Vergleichende Morphologie des Schmetterlingsrüssels und seiner Sensillen — Ein Beitrag zur phylogenetischen Systematik der Papilionoidea (Insecta, Lepidoptera)

Comparative morphology of the butterfly proboscis and its sensilla — a contribution to the phylogenetic systematics of Papilionoidea (Insecta, Lepidoptera) The morphology of the proboscis was investigated in more than 70 European representatives of Papilionoidea using light microscopy and scanning electron microscopy. The composition of the proboscis wall, its surface structures, as well as the shape and distribution of the different types of sensilla are compared. Special attention is given to the tip region and the diversity of the sensilla styloconica. Plesiomorphic features of the proboscis of Papilionoidea were found to include vertically extended exocuticular ribs composing the galeal wall, cuticular spines restricted to the ventral side of the proximal galea, and two rows of fluted sensilla styloconica restricted to the tip region. Apomorphic features of the proboscis in Papilionidae are three rows of small sensilla styloconica. The presence of cuticular spines all over the galeae was identified as an autapomorphy of Pieridae. Possible apomorphies of Nymphalidae are oblique exocuticular ribs of the galeal wall and the great number and length of the sensilla styloconica (significant at p < 0.01, t-test). A possible synapomorphy of Lycaenidae and Riodinidae are cuticlar spines up to the distal galeae. Distinct transformation series of sensilla styloconica give evidence that divergent evolutionary trends led from fluted shafts to a multitude of other shapes in Papilionidae, Nymphalidae (sensu lato), and Lycaenidae. Long smooth-shafted, club-shaped sensilla styloconica, bearing apical spines, are found in Nymphalinae, Apaturinae and Limenitidinae. Highly derived sensilla styloconica evolved in Heliconiinae and Melitaeini, which are arranged in only one row in both taxa. Their shafts are smooth, flattened and bear an excentral sensory cone. Further apomorphic character states are dented flutes which evolved several times, independently from each other in Satyrinae, Lycaeninae and Riodinidae. The results are discussed in a systematical context and provide the basis for a better understanding of the function of different morphological structures of the proboscis in feeding.     

The male reproductive system of Zorotypus caudelli Karny (Zoraptera): Sperm structure and spermiogenesis

Considering the overall uniformity of the morphology of Zoraptera, the structural diversity of the male genital system is remarkable. Structures related to the male reproductive system of Zorotypus caudelli differ profoundly from those of Zorotypus hubbardi. The testes are elongated rather than spherical, the seminal vesicle is apparently absent, and the deferent ducts are very long. A feature shared by these two species and other zorapterans examined is that the two accessory glands are closely adherent to each other and form a single large structure, from which the ejaculatory duct originates. This is a potential zorapteran autapomorphy. Another feature possibly present in the groundplan of the order is the strong elongation of the sperm cells. This may be connected with a reproductive strategy of males trying to avoid re-mating of females with other males after the first copulation. The extremely long and coiled spermathecal duct of Z. caudelli and other zorapteran species is possibly correlated with the sperm elongation, and both features combined may result in a sexual isolating mechanism. The short duration of mating of Zorotypus barberi and Zorotypus gurneyi suggests that the male introduces sperm into the female tract up to the opening of the spermathecal duct using their long coiled aedeagus. A thick glycocalyx around the sperm in the distal part of the deferent ducts probably protects the sperm cells during their forward progression towards the long spermathecal duct, and is removed when they reach the apical receptacle. The spermatogenesis of Z. caudelli follows a pattern commonly found in insects, but differs distinctly from that of Z. hubbardi in the number of spermatids in each sperm cyst. An unusual and possibly autapomorphic feature of Z. caudelli is a disconnection of sub-tubules A and B at the level of microtubule doublets 1 and 6 of the mature sperm cells. It is conceivable that this results in a shorter period of sperm motility. The character combination found in different zorapteran species supports the view that the sperm, a very compact functional unit, does not evolve as a unit, but like in other more complex body regions, sperm components can also be modified independently from each other. This results in different mosaic patterns of plesiomorphic and derived features in a very compact entity in different species of the very small and otherwise uniform order Zoraptera. In Z. caudelli, for instance, the bi-layered acrosome and small accessory bodies are plesiomorphic states among several others, whereas the mitochondrial derivatives and the elongate nucleus are apparently derived conditions. Other combinations likely occur in other zorapteran species. Only few but noteworthy sperm characters indicate possible phylogenetic affinities of Zoraptera. A possible synapomorphic feature, the presence of dense laminae radiating in a cartwheel array between neighbouring centriolar triplets, is shared with Phasmatodea and Embioptera. Another potential synapomorphy shared with Phasmatodea is the presence of 17 protofilaments in the tubular wall of the outer accessory microtubules.     

