Atrophaneura Alcinous Research Articles

Asymmetric interactions between two butterfly species mediated by food demand.

Recent studies on insect interactions on plants have revealed that herbivorous insects indirectly interact with each other through changes in plant traits following herbivory. However, less attention has been given to plant biomass relative to plant quality in relation to indirect interactions among herbivores. We explored the extent to which the larval food demand of two specialist butterflies (Sericinus montela and Atrophaneura alcinous) explains their interaction on a host plant, Aristolochia debilis. A laboratory experiment showed that plant mass consumption by A. alcinous larvae was 2.6 times greater than that by S. montela. We predicted that A. alcinous, which requires more food, is more vulnerable to food shortages than S. montela. In a cage experiment, an asymmetric interspecific interaction was detected between the two specialist butterflies; S. montela larval density significantly decreased the survival and prolonged the development time of A. alcinous, but A. alcinous density affected neither the survival nor the development time of S. montela. The prediction based on the food requirement was partly supported by the fact that increasing A. alcinous density likely caused a food shortage, which more negatively affected A. alcinous survival than S. montela survival. Conversely, increasing the density of S. montela did not reduce the remaining food quantity, suggesting that the negative effect of S. montela density on A. alcinous was unlikely to be due to food shortage. Although aristolochic acid I, a defensive chemical specific to Aristolochia plants, did not influence the food consumption or growth of either butterfly larva, unmeasured attributes of plant quality may have mediated an indirect interaction between the two butterflies. Consequently, our study suggests that not only the quality but also the quantity of plants should be considered to fully understand the characteristics, such as symmetry, of interspecific interactions among herbivorous insects on the same host plant.

Electrophysiological and Morphological Characterization of Contact Chemosensilla in Adults and Larvae of the Butterfly, Atrophaneura alcinous

Simple SummaryIn butterflies, oviposition and feeding are induced by the recognition of host plant compounds with contact sensory organs called chemosensilla. Oviposition behavior of the butterfly Atrophaneura alcinous involves detection of two plant compounds—an alkaloid and a monosaccharide—using tarsal contact chemosensilla of the foreleg. Feeding behavior of the adults and larvae involves detection of sucrose, sodium chloride, and other chemicals. We examined the distribution and electrophysiological response of contact chemosensilla in A. alcinous adults and larvae. In adults, tarsal contact chemosensilla were classified into long- and short-type sensilla. Long-type sensilla were distributed widely much more in females than in males, whereas short-type sensilla were found at the edge of the tarsi in a similar manner in both sexes. Long- and short-type sensilla showed different responses to methanol extracts of host plant compounds. Our findings suggest that the adult butterfly discriminates between host plant compounds using long-type sensilla during oviposition and recognizes the taste using short-type sensilla. In the larval mouthparts, contact chemosensilla were present on the maxillary galea and epipharynx. Electrophysiological responses of chemosensilla suggest that larvae can discriminate between host plant compounds.Distribution and electrophysiological responses of contact chemosensilla were examined in the Aristolochiaceae-feeding butterfly Atrophaneura alcinous. In adult butterflies, tarsal contact chemosensilla of the foreleg were classified into two groups based on length: long- and short-type sensilla. Long-type sensilla were distributed much more widely in females than in males, whereas short-type sensilla were found at the edge of the tarsi in a similar manner in both sexes. Taste responses of the long- and short-type sensilla to methanol extracts of Aristolochia debilis and Citrus spp. were recorded. Aristolochia debilis extracts evoked spikes with different amplitudes, whereas Citrus spp. extracts evoked spikes with a single amplitude in the long-type sensilla. Short-type sensilla did not respond to either extract. Moreover, we recorded responses to different concentrations of sucrose and NaCl. Results suggest that adult butterflies can discriminate the taste of host plant components from other chemicals using long-type sensilla during oviposition and may recognize diets containing sugar and salts during feeding using short-type sensilla. In the larval mouthparts, there were lateral and medial styloconic sensilla on the maxillary galea and epipharyngeal sensillum on the epipharynx. Electrophysiological responses of these sensilla suggest that larvae can discriminate between host plant compounds.

