent effect of larval feeding treatment. Realized fecundity decreased with decreasing adult survival, and was not affected by body mass or larval feeding treatment. Adult survival was lower in insects subjected to larval semi-starvation, with no effect of body mass. In contrast, previous laboratory studies on adult nectar restriction showed that adult survival was not affected by such stress, whereas fecundity was reduced in direct 11 proportion to the reduction of adult food. We thus see a direct impact of larval dietary restriction on survival, whereas fecundity is affected by adult dietary restriction, a pattern reminiscent of a survival/reproduction trade-off, but across a developmental boundary. The data, in combination with previous work, thus provide a picture of the intra-specific response of a suite of traits to ecological stress.

Animal Nutritional Physiological Phenomena↗

Differential utilization of pyrrolizidine alkaloids by males of a danaid butterfly, Parantica sita, for the production of danaidone in the alar scent organ.

Males of the chestnut tiger butterfly, Parantica sita, secrete danaidone as a major component from the alar androconial organ (sex brand). Since danaidone has been postulated to be derived from various pyrrolizidine alkaloids (PAs), which males ingest as adults from PA-containing plants, we conducted oral administration tests of several PAs to examine their availability for danaidone production by P. sita males. Males fed with a mixture of intermedine (80%) and lycopsamine (20%) produced danaidone at an average of 25.7 microg per individual, which was comparable to that found in field-caught males. In contrast, a smaller amount of danaidone (5.7 to 7.0 microg/ male) was formed when males ingested retronecine or heliotrine, and those fed with an HCI salt of monocrotaline or retrorsine produced only traces of danaidone (<0.5 microg/male). In addition, males showed a strong feeding response to intermedine/lycopsamine, whereas the other PAs elicited no positive feeding behavior. These results indicate that, unlike the arctiid moths, P. sita males can only successfully convert limited chemical types of PAs into danaidone, and further suggest that in the field, males selectively ingest particular PAs that are readily transformable into danaidone.

Animals↗

The effects of ants on the entomophagous butterfly caterpillar Feniseca tarquinius, and the putative role of chemical camouflage in the Feniseca-ant interaction.

Butterfly caterpillars in the lycaenid subfamily Miletinae are predators of ant-tended Homoptera, yet they lack specialized secretory and call-production organs crucial to ant association in other lycaenids. Here, we address the question of how miletine caterpillars have invaded the ant-Homoptera symbiosis through a study of the only New World miletine, Feniseca tarquinius, a predator of the wooly aphid Prociphilus tesselatus. Previous interpretations have suggested that F. tarquinius and other miletine caterpillars avoid ant aggression by concealing themselves under silken webs. In contrast, our field data indicate that F. tarquinius caterpillars are less likely to be concealed in the presence of the ants Camponotus pennsylvanicus and Formica obscuriventris than in the absence of ants, although caterpillar and ant behaviors vary between years. Chemical analysis and behavioral assays suggest that chemical camouflage, not physical concealment, is responsible for the ants' failure to detect and remove F. tarquinius caterpillars from aphid colonies. Analyses by gas chromatography indicate that the cuticular lipid composition of caterpillars are similar to that of their aphid prey, although it varies with prey species. Behavioral assays confirm that solvent extracts of F. tarquinius caterpillars and P. tesselatus aphids evoke similar behavioral responses in C. pennsylvanicus ants. Chemical camouflage is well known in social parasites of ants, but the present study represents one of a few documented cases where chemical deceit is important to interactions with ants outside the nest.

Animals↗

Color and spatial antagonism in the visual system of the larval swallowtail butterfly.

Many neurons exhibiting an antagonistic interaction between signals from one particular stemma (center) and neighboring stemmata (surround) were found in the second optic neuropil (medulla) of the larval swallowtail butterfly. Furthermore, most of the neurons showed a color opponent interaction in the center: combinations of opponent colors were blue vs green, UV vs green, and blue vs UV + green. Thus, photoreceptor signals are integrated into chromatically and spatially antagonistic mechanisms, even in the invertebrate.

