Venom gland of the digger wasp Liris niger: morphology, ultrastructure, age-related changes and biochemical aspects

Females of the parasitoid digger wasp species Liris niger hunt crickets as food for their future brood. The wasps paralyse the prey by injecting their venom directly into the CNS. The venom is produced in a gland consisting of two ramified glandular tubules terminating in a common reservoir. The reservoir contents enter the sting bulb via a ductus venatus. Secretory units of dermal gland type III line the two free gland tubules, the afferent ducts to the reservoir and the cap region within the reservoir. Secretion products of tubules reach the reservoir through the cuticle-lined central funnel. Secretory cells in the distal and middle parts of the tubules contain extensive rough endoplasmic reticulum and numerous electron-dense vesicles, whereas secretory cells of the afferent ducts and the cap region of the reservoir lack electron-dense vesicles and the endoplasmic reticulum is poorly developed. The secretory apparatus undergoes age-related changes. The secretory units in the venom gland tubules and inside the reservoir complete differentiation 1 day after imaginal ecdysis. After 30 days, massive autolytic processes occur in the secretory cells and in the epithelial cells of the reservoir. Analysis of the polypeptide composition demonstrates that the venom reservoir contains numerous proteins ranging from 3.4 to 200 kDa. A dominant component is a glycoprotein of about 90 kDa. In contrast the polypeptide composition of Dufour's gland is completely different and contains no glycoproteins. Comparison of the venom reservoir contents with the polypeptide pattern of venom droplets reveals that all of the major proteinaceous constituents become secreted. Thus the secreted venom contains exclusively proteins present in the soluble contents of the venom gland.     

黄腹潜蝇茧蜂寄生因子的特性及其对寄主的生理效应

初步研究了黄腹潜蝇茧蜂Opius caricivorae Fischer寄生因子的特性及其对寄主美洲斑潜蝇Liriomyza sativae Blanchard幼虫的生理效应。黄腹潜蝇茧蜂携带的主要因子是毒液。毒液器官是由一个土黄色的锥形毒囊和7个透明的椭圆形的毒腺及导管构成的;毒液的电泳图谱显示约有12条蛋白带,其中绝大多数低于100 kD,含量最高的3条蛋白带为43.5、25.9和20.1 kD;杜氏腺约有15条左右蛋白质条带,其中有5条含量很高（121.4、77.0、51.5、42.7和36.5 kD）。通过透射电镜观察,在黄腹潜蝇茧蜂毒腺分泌细胞和卵巢表皮细胞中新发现存在一种类病毒颗粒,这些球状颗粒直径大约为50 nm。雌蜂经Co60辐射处理后再寄生（即假寄生）3龄寄主幼虫,被寄生后的寄主依然能正常化蛹,但不能羽化;7 h后寄生体壁开始出现红斑;脂肪体形态结构无显著变化;绝大多数的蜂卵没有被包囊。推测在正常寄生的情况下可能是毒液抑制了寄主的包囊作用,而新发现的类病毒颗粒是否参与了这一过程目前还不清楚。     

Comparative morphology of the venom gland and reservoir in opiine and alysiine braconid wasps (Insecta, Hymenoptera, Braconidae)

Detailed venom reservoir and venom gland intima morphology was investigated in representatives of 178 species and 76 genera of Braconidae belonging to the subfamilies Opiinae (23 genera and 69 species) and Alysiinae (36 genera and 67 species of Alysiini, and 17 genera and 42 species of Dacnusini). The presence of an unsculptured sack–like expansion of the anterior end of the reservoir suggests a relationship between some members of the alysiine tribe Dacnusini, viz. Exotela and Chorebus , and the genera Oenonogaster and Glyphogaster , both of which are currently placed in the Alysiini. The same feature also suggests that Exotela, Chorebus, Oenonogaster and Glyphogaster may be derived from the Opiinae separately from other Alysiinae. Other dacnusines examined are indicated as forming a monophyletic taxon on the basis of their reduced and narrow reservoirs. An anterior insertion of the venom glands on to the reservoir generally supports the Phaenocarpa group of alysiine genera, with some modification, whereas the presence of a discrete, narrow posterior extension of the reservoir represents a synapomorphy for members of the Aspilota group of alysiine genera. The phylogenetic inferences from this venom apparatus study are discussed in the light of the biology of the taxa concerned. Twenty-one new combinations are made in the Opiinae and seven in the Alysiinae.     

