The antennae and antennal sensilla of the giant looper,Boarmia (Ascotis) selenaria SchiffermÜller (Lepidoptera,Geometridae)*

The general morphology of the antennae and the antennal sensilla of both sexes of Boarmia (Ascotis) selenaria SchiffermÜller are described. The following types of sensilla were found: Bohm bristles and sensilla chaetica, trichodea, coeloconica, auricillica and styloconica. Antennal sexual dimorphism is marked, but mainly confined to the presence of long sensilla trichodea in males and the absence in females. The sensilla trichodea are the most abundant sensillum type.     

Developmental polymorphism in a Newfoundland population of the hemlock looper, Lambdina fiscellaria (Lepidoptera: Geometridae)

The hemlock looper [Lambdina fiscellaria (Guenée)], a widespread and highly polyphagous Geometridae, is considered one of the most economically important defoliators of North American coniferous forests. Variations in the number of larval instars between geographic populations of this species have been previously reported in the literature. However, whether such developmental polymorphism occurs within a given population is unknown. In this study, we report the presence of both four and five larval instar individuals within a population of hemlock looper in Newfoundland when reared on balsam fir. For both sexes, the majority of individuals reared on balsam fir shoots went through four larval instars, but more than one third of the females (35.3%) went through five larval instars. Females with four larval instars developed faster and had smaller pupal weight than females with five larval instars. However, a growth-related index (weight gain per unit of time) was similar for the two ecotypes (four or five larval instars). No significant difference was observed between the two ecotypes in terms of reproductive capacity (fecundity and egg size). We also found significant differences in life history traits between males and females. Results indicate that developmental polymorphism, in this case, the variation in the number of larval instars, might provide some adaptive attributes that allowed exploitation of a broader ecological niche.     

Structure and function of antennal lobe neurons in the male turnip moth,Agrotis segetum (Lepidoptera: Noctuidae)

Interneurons with dendritic branches in the antennal lobe of the male turnip moth, Agrotis segetum (Schiff., Lepidoptera: Noctuidae), were investigated with intracellular recording and staining methods. Seventeen projection neurons that transmit information from the antennal lobe to higher centers in the brain displayed dendritic arbors in the male specific macroglomerular complex (MGC) and responded to chemical components of the female sex pheromone used in species-specific sexual communication. Most of the projection neurons responded to several of the pheromone components tested, and a precise correlation between the location of the dendritic arborization and the physiological response could not be demonstrated. One MGC-projection neuron fit the definition of blend specialist. It did not respond to the individual components of the behaviorally active pheromone blend, but showed a strong response to the components when combined in the species-specific blend. Some of the projection neurons also showed clear responses to phenylacetaldehyde, a flower-produced compound and/or to (E)-2-hexenal, a common green-leaf volatile. In eight neurons, the axonal projection could be followed to the calyces of the mushroom body, and subsequently to the inferior lateral protocerebrum.Four local interneurons were characterized both morphologically and physiologically. Each neuron arborized extensively throughout the antennal lobe, and each responded to one or several of the pheromone compounds, and/or to one or both of the plant-produced compounds. One of the local interneurons responded exclusively to the pheromone blend, but not to the individual components.Abbreviations AL antennal lobe - AN antennal nerve - CB cell body - E2H (E)-2-hexenal - IACT inner antennocerebral tract - ILPR inferior lateral protocerebrum - LH lateral horn of the protocerebrum - LN local interneuron - MB mushroom body - MGC macroglomerular complex - OACT outer antennocerebral tract - PAA phenylacetaldehyde - PN projection interneuron - RN receptor neuron - Z5-10:OAc (Z)-5-decenyl acetate - Z5-10:OH (Z)-5-decenol - Z5-12:OAc (Z)-5-dodecenyl acetate - Z7-12:OAc (Z)-7-dodecenyl acetate - Z9-14:OAc (Z)-9-tetradecenyl acetate     

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.     

cDNA cloning and in situ hybridization of Delta11-desaturase, a key enzyme of pheromone biosynthesis in Ostrinia scapulalis (Lepidoptera: Crambidae)

