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Paul Feeny - One of the best experts on this subject based on the ideXlab platform.
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effects of contact chemistry and host plant experience in the oviposition behaviour of the eastern black Swallowtail Butterfly
Animal Behaviour, 2005Co-Authors: Cheryl A Heinz, Paul FeenyAbstract:Papilio polyxenes, the eastern black Swallowtail Butterfly, is a specialist on members of the carrot family, Apiaceae. Females choose food plants for their larvae, laying eggs singly on acceptable host plants, which vary considerably visually and chemically. To allow for fidelity of these vagile females to a single family of host plants in the absence of information transfer across life stages, the response to at least one type of host plant cue, be it visual or chemical, must be innate. Although much work has been done to identify the plant chemicals used by ovipositing Swallowtails to recognize their host plants after landing, it has been assumed rather than demonstrated that the response to this cue is innate. We conducted two experiments to test the nature of the black Swallowtail's response to contact chemical host plant cues, and a third to examine the role of the larval host plant in adult behaviour. The results of the first experiment verified that females without host plant experience respond specifically to cues from the host plant, Daucus carota, with typical abdomen curling, and not to the nonhost fava bean, Vicia faba. The nondeterrency of bean was also verified. The second experiment demonstrated that this response was unaltered by specific host plant experience: females responded to extracts of hosts, carrot and poison hemlock, Conium maculatum, differentially, but without regard to the plant previously experienced. We also examined effects of deprivation and age in the first experiment, with each having a significant effect; these were controlled for in the second experiment. Finally, feeding on a host plant as a larva did not increase the preference for that plant as an adult. This is the first demonstration that the response to these contact chemical cues is not only innate but unaltered by specific host plant experience.
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Host-Plant Chemistry Influences Oviposition Choice of the Spicebush Swallowtail Butterfly
Journal of Chemical Ecology, 1999Co-Authors: Maureen Carter, Paul FeenyAbstract:The spicebush Swallowtail, Papilio troilus (L.), lays its eggs on plants in the family Lauraceae. Sassafras [ Sassafras albidum (Nutt.) Nees], spicebush [ Lindera benzoin (L.) Spreng.], redbay ( Persea borbonia (L.)] and camphortree [ Cinnamomum camphora (Nees) Eberm.] are four of its known host plants. In one-choice tests, free-flying spicebush Swallowtail females laid eggs on chemical extracts of the leaves of each of these four hosts. In two-choice experiments, females always preferred to oviposit on an extract of sassafras compared to extracts of the other three hosts. It was shown for spicebush extract that this response was not due to oviposition experience. Previously we had identified one of the host plant chemicals acting as an oviposition stimulant in sassafras extract as 3-caffeoyl- muco -quinic acid (3-C m QA). Extracts of the other three hosts did not contain this compound. The addition of 3-C m QA alone to spicebush extract did not increase oviposition activity. It did, however, increase discrimination between hosts and nonhosts. When a fraction of sassafras extract containing 3-C m QA and other synergistic stimulants was added to spicebush extract, preference for sassafras extract was no longer recorded. These results show existing differences in oviposition chemistry among host plants of the spicebush Swallowtail and how these differences can influence oviposition choice in bioassay experiments.
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oviposition stimulant for the zebra Swallowtail Butterfly eurytides marcellus from the foliage of pawpaw asimina triloba
Chemoecology, 1998Co-Authors: Meena Haribal, Paul FeenyAbstract:We have isolated a caffeoylcyclohexane-1-carboxylic acid derivative, 3-caffeoyl-muco-quinic acid (3-CmQA), as a contact oviposition stimulant for the zebra Swallowtail Butterfly, Eruytides marcellus (Papilionidae), from the foliage of its primary host plant, Asimina triloba (Annonaceae). This compound alone was as active in stimulating oviposition by females as were the parent ethanolic plant extract and the host plant itself. Other tested isomers of 3-CmQA, including 5-caffeoylquinic acid (5-CQA or trans-chlorogenic acid), were inactive. We found, however, that experienced female butterflies responded strongly to host volatiles, which enhanced landing rates and hence oviposition.¶ This is the first report of an oviposition stimulant for a Swallowtail Butterfly of the tribe Graphiini. We found 3-CmQA to be the major caffeoylcyclohexane-1-carboxylic acid isomer in plants of the genus Asimina. These plants lack appreciable amounts of 5-CQA, which has been shown previously to be one of the oviposition stimulants for certain Rutaceae- or Apiaceae-feeding Swallowtails of the related tribe Papilionini.¶ Our findings, along with earlier results from the tribes Troidini and Papilionini, suggest that responses by Swallowtails to hydroxycinnamic acid derivatives as oviposition cues date back at least to the ancestor of the subfamily Papilioninae.
