Parameters influencing reversal of segment sequence in posterior egg fragments ofCallosobruchus (Coleoptera)

Summary The double abdomen type of embryonic segment pattern can develop in posterior fragments ofCallosobruchus eggs. In this type of pattern, a series of posterior segments is joined in reversed polarity to an equal set from the original pattern persisting in normal polarity. Reversed and non-reversed sets are fused in a plane of mirror symmetry, which shows in the larval cuticle as a symmetry line. This line may be located anywhere in the posterior thorax or the anterior abdomen. The reversed abdomen may be incomplete caudally due to secondary causes. Polarity reversal and concomitant double abdomen formation occurred only when temporary constriction was terminated before cellularization of the blastoderm, and only when the anterior fragment was degenerating. Maximum reversal frequency was 94% of analyzable posterior partial larvae when the constriction was applied slightly anterior to the middle of the egg when the egg contained 4–32 nuclei. Reversal was often restricted to longitudinal strips of the larval cuticle. The longitudinal borderlines between the reversed and the non-reversed strips ran predominantly along the larval midlines. Such borderlines probably existed in the blastoderm anywhere around its circumference, but borderlines in the future mesoderm and serosa would be internalized during gastrulation and dorsal closure, respectively, and the embryonic midlines would then become secondary borderlines visible in the larval cuticle. If a morphogen is involved in segment pattern formation, its transport in the egg must be polarized longitudinally in order to account for reversals restricted to longitudinal cuticular strips.     

Pattern formation in fragmented eggs of the short germ insect Schistocerca gregaria

Summary The effect of transverse fragmentation on the segment pattern of the short germ embryo of the locust Schistocerca gregaria has been investigated at two stages subsequent to the formation of the germ anlage. Following fragmentation both anterior and posterior partial embryos were observed, although rarely in a single egg. Anterior partial patterns usually terminated with a segment visible at the time of fragmentation or with the next segment due to appear. Posterior partial patterns began with a wide range of segments depending on the level of fragmentation.Anterior and posterior partial patterns developing in a single egg were usually not complementary and the segments missing sometimes included some segments visible when the embryo was fragmented. Non-complementary patterns resulted following fragmentation in all regions, while complementary patterns only occurred after fragmentation in the visibly-segmented region.The results suggest that following fragmentation isolated posterior portions of the embryo continue to form segments, while isolated anterior regions usually do not. This effect could result from variable damage to an existing pattern of unequally-sized segment primordia, or from the disruption of a process of sequential segmentation in the elongating posterior region of the embryo. The results are broadly compatible with the progress zone model proposed by Summerbell et al. (1973).     

Development of the embryonic segment pattern in the pea-beetle Callosobruchus maculatus. Supracellular,cellular and molecular aspects

Summary

This review deals with the question of how cells in the early embryo of the pea-beetle differentiate into a sequential pattern of segments. Anterior and posterior fragments of an egg have different options for development depending on whether they are exposed, before cellularization, to decaying ooplasm in the complementary fragment. Without such exposure all fragments produce fewer segments than corresponding fragments obtained at cellularization. With exposure a fraction of anterior and posterior fragments produces considerably more segments than corresponding fragments obtained at cellularization. In addition, posterior fragments are uniquely different from anterior ones in that they also produce reversal of segment sequence which can be restricted to longitudinal strips of the larval cuticle.

The difference in reaction to decaying ooplasm between anterior and posterior fragments suggests an asymmetry in the control of metamerization. Lateral inhibition by an asymmetric gradient of a diffusible morphogen can describe these observations [18] except for the restriction of reversal to longitudinal strips. The latter requires either that morphogen transport be polarized, possibly by a voltage gradient in the egg, or that the interpretation of cell position is polarized. The induction of double abdomens with UV-light and RNase suggests that RNA is part of the control mechanism. This and strip-restricted reversal are features shared by eggs of Coleoptera and Diptera.     