Animals↗

Patterning during pupal commitment of the epidermis in the butterfly, Precis coenia: the role of intercellular communication.

The commitment of cells to pupal development in the larvae of holometabolous insects can be prevented by treatment with juvenile hormone (JH) or a JH mimic during a critical period early in the last larval instar. By treating larvae of different ages with a JH mimic, pupal commitment of the epidermis of the butterfly, Precis coenia, was found to occur in a strict temporal and spatial progression, which was serially homologous and occurred independently in each segment. The mechanism underlying this sequence of pupal commitment was examined by cauterizing regions of the epidermis to observe the effects of local ablation on the pattern of pupal commitment revealed by treatment with the JH mimic. Cautery of the segmental site of origin of pupal commitment, the dorsal midline, suppressed pupal commitment in the rest of the operated segment, indicating that the midline has a special effect on commitment of the rest of the segment. Cautery off the midline produced asymmetries in the pattern of pupal commitment; when placed close to the midline, such cauteries prevented pupal commitment in the region "downstream" of the cautery, suggesting that a signal (diffusible or transducible) emanates from the midline. Finally, cautery of a circle around the midline inhibited pupal commitment only outside the circle, showing that cautery could act as a barrier to the passage of a signal coming from the midline. These results suggest that inductive as well as hormonal signals are involved in the regulation of pupal commitment in the epidermis of the lepidopteran, P. coenia.

Animals↗

Quinone methide as a reactive intermediate formed during the biosynthesis of papiliochrome II, a yellow wing pigment of papilionid butterflies.

Mushroom tyrosinase and the recently identified, 4-alkyl-o-benzoquinone: 2-hydroxy-p-quinone methide isomerase were used to investigate the mechanism of biosynthesis of papiliochrome II pigment found in the yellow scales of the papilionid butterflies. Incubation of N-beta-alanyldopamine (NBAD) and L-kynurenine with mushroom tyrosinase resulted in the formation of adducts tentatively characterized as NBAD quinone-L-kynurenine adducts. If quinone isomerase was included in this reaction mixture, the formation of two new products could be witnessed. These two products exhibited the same retention time and the same UV and visible spectral properties as those of papiliochrome II diastereoisomers. Since quinone isomerase catalyzes the conversion of quinones to quinone methides, the above studies indicate that papiliochrome II biosynthesis involves non-enzymatic and hence non-stereoselective condensation of enzymatically generated NBAD quinone methide with L-kynurenine.

Animals↗

Oviposition stimulants for the monarch butterfly: flavonol glycosides from Asclepias curassavica.

The monarch butterfly, Danaus plexippus oviposits on milkweed plants, primarily within the Asclepiadaceae. Oviposition stimulants responsible for host plant recognition were isolated from Asclepias curassavica. Six flavonoid glycosides-quercetin 3-O-(2",6"-alpha-L-dirhamnopyranosyl)-beta-D-galactopyranoside, quercetin 3-O-beta-D-glucopyranosyl-(1-->6)-beta-D-galactopyranoside, quercetin 3-O-(2"-O-alpha-L-rhamnopyranosyl)-beta-D-galactopyranoside, quercetin 3-O-alpha-L-rhamnopyranosyl-(1-->6)-beta-D-glucopyranoside, quercetin 3-O-beta-D-galactopyranoside, quercetin 3-O-beta-D-glucopyranoside, and an unidentified flavonoid mixture were isolated and characterized from this plant. An additional glycoside, possibly quercetin 3-O-(2",6"-alpha-L-dirhamnopyranosyl)-beta-D-glucopyranoside, which could not be separated from the first triglycoside, was also found in some batches of plant extract. The two dirhamnosyl glycosides, the glucosylgalactose and the rutinoside were found to be active as oviposition stimulants at 0.5 g leaf equivalents.