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oviposition stimulant for the zebra Swallowtail Butterfly eurytides marcellus from the foliage of pawpaw asimina triloba
Chemoecology, 1998Co-Authors: Meena Haribal, Paul FeenyAbstract:We have isolated a caffeoylcyclohexane-1-carboxylic acid derivative, 3-caffeoyl-muco-quinic acid (3-CmQA), as a contact oviposition stimulant for the zebra Swallowtail Butterfly, Eruytides marcellus (Papilionidae), from the foliage of its primary host plant, Asimina triloba (Annonaceae). This compound alone was as active in stimulating oviposition by females as were the parent ethanolic plant extract and the host plant itself. Other tested isomers of 3-CmQA, including 5-caffeoylquinic acid (5-CQA or trans-chlorogenic acid), were inactive. We found, however, that experienced female butterflies responded strongly to host volatiles, which enhanced landing rates and hence oviposition.¶ This is the first report of an oviposition stimulant for a Swallowtail Butterfly of the tribe Graphiini. We found 3-CmQA to be the major caffeoylcyclohexane-1-carboxylic acid isomer in plants of the genus Asimina. These plants lack appreciable amounts of 5-CQA, which has been shown previously to be one of the oviposition stimulants for certain Rutaceae- or Apiaceae-feeding Swallowtails of the related tribe Papilionini.¶ Our findings, along with earlier results from the tribes Troidini and Papilionini, suggest that responses by Swallowtails to hydroxycinnamic acid derivatives as oviposition cues date back at least to the ancestor of the subfamily Papilioninae.
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a caffeoylcyclohexane 1 carboxylic acid derivative from asimina triloba
Phytochemistry, 1998Co-Authors: Meena Haribal, Paul Feeny, Cathy C LesterAbstract:Abstract Three caffeoyl derivatives of 1,3,4,5-tetrahydroxycyclohexane 1-carboxylic acid, one of them an oviposition stimulant for the zebra Swallowtail Butterfly, Eurytides marcellus , were isolated from Asimina triloba . The structure of the active isomer, 3-caffeoyl- muco -quinic acid, was determined by extensive NMR studies.
Ritsuo Nishida - One of the best experts on this subject based on the ideXlab platform.
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Larval Feeding Stimulants for a Rutaceae-Feeding Swallowtail Butterfly, Papilio xuthus L. in Citrus unshiu Leaves
Journal of Chemical Ecology, 2011Co-Authors: Toshihiro Murata, Naoki Mori, Ritsuo NishidaAbstract:Larvae of a Swallowtail Butterfly, Papilio xuthus L., feed exclusively on plants of the family Rutaceae, including various Citrus crops. Larvae were strongly stimulated to feed on paper strips impregnated with ethanolic extracts of host-plant leaves. Stimulation of feeding on extracts of Citrus unshiu leaves required a mixture of chemicals including sugars ( d -glucose, d -fructose, and d -sucrose), a betaine [(−)-stachydrine], a cyclic peptide (citrusin I), a polymethoxyflavone (isosinensetin), and the lipids 1-linolenoylglycerol, 1-linoleoylglycerol, 1-octadecenoylglycerol, 1-stearoylglycerol, and 1,2-dilinolenoyl-3-galactosyl-sn-glycerol. When these compounds were assayed individually, few larvae consumed test strips. However, larvae readily chewed the test strips treated with a mixture of all compounds, indicating that host recognition by P. xuthus larvae is mediated by a specific combination of both primary and secondary substances. Comparison of 11 stimulant components with 10 compounds from C. unshiu leaves previously reported as stimulant components for oviposition by P. xuthus adult females revealed only one compound, stachydrine, as an ingredient in common. While the larval feeding-stimulant mixture is dominated by nutrients and other compounds of general significance for primary metabolism, the component oviposition stimulants are mostly secondary substances, including flavonoid glycosides, protoalkaloids, a cyclitol, and a betaine, that have restricted distributions in plants. Reliance by adult females on unique profiles of secondary compounds presumably reflects the need to locate and recognize specific host-plant species within a diverse flora. Since the initial host choice for the larvae is made typically by the ovipositing female, however, unique secondary compounds may be less important for larval feeding than are compounds useful for indicating food and microhabitat quality once on the host plant.