Double abdomen induction by UV in Bradysia tritici (syn. Sciara ocellaris,Sciaridae): sensitive stages and conditions for photoreversal

Summary The pattern anomaly double abdomen was induced in embryos of Bradysia tritici (syn. Sciara ocellaris) by irradiation of the anterior egg pole with far UV (254 or 285 nm) using low UV fluences. The maximum yield of 18% of double abdomens was obtained when 2.5 h embryos were irradiated (late intravitelline cleavage stage); earlier irradiation failed to yield double abdomens, as did irradiations after the early syncytial blastoderm stage. Exposing irradiated embryos to photoreverting light (366 nm) reduced the yield of malformations. Most double abdomens were symmetrical and the number of segments ranged from 3 to 8 in each set, with the mean value at 6.4 segments.     

The ultrastructural organization of the isolated cortex in eggs ofNassarius reticulatus (Mollusca)

Summary We have studied the organization of the cortex in fertilized eggs ofNassarius reticulatus by examining rotary-shadowed whole mounts of isolated cortices in the transmission electron microscope. The following components were distinguished: (a) the plasma membrane, with clathrin-coated areas and coated pits, (b) microfilaments and microtubules, and (c) a tubulovesicular network of endoplasmic reticulum. Microfilaments were identified by labeling with heavy meromyosin, and microtubules with a monoclonal anti-tubulin antibody, using both immunofluorescence microscopy and immunogold labeling for transmission electron microscopy. The microfilaments are organized in a network parallel to and closely associated with the plasma membrane, with typical Y- and X-shaped intersections. The endoplasmic reticulum is associated with this microfilamentous lattice. The microtubules also run parallel to the plasma membrane, but they are located at a greater distance, as can be inferred from stereo images. In the uncleaved egg, numerous microtubules are present in the egg cortex. Shortly before polar lobe formation, at the onset of mitosis, the microtubules disappear almost entirely. They reappear again at the end of first cleavage, as the polar lobe is being resorbed. The synthesis of cortical microtubules at this stage appears to depend on the presence of microtubule-organizing centers in the animal hemisphere of the egg, since microtubules do not reappear in isolated polar lobes. Clathrin-coated areas are present in both the animal and vegetal hemisphere before polar lobe formation. During mitosis, the clathrin-coated plaques and pits are found almost exclusively in the animal hemisphere. After resorption of the polar lobe, at the two-cell stage, no clathrin-coated areas were found at all.     

Expression patterns of twist and snail in Tribolium (Coleoptera) suggest a homologous formation of mesoderm in long and short germ band insects

The mesodermal region in Drosophila is determined by a maternally derived morphogenetic gradient system which specifies the different cell fates along the dorsoventral axis, including the prospective mesodermal cells at the ventral side of the embryo. There are at least two zygotic target genes, twist and snail, which are required for mesoderm formation in Drosophila. To analyze whether a similar mode of mesoderm specification might also apply to short germ band insect embryos, we have cloned twist and snail- related gene fragments from the flour beetle Tri-bolium and have analyzed their expression pattern. Both genes are expressed in a ventral stripe at early blastoderm stage, which is restricted to the region of the developing germ rudiment. The cells expressing the two genes are those that invaginate during gastrulation, indicating that the early stages of mesoderm specification are indeed very similar between the two species. Interestingly, both genes are also expressed during germband extension in a subregion of the growth zone of the embryo which forms the mesodermal cells. This suggests that the expression of the two genes is required for mesoderm formation both at early blastoderm stage and during germband elongation until the end of the segmental growth process. © 1994 Wiley-Liss, Inc.     

Regional distribution of three ultrastructural retinula types in the retina of Cataglyphis bicolor Fabr. (Formicidae,Hymenoptera)

Summary The retina of Cataglyphis bicolor was investigated by electron microscopy. Three types of structurally distinct retinulae were found and mapped throughout the compound eye: Type I is composed of four unpigmented thin cells, four larger pigmented cells as well as a basal ninth cell. Its rhabdom possesses a round cross section and four microvilli directions. This type occupies most of the dorsal two-thirds of the retina. Type II consists of two thin cells, two intermediate cells and four large cells. A basal ninth cell is also present; the rhabdom is as in type I. Type II retinulae are located in the ventral third of the retina. Type III ommatidia are unique within the Hymenoptera: there are four large pigmented cells, four thinner unpigmented cells and a basal ninth cell. The rhabdom, however, has a dumb-bell shaped cross section; two small cells lie at its opposed extremities and the remaining six cells have mutually perpendicular microvilli orientations. This type of retinula is found at the dorso-medial eye margin. Serial sectioning in this region revealed a conical shaped rhabdom without any torsion along the longitudinal axis. The rhabdomere cross section was calculated from distal and proximal thin sections. Angular statistics were applied to the microvilli directions of all three ommatidial types to determine the degree of order. A possible functional significance of the structural specializations of the different eye regions is discussed.Supported by Swiss National Science Foundation, Grant No. 3.814.72 awarded to Prof. Dr. R. Wehner. This work is part of a Ph. D. thesis. I wish to thank Prof. Dr. R. Wehner for continuous support and my colleagues Dr. P. Duelli and Dr. E. Meyer for a fruitful collaboration     