Animals↗

Synthesis of 3-hydroxyretinal in the cytosol of the butterfly compound eye.

The metabolism of 3-hydroxyretinoids in the cytosol of the compound eyes of a species of butterfly, Papilio xuthus, was investigated. The cytosol was found to contain 25-30% of the total 3-hydroxyretinal and 70-82% of the total 3-hydroxyretinol in the eye. These percentages of 3-hydroxyretinoids in the cytosol were found to be constant regardless of whether the eyes are light-adapted or dark-adapted. 3-Hydroxyretinal can be newly synthesized in the cytosol of light-adapted eyes. Blue light specifically increases the amount of 11-cis and all-trans 3-hydroxyretinal ca 2.5 and 1.8 times respectively, compared to pre-irradiation. When 3-hydroxyretinal was synthesized, 3-hydroxyretinol was decreased or disappeared in the cytosol. When retinol (non-native chemical) was added to the cytosol, it was converted into retinal. This result indicates that an oxidative system exists in the compound eye which can convert 3-hydroxyretinol to 3-hydroxyretinal.

Adaptation, Ocular↗

Light suppresses the activity of serotonin-immunoreactive neurons in the optic lobe of the swallowtail butterfly.

Intracellular recordings and stainings were made from 5-HT-immunoreactive neurons with a large soma at the anterior edge of the second optic neuropile of the swallowtail butterfly Papilio xuthus. The neurons have a dendritic arborization in the third optic neuropile and project axonal processes to the first optic neuropile and the dorsal protocerebrum. The neurons discharged impulses in the dark and the impulse discharge was inhibited by illumination of the ipsilateral eye. The anatomical and physiological profiles suggest that the neurons may have a role in controlling the light-dark adaptation or the overall activity of the optic lobe and the midbrain.

Animals↗

The pheromone system of the male danaine butterfly, Idea leuconoe.

Male Idea leuconoe butterflies release a complex mixture of volatiles from their pheromone glands (hairpencils) during courtship. The pheromone components geranyl methyl thioether (2), viridifloric beta-lactone (3) and 6-hydroxy-4-dodecanolide (10) have been synthesized for the first time. Therefore, the structural assignment of these new natural products could be proved. Related 7-hydroxy-5-alkanoides are also present in the extract. The volatiles are embedded in a lipidic, matrix with more than 150 components. This matrix consists of alkanes, alkenes, 2,5-dialkyltetrahydrofurans, secondary alkanols and alkenols as well as alkanones and alkenones. Several regioisomers of the oxidized hydrocarbons occur. The elucidation of double bond positions has been performed by MS using DMDS adducts.

Animals↗

Characterization and cDNA cloning of hinnavin II, a cecropin family antibacterial peptide from the cabbage butterfly, Artogeia rapae.

Hinnavins, together with lysozymes, are the main types of antibacterial peptides/proteins previously isolated from the larval haemolymph of the cabbage butterfly, Artogeia rapae as part of the humoral immune response to a bacterial invasion. One of these antibacterial peptides, named hinnavin II, was purified and characterized after cDNA cloning. The purified hinnavin II was more active against Gram negative than against Gram positive bacteria. Hinnavin II also showed a powerful synergistic effect on the inhibition of bacterial growth with purified lysozyme. The cDNA has a total length of 186 bp with a 114 coding region. The deduced protein sequence contains 38 amino acids with a coding capacity of 4142.8 Da. The result of a multiple sequence alignment and phylogenetic analysis with Clustal W indicated that mature hinnavin II showed an approximately 78.9% amino acid sequence identity with cecropin A and originated from a group containing mostly lepidopteran cecropins.

Amino Acid Sequence↗

Pattern formation: a focus on notch in butterfly eyespots.

New observations of early and dynamic expression of Notch in developing lepidopteran wings suggests that this signalling pathway may function in defining the central focus that will specify the butterfly eyespot colour pattern.