Female sex pheromones are considered to be produced in a "pheromone gland" located in the terminal abdominal segments (8th-10th, TAS) of a moth; however, in many moth species, the cells that produce pheromones have not actually been specified. We investigated cells in the TAS that synthesize pheromones in the adzuki bean borer Ostrinia scapulalis, by locating pheromones and their precursors, and mRNA for Delta11-desaturase, a key enzyme in pheromone biosynthesis. We demonstrated that the pheromone components, (E)-11- and (Z)-11-tetradecenyl acetates, and their fatty acyl precursors were specifically contained in the dorsal part of the TAS. A cDNA (OscaZ/E11) that encodes a Delta11-desaturase was cloned from the TAS. RT-PCR and in situ hybridization unequivocally showed that OscaZ/E11 is specifically expressed in the modified epidermal cells located at the dorsal end of the 8th-9th intersegmental membrane.     

Reduction of the response to sex pheromone in the oriental fruit moth,Grapholita molesta (Lepidoptera: Tortricidae) following successive pheromonal exposures

The effects of prior pheromonal experience upon the pheromone- mediated upwind flight response was examined in the oriental fruit moth, Grapholita molesta(Busck) (Lepidoptera: Tortricidae). Adult male G. molestawere subjected to a parallel series of staggered and repeated pheromonal exposures in a sustained-flight wind tunnel. Levels of response to pheromone in male G. molestasignificantly decreased in a (a) rectilinear function with increased ages of individuals, (b) logarithmic function of successive trials, and (c) steeper logarithmic function of successive trials with increased dosage of sex pheromone. The baseline levels of responding were not affected by either the (a) dosage of sex pheromone, (b) posteclosion ages of individuals for their initial exposures once the main effect of age itself was estimated, (c) elapsed time in hours between trials, or (d) discrete days of testing as integral intervals, disregarding hours within days.     

Some factors that affect the annual cycle of the winter moth, Operophtera brumata (L.) (Lepidoptera: Geometridae) in western Europe

Egg development rate in the winter moth (Operopbtera brumata (L.)) is increased by higher temperatures between at least 6.0° C and 18° C. Pupal development rate is, in general, reduced by higher temperatures. Therefore, lower temperatures in northern localities and at high altitudes in Europe increase the duration of the egg stage and decrease that of the pupa. Inherent differences in the eggs and pupae from different localities increase the effect of temperature differences.

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Resume La température affecte directement le taux de développement des oeufs et inversement celui des pupes chez la phalène hiémale, Operopbtera brumata (L.). Dans le nord de l'Europe, une température relativement basse a pour effet de prolonger la durée du stade de l'oeuf et de raccourcir proportionnellement celle du stade de la pupe, tandis que dans le sud de l'Europe avec de plus hautes températures, l'effet est contraire. Les différences inhérentes aux oeufs et aux pupes dans diverses localités augmentent l'effet des différences de température.

     

Ecdysone and a dietary alkaloid interact in the development of the pheromone gland of a male moth (Creatonotos,Lepidoptera: Arctiidae)

Summary Hair-covered scent organs of the male arctiid moth Creatonotos produce and dissipate the volatile pheromone hydroxydanaidal. The biosynthesis of this substance depends quantitatively upon the uptake of pyrrolizidine alkaloids (PA) with the larval foodplant. The size of the tubular, eversible scent organ (corema) is also positively correlated with the ingested amount of the same alkaloid, which acts like a specific growth factor. After an assessment of the corema normogenesis by Rick-Wagner (PhD thesis, University of Cologne, 1986) we injected PA into PA-free raised larvae, prepupae, and pupae. We found that the PA competence (sensitivity) of the corema anläge terminates with the first prepupal day. Ecdysone titer determinations (radioimmunoassay) are in agreement with those in other moth species. Ligated (ecdysone-free) pupal abdomina never developed imaginal structures, with or without earlier PA application. Ecdysone injection into ligated pupal abdomina of PA-fed specimens initiated the development of imaginal structures and also of coremata of more than (ecdysone-free) control size. Pupal abdomina without PA pre-treatment only developed very small coremata. With these experiments we have separated and identified two morphogenetic control agents of corema development: the dietary PA specifies the size of the organ whereas ecdysone induces the anlage to proliferate within these PA-dependent ranges and to differentiate adult structures, as it does with other imaginal anlagen.     