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Hydroxybenzoic Acid Derivatives in a Nonhost Rutaceous Plant, Orixa japonica, Deter Both Oviposition and Larval Feeding in a Rutaceae-Feeding Swallowtail Butterfly, Papilio xuthus L.
Journal of Chemical Ecology, 2004Co-Authors: Yasumasa Kuwahara, Ritsuo NishidaAbstract:A Rutaceae-feeding Swallowtail Butterfly, Papilio xuthus L., feeds on various rutaceous plants but always rejects Orixa japonica Thunb. (Rutaceae). Females were strongly deterred from laying eggs by a methanolic extract of O . japonica leaves. Larvae also rejected a diet leaf medium impregnated with O . japonica leaf extracts. Several components in the water-soluble fraction of the leaf extract were found to deter both oviposition and feeding responses. Two major deterrent compounds were characterized as 5-{[2- O -(β-D-apiofuranosyl)-β-D-glucopyranosyl]oxy}-2-hydroxybenzoic acid and a disyringoyl aldaric acid. These compounds induced potent deterrence of both oviposition and larval feeding by P . xuthus , which suggests a congruent chemosensory mechanism of allomonal chemicals acting on both female tarsal chemoreceptors and larval maxillary taste receptors.
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a dihydroxy γ lactone as an oviposition stimulant for the Swallowtail Butterfly papilio bianor from the rutaceous plant orixa japonica
Bioscience Biotechnology and Biochemistry, 2000Co-Authors: Hajime Ono, Ritsuo Nishida, Yasumasa KuwaharaAbstract:The oviposition response of the Rutaceae-feeding Swallowtail Butterfly, Papilio bianor, was induced by a methanolic extract from leaves of its major host, Orixa japonica. Several components were responsible for this oviposition response. One of the stimulants was isolated and identified as (-)-2-C-methyl-D-erythrono-1,4-lactone. The compound was inactive alone, but elicited oviposition behavior when mixed with other fractions.
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oviposition stimulant for a rutaceae feeding Swallowtail Butterfly papilio bianor lepidoptera papilionidae hydroxycinnamic acid derivative from orixa japonica
Applied Entomology and Zoology, 2000Co-Authors: Hajime Ono, Ritsuo Nishida, Yasumasa KuwaharaAbstract:The oviposition response of a Swallowtail Butterfly, Papilio bianor, was elicited by a methanolic extract from leaves of its host plant Orixa japonica (Rutaceae). Reverse phase ODS column chromatography of a water soluble layer separated two active fractions, eluted with 1% acetic acid in water and 40% methanol +1% acetic acid in water. A major active stimulant in the latter eluate was isolated and identified as (−)-4-(E)-caffeoyl-L-threonic acid. This compound was found to be active alone (43% oviposition response at 0.1 g leaf equivalent per 10 cm2 filter paper) and there was no significant synergistic effect of the compound on other active fractions. Hydroxycinnamic acid derivatives appear to be utilized widely as oviposition cues among the Papilionidae.