Animals↗

Competing selfish genetic elements in the butterfly Hypolimnas bolina.

Maternally inherited selfish genetic elements are common in animals . Whereas host genetics and ecology are recognized as factors that may limit the incidence of these parasites , theory suggests one further factor-interference with other selfish elements-that could affect their prevalence . In this paper, we show that spatial heterogeneity in the occurrence of the male-killing Wolbachia wBol1 in the tropical butterfly Hypolimnas bolina is caused by a second infection that can exclude the male-killer. We first provide evidence of a second Wolbachia strain, wBol2, present in most populations that do not carry the male-killer but rare or absent when the male-killer is present. Crossing data indicate that wBol2 in males induces cytoplasmic incompatibility to both uninfected and wBol1-infected females. The wBol2 infection can therefore not only spread through uninfected populations but also resist invasion by wBol1. Thus, we provide empirical support for the hypothesis that the incidence of particular selfish genetic elements can limit the presence of competing types.

Animals↗

Molecular systematics and evolution of the "Apollo" butterflies of the genus Parnassius (Lepidoptera: Papilionidae) based on mitochondrial DNA sequence data.

Sequences of 777 bp of mtDNA-ND5 locus were determined in order to shed light on the molecular systematics and evolution of the "Apollo" butterflies. Examined were nearly all of about 50 species of the genus Parnassius, together with seven species of the allied genera in the subfamily Parnassiinae (Papilionidae). The NJ and the MP phylogenetic trees show that the "Apollos" constitute a monophyletic group, comprising a number of cluster groups probably reflecting a relatively rapid radiation in evolution. The clusters of species-groups denoted I-VIII correspond to those species-groups recognized on the basis of morphological characters. Our findings will also help understand the biological relationships among several species or subspecies on which the classical taxonomy is in dispute. The unexpected finding is that among the samples of allied genera compared, Hypermnestra helios appears to be the most closely related to the "Apollos", despite morphological and behavioral dissimilarity. Furthermore, in contrast to the previous higher taxonomy, Archon apollinus which is classified in the tribe Parnassiini was found genetically closer to the tribe Zerynthiini, raising a taxonomic controversy.

Animals↗

Expression of one isoform of GTP cyclohydrolase I coincides with the larval black markings of the swallowtail butterfly, Papilio xuthus.

The larva of the swallowtail butterfly Papilio xuthus changes its body markings during the fourth ecdysis. We found that stage-specific cuticular black markings are mainly regulated by co-localization of two melanin synthesis enzymes; tyrosine hydroxylase (TH) and dopa decarboxylase (DDC). TH converts tyrosine to dihydroxyphenylalanine (dopa), and tyrosine itself is converted from phenylalanine by phenylalanine hydroxylase (PAH). Guanosine triphosphate cyclohydrolase I (GTPCHI) is essential for the synthesis of tetrahydrobiopterin (BH4) that is a cofactor of TH and PAH. In this report, we found that a GTPCHI inhibitor prevents pigmentation in cultured integuments, suggesting that the GTPCHI activity is also involved in cuticle pigmentation. We have cloned GTPCHI and PAH cDNAs from P. xuthus and investigated their spatial expression patterns in epidermis by whole-mount in situ hybridization. There are two isoforms of GTPCHI in larval epidermis (GTPCHIa and GTPCHIb). GTPCHIa is expressed at the black markings of the subsequent instar, similar to TH, whereas GTPCHIb is expressed uniformly, similar to PAH. This suggests that the region-specific expression of GTPCHIa supplies sufficient BH(4) reinforcing the TH activity in black marking area. Our results imply that larval markings are regulated by not only melanin synthesis enzymes but also the cofactor supplying enzyme.

Amino Acid Sequence↗

Two different sensory mechanisms for the control of pupal protective coloration in butterflies.