Germ-cell cluster formation in the telotrophic meroistic ovary of <Emphasis Type="Italic">Tribolium castaneum</Emphasis> (Coleoptera,Polyphaga, Tenebrionidae) and its implication on insect phylogeny

Tribolium castaneum has telotrophic meroistic ovarioles of the Polyphaga type. During larval stages, germ cells multiply in a first mitotic cycle forming many small, irregularly branched germ-cell clusters which colonize between the anterior and posterior somatic tissues in each ovariole. Because germ-cell multiplication is accompanied by cluster splitting, we assume a very low number of germ cells per ovariole at the beginning of ovariole development. In the late larval and early pupal stages, we found programmed cell death of germ-cell clusters that are located in anterior and middle regions of the ovarioles. Only those clusters survive that rest on posterior somatic tissue. The germ cells that are in direct contact with posterior somatic cells transform into morphologically distinct pro-oocytes. Intercellular bridges interconnecting pro-oocytes are located posteriorly and are filled with fusomes that regularly fuse to form polyfusomes. Intercellular bridges connecting pro-oocytes to pro-nurse cells are always positioned anteriorly and contain small fusomal plugs. During pupal stages, a second wave of metasynchronous mitoses is initiated by the pro-oocytes, leading to linear subclusters with few bifurcations. We assume that the pro-oocytes together with posterior somatic cells build the center of determination and differentiation of germ cells throughout the larval, pupal, and adult stages. The early developmental pattern of germ-cell multiplication is highly similar to the events known from the telotrophic ovary of the Sialis type. We conclude that among the common ancestors of Neuropterida and Coleoptera, a telotrophic meroistic ovary of the Sialis type evolved, which still exists in Sialidae, Raphidioptera, and a myxophagan Coleoptera family, the Hydroscaphidae. Consequently, the telotrophic ovary of the Polyphaga type evolved from the Sialis type. Electronic supplementary material Supplementary material is available in the online version of this article at and is accessible for authorized users.     

Constancy of ascending projections in the metamorphosing brain of the meal-beetle Tenebrio molitor L. (Insecta: Coleoptera)

Summary The fate of ascending projections of thoracic interneurons in the metamorphosing brain of Tenebrio molitor is described. Persistent brain neurons were identified and their fate is described during metamorphosis. The projection sites of ascending elements are invariable throughout metamorphosis both in quantitative and in qualitative terms. Some of these ascending neurons are serotonin-immunoreactive and this set of neurons maintains a constant projection site within the metamorphosing brain. The alterations in the projection sites of these and other ascending neurons in the ventral nerve cord were analysed experimentally. The central projection sites of these persistent ascending neurons are not important for the maintenance of their nerve cord projections throughout metamorphosis. Experimental deletion of ascending neurons which project into the suboesophageal ganglion varies the shape of persistent central neurons.     

Lehr's fields of campaniform sensilla in beetles (Coleoptera): Functional morphology. II. Wing reduction and the sensory field

Loss of the flight ability and wing reduction has been reported for many taxa of Coleoptera. If elytra are closed, their roots are clenched between the tergum and the pleuron, forces applied to the elytra can not be transmitted to the field of campaniform sensilla situated on the root. That is why it is plausible to assume that the field becomes redundant in non-flying beetles. We examined the relationships between the hind wing reduction and characters of this mechanosensory field in beetles of six families. We measured the size of the elytron, that of the hind wing and counted the number of sensilla in the sensory field. Mesopterous non-flying beetles retain one half to one third of sensilla present in macropterous species of the same body size. Further reduction of the sensory field in brachypterous species is obvious, but sensilla are still present in insects with strongly reduced wings, as long as their elytra are separable and mesothoracic axillaries are present. Complete loss of sensilla coincides with the existence of a permanent sutural lock. However, some beetles with permanently locked elytra and absence of axillaries still retain few campaniform sensilla. A very special case of an extreme wing modification in feather-wing beetles is considered. No sensilla were revealed either on the root of the elytron or on the basal segment of such fringed wings in flying ptiliid species.     