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oviposition stimulant of a zeryntiine Swallowtail Butterfly luehdorfia japonica
Phytochemistry, 1994Co-Authors: Ritsuo NishidaAbstract:Abstract Luehdorfia japonica Leech (Papilionidae, tribe Zerynthiini) is a Swallowtail Butterfly whose larvae feed exclusively on the plant genus Heterotropa (Aristolochiaceae). Luehdorfia japonica females were strongly stimulated to lay eggs on filter paper treated with a methanolic extract of the host plant leaves. The oviposition stimulant factor was composed of several water-soluble components. One of the oviposition stimulants was isolated from the leaves of Heterotropa aspera and identified as isorhamnetin 3-O-glucosyl-(1→6)-galactoside-7-O-glucoside. This compound was inactive alone but induced the specific oviposition response from L. japonica females when tested as a mixture with other unidentified components.
Haruhiko Fujiwara - One of the best experts on this subject based on the ideXlab platform.
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molecular mechanisms of larval color pattern switch in the Swallowtail Butterfly
2017Co-Authors: Hongyuan Jin, Haruhiko FujiwaraAbstract:In lepidopterans (butterflies and moths), larval body color pattern, which is an important mimicry trait involved in prey–predator interactions, presents a great diversity of pigmentation and patterning. Unlike wing patterns, larval body color patterns can switch during development with larval molting. For example, in the Asian Swallowtail Butterfly Papilio xuthus, a younger larva (first–fourth instar) has a white/black color pattern that mimics bird droppings, whereas the final instar (fifth) larva drastically changes to a greenish pattern that provides camouflage on plants. Insect mimicry has interested scientists and the public since Darwin’s era. Broadly, mimicry is an antipredation strategy whereby one creature’s color, shape, or behavior resembles another creature or object. In this review, I address basic knowledge about larval cuticular pigmentation and advanced understanding of its regulatory mechanism in P. xuthus; I also discuss larval body color patterns among members of the genus Papilio, followed by conclusions and prospects for further research.
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comprehensive microarray based analysis for stage specific larval camouflage pattern associated genes in the Swallowtail Butterfly papilio xuthus
BMC Biology, 2012Co-Authors: Ryo Futahashi, Hiroko Shirataki, Takanori Narita, Kazuei Mita, Haruhiko FujiwaraAbstract:Body coloration is an ecologically important trait that is often involved in prey-predator interactions through mimicry and crypsis. Although this subject has attracted the interest of biologists and the general public, our scientific knowledge on the subject remains fragmentary. In the caterpillar of the Swallowtail Butterfly Papilio xuthus, spectacular changes in the color pattern are observed; the insect mimics bird droppings (mimetic pattern) as a young larva, and switches to a green camouflage coloration (cryptic pattern) in the final instar. Despite the wide variety and significance of larval color patterns, few studies have been conducted at a molecular level compared with the number of studies on adult Butterfly wing patterns. To obtain a catalog of genes involved in larval mimetic and cryptic pattern formation, we constructed expressed sequence tag (EST) libraries of larval epidermis for P. xuthus, and P. polytes that contained 20,736 and 5,376 clones, respectively, representing one of the largest collections available in butterflies. A comparison with silkworm epidermal EST information revealed the high expression of putative blue and yellow pigment-binding proteins in Papilio species. We also designed a microarray from the EST dataset information, analyzed more than five stages each for six markings, and confirmed spatial expression patterns by whole-mount in situ hybridization. Hence, we succeeded in elucidating many novel marking-specific genes for mimetic and cryptic pattern formation, including pigment-binding protein genes, the melanin-associated gene yellow-h3, the ecdysteroid synthesis enzyme gene 3-dehydroecdysone 3b-reductase, and Papilio-specific genes. We also found many cuticular protein genes with marking specificity that may be associated with the unique surface nanostructure of the markings. Furthermore, we identified two transcription factors, spalt and ecdysteroid signal-related E75, as genes expressed in larval eyespot markings. This finding suggests that E75 is a strong candidate mediator of the hormone-dependent coordination of larval pattern formation. This study is one of the most comprehensive molecular analyses of complicated morphological features, and it will serve as a new resource for studying insect mimetic and cryptic pattern formation in general. The wide variety of marking-associated genes (both regulatory and structural genes) identified by our screening indicates that a similar strategy will be effective for understanding other complex traits.