Comparative morphology of spermatozoa and reproductive systems of zorapteran species from different world regions (Insecta,Zoraptera)

The male and female reproductive apparatus of Zorotypus magnicaudelli (Malaysia), Zorotypus huxleyi (Ecuador) and Zorotypus weidneri (Brazil) were examined and documented in detail. The genital apparatus and sperm of the three species show only minor differences. The testes are larger in Z. magnicaudelli. Z. huxleyi lacks the helical appendage in the accessory glands. A long cuticular flagellum is present in Z. magnicaudelli and in the previously studied Zorotypus caudelli like in several other species, whereas it is absent in Z. weidneri, Z. huxleyi, Zorotypus hubbardi, Zorotypus impolitus and Zorotypus guineensis. Characteristic features of the very similar sperm are the presence of: a) two dense arches above the axoneme; b) a 9 + 9+2 axoneme with detached subtubules A and B of doublets 1 and 6; c) the axonemal end degenerating with enlarging accessory tubules; d) accessory tubules with 17 protofilaments; e) three accessory bodies beneath the axoneme; and f) two mitochondrial derivatives of equal shape. The first characteristic (a) is unknown outside of Zoraptera and possibly autapomorphic. The sperm structure differs distinctly in Z. impolitus and Z. hubbardi, which produce giant sperm and possess a huge spermatheca. The presence of the same sperm type in species either provided with a sclerotized coiled flagellum in males or lacking this structure indicates that a different organization of the genital apparatus does not necessarily affect the sperm structure. The flagellum and its pouch has probably evolved within Zoraptera, but it cannot be excluded that it is a groundplan feature and was reduced several times. The fossil evidence and our findings suggest that distinct modifications in the genital apparatus occurred before the fragmentation of the Gondwanan landmass in the middle Cretaceous.     

Morphology of the male reproductive system and sperm ultrastructure of the egg parasitoid Gryon pennsylvanicum (Ashmead) (Hymenoptera,Platygastridae)

Gryon pennsylvanicum is a platygastrid hymenopteran that has lately received increasing attention in Europe due to its possible use in biological control of the conifer seed bug pest Leptoglossus occidentalis. Here the male reproductive system and the spermatogenesis of this species, along with those of Gryon muscaeformis, are examined ultrastructurally for the first time. The male genital system is formed by a pair of testes, each containing only one follicle, a pair of accessory glands and deferent ducts connected to a single ejaculatory duct. All the stages of spermatogenesis are described in detail. Characteristic features of the Gryon spp. sperm, which are 100 μm long, are the presence of a polygonal nucleus, only one mitochondrial derivative, the occurrence of the centriole adjunct and a typical insect 9 + 9 + 2 flagellar axoneme. The single derivative, however, results from a process in which one of the two mitochondria is lost during spermiogenesis. Unlike in other insects, two centrioles occur in spermatids as a consequence of the ameiotic parthenogenesis. These characteristics stand as a valuable tool for phylogenetic inferences. Furthermore this study suggests a useful strategy for laboratory mass rearing.     