Venom gland of the ichneumonid Diadromus collaris： morphology, ultrastructure and age-related changes

Females of the solitary parasitoid Diadromus collaris （Insecta： Hymenoptera： Ichneumonidae） lay eggs in the pupae of Plutella xylostella （Lepidoptera： Plutellidae）, and the venom is synchronously injected into hosts. The venom apparatus consists of two glandular tubules terminating in a common reservoir, A ductule connects the reservoir with the sting apparatus, by which the reservoir content enters the latter. Secretory units line the two glandular tubules. All secretory cells belong to dermal gland type Ⅲ. Dermal gland cells in glandular tubules are more abundant and developed than those in the reservoir. There are extensive rough endoplasmic reticulum and electrondense vesicles, and the microvilli are well developed. By the cuticle-lined central funnel secretion products of secretory units reach the reservoir. Moreover, the secretory apparatus undergoes age-related changes. The secretory units in the venom gland are better developed and more vigorous 7 days after eclosion than those 1 day after eclosion; autolytic processes occur 15 days after eclosion, and the tissue of the reservoir is more prostrate 15 day after eclosion than those 1 day after eclosion. The ovipostion peak of this parasitoid, about 3-7 days after eclosion, corresponds with the period when the venom gland is highly developed in the life span of the wasp.     

Morphology and ultrastructure of a secretory region enclosed by the venom reservoir in social wasps (Insecta,Hymenoptera)

The morphology and ultrastructure of the convoluted gland inside the venom reservoir of four species of social Vespidae are described. The cells of the venom gland (including the convoluted gland) can be divided into six groups: (1) epithelial cells, (2) glandular cells with the end apparatus secreting into the tubule inside the convoluted gland (internal or embedded tubule), (3) a continuous arrangement of glandular cells with the end apparatus secreting directly into the venom reservoir, (4) glandular cells that are loosely dispersed along the tubule lumen between the free tubules and the embedded tubule of the convoluted gland, (5) secretory cells of the free tubules and (6) duct cells. One kind of secretory cell, hitherto unknown and described in this paper (group 3), is characterized by the presence of a well-developed end apparatus, usually with enlarged extracellular spaces, but lacking the normally associated duct cells. The secretory cells contain several stacks of granular endoplasmic reticulum, but these are mainly concentrated in the middle of the cell. The basal half of the cells contains many lipid droplets. Although the function of the convoluted gland is not yet understood, an hypothesis is related to what is known of the function of reservoir secretory cells in solitary wasps. All wasp species studied showed the same organization of the convoluted gland, which clearly distinguishes their venom gland from that of Sphecidae.     

Virus-like particles in venom of Meteorus pulchricornis induce host hemocyte apoptosis

Ultrastructural studies on the reproductive tract and venom apparatus of a female braconid, Meteorus pulchricornis, revealed that the parasitoid lacks the calyx region in its oviduct, but possesses a venom gland with two venom gland filaments and a venom reservoir filled with white and cloudy fluid. Its venom gland cell is concaved and has a lumen filled with numerous granules. Transmisson electron microscopic (TEM) observation revealed that virus-like particles (VLPs) were produced in venom gland cells. The virus-like particle observed in M. pulchricornis (MpVLP) is composed of membranous envelopes with two different parts: a high-density core and a whitish low-density part. The VLPs of M. pulchricornis is also found assembling ultimately in the lumen of venom gland cell. Microvilli were found thrusting into the lumen of the venom gland cell and seem to aid in driving the matured MpVLPs to the common duct of the venom gland filament. Injection of MpVLPs into non-parasitized Pseudaletia separata hosts induced apoptosis in hemocytes, particularly granulocytes (GRs). Rate of apoptosis induced in GRs peaked 48h after VLP injection. While a large part of the GR population collapsed due to apoptosis caused by MpVLPs, the plasmatocyte population was minimally affected. The capacity of MpVLPs to cause apoptosis in host's hemocytes was further demonstrated by a decrease ( approximately 10-fold) in ability of host hemocytes to encapsulate fluorescent latex beads when MpVLPs were present. Apparently, the reduced encapsulation ability was due to a decrease in the GR population resulting from MpVLP-induced apoptosis.     