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identification of stage specific larval camouflage associated genes in the Swallowtail Butterfly papilio xuthus
Development Genes and Evolution, 2008Co-Authors: Ryo Futahashi, Haruhiko FujiwaraAbstract:The Swallowtail Butterfly Papilio xuthus changes its larval body pattern dramatically during the fourth ecdysis. The penultimate (fourth) instar larvae mimic bird droppings that have many tubercle structures on their surface, whereas the final (fifth) instar larvae have a green camouflage color. To identify the genes involved in stage-specific larval mimicry markings, we compared the epidermal messenger ribonucleic acid expression between the third and fourth molts of P. xuthus using a complementary deoxyribonucleic acid subtraction method. After analyzing 2,072 clones from two subtractive libraries, we obtained 31 and 64 candidate genes for final (Fsg) or penultimate instar-specific genes (Psg), respectively. The expression pattern of each gene was examined by reverse transcriptase polymerase chain reaction and in situ hybridization. Among Fsgs, Fsg02 and Fsg20 were expressed in the presumptive green region only during the fourth molt, suggesting that these two genes were correlated with green coloration in the final instar. Among Psgs, 11 cuticular protein genes were expressed specifically in the tubercle structures during the third molt. These genes are likely to be involved in the formation of the unique tubercle structures observed in the juvenile instar. We found that genes with similar expression patterns do not necessarily share the same protein motifs and vice versa. This study provides novel molecular markers and insights into the molecular mechanisms of the larval color pattern and body shape.
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juvenile hormone regulates Butterfly larval pattern switches
Science, 2008Co-Authors: Ryo Futahashi, Haruhiko FujiwaraAbstract:Insect color patterns can be very diverse. This variation is also seen among many larval instar stages, which can take on vastly different phenotypes. Young caterpillars of the Swallowtail Butterfly, Papilio xuthus, are mimics of bird droppings, whereas the fifth larval instar is camouflaged among the leaves of host plants (cryptic pattern). We find that juvenile hormone (JH) titers decrease during the fourth larval instar. Furthermore, treatment with JH analog at the beginning of the fourth instar stage resulted in reproducing the mimetic pattern instead of the usual cryptic one and likewise altered gene expression patterns to that associated with the mimetic pattern. These findings suggest that JH regulates the progressive larval pattern switch of this insect.
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regulation of 20 hydroxyecdysone on the larval pigmentation and the expression of melanin synthesis enzymes and yellow gene of the Swallowtail Butterfly papilio xuthus
Insect Biochemistry and Molecular Biology, 2007Co-Authors: Ryo Futahashi, Haruhiko FujiwaraAbstract:The Swallowtail Butterfly, Papilio xuthus, changes its larval body pattern dramatically during the fourth ecdysis. Cuticular pigmentation occurs with precise timing just before ecdysis. We previously found that the cuticular pigmentation was regulated by three melanin synthesis genes, tyrosine hydroxylase (TH), dopa decarboxylase (DDC), and ebony. We discovered that yellow is strongly expressed in the presumptive black markings earlier than TH and DDC. Because the ecdysis is triggered by 20-hydroxyecdysone (20E), the effects of 20E on the pigmentation and expression of the melanin synthesis genes were examined. Here, we established a method for the topical application of 20E to molting specimens, so that 20E has only a partial effect, resulting in successful ecdysis. When we applied 20E during the mid-phase of the molting period, when the 20E titer is declining, cuticular pigmentation was completely inhibited. The cessation of hormonal treatments caused delayed pigmentation. yellow expression was promoted by a high titer of 20E, whereas the expression of TH, DDC, and ebony was suppressed, suggesting that a decline in the 20E concentration is necessary for the induction of the expression of the latter three genes. These results indicate that cuticular pigmentation is controlled by the exposure to 20E and its removal.
Kentaro Arikawa - One of the best experts on this subject based on the ideXlab platform.