The fine structure of the female reproductive system of Zorotypus caudelli Karny (Zoraptera)

The general structure of the female genital system of Zorotypus caudelli is described. The ovarioles are of the panoistic type. Due to the reduction of the envelope (tunica externa) the ovarioles are in direct contact with the hemolymph like in some other insect groups, Plecoptera included. The calices are much larger in Z. caudelli then in Zorotypus hubbardi and their epithelial cells produce large amounts of secretions, probably protecting the surface of the eggs deposited on the substrate. Eggs taken from the calyx bear a series of long fringes, which are missing in the eggs found in the ovariole, and in other zorapteran species. The long sperm of Z. caudelli and the long spermathecal duct are likely related to a sexual isolating mechanism (cryptic female choice), impeding female re-mating. The apical receptacle and the spermathecal duct - both of ectodermal origin - consist of three cell types. In addition to the cells beneath the cuticle lining the lumen, two other cell types are visible: secretory and canal cells. The cytoplasm of the former is rich in rough endoplasmic reticulum cisterns and Golgi complexes, which produce numerous discrete dense secretory bodies. These products are released into the receiving canal crossing the extracellular cavity of secretory cells, extending over a series of long microvilli. The secretion is transported towards the lumen of the apical receptacle of the spermatheca or to that of the spermathecal duct by a connecting canal formed by the canal cells. It is enriched by material produced by the slender canal cells. Before mating, the sperm cells are enveloped by a thick glycocalyx produced at the level of the male accessory glands, but it is absent when they have reached the apical receptacle, and also in the spermathecal duct lumen. It is likely removed by secretions of the spermatheca. The eggs are fertilized at the level of the common oviduct where the spermathecal duct opens. Two micropyles at the dorsal side of the equator level possibly facilitate fertilization. The presence of these two micropyles is a presumably derived feature shared with Phasmatodea. The fine structure of the female reproductive system of Z. caudelli does not allow to assess the phylogenetic position at the present stage of knowledge. The enlarged calyx and the temporary presence of long fringes on the eggs are potential autapomorphies of Z. caudelli or may indicate relationships with other Zorotypus species.     

The Nature of Quantitative Genetic Variation in Drosophila. III. Mechanism of Dosage Compensation for Sex-Linked Abdominal Bristle Polygenes

Seventeen lines, each homozygous for a different X chromosome but all with a common autosomal genetic background, were constructed and assayed for abdominal bristle number to determine whether dosage compensation operates for sex-linked genes affecting this character.—The regression coefficient of male mean on female mean using a logarithmic scale was 0.90 ± 0.13 and the genetic regression coefficient 0.92, neither differing significantly from unity. The genetic components of variance in males and females were also very similar (0.000234 or 0.000228, respectively). These results indicate that dosage compensation is complete (or nearly so) for sex-linked genes affecting this character. The bristle scores of females did not differ in reciprocal crosses between these lines, thus dosage compensation does not operate by paternal X inactivation.—The question of an adequate scale for abdominal bristle number had to be examined during the study. A logarithmic scale appeared to be adequate for both genotypic and environmental differences.     

Variation in flight duration among individual Tetraopes beetles: Implications for studies of insect flight

Milkweed beetles, Tetraopes tetraophalmus (Forster) (Cerambycidae), were flight tested three times weekly throughout their lives. Flight durations peaked early in life and then declined rapidly with age. Significant variation existed (1) between individuals, with some flying for long periods of time, others for only a few seconds, and (2) within individuals, with some flying for long periods on some test days and very briefly or not at all on other days. Long and short fliers were indistinguishable on the basis of size, sex, or lifespan. The data show that studies of insect flight will underestimate the number of long fliers in a population by as much as 50% or more unless individuals are flight tested more than once.     

Organometallic coordination polymers: Sn(IV) derivatives with the bis(triazolyl)methane ligand