The photomechanic infrared receptor for the detection of forest fires in the beetle Melanophila acuminata (Coleoptera: Buprestidae)

We recorded from single units of individual sensilla of the thoracic infrared (IR) pit organs of Melanophila acuminata. When the organ was stimulated with a thermal radiator whose emission spectrum was similar to that of a typical forest fire, units responded phasically with up to seven spikes within 30–40 ms at a radiation power of 24 mW cm⁻². In the experiments all wavelengths shorter than 1.6 μm were excluded by a longpass IR filter. Response latencies were about 4 ms and initial impulse frequencies were up to 250 impulses per second (ips). A single spike could be generated even when stimulus duration was only 2 ms. Reduction of total radiation power from 24 mW cm⁻² to 5 mW cm⁻² resulted in increased response latencies of 5–6 ms and the occurrence of only two to three spikes. Initial impulse frequencies decreased to 125 ips. According to our physiological results and calculations, Melanophila should be able to detect a 10-hectare fire from a distance of 12 km. Mechanical stimuli also evoked responses of the IR sensilla. All present morphological and physiological findings lead to the conclusion that the IR receptors of Melanophila must function by means of a hitherto undescribed photomechanic mechanism. Accepted: 1 November 1997     

Effect of food shortage and temperature on oxygen consumption in the lesser mealworm, Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae)

Abstract.  Temperature and food availability are limiting factors for the establishment of tropical insects in temperate countries. In the alien pest beetle, Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae), starvation and temperature have a significant impact on metabolic rate with oxygen consumption ranging from 0.5 µmol/g fresh mass (FM)/h at 12 °C to 3.4 µmol/g FM/h at 24 °C. At 12 °C, oxygen consumption decreased continuously during an entire period of starvation. However, at 16, 20 and 24 °C, beetles display a marked hyperactivity that leads to an increase in the oxygen consumption level during the first week of starvation, followed by a steep decrease until the end of the starvation period. Oxygen consumption either does not decline in fed beetles (observed at higher temperatures) or declines at a much shallower rate than in starved beetles (observed at cooler temperatures). During the first week of refeeding, Oxygen consumption rose steeply at 16, 20 and 24 °C before levelling off to the initial value ( t ₀). At 12 °C, no compensation process was observed during recovery. This study reveals that an important threshold in the biology of A. diaperinus lies between 12 and 16 °C, leading to the onset of reduced locomotor activity and the promotion of survival to the detriment of reproduction. This 'sit and wait' behaviour is proposed as an adaptive strategy (i.e. inactivity and lower oxygen consumption coupled with low energetic requirements and high recovery abilities). Such behaviour and the observed hyperactivity were rarely described in insects before the present study. Together, the previous and present results suggest that A. diaperinus populations are likely maintained in temperate regions by immigration from warmer situations.     

Peptidic soil components are a major dietary resource for the humivorous larvae of Pachnoda spp. (Coleoptera: Scarabaeidae)

Humivorous scarab beetle larvae can thrive exclusively on soil organic matter. Feeding experiments have revealed that the larva of Pachnoda ephippiata mineralizes all major humus components except the polyphenolic fraction. High proteolytic activity in the alkaline midgut fluid and an enormous ammonia production during gut passage suggested that peptidic soil components are an important dietary resource for the larva. By comparing acid-hydrolyzable amino acids in food soil and feces, we showed that a significant fraction of the peptides in soil are removed during gut passage. This agrees well with the high concentrations of free amino acids found the midgut section. Incubation experiments revealed the presence of substantial particle-associated proteolytic activity also in the hindgut, most probably due to microbial activity. High rates of ammonia formation in hindgut homogenates and the conversion of radiolabeled amino acids to acetate and propionate indicated that microbial fermentations of soil peptides play an important role in the hindgut. This was corroborated by viable counts of amino-acid-fermenting bacteria, which formed a substantial fraction of the hindgut microbiota. A complete inventory of organic and inorganic nitrogen species before, during, and after gut passage revealed the formation of nitrite and nitrate in midgut and hindgut, and a substantial nitrogen deficit in the feces, suggesting that part of the ammonia formed by mineralization is subjected to oxidation and subsequent denitrification to N2. Together, the results strongly support the hypothesis that peptidic soil components form a major energy and nutrient source for humivorous insects, supplying the animal with microbial fermentation products and essential amino acids.     