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combined pigmentary and structural effects tune wing scale coloration to color vision in the Swallowtail Butterfly papilio xuthus
Zoological letters, 2015Co-Authors: Doekele G Stavenga, Atsuko Matsushita, Kentaro ArikawaAbstract:Butterflies have well-developed color vision, presumably optimally tuned to the detection of conspecifics by their wing coloration. Here we investigated the pigmentary and structural basis of the wing colors in the Japanese yellow Swallowtail Butterfly, Papilio xuthus, applying spectrophotometry, scatterometry, light and electron microscopy, and optical modeling. The about flat lower lamina of the wing scales plays a crucial role in wing coloration. In the cream, orange and black scales, the lower lamina is a thin film with thickness characteristically depending on the scale type. The thin film acts as an interference reflector, causing a structural color that is spectrally filtered by the scale’s pigment. In the cream and orange scales, papiliochrome pigment is concentrated in the ridges and crossribs of the elaborate upper lamina. In the black scales the upper lamina contains melanin. The blue scales are unpigmented and their structure differs strongly from those of the pigmented scales. The distinct blue color is created by the combination of an optical multilayer in the lower lamina and a fine-structured upper lamina. The structural and pigmentary scale properties are spectrally closely related, suggesting that they are under genetic control of the same key enzymes. The wing reflectance spectra resulting from the tapestry of scales are well discriminable by the Papilio color vision system.
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Simultaneous color contrast in the foraging Swallowtail Butterfly, Papilio xuthus
Journal of Experimental Biology, 2008Co-Authors: M Kinoshita, Y Takahashi, Kentaro ArikawaAbstract:This study demonstrates that the color vision of foraging Japanese yellow Swallowtail butterflies, Papilio xuthus, involves simultaneous color contrast. We trained newly emerged Papilio to select a disk of pale green among a set of differently colored disks presented on a black background. When the same set of disks was presented on blue background, the pale green-trained butterflies selected blue-green. The difference in spectra between pale green and blue green was similar to the spectrum of yellow for human vision, suggesting that blue induces yellow. Similarly, the pale green-trained Papilio selected a more bluish spring green on yellow background. We also trained Papilio with orange disks and tested on a green and violet background. The results showed that green induced violet and vice versa. Taken together, we concluded that simultaneous color contrast of Papilio is similar to the effect of complementary colors in human color vision.
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color discrimination at the spatial resolution limit in a Swallowtail Butterfly papilio xuthus
The Journal of Experimental Biology, 2006Co-Authors: Yuichi Takeuchi, Kentaro Arikawa, Michiyo KinoshitaAbstract:Spatial resolution of insect compound eyes is much coarser than that of humans: a single pixel of the human visual system covers about 0.008° whereas that of diurnal insects is typically about 1.0°. Anatomically, the pixels correspond to single cone outer segments in humans and to single rhabdoms in insects. Although an outer segment and a rhabdom are equivalent organelles containing visual pigment molecules, they are strikingly different in spectral terms. The cone outer segment is the photoreceptor cell part that expresses a single type of visual pigment, and is therefore monochromatic. On the other hand, a rhabdom is composed of several photoreceptor cells with different spectral sensitivities and is therefore polychromatic. The polychromatic organization of the rhabdom suggests that insects can resolve wavelength information in a single pixel, which is an ability that humans do not have. We first trained the Japanese yellow Swallowtail Butterfly Papilio xuthus to feed on sucrose solution at a paper disk of certain color. We then let the trained butterflies discriminate disks of the training color and grey disks each presented in a Y-maze apparatus. Papilio correctly selected the colored disk when the visual angle was greater than 1.18° for blue, 1.53° for green or 0.96° for red: they appeared to see colors in single pixels to some extent. This ability may compensate their rather low spatial resolution. Summary
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color discrimination at the spatial resolution limit in a Swallowtail Butterfly papilio xuthus
The Journal of Experimental Biology, 2006Co-Authors: Yuichi Takeuchi, Kentaro Arikawa, Michiyo KinoshitaAbstract:Spatial resolution of insect compound eyes is much coarser than that of humans: a single pixel of the human visual system covers about 0.008 degrees whereas that of diurnal insects is typically about 1.0 degrees . Anatomically, the pixels correspond to single cone outer segments in humans and to single rhabdoms in insects. Although an outer segment and a rhabdom are equivalent organelles containing visual pigment molecules, they are strikingly different in spectral terms. The cone outer segment is the photoreceptor cell part that expresses a single type of visual pigment, and is therefore monochromatic. On the other hand, a rhabdom is composed of several photoreceptor cells with different spectral sensitivities and is therefore polychromatic. The polychromatic organization of the rhabdom suggests that insects can resolve wavelength information in a single pixel, which is an ability that humans do not have. We first trained the Japanese yellow Swallowtail Butterfly Papilio xuthus to feed on sucrose solution at a paper disk of certain color. We then let the trained butterflies discriminate disks of the training color and grey disks each presented in a Y-maze apparatus. Papilio correctly selected the colored disk when the visual angle was greater than 1.18 degrees for blue, 1.53 degrees for green or 0.96 degrees for red: they appeared to see colors in single pixels to some extent. This ability may compensate their rather low spatial resolution.