A phylogenetic analysis of higher-level gall wasp relationships (Hymenoptera: Cynipidae)

We present the most comprehensive analysis of higher-level relationships in gall wasps conducted thus far. The analysis was based on detailed study of the skeletal morphology of adults, resulting in 164 phylogenetically informative characters, complemented with a few biological characters. Thirty-seven cynipid species from thirty-one genera, including four genera of the apparently monophyletic Cynipini and almost all of the genera in the other tribes, were examined. The outgroup included exemplar species from three successively more distant cynipoid families: Figitidae (the sister group of the Cynipidae), Liopteridae and Ibaliidae. There was considerable homoplasy in the data, but many groupings in the shortest tree were nonetheless well supported, as indicated by bootstrap proportions and decay indices. Partitioning of the data suggested that the high level of homoplasy is characteristic of the Cynipidae and not the result of the amount of available phylogenetically conservative characters being exhausted. The analysis supported the monophyly of the Cynipini (oak gall wasps) which, together with the Rhoditini (the rose gall wasps), Eschatocerini and Pediaspidini formed a larger monophyletic group of gall inducers restricted to woody representatives of the eudicot subclass Rosidae. The inquilines (Synergini) were indicated to be monophyletic, whereas the Aylacini, primarily herb gall inducers, appeared as a paraphyletic assemblage of basal cynipid groups. The shortest tree suggests that the Cynipidae can be divided into three major lineages: one including the inquilines, the Aylacini genera associated with Rosaceae, and Liposthenes ; one consisting entirely of Aylacini genera, among them Aulacidea , Isocolus and Neaylax ; and one comprising the woody rosid gallers (the oak and rose gall wasps and allies), the Phanacis-Timaspis complex and the Aylacini genera associated with Papaveraceae.     

Morphology and ultrastructure of the convoluted gland in the ant Dinoponera australis (Hymenoptera: Formicidae)

The morphology and fine structure of the convoluted gland inside the venom reservoir of the ponerine ant Dinoponera australis (Hymenoptera: Formicidae) are described. The cells of the convoluted gland can be divided into 3 major groups: (1) epithelial cells, (2) glandular cells with end apparatus secreting into the tubule inside the convoluted gland, and (3) glandular cells with end apparatus secreting directly into the venom reservoir. A fourth group of cells belonging to the venom gland of this ant is also discussed as (4) secretory cells of the free tubule (not a part of the convoluted gland). The epithelial cells in the convoluted gland do not have many organelles. Most cells of group 3 are characterized by numerous mitochondria. In some of these cells, the mitochondria possess tubular cristae. Tubule cells of group 2 inside the convoluted gland, possess little rough endoplasmic reticulum when compared with cells of group 4, situated in the free tubule.     

腰带长体茧蜂毒液器官和卵巢的形态学及其超微结构

应用超薄切片和电镜技术,观察内寄生蜂腰带长体茧蜂Macrocentrus cingulum Brischke毒液器官和卵巢的形态结构。腰带长体茧蜂毒液器官由1个毒囊和2条毒腺组成,毒腺接于毒囊的顶端。毒腺由单层分泌细胞、退化的外胚层细胞和环腔的内膜构成,分泌细胞主要由1个明显的细胞核和1个较大囊状细胞器构成,囊状细胞器的功能是分泌毒液。毒囊由肌肉鞘和扁平细胞层构成,但没有分泌细胞。腰带长体茧蜂卵巢1对,每个卵巢由10条左右卵巢小管组成,与侧输卵管相接处略微膨大形成卵巢萼区。2条侧输卵管在产卵管基部会合形成1条总输卵管与产卵管相接。毒液器官通过毒囊的毒液导管附着在总输卵管上。对寄生蜂毒液器官的生物学、细胞学及在分类进化上的意义进行研究。     

管氏肿腿蜂毒液器官超微结构观察

应用透射电镜技术,观察了管氏肿腿蜂Scleroderma guani毒液器官的超微结构.毒腺由基膜层、分泌细胞层、导管细胞层和内膜层构成,分泌细胞内含内质网、末端附器、分泌囊泡、分泌颗粒、液泡等细胞器,其内合成的毒液由末端附器输送至毒腺的腔体.毒囊由肌肉鞘层、上皮细胞层和内膜层组成,肌肉鞘内的肌纤丝规则排列不交错,上皮细胞层内细胞器稀少,内膜层呈波浪状均匀加厚.     