Superhydrophobic cuticle with a "pinning effect" in the larvae of the iris sawfly, Rhadinoceraea micans (Hymenoptera, Tenthredinidae)

The integument of some sawfly larvae can be easily damaged causing haemolymph bleeding (easy bleeding phenomenon). In the present study, we investigated intact cuticle, cuticle without wax coverage and cuticle replicas of sawfly larvae by using cryo-scanning electron microscopy and contact angle (CA) measurements. The easy bleeder Rhadinoceraea micans was compared to the non-easy bleeder Nematus pavidus (both Tenthredinidae), and the examination showed that the surface structures of R. micans were hierarchically arranged, whereas the cuticle surface of N. pavidus appeared much smoother. The intact cuticle proved to be superhydrophobic in R. micans (CAs: 156° dorsally, 120° ventrally), but not in N. pavidus (CAs: 67° dorsally, 47° ventrally). The wettability of R. micans increased significantly after the waxes had been peeled off. Replicas of the intact cuticle surface were hardly wettable in both species. It is suggested that wax crystals in combination with cuticle microsculptures are the most important features for rendering the integument of the easy bleeder species superhydrophobic. The wax-free tips of the cuticle microsculptures in R. micans are sites where haemolymph droplets can adhere. The integument surface of easy bleeders is assumed to be adapted to their particular defence strategy, simultaneously contributing to the maintenance and "reuse" of released haemolymph droplets on the superhydrophobic surface due to this "pinning effect".     

Presence of the storage seed protein vicilin in internal organs of larval Callosobruchus maculatus (Coleoptera: Bruchidae)

Variant vicilins (7S storage globulins) of cowpea seeds (Vigna unguiculata) are considered as the main resistance factor present in some African genotypes against the bruchid Callosobruchus maculatus. It has been suggested that the toxic properties of vicilins may be related to their recognition and interaction with glycoproteins and other membrane constituents along the digestive tract of the insect. However, the possibility of a systemic effect has not yet been investigated. The objective of this work was to study the fate of 7S storage globulins of V. unguiculata in several organs of larvae of the cowpea weevil C. maculatus. Results demonstrated binding of vicilins to brush border membrane vesicles, suggesting the existence of specific receptors. Vicilins were detected in the haemolymph, in the midgut, and in internal organs, such as fat body and malpighian tubules. There is evidence of accumulation of vicilins in the fat body of both larvae and adults. The absorption of vicilins and their presence in insect tissues parallels classical sequestration of secondary compounds.     

Variation in number and differentiation of the abdominal infrared receptors in the Australian 'fire-beetle' Merimna atrata (Coleoptera, Buprestidae)

Most individuals of the Australian ‘fire-beetle’ Merimna atrata have two pairs of IR receptors which are located ventrolaterally on the second and third abdominal sternite. An IR receptor consists of a specialized IR absorbing area, which is innervated by a neural complex. This complex contains one thermoreceptive multipolar neuron with a unique terminal dendritic mass (TDM) and two scolopidia and was termed ‘sensory complex’. However, also individuals with one pair of IR receptors on the second sternite and beetles with three pairs on the second, third, and fourth sternites were found. Additionally, beetles having one or two pairs of IR receptors may have preliminary stages of IR receptors on the third and fourth sternite, respectively. We found two kinds of preliminary stages, both of which are characterized by a much less pronounced absorbing area. In all five abdominal sternites segmental nerves are attached to the cuticle with a neural complex. Investigation of complexes of non-IR sternites suggests that the sensory cells inside the sensory complex of an IR receptor have developed from common internal stretch receptors. From our results it can be hypothesized that the IR sensory system in Merimna atrata has not yet reached a stage, which can be regarded as evolutionary stable.     