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colour constancy of the Swallowtail Butterfly papilio xuthus
The Journal of Experimental Biology, 2000Co-Authors: Michiyo Kinoshita, Kentaro ArikawaAbstract:We have recently shown that the Japanese yellow Swallowtail Butterfly Papilio xuthus uses colour vision when searching for food. In the field, these butterflies feed on nectar provided by flowers of various colours not only in direct sunlight but also in shaded places and on cloudy days, suggesting that they have colour constancy. Here, we tested this hypothesis. We trained newly emerged Papilio xuthus to feed on sucrose solution on a paper patch of a certain colour under white illumination. The butterflies were then tested under both white and coloured illumination. Under white illumination, yellow- and red-trained butterflies selected the correctly coloured patch from a four-colour pattern and from a colour Mondrian collage. Under four different colours of illumination, we obtained results that were fundamentally similar to those under white illumination. Moreover, we performed critical tests using sets of two similar colours, which were also correctly discriminated by trained butterflies under coloured illumination. Taken together, we conclude that the Butterfly Papilio xuthus exhibits some degree of colour constancy when searching for food.
Ryo Futahashi - One of the best experts on this subject based on the ideXlab platform.
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comprehensive microarray based analysis for stage specific larval camouflage pattern associated genes in the Swallowtail Butterfly papilio xuthus
BMC Biology, 2012Co-Authors: Ryo Futahashi, Hiroko Shirataki, Takanori Narita, Kazuei Mita, Haruhiko FujiwaraAbstract:Body coloration is an ecologically important trait that is often involved in prey-predator interactions through mimicry and crypsis. Although this subject has attracted the interest of biologists and the general public, our scientific knowledge on the subject remains fragmentary. In the caterpillar of the Swallowtail Butterfly Papilio xuthus, spectacular changes in the color pattern are observed; the insect mimics bird droppings (mimetic pattern) as a young larva, and switches to a green camouflage coloration (cryptic pattern) in the final instar. Despite the wide variety and significance of larval color patterns, few studies have been conducted at a molecular level compared with the number of studies on adult Butterfly wing patterns. To obtain a catalog of genes involved in larval mimetic and cryptic pattern formation, we constructed expressed sequence tag (EST) libraries of larval epidermis for P. xuthus, and P. polytes that contained 20,736 and 5,376 clones, respectively, representing one of the largest collections available in butterflies. A comparison with silkworm epidermal EST information revealed the high expression of putative blue and yellow pigment-binding proteins in Papilio species. We also designed a microarray from the EST dataset information, analyzed more than five stages each for six markings, and confirmed spatial expression patterns by whole-mount in situ hybridization. Hence, we succeeded in elucidating many novel marking-specific genes for mimetic and cryptic pattern formation, including pigment-binding protein genes, the melanin-associated gene yellow-h3, the ecdysteroid synthesis enzyme gene 3-dehydroecdysone 3b-reductase, and Papilio-specific genes. We also found many cuticular protein genes with marking specificity that may be associated with the unique surface nanostructure of the markings. Furthermore, we identified two transcription factors, spalt and ecdysteroid signal-related E75, as genes expressed in larval eyespot markings. This finding suggests that E75 is a strong candidate mediator of the hormone-dependent coordination of larval pattern formation. This study is one of the most comprehensive molecular analyses of complicated morphological features, and it will serve as a new resource for studying insect mimetic and cryptic pattern formation in general. The wide variety of marking-associated genes (both regulatory and structural genes) identified by our screening indicates that a similar strategy will be effective for understanding other complex traits.