Interpopulational variation in pupal diapause of the butterfly Atrophaneura alcinous (Klug) (Lepidoptera, Papilionidae) in the Kanto District, eastern Japan

Interpopulational variation in pupal diapause of the butterfly Atrophaneura alcinous (Klug) (Lepidoptera, Papilionidae) in the Kanto District, eastern Japan

Larval growth and the effect of temperature on head capsule size in Atrophaneura alcinous (Lepidoptera: Papilionidae)

The size of head capsule is one of the most important factors for identifying developmental stage. In order to understand the developmental characteristics of the butterfly Atrophaneura alcinous, we examined fluctuations in larval head capsule size under three different temperature conditions (20, 25, and 30°C) and 60% humidity. As a result, larvae developed to the fifth instar at all three temperatures. The head capsule size of larvae tended to be larger at the lowest temperature and smaller at the highest temperature. The development rate showed a regular change, consistent with Dyar`s rule regarding head capsule size development. Furthermore, the development of head capsule size was found to correspond to a second degree polynomial regression better than to a linear regression. On the basis of these results, it would appear to be possible to perform an accurate assessment of instar status during the development of A. alcinous.

How do two specialist butterflies determine growth and biomass of a shared host plant?

AbstractAlthough insect herbivory can modify subsequent quantity and quality of their host plants, change in plant quantity following herbivory has received less attention than plant quality. In particular, little is known about how previous herbivore damage determines plant growth and biomass in an insect species‐specific manner. We explored whether herbivore species‐specific food demand influences plant growth and biomass. To do this, we conducted a series of experiments and field survey using two specialist butterflies, Sericinus montela and Atrophaneura alcinous, and their host plant, Aristolochia debilis. It is known that A. alcinous larva requires four times more food resources to fulfill its development than S. montela larva. Despite that A. alcinous larvae imposed greater damage on plants than S. montela larvae, plant growth did not differ due to herbivory by these species both in single and multiple herbivory events. On the other hand, total aboveground biomass of the plants was reduced more by A. alcinous than S. montela feeding regardless of the number of herbivory events. Feeding on plants with a history of previous herbivory neither decreased nor increased larval growth. Our results suggest that food demand of the two butterfly species determined subsequent plant biomass, although the plant response may depend on tolerance of the host plant (i.e., ability to compensate for herbivore damage). Such difference in the effects of different herbivore species on host plant biomass is more likely to occur than previously thought, because food demand differs in most herbivore species sharing a host plant.

The transfer of titanium dioxide nanoparticles from the host plant to butterfly larvae through a food chain.

This study aimed to examine the transfer of nanoparticles within a terrestrial food chain. Oviposited eggs of the swallowtail butterfly (Atrophaneura alcinous) were hatched on the leaves of the host plant (Aristolochia debilis), and the root stock and root hairs were submerged in a suspension of 10 μg/ml titanium dioxide nanoparticles (TiO2-NPs) in a 100 ml bottle. The presence of TiO2-NPs in the veins of the leaves was confirmed by X-ray analytical microscopy (X-ray AM). The hatched 1st instar larvae fed on the leaves to moult into 2nd instar larvae. Small agglomerates of TiO2-NPs less than 150 nm in diameter were identified in the vascular tissue of the exposed plant, the midgut and the excreta of the larvae by transmission electron microscopy. The image of Ti elemental mapping by X-ray AM was analysed with the quantitative spatial information mapping (QSIM) technique. The results demonstrated that TiO2-NPs were transferred from the plant to the larvae and they were disseminated throughout the environment via larval excreta.