Comparative morphology of the mesosomal 'gland' in male large carpenter bees (Apidae: Xylocopini)

The gross morphology of the reservoir of the mesosomal gland of male large carpenter bees (tribe Xylocopini) was examined in 46 taxa (species and subspecies) in the three genera of the tribe. Males of all but six species of Xylocopa examined possess a cuticle-lined invagination that originates along the suture between the metanotum and propodeum. Size of the reservoir and arrangement of the tubules are consistent within subgenera (with a single exception) but variable among most subgenera. The morphological extremes range from a simple, short invagination that opens medially to a complex set of convoluted tubules that open from paired, laterally-positioned openings and extend forward to the metanotum or scutellum and posteriorly to the propodeal pit. Comparisons of mesosomal shapes between the sexes or between males in taxa with and without this reservoir reveal that structural changes of the posterior elements of the mesosoma are associated with the presence of the large reservoirs. In turn, reservoir size is predictive of male reproductive behaviour. The probable mode of gland function, the potential use and limitations of this structure as a phylogenetic character and the relationship of gland reservoir size to the diversity of mate-locating behaviours found in the Xylocopini are discussed.     

A failed attempt to demonstrate pheromone communication in archaic moths of the genus Sabatinca Walker (Lepidoptera, Micropterigidae)

Data on pheromones of phylogenetically archaic moths are very scant, despite being needed to understand the life history evolution and phylogeny of the basal lineages of Lepidoptera. Two New Zealand micropterigid moths, Sabatinca (Palaeomicra) chalcophanes Meyr. and Sabatinca (s.str.) demissa Philp., were investigated for pheromone communication. Consistent negative results were demonstrated by (i) behavioural observations, (ii) exposure of blends containing active components of pheromones identified in caddis-flies and archaic moths, (iii) exposure of caged males and females in sticky traps, and (iv) detection of male and female antennal responses to both male and female extracts. Since no data indicated the presence of long-distance sex pheromones in Sabatinca , we concluded that these moths use visual clues to meet sexual partners. The secondary lack of pheromone communication in Micropterigidae suggests a high degree of ecological specialization in this plylogenetically most archaic moth family.     

Responses to sex pheromone and plant odours by olfactory receptor neurons housed in sensilla auricillica of the codling moth, Cydia pomonella (Lepidoptera: Tortricidae)

Antennal olfactory receptor neurons located in a limited number of two types of sensilla auricillica, the rabbit-eared shoehorn and the regular shoehorn, located on the 5-30 flagellomere of the codling moth, Cydia pomonella, antenna were screened for selectivity to 11 plant compounds, the major sex pheromone component, three minor pheromone components and one behavioural antagonist. Both types of sensilla housed at least three neurons characterised by different action potential amplitudes. Neurons in both males and females responded to the plant compounds, ethyl (E,Z)-2,4-decadienoate, (+/-)-linalool, (E)-ss-farnesene, hexanol, (Z)-3-hexenyl acetate, 4,8-dimethyl-1,3,(E)7-nonatriene, nonanol, the major pheromone component codlemone [(E,E)-8,10-dodecadienol] and the minor pheromone component tetradecanol. Additionally, (E,E)-alpha-farnesene and (Z)-3-hexenol elicited responses specifically in female neurons, whereas (E,E)-farnesol elicited a specific response in a male neuron. Neurons responded to 1-3 odorants, with sometimes overlapping response spectra. A scanning electron microscopic study of the antennae of both sexes supported an earlier study, apart from that long s. trichodea were present in a wreath at the proximal margin of the flagellomere and in addition evenly distributed over the remaining surface, and a previously non-described sensillum type with external basiconic features was revealed, distributed on the proximal and medial region of the flagellomeres.     

No inhibitory effect on receptor neurone activity by sulphur analogues of the sex pheromone component (Z)-5-decenyl acetate in the turnip moth, Agrofis segetum (Lepidoptera: Noctuidae)

Abstract. Olfactory responses from the entire antenna and from single antennal sensilla of the male turnip moth, Agrotis segetum (Lepidoptea: Noctuidae Schiff.), were recorded after stimulation of the antenna with the sex pheromone component, (Z)-5-decenyl acetate (Z5-10:OAc), and three sulphur analogues of this compound. Adaptation of olfactory receptor neurones tuned to Z5-10:OAc was investigated after pre-exposure of these receptor neurones to the key stimulus and to the three sulphur analogues. Both electro-antennographic and single sensillum recordings revealed that the sulphur analogues had a significantly decreased effect compared to the natural stimulus. The pre-exposure experiments demonstrated that no further inhibition of neural activity was observed than could be expected from receptor neurone adaptation. Earlier reports, describing sulphur analogues as possible hyperagonists acting on moth pheromone receptor neurones, are not supported by the present study.     