Morphological,ultrastructural, histochemical and electrophoretic studies on the venom system of Nasonia vitripennis walker (hymenoptera: Pteromalidae)

The venom system of Nasonia vitripennis is well-developed and composed of an unbranched acid gland and associated reservoir. Fine-structural, histochemical and electrophoretic studies indicate that the venom is produced by two protein-secreting epithelia. The bulk of the venom is synthesised in the columnar cells of the acid gland and discharged via “vesicular organelles” and the efferent ductular system into the lumen of the reservoir. The acid gland also contains squamous chitogenous cells, situated either around the central lumen or interposed between the bases of the columnar cells. Once within the reservoir, the venom is probably activated by enzymatic secretions from the reservoir secretory cells. Each of these cells has a “vesicular organelle” but, in contrast to the columnar cells of the acid gland, the cytoplasm contains a preponderance of free ribosomes, and protein segregation apparently occurs outside the Golgi complexes. The venom is expelled through the efferent discharge duct by muscular contractions, which open the duct lumen and bring it into contact with the funnel of the ovipositor. Excessive distortion of the duct is prevented by a massive ventral ligament.     

Diverse Filters to Sense: Great Variability of Antennal Morphology and Sensillar Equipment in Gall-Wasps (Hymenoptera: Cynipidae)

Comparative studies on antennal sensillar equipment in insects are largely lacking, despite their potential to provide insights into both ecological and phylogenetic relationships. Here we present the first comparative study on antennal morphology and sensillar equipment in female Cynipoidea (Hymenoptera), a large and diverse group of wasps, with special reference to the so-called gall-wasps (Cynipidae). A SEM analysis was conducted on 51 species from all extant cynipoid families and all cynipid tribes, and spanning all known life-histories in the superfamily (gall-inducers, gall-inquilines, and non-gall associated parasitoids). The generally filiform, rarely clavate, antennal flagellum of Cynipoidea harbours overall 12 types of sensilla: s. placoidea (SP), two types of s. coeloconica (SCo-A, SCo-B), s. campaniformia (SCa), s. basiconica (SB), five types of s. trichoidea (ST-A, B, C, D, E), large disc sensilla (LDS) and large volcano sensilla (LVS). We found a great variability in sensillar equipment both among and within lineages. However, few traits seem to be unique to specific cynipid tribes. Paraulacini are, for example, distinctive in having apical LVS; Pediaspidini are unique in having ≥3 rows of SP, each including 6–8 sensilla per flagellomere, and up to 7 SCo-A in a single flagellomere; Eschatocerini have by far the largest SCo-A. Overall, our data preliminarily suggest a tendency to decreased numbers of SP rows per flagellomere and increased relative size of SCo-A during cynipoid evolution. Furthermore, SCo-A size seems to be higher in species inducing galls in trees than in those inducing galls in herbs. On the other hand, ST seem to be more abundant on the antennae of herb-gallers than wood-gallers. The antennal morphology and sensillar equipment in Cynipoidea are the complex results of different interacting pressures that need further investigations to be clarified.     

Comparative morphology of terminal-instar larvae of Cynipoidea: phylogenetic implications

We describe the external morphology of the terminal-instar larvae of 30 species of Cynipoidea (Hymenoptera), with special reference to the head capsule and mouthparts. Twenty-five of the species belong to the Cynipidae and are gall inducers or phytophagous inquilines (guests) in galls, while five represent different insect-parasitic lineages of the Cynipoidea. Although we find only limited variation in body shape, the head sclerites and mandibles offer many characters of potential phylogenetic value. For instance, the mandibles of the parasitoids have one large pointed tooth, with several smaller dents along the inner margin in core figitids, whereas the phytophagous gall inducers and inquilines have mandibles with two or three blunt teeth of subequal size. The mandibles of inquiline larvae are unique in being covered by vertical striations and in having a dominating, broad second tooth. We summarize the qualitative variation among the studied terminal-instar larvae in terms of 33 morphological characters and one life-history trait and examine the phylogenetic implications of these data by running parsimony analyses under uniform character weights and under implied weights (Goloboff weights). The analysis under uniform weights is poorly resolved but the relationships suggested by the implied-weights analysis are largely congruent with previous analyses of adult morphology and molecular data. The larval data support inclusion of the genus Liposthenes in the Neaylax – Isocolus clade, in