The butterflies Graphium sarpedon nipponum Fruhstorfer and Papilio xuthus Linné show pupal protective color polymorphism, but the two species appear to have different sensory mechanisms for determining pupal coloration. When light was of sufficient illumination, the larvae of Graphium sarpedon became bright yellowish green pupae on white pupation boards and reddish brown pupae on black pupation boards. The pupal coloration thus strongly depended on the brightness of the pupation site. In addition, larvae became bright yellowish green pupae in complete darkness. From these results, measurement of the illumination suggested that pupal color is determined by the illuminant difference between incidence light from the dorsal direction and ventral light from a paper board; i.e., the sum of the reflected light of the board plus the penetrated light passing through the board. The illuminant difference required for reddish brown coloration was 40 lux or more. The optical signals received through the stemmata during a critical period before formation of the thorax garter (band string) were important for coloration. By contrast, in Papilio xuthus, successive tactile signals from a rough surfaced pupation site during a critical period before and after formation of the garter were important for determining brown pupal coloration.

Animals↗

Interactions of environmental factors influencing pupal coloration in swallowtail butterfly Papilio xuthus.

The swallowtail butterfly Papilio xuthus Linné [Lepidoptera: Papilionidae] exhibits pupal protective color polyphenism. Interactions of various environmental factors on pupal coloration were analyzed in non-diapausing individuals. Under sufficient light (200lux), most pupating larvae became green pupae when the surface of the pupation site was smooth, while they became brown when the surface was rough. Tactile signals are the positive environmental factors causing induction of the brown pupal coloration. In dark boxes, the induction of the brown pupal coloration was easily induced even on a smooth surface, suggesting that light suppresses induction of brown coloration. Different colors of pupation sites did not affect pupal coloration under sufficient light. Environmental factors received during a critical period both before girdling and after girdling affected pupal coloration. When tactile signals received from rough surfaces reach threshold levels during pupation, brown pupal coloration is determined. Larvae reared under a daily periodicity of natural light formed a girdle at midnight, subsequently, the prepupae received strong daylight the following day. Under natural light most larvae produced brown pupae on rough surfaces and green pupae on smooth surfaces.

Animals↗

Major components in the hairpencil secretion of a butterfly, Euploea mulciber (Lepidoptera, Danaidae): their origins and male behavioral responses to pyrrolizidine alkaloids.

Two compounds, 9,10-epoxytetrahydroedulan (ET) and viridiflorine beta-lactone (VL), were identified as major components from the hairpencils of field-caught males of a danaid butterfly, Euploea mulciber. By contrast, laboratory-reared males entirely lacked VL, but possessed a significant quantity of ET. Various feeding experiments with larvae and indoor adult males strongly suggested that ET is biosynthesized de novo only after eclosion from nutrients ingested during the larval development. Since VL was suspected to be derived from pyrrolizidine alkaloids (PAs) acquired as an adult, tests for feeding response to and oral administration of four PAs (a 4:1 mixture of intermedine/lycopsamine, heliotrine, monocrotaline, and retronecine) were conducted. When the tarsi or proboscis were stimulated with PA solutions, males showed positive feeding responses (proboscis extension and sucking movements) to intermedine/lycopsamine, heliotrine, and retronecine in decreasing order of responsiveness, thereby providing evidence that male adults are endowed with taste receptor(s) specific to PAs on the legs as well as on the proboscis. Differently from gustatory responsiveness, only males fed with intermedine/lycopsamine produced a significant quantity of VL (ca. 35 microg/male), whereas those that ingested heliotrine or monocrotaline hydrochloride produced traces of VL (<0.18 microg/male). Uptake of retronecine did not lead to VL formation at all. In behavioral bioassays to test the attractivity of PAs to males, all individuals tested were attracted exclusively to intermedine/lycopsamine. This shows that certain PA(s) per se serve as attractant(s) for males in locating PA sources, and further suggests that in the field, males will seek particular PA(s) that are indispensable as precursors for the efficient biosynthesis of VL.

Animal Communication↗