Several alkyl- or aryl-tin(IV) halides of general formula SnR_nX_4−n (n = 0-2; R = Me, Et, ⁿBu, ^tBu, Ph; X = Cl, Br), possessing Lewis acidic character, have been reacted with the polydentate N-donor ligand bis(1,2,4-triazolyl)methane (Btm), affording Btm(SnR_nCl_4−n) complexes. (Btm)₂SnⁿBu₂Br₂ and (Btm)SnⁿBu₂(NO₃)₂ are also reported. These materials were characterized by elemental analyses, IR and ¹H (and, in selected cases, ¹¹⁹Sn) NMR spectroscopy, and, were possible, ab initio X-ray powder diffraction methods. The crystal structures determined by the latter method showed that Btm ligands, in the exobidentate mode, link Sn(IV) fragments which lie 9.5-11.2 Å apart (depending on the Btm conformation and on the local metal stereochemistry), in one-dimensional chains packed in parallel bundles. The main geometrical features of these 1D polymers are compared with those of the bis(imidazolyl)methane complexes and of the known Btm derivative, Btm(Ph₂SnBr₂). Interestingly, the expected isomorphous structures for selected couples was not found, as if very subtle energetic differences were driving the crystallization of these species into different structure types.     

Toward reconstructing the evolution of advanced moths and butterflies (Lepidoptera: Ditrysia): an initial molecular study

Background

In the mega-diverse insect order Lepidoptera (butterflies and moths; 165,000 described species), deeper relationships are little understood within the clade Ditrysia, to which 98% of the species belong. To begin addressing this problem, we tested the ability of five protein-coding nuclear genes (6.7 kb total), and character subsets therein, to resolve relationships among 123 species representing 27 (of 33) superfamilies and 55 (of 100) families of Ditrysia under maximum likelihood analysis.

Results

Our trees show broad concordance with previous morphological hypotheses of ditrysian phylogeny, although most relationships among superfamilies are weakly supported. There are also notable surprises, such as a consistently closer relationship of Pyraloidea than of butterflies to most Macrolepidoptera. Monophyly is significantly rejected by one or more character sets for the putative clades Macrolepidoptera as currently defined (P < 0.05) and Macrolepidoptera excluding Noctuoidea and Bombycoidea sensu lato (P ≤ 0.005), and nearly so for the superfamily Drepanoidea as currently defined (P < 0.08). Superfamilies are typically recovered or nearly so, but usually without strong support. Relationships within superfamilies and families, however, are often robustly resolved. We provide some of the first strong molecular evidence on deeper splits within Pyraloidea, Tortricoidea, Geometroidea, Noctuoidea and others. Separate analyses of mostly synonymous versus non-synonymous character sets revealed notable differences (though not strong conflict), including a marked influence of compositional heterogeneity on apparent signal in the third codon position (nt3). As available model partitioning methods cannot correct for this variation, we assessed overall phylogeny resolution through separate examination of trees from each character set. Exploration of "tree space" with GARLI, using grid computing, showed that hundreds of searches are typically needed to find the best-feasible phylogeny estimate for these data.

Conclusion

Our results (a) corroborate the broad outlines of the current working phylogenetic hypothesis for Ditrysia, (b) demonstrate that some prominent features of that hypothesis, including the position of the butterflies, need revision, and (c) resolve the majority of family and subfamily relationships within superfamilies as thus far sampled. Much further gene and taxon sampling will be needed, however, to strongly resolve individual deeper nodes.     

Role of galeal sensilla in host recognition and feeding behaviour of the Colorado potato beetle

Abstract. Close-range interactions with plants and the early stages of feeding behaviour of adult Colorado potato beetles, Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae), were studied using beetles with and without various mouthpart sensilla. Representative host, potato (Solarium tuberosum) , and non-host, tomato (Lycopersicon esculentum) , leaves were treated very differently by beetles with and without their galeal or palpal sensilla. Galeal sensilla were particularly important for beetles interacting with these two plants. Compared with normal beetles, fewer beetles without galeae fed on potato leaves, and those that did feed spent more time assessing the plant and took more bites before feeding. On tomtato, more beetles without galeae fed, spent less time assessing the plant and took fewer bites before feeding. Beetles without galeae also had lower consumption rates on potato and higher consumption rates on tomato. Beetles were very reluctant to feed on tomato if the galeae were present so removal of maxillary sensilla was studied only on potato. Without the sensilla on the maxillary palpi only the time between first touch and first bite was affected (lengthened). The importance of galeal sensilla in host recognition by this species and the implications for chemosensory studies are discussed here and in a companion paper.     