Effects of Temperature on the Development of Chinese Windmill Butterfly, Atrophaneura alcinous (Lepidoptera: Papilionidae)

The Chinese windmill butterfly, Atrophaneura alcinous, is an important butterfly for exhibition in butterfly garden. The objective of this study was to determine the effect of temperature on A. alcinous in the laboratory. Development of A. alcinous reared on leaves of Aristolochia contorta was investigated at five constant the laboratory condition (20, 22.5, 25, 27.5 and <TEX>$30^{\circ}C$</TEX>) and at relative humidity of 60% with a photoperiod of 14:10 (L:D). Temperatures have been suggested as an important determinant of developmental rate, lifespan and mortality in invertebrates. As the temperature increased, the length of the developmental period gradually decreased. The developmental time (pupation) from egg hatching to pupation was respectively 25.8, 23.6, 19.6, 15.5, and 12.9 days at the temperatures of 20, 22.5, 25, 27.5 and <TEX>$30^{\circ}C$</TEX>. And pupation was respectively 40.0, 30.0, 63.4, 50.0, 23.3% at the temperatures of 20, 22.5, 25, 27.5 and <TEX>$30^{\circ}C$</TEX>. The developmental threshold temperature estimated for egg-to-pupae was 10.8, with a thermal constant of 230.4 degree-days. Therefore, the optimal developmental temperature for A. alninous was determined to be <TEX>$25^{\circ}C$</TEX>. To compare the effects of the total duration of chilling on the termination of diapause, larvae were subjected to a temperature of <TEX>$8^{\circ}C$</TEX> from 60 to 120 days. The rate of termination of diapause was significantly higher at 60 days compared to other incubation period.

An oviposition stimulant binding protein in a butterfly

Oviposition is evoked by plant compounds, which are recognized by chemoreceptive organs of insects. The swallowtail butterfly, Atrophaneura alcinous, oviposits its eggs on the host plant, Aristolochia debilis, in the presence of only two stimulating compounds: an alkaloid, aristolochic acid, and a monosaccharide, sequoyitol. In our previous study, a unique protein of 23 kDa [Oviposition stimulant(s) binding protein (OSBP)] was found in the forelegs of female, but not male, A. alcinous. The electrophysiological response of A. alcinous to an extract of A. debilis was depressed by the presence of OSBP antiserum, suggesting that OSBP presumably binds to oviposition stimulant(s). We show here, using a highly sensitive fluorescence micro-binding assay that native OSBP binds to a main oviposition stimulant, aristolochic acid, from its host plant, A. debilis. Three-dimensional molecular modeling studies also gave a reasonable structure for the OSBP / aristolochic acid complex. This is the first report of a native chemoreceptive protein binding to an oviposition stimulant ligand in insects.

Environmental Factors Affecting Black/White Coloration of the Silken Girdle in the Swallowtail Butterfly, Atrophaneura alcinous (Lepidoptera: Papilionidae)

The silken girdles of pupae of the swallowtail butterfly Atrophaneura alcinous show black and white color diphenism. Field observations revealed that all pupae observed on non-food plants and the leaves and stems of the larval food plant Aristolochia debilis were classified as a silken girdle of a black type, while a large portion of pupae pupating on the twigs and trunks of cherry trees in close proximity to A. debilis were classified as a silken girdle of a black type. Additionally, all pupae observed on the surfaces of artificial objects in areas where there are no surrounding plants or trees were classified as a silken girdle of a white type. We demonstrated the effect of day length and the texture, light, plant odor and humidity of pupation sites on the coloration of the silken girdle in A. alcinous. Regardless of long-day or short-day day length conditions, light conditions of constant light or dark, or the presence of a plant odor of A. debilis as environmental cues, all larvae placed at over 80% relative humidity (R.H.) developed into pupae with a silken girdle of a black type. However, all larvae developed into pupae with a silken girdle of a white type when R.H. was below 75%. Furthermore, when pupae with a silken girdle of a white type were transferred to conditions of 90% R.H. within 24 hr of pupation, the white color of the silken girdle changed into a black type within 24 hr of the transfer. The present data suggest that the induction of a black coloration of the silken girdle in A. alcinous requires a R.H. of approximately 80% or more as an environmental factor.