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identification of stage specific larval camouflage associated genes in the Swallowtail Butterfly papilio xuthus
Development Genes and Evolution, 2008Co-Authors: Ryo Futahashi, Haruhiko FujiwaraAbstract:The Swallowtail Butterfly Papilio xuthus changes its larval body pattern dramatically during the fourth ecdysis. The penultimate (fourth) instar larvae mimic bird droppings that have many tubercle structures on their surface, whereas the final (fifth) instar larvae have a green camouflage color. To identify the genes involved in stage-specific larval mimicry markings, we compared the epidermal messenger ribonucleic acid expression between the third and fourth molts of P. xuthus using a complementary deoxyribonucleic acid subtraction method. After analyzing 2,072 clones from two subtractive libraries, we obtained 31 and 64 candidate genes for final (Fsg) or penultimate instar-specific genes (Psg), respectively. The expression pattern of each gene was examined by reverse transcriptase polymerase chain reaction and in situ hybridization. Among Fsgs, Fsg02 and Fsg20 were expressed in the presumptive green region only during the fourth molt, suggesting that these two genes were correlated with green coloration in the final instar. Among Psgs, 11 cuticular protein genes were expressed specifically in the tubercle structures during the third molt. These genes are likely to be involved in the formation of the unique tubercle structures observed in the juvenile instar. We found that genes with similar expression patterns do not necessarily share the same protein motifs and vice versa. This study provides novel molecular markers and insights into the molecular mechanisms of the larval color pattern and body shape.
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juvenile hormone regulates Butterfly larval pattern switches
Science, 2008Co-Authors: Ryo Futahashi, Haruhiko FujiwaraAbstract:Insect color patterns can be very diverse. This variation is also seen among many larval instar stages, which can take on vastly different phenotypes. Young caterpillars of the Swallowtail Butterfly, Papilio xuthus, are mimics of bird droppings, whereas the fifth larval instar is camouflaged among the leaves of host plants (cryptic pattern). We find that juvenile hormone (JH) titers decrease during the fourth larval instar. Furthermore, treatment with JH analog at the beginning of the fourth instar stage resulted in reproducing the mimetic pattern instead of the usual cryptic one and likewise altered gene expression patterns to that associated with the mimetic pattern. These findings suggest that JH regulates the progressive larval pattern switch of this insect.
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regulation of 20 hydroxyecdysone on the larval pigmentation and the expression of melanin synthesis enzymes and yellow gene of the Swallowtail Butterfly papilio xuthus
Insect Biochemistry and Molecular Biology, 2007Co-Authors: Ryo Futahashi, Haruhiko FujiwaraAbstract:The Swallowtail Butterfly, Papilio xuthus, changes its larval body pattern dramatically during the fourth ecdysis. Cuticular pigmentation occurs with precise timing just before ecdysis. We previously found that the cuticular pigmentation was regulated by three melanin synthesis genes, tyrosine hydroxylase (TH), dopa decarboxylase (DDC), and ebony. We discovered that yellow is strongly expressed in the presumptive black markings earlier than TH and DDC. Because the ecdysis is triggered by 20-hydroxyecdysone (20E), the effects of 20E on the pigmentation and expression of the melanin synthesis genes were examined. Here, we established a method for the topical application of 20E to molting specimens, so that 20E has only a partial effect, resulting in successful ecdysis. When we applied 20E during the mid-phase of the molting period, when the 20E titer is declining, cuticular pigmentation was completely inhibited. The cessation of hormonal treatments caused delayed pigmentation. yellow expression was promoted by a high titer of 20E, whereas the expression of TH, DDC, and ebony was suppressed, suggesting that a decline in the 20E concentration is necessary for the induction of the expression of the latter three genes. These results indicate that cuticular pigmentation is controlled by the exposure to 20E and its removal.
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expression of one isoform of gtp cyclohydrolase i coincides with the larval black markings of the Swallowtail Butterfly papilio xuthus
Insect Biochemistry and Molecular Biology, 2006Co-Authors: Ryo Futahashi, Haruhiko FujiwaraAbstract:Abstract 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 BH4 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.