Two new species and a new subgenus of toothed Brachyhypopomus electric knifefishes (Gymnotiformes,Hypopomidae) from the central Amazon and considerations pertaining to the evolution of a monophasic electric organ discharge

We describe two new, closely related species of toothed Brachyhypopomus (Hypopomidae: Gymnotiformes: Teleostei) from the central Amazon basin and create a new subgenus for them. Odontohypopomus, new subgenus of Brachyhypopomus, is diagnosed by (1) small teeth present on premaxillae; (2) medialmost two branchiostegal rays thin with blades oriented more vertically than remaining three rays; (3) background color in life (and to lesser extent in preservation) distinctly yellowish with head and sides peppered with small, widely spaced, very dark brown stellate chromatophores that greatly contrast with light background coloration; (4) a dark blotch or bar of subcutaneous pigment below the eye; (5) electric organ discharge waveform of very long duration (head-positive phase approx. 2 milliseconds or longer, head-negative phase shorter or absent) and slow pulse repetition rate (3–16 Hz). The type species of the new subgenus, Brachyhypopomus (Odontohypopomus) walteri sp. n., is diagnosed by the following additional character states: (1) subcutaneous dark pigment at base of orbit particularly prominent, (2) body semi-translucent and nearly bright yellow background coloration in life, (3) a biphasic electric organ discharge (EOD) waveform of very long duration (between 3.5 and 4 milliseconds at 25° C) with head-positive first phase significantly longer than second head-negative phase in both sexes. Brachyhypopomus (Odontohypopomus) bennetti sp. n. is diagnosed by two character states in addition to those used to diagnose the subgenus Odontohypopomus: (1) a deep electric organ, visible as large semi-transparent area, occupying approximately 14–17% body depth directly posterior to the abdominal cavity in combination with a short, but deep, caudal filament, and (2) a monophasic, head-positive EOD waveform, approximately 2.1 milliseconds in duration in both sexes. These are the only described rhamphichthyoid gymnotiforms with oral teeth, and Brachyhypopomus bennetti is the first Brachyhypopomus reported to have a monophasic (head-positive) EOD waveform. Unlike biphasic species, the waveform of its EOD is largely unaffected by tail damage from predators. Such injuries are common among specimens in our collections. This species’ preference for floating meadow habitat along the major channels of the Amazon River basin may put it at particularly high risk of predation and “tail grazing.”     

The role of endogenous antifreeze protein enhancers in the hemolymph thermal hysteresis activity of the beetle Dendroides canadensis

Antifreeze proteins (AFPs) lower the freezing point of water by a non-colligative mechanism, but do not lower the melting point, therefore producing a difference between the freezing and melting points termed thermal hysteresis. Thermal hysteresis activity (THA) of AFPs from overwintering larvae of the beetle Dendroides canadensis is dependent upon AFP concentration and the presence of enhancers of THA which may be either other proteins or low molecular mass enhancers. The purpose of this study was to determine the relative contributions of endogenous enhancers in winter D. canadensis hemolymph.Winter hemolymph collected over four successive winters (1997-1998 to 2000-2001) was tested. The first three of these winters were the warmest on record in this area, while December of the final year was the coldest on record. Protein and low molecular mass enhancers raised hemolymph THA 60-97% and 35-55%, respectively, based on hemolymph with peak THA for each year collected over the four successive winters. However, the hemolymph AFPs were not maximally enhanced since addition of the potent enhancer citrate (at non-physiologically high levels) resulted in large increases in THA. ¹³NMR showed that glycerol was the only low molecular mass solute present in sufficiently high concentrations in the hemolymph to function as an enhancer. Maximum THA appears to be ∼8.5 °C.     