Geographic variation in photoperiodic response for the induction of pupal diapause in the Aristolochia-feeding butterfly Atrophaneura alcinous

The photoperiodic response curve for pupal diapause induction of the butterfly Atrophaneura alcinous was determined in several temperate and subtropical populations of the Japan Archipelago at 20°C, using Aristolocia debilis leaves as larval food. Altitude and host plant use also differed among them. For the temperate populations of Kiire, Kashihara, Fuchu, Yokosuka and Yamagata, the critical photoperiod increased with latitude, and the subtropical Ishigaki population showed the shortest critical photoperiod. Further, a change in the shape of photoperiodic response curve, but no shift in the critical photoperiod, was found in a mountain population of Kiire, giving a high incidence of diapause in long photoperiods. These results are discussed in relation to latitude, altitude, and host plant.

Seasonal occurrence of the Aristolochia-feeding butterfly Atrophaneura alcinous (Lepidoptera, Papilionidae) : Comparison between the lowland and mountain populations

Seasonal occurrence of the Aristolochia-feeding butterfly Atrophaneura alcinous (Lepidoptera, Papilionidae) : Comparison between the lowland and mountain populations

A putative binding protein for lipophilic substances related to butterfly oviposition

A unique protein of 23 kDa (Jf23) was found in the tarsus of the female swallowtail butterfly, Atrophaneura alcinous. Jf23 has 38% identity with a bilin-binding protein, which was found in the cabbage butterfly, Pieris brassicae, and which has two consensus sequences in common with the members of the lipocalin family, suggesting that it is a binding protein for lipophilic ligands. Western blot analysis showed that Jf23 was expressed only in the female, and not in the male. Electrophysiological response of the female tarsi was stimulated by methanolic extract of their host plant, Dutchman’s pipe ( Aristolochia debilis). The stimulated response was depressed by the presence of Jf23 antiserum. These results suggest that Jf23 is one of the chemosensory signaling proteins, which plays one or more roles in female butterfly oviposition.

Morphological evidence that spermatogenic cells undergo apoptosis within testis of diapausing swallowtail pupa (Atrophaneura alcinous)

In many lepidopteran species, degeneration of germ cells occurs in the testis of diapausing pupa. We demonstrated that degenerating cells clearly exhibit apoptotic characteristics by supravital staining, TUNEL assay and ultrastructural observations. These characterizations allowed us to identify several steps of spermatocyte degeneration, segregation of nuclear chromatin, nuclear fragmentation, cytoplasmic condensation and phagocytosis by cyst cells. A cluster of TUNEL-positive cells were observed under the light microscope which coincided with a cyst of degenerating spermatocytes revealed under the electron microscope. Secondary spermatocytes in pupal testis undergo apoptotic cell death during diapause. Therefore, neither cysts of spermatids nor sperm bundles exist in testis of diapausing pupa. Our observations also indicate that spermatogenic cells undergo phagocytosis of cyst cells as the diapause period increased. This phenomenon seems to be associated with the seasonal reproductive strategy. J. Exp. Zool. 283:194–201, 1999. © 1999 Wiley-Liss, Inc.

Morphological changes in eupyrene and apyrene spermatozoa in the reproductive tract of the male butterfly Atrophaneura alcinous Klug

Summary Eupyrene and apyrene spermatozoa are contained in separate cysts in the testis of the butterfly Atrophaneura alcinous. Spermatozoa of both types from various parts of the male reproductive tract were examined with particular reference to their morphological characteristics. All spermatozoa collected from the vas deferens and the vesicula seminalis were found to be immotile under a dissecting microscope. No spermatozoa of either type were recognized in any part of the ejaculatory duct. Within the testis, eupyrene spermatozoa are present in bundles and each spermatozoon has a slender nucleus with an acrosome and a long flagellum containing mitochondrial derivatives. Two kinds of appendages, lacinate and reticular, are present on the surface of the sperm membrane. They are replaced with an extracellular sheath during passage through the vas deferens. In contrast, apyrene spermatozoa have neither nucleus nor acrosome, whereas a cup-shaped structure was found at the sperm tip instead of the acrosome. Unlike eupyrene spermatozoa, they are surrounded by a concentric sheath outside the sperm membrane in the vas deferens. Individual apyrene spermatozoa and coiled bundles of eupyrene spermatozoa were both found to accumulate in the vesicula seminalis before mating. These morphological changes during passage through the male reproductive tract suggests the occurrence of a kind of maturation and capacitation process reminiscent of mammalian spermatozoa.