The Role of Olfactory Cues in Short-Range Mate Finding by the Emerald Ash Borer, Agrilus planipennis Fairmaire (Coleoptera: Buprestidae)

We investigated the relative importance of olfaction versus vision in the mate-finding behavior of Agrilus planipennis. When coupled in male–female, male–male and female–female pairs, attempts to mate occurred only in the male–female pairs, suggesting that beetles can identify the opposite sex before attempting to mate. In a set of sensory deprivation experiments with male–female pairs, we evaluated whether males could find females when deprived of their sense of olfaction, vision or both. Males whose antennae were blocked with model paint took significantly longer to find females and spent less time in copula compared to untreated males. Males whose eyes were similarly blocked did not differ in their mate finding capacity compared to untreated males. In a third experiment that compared both olfaction and vision, olfactorily impaired beetles never mated whereas the mate finding potential of visually impaired beetles did not differ from that of untreated beetles. Our results indicate that males can identify females before coming into physical contact with them, and that at short range (≤5 cm), volatile cues detected by olfaction are involved in mate finding by A. planipennis.     

The pregenital abdominal musculature in phasmids and its implications for the basal phylogeny of Phasmatodea (Insecta: Polyneoptera)

Recently several conflicting hypotheses concerning the basal phylogenetic relationships within the Phasmatodea (stick and leaf insects) have emerged. In previous studies, musculature of the abdomen proved to be quite informative for identifying basal taxa among Phasmatodea and led to conclusions regarding the basal splitting events within the group. However, this character complex was not studied thoroughly for a representative number of species, and usually muscle innervation was omitted. In the present study the musculature and nerve topography of mid-abdominal segments in both sexes of seven phasmid species are described and compared in detail for the first time including all putative basal taxa, e.g. members of Timema, Agathemera, Phylliinae, Aschiphasmatinae and Heteropteryginae. The ground pattern of the muscle and nerve arrangement of mid-abdominal segments, i.e. of those not modified due to association with the thorax or genitalia, is reconstructed. In Timema, the inner ventral longitudinal muscles are present, whereas they are lost in all remaining Phasmatodea (Euphasmatodea). The ventral longitudinal muscles in the abdomen of Agathemera, which span the whole length of each segment, do not represent the plesiomorphic condition as previously assumed, but might be a result of secondary elongation of the external ventral longitudinal muscles. Sexual dimorphism, common within the Phasmatodea, also applies to the muscle arrangement in the abdomen of some species. Only in the females of Haaniella dehaanii (Heteropteryginae) and Phyllium celebicum (Phylliinae) the ventral external longitudinal muscles are elongated and span the length of the whole segment, possibly as a result of convergent evolution.     

Inhibitory effect of genital photoreceptors on the activity of abdominal motoneurones in Papilio xuthus (Lepidoptera,Papilionidae)

The inhibitory effect of butterfly genital photoreceptors on the activities of abdominal motoneurones is described. In two (N1, N3) of the six lateral nerves (N1–6) belonging to the last abdominal ganglion, spontaneous motoneurone activity was inhibited by illumination of the genital photoreceptors. N1 and N3 innervate the ventral longitudinal muscles. N2 and N4, which supply branches to the spiracular muscles, were not inhibited. The results allow some of the properties of the circuits in the abdominal nervous system supplied by the genital photoreceptors to be inferred. Some possible functions of the photoreceptors are discussed.     

The eyes of a patrolling butterfly: visual field and eye structure in the Orange Sulphur, Colias eurytheme (Lepidoptera, Pieridae)

Sensory information plays a critical role in determining an animal's behavior on both proximate and evolutionary timescales. Butterflies, like many other insects, use vision extensively over their lifetimes, and yet relatively little work has been published to date on their visual capabilities. We describe the visual system of a pierid butterfly, Colias eurytheme, with the ultimate goal of better understanding its role in shaping the behavior of this animal. We made several measurements: visual field dimensions, eye surface area, interommatidial angle (Deltaphi), facet diameter (D), and eye parameter (p). C. eurytheme had a large visual field and considerable regional variation in visual acuity, as inferred by Deltaphi and D. When compared to females, males had larger eye surface areas, smaller Deltaphi, and larger D in all regions except ventrally. Both sexes had proportionally large eye surface areas compared to other butterflies. Minimum p in males was small, indicating that some regions of their eyes may operate close to the diffraction limit. Finally, we found that both eye surface area and D scaled positively, but with negative allometry to body size. We discuss the relevance of these visual characteristics to the biology and behavior of C. eurytheme.     