Efficiency of biomass transfer and the stability of model food-webs

Low efficiency of biomass transfer between trophic levels was found to enhance the stability of a model ecosystem in Lotka-Volterra form, consisting of six basal species, three primary consumers and an omnivore (De Angelis, 1975). The present paper extends De Angelis's result to a more general class of models, varying in species number, connectance and precise features of web design. Whilst the efficiency of biomass transfer does influence stability, it does not do so in the relatively simple way proposed by De Angelis. Three ranges of maximum efficiency of biomass transfer, γ_max, were employed, biologically probable (1 > γ_max > 0· 1), possible to very unlikely (0 · 1 > γ_max>0 · 01) and highly improbable (0.01> γ_max> 0· 001). Over most web configurations, in biologically probable regions of ymax, increasing connectance decreased the probability of webstability—the opposite of De Angelis's prediction. Increasing connectance only enhanced stability in biologically unrealistic regions of γ_max. Stability was also markedly influenced in these models by the number of component species and trophic levels in the web, features not evaluated by De Angelis. Low efficiency of biomass transfer enhanced stability in some types of web, but not in others.     

Pollination ecology of Acacia gerrardii Benth. (Fabaceae: Mimosoideae) under extremely hot-dry conditions

Talh trees (Acacia gerrardii Benth.) are acacias that are native to the arid and semiarid Africa and west Asia. We investigated the flowering biology, pod set and flower visitors of Talh and discussed the role of these visitors in pollen transfer. The Talh trees blossomed laterally on the nodes of one-year-old twigs. Each node produced 21 flower buds seasonally. Each flower bud opened to a flower head (FH) of 60 florets. The bagged FHs podded significantly (p ⩽ 0.05) less than did the unbagged FHs. The FHs were visited by 31 insect species (25 genera, 16 families and 5 orders). The major taxa were honeybees, megachilids, butterflies, ants, beetles and thrips. Each of honeybees, megachilids and beetles showed a significant (p ⩽ 0.05) hourly pattern, while each of butterflies, ants and thrips had no hourly pattern (p > 0.05). Furthermore, some birds and mammals touched the Talh FHs. Talh trees evolved a mass flowering behavior to face pre- and post-flowering obstacles. Megachilids seemed to play the major effort of zoophily because of their relatively high numbers of individuals and species and their effective movement behavior on the FH surface. Nevertheless, honeybees and other insects and vertebrate taxa also contributed to the pollen transfer. These results greatly contribute to our understanding of the pollination ecology of acacias, especially Arabian acacias.     

Patch-clamp analysis of voltage- and ligand-activated whole-cell currents in freshly dissociated neurons of the locust optic lobe

Whole-cell patch-clamp recordings were obtained from 116 freshly dissociated neuronal somata from the optic lobe of adult locusts (Schistocerca gregaria). Prerequisites were a papain treatment and the directed transfer of somata to the recording chamber by dabbing. Of the recorded somata, 65 were from lamina and 51 from other optic lobe neurons. All somata supported voltage-activated outward currents and some (24% of optic lobe, 3% of lamina neurons) also fast inward currents. Most lamina neurons supported an outward current that activated (V _1/2=−8.5 mV) and inactivated rapidly and a sustained outward current. Some lamina and most optic lobe neurons expressed only a sustained outward current (V _1/2=−9.4 mV). GABA and histamine elicited inward currents at negative holding potentials. Most optic lobe (95%) but only 18% of lamina neurons showed a γ-aminobutyric acid (GABA) current, whereas a similar percentage of optic lobe (50%) and lamina neurons (67%) expressed a histamine current. Both currents reversed near the chloride equilibrium potential, were reversibly reduced by picrotoxin, and did not show rundown. Thus, they likely represent chloride currents mediated by ionotropic receptors. Our data indicate that the lamina neurons recorded mainly represent monopolar cells postsynaptic to histaminergic photoreceptors. The optic lobe neurons, on which GABA and histamine apparently act as inhibitory neurotransmitters, are more heterogeneous. Accepted: 30 November 1997     

Regulation of cuticular hydrocarbon profile maturation by Drosophila tanning hormone,bursicon, and its interaction with desaturase activity