Lifetime mating success of males in a natural population of the papilionid butterfly, Atrophaneura alcinous (Lepidoptera: Papilionidae)

SummaryLifetime mating success of males in a natural population of the papilionid butterfly, Atrophaneura alcinous, was investigated and causes of the variation were examined. The most successful males mated with 5 females, whereas about 73% of the males failed to mate. Body size of males was not correlated with their eclosion date, longevity and lifetime mating success. There was no trade‐off between mating success and longevity, and long‐lived males had a disproportionately high mating success. Although number of available females per male per day was not variable among males with different longevities, long‐lived males had higher mating efficiency. Time interval between matings by non‐virgin males was shorter than that from eclosion to the first mating. High lifetime mating success of long‐lived males was strongly related to their mating experience, not to their age per se.

Effectiveness of the mating plug in Atrophaneura alcinous (Lepidoptera : Papilionidae)

At mating, males of Atrophaneura alcinous plug the female copulatory duct with a secretory material, but some females copulate several times and can receive spermatophores from different males up to three times. Remating by plugged females mostly takes place shortly after the first mating, when the freshly formed first plug has not completely hardened. A freshly formed plug can be pressed aside or tunneled by the male aedeagus, and thus intromission and subsequent spermatophore transfer are sometimes possible. This is not always the case, however, and larger plugs are more likely to prevent aedeagus intrusion even if they are not completely hardened. Also, a few females remated a day after the first mating or later, when the first plug is already very hard. Completely hardened plugs are more effective than freshly formed ones; later mating males can thrust the aedeagus into the ductus bursae only when the plug is less than ca 1.2 mg (dry weight). The occurrence of female re-mating in plug-forming species thus does not necessarily mean that the plug is ineffective.

Ecological adaptation of an Aristolochiaceae-feeding swallowtail butterfly,Atrophaneura alcinous, to aristolochic acids

Seven analogs of aristolochic acids (I, II, III, B, C, D, and E) were isolated from the leaves ofAristolochia debilis and characterized as the larval feeding stimulants of an Aristolochiaceae-feeding swallowtail butterfly,Atrophaneura alcinous. Aristolochic acids showed synergistic activity in combination with the water-soluble components in the leaf extract. Aristolochic acids were detected in the body tissues and specialized organs throughout all life stages ofA. alcinous. Larval osmeterial fluid contained aristolochic acids as high as 2% of the secretions, the compositions of which were similar to that found in the leaf extracts. In contrast, the eggs selectively contained aristolochic acids I and II. These two acids were present both in the egg yolk and egg coating material as well as in the collateral glands (glandulae sebaceae) of the adult females. The cannibalistic activity of the larvae against the eggs and pupae seemed to be triggered by aristolochic acids present on the egg surface and pupal cuticle, suggesting a possible adaptive mechanism in this species. Aristolocic acid I deterred feeding of tree sparrows, which suggested a defensive role against vertebrate predators.

Oviposition stimulants of an Aristolochiaceae-feeding swallowtail butterfly,Atrophaneura alcinous

Oviposition stimulants of an Aristolochiaceae-feeding swallowtail butterfly,Atrophaneura alcinous, were isolated from the leaves ofAristolochla debilis and characterized as a mixture of aristolochic acids and sequoyitol. An artificial blend of these components applied to filter paper induced a significant oviposition response by the female butterflies, identical to that elicited by intact leaves of theAristolochia host plant.