The potential of the fungus, Muscodor albus, as a microbial control agent of potato tuber moth (Lepidoptera: Gelechiidae) in stored potatoes

Potato tuber moth (PTM), Phthorimaea operculella, is a serious pest of stored potato in most countries where potatoes are grown. Entomopathogens offer promise as alternatives to broad spectrum insecticides for management of this pest. The fungus Muscodor albus, which produces a mixture of antimicrobial volatile organic chemicals, was tested for its insecticidal activity against PTM. Adults and neonate larvae were exposed to volatiles generated by 15 or 30 g of M. albus rye grain culture plus water for 72 h in hermetically sealed 28.3 L chambers at 24 degrees C. Mean percent mortalities in adult moths exposed to 0, 15, and 30 g of fungal formulation were 0.9, 84.6, and 90.6%, respectively. Development to the pupal stage of PTM that were exposed as neonate larvae to 15 or 30 of M. albus culture was reduced by 61.8 and 72.8%, respectively, relative to controls.     

Catches of vine bud moth Theresimima ampellophaga (Lep., Zygaenidae: Procridinae) males in pheromone traps: effect of the purity and age of baits, design, colour and height of the traps, and daily sexual activity of males

Pure (2R)‐butyl (7Z)‐tetradecenoate, as well as racemic 2‐butyl (7Z)‐tetradecenoate, in a dose of 100 μg (calculated for the active (2R)‐enantiomer) applied onto serum bottle caps of grey rubber, were an effective pheromone bait for Theresimima ampellophaga (Bayle‐Barelle, 1808) (Lepidoptera: Zygaenidae: Procridinae). This bait remained active for longer than one full flight of the pest in the regions with one generation per year. Colourless transparent as well as red and yellow sticky traps were the cheapest and most simple design for trapping T. ampellophaga, while green and blue traps performed worse. Among the traps tested, VARL (CSALOMON^®) funnel traps had the highest capture ability for the pest. Traps had to be mounted at least 1.0–2.0 m above ground level. T. ampellophaga males flew to a source of sex pheromone all day long with a main peak between 07.00 and 09.00 hours and a much smaller one between 19.00 and 21.00 hours.     

Subcellular localization of the fatty acyl reductase involved in pheromone biosynthesis in the tobacco budworm,Heliothis virescens (Noctuidae: Lepidoptera)

Sex pheromone components are produced in specialized glands of female moths via well-characterized biosynthetic pathways, where a Fatty Acyl Reductase (FAR) is often essential for producing the specific ratio of the different pheromone components. The subcellular localization and membrane topology of FARs is important for understanding how pheromones are synthesized and exported to the exterior for release. We investigated the subcellular localization of HvFAR from the noctuid moth Heliothis virescens by producing recombinant fusion proteins with green fluorescent protein (GFP) in yeast. A C-terminally tagged construct was localized to the endoplasmic reticulum (ER) and retained full reductive activity on a broad range of saturated and unsaturated fatty acyl precursors. In contrast, an N-terminally-tagged construct was poorly expressed in the cytoplasm and was not enzymatically active, indicating that HvFAR requires a free N-terminal for both proper targeting and catalytic activity. A series of truncations of the N-and C-termini of HvFAR was conducted based on in silico-predicted hydrophobic domains and transmembrane regions. The N-terminally truncated protein was found in the cytoplasm and did not retain activity, emphasizing the importance of the N-terminal for FAR function. In addition, the orientation in the membrane of the C-terminus-tagged HvFAR-GFP construct was analyzed using a fluorescence protease protection (FPP) assay, implying that the C-terminal of HvFAR is orientated towards the cytoplasm. These results, together with previous data on the localization of desaturases, confirm the importance of the ER as a subcellular site of pheromone production.