Shortly after emergence the exoskeleton (cuticle) of adult insects is rapidly expanded, hardened (sclerotized), and pigmented (melanized). In parallel with this process, the oenocytes, which are large polyploid cells located below the abdominal epidermis, secrete onto the cuticle a cocktail of cuticular hydrocarbons (CHs) and waxes. These improve the waterproofing of the cuticle, and also provide important chemosensory and pheromonal cues linked with gender, age, and species differentiation. The hardening and pigmentation of the new cuticle are controlled by the neurohormone, bursicon, and its receptor, encoded by the DLGR2 receptor, rickets (rk); by contrast, little is known about the timecourse of changes in CH profile and about the role of bursicon in this process. Here we show in Drosophila that rk function is also required for the normal maturation of the fly's CH profile, with flies mutant for rk function showing dramatically elevated levels of CHs. Interestingly, this effect is mostly abrogated by mutations in the Δ9 desaturase encoded by the desaturase1 gene, which introduces a first double bond into elongated fatty-acid chains, suggesting that desaturase1 acts downstream of rk. In addition, flies mutant for rk showed changes in the absolute and relative levels of specific 7-monoenes (in males) and 7,11-dienes (in females). The fact that these differences in CH amounts were obtained using extractions of very different durations suggests that the particular CH profile of flies mutant for rk is not simply due to their unsclerotized cuticle but that bursicon may be involved in the process of CH biosynthesis itself.     

FISH identification of Helicoverpa armigera and Mamestra brassicae chromosomes by BAC and fosmid probes

Since the Bombyx mori genome sequence was published, conserved synteny between B. mori and some other lepidopteran species has been revealed by either FISH (fluorescence in situ hybridization) with BAC (bacterial artificial chromosome) probes or linkage analysis. However, no species belonging to the Noctuidae, the largest lepidopteran family which includes serious polyphagous pests, has been analyzed so far with respect to genome-wide conserved synteny and gene order. For that purpose, we selected the noctuid species Helicoverpa armigera and Mamestra brassicae, both with n = 31 chromosomes. Gene-defined fosmid clones from M. brassicae and BAC clones from a closely related species of H. armigera, Heliothis virescens, were used for a FISH analysis on pachytene chromosomes. We recognized all H. armigera chromosomes from specific cross-hybridization signals of 146 BAC probes. With 100 fosmid clones we identified and characterized all 31 bivalents of M. brassicae. Synteny and gene order were well conserved between the two noctuid species. The comparison with the model species B. mori (n = 28) showed the same phenomenon for 25 of the 28 chromosomes. Three chromosomes (#11, #23 and #24) had two counterparts each in H. armigera and M. brassicae. Since n = 31 is the modal chromosome number in Lepidoptera, the noctuid chromosomes probably represent an ancestral genome organization of Lepidoptera. This is the first identification of a full karyotype in Lepidoptera by means of BAC cross-hybridization between species. The technique shows the potential to expand the range of analyzed species efficiently.     

A receptor and binding protein interplay in the detection of a distinct pheromone component in the silkmoth Antheraea polyphemus

Male moths respond to conspecific female-released pheromones with remarkable sensitivity and specificity, due to highly specialized chemosensory neurons in their antennae. In Antheraea silkmoths, three types of sensory neurons have been described, each responsive to one of three pheromone components. Since also three different pheromone binding proteins (PBPs) have been identified, the antenna of Antheraea seems to provide a unique model system for detailed analyzes of the interplay between the various elements underlying pheromone reception. Efforts to identify pheromone receptors of Antheraea polyphemus have led to the identification of a candidate pheromone receptor (ApolOR1). This receptor was found predominantly expressed in male antennae, specifically in neurons located beneath pheromone-sensitive sensilla trichodea. The ApolOR1-expressing cells were found to be surrounded by supporting cells co-expressing all three ApolPBPs. The response spectrum of ApolOR1 was assessed by means of calcium imaging using HEK293-cells stably expressing the receptor. It was found that at nanomolar concentrations ApolOR1-cells responded to all three pheromones when the compounds were solubilized by DMSO and also when DMSO was substituted by one of the three PBPs. However, at picomolar concentrations, cells responded only in the presence of the subtype ApolPBP2 and the pheromone (E,Z)-6,11-hexadecadienal. These results are indicative of a specific interplay of a distinct pheromone component with an appropriate binding protein and its related receptor subtype, which may be considered as basis for the remarkable sensitivity and specificity of the pheromone detection system.