Lehr's fields of campaniform sensilla in beetles (Coleoptera): Functional morphology. III. Modification of elytral mobility or shape in flying beetles

Some flying beetles have peculiar functional properties of their elytra, if compared with the vast majority of beetles. A “typical” beetle covers its pterothorax and the abdomen from above with closed elytra and links closed elytra together along the sutural edges. In the open state during flight, the sutural edges diverge much more than by 90°. Several beetles of unrelated taxa spread wings through lateral incisions on the elytra and turn the elytron during opening about 10–12° (Cetoniini, Scarabaeus, Gymnopleurus) or elevate their elytra without partition (Sisyphus, Tragocerus). The number of campaniform sensilla in their elytral sensory field is diminished in comparison with beetles of closely related taxa lacking that incision. Elytra are very short in rove beetles and in long-horn beetles Necydalini. The abundance of sensilla in brachyelytrous long-horn beetles Necydalini does not decrease in comparison with macroelytrous Cerambycinae. Strong reduction of the sensory field was found in brachyelytrous Staphylinidae. Lastly, there are beetles lacking the linkage of the elytra down the sutural edge (stenoelytry). Effects of stenoelytry were also not uniform: Oedemera and flying Meloidae have the normal amount of sensilla with respect to their body size, whereas the sensory field in the stenoelytrous Eulosia bombyliformis is 5–6 times less than in chafers of the same size but with normally linking broad elytra.     

Evolution of clutch size in insects. II. A test of static optimality models using the beetle Callosobruchus maculatus (Coleoptera: Bruchidae)

Wilson and Lessells (1993) analysed the effect of constraint assumptions on the predictions of static optimality models for insect clutch size. They concluded that the models could be reliably distinguished between (and hence the main constraints identified) only after precise quantitative predictions had been examined. The present paper describes a series of laboratory experiments, using the bruchid beetle Callosobruchus maculatus, that allow these quantitative predictions to be made and tested. Experiments in which female encounter rate with hosts was altered gave qualitative support for 3 out of 6 basic (single oviposition) models, but the quantitative fit of them all was poor. However, when the (a priori) condition was included in these models that several other females would oviposit on the same hosts (the multiple oviposition models), the time limiting multiple oviposition model alone produced quantitative predictions that were supported by observations. In other words, the results suggest that the main constraints on bruchid oviposition behaviour are the amount of time available for laying eggs and the number of other females ovipositing. However, additional qualitative predictions indicate that the number of eggs available to the female may also constrain clutch size evolutionarily. The usefulness of static optimality models for examining clutch size decisions in insects is discussed in the context of these results.     

Host location in Oomyzus gallerucae (Hymenoptera: Eulophidae), an egg parasitoid of the elm leaf beetle Xanthogaleruca luteola (Coleoptera: Chrysomelidae)

Eggs of the elm leaf beetle Xanthogaleruca luteola are often heavily attacked by the chalcidoid wasp Oomyzus gallerucae. We studied the chemical signals mediating interactions between the egg parasitoid, its host, and the plant Ulmus campestris. Olfactometer bioassays with O. gallerucae showed that volatiles of the host-plant complex attract the parasitoid. In order to determine the source of attractive volatiles within this host-plant-complex, we tested separately the effect of odours of eggs, gravid elm leaf beetle females, faeces of the beetles and elm twigs (with undamaged leaves and leaves damaged either mechanically or by feeding of the beetles). Odours of faeces of the elm leaf beetle were attractive, whereas neither volatiles from eggs nor from gravid females acted as attractants. Volatiles from undamaged or damaged plants did not elicit a positive reaction in O. gallerucae, whereas volatiles from feeding-damaged plants onto which host eggs had been deposited were attractive. This latter result suggests that it is not feeding but deposition of host eggs onto elm leaves that induces the production of plant volatiles attractive to the egg parasitoid. Investigations of the search patterns of O. gallerucae within the habitat by laboratory bioassays revealed that the egg parasitoid encounters host eggs by chance. Contact kairomones from faeces were demonstrated to be important in microhabitat acceptance, while contact kairomones isolated from the host eggs are relevant for host recognition. Received: 12 February 1997 / Accepted: 29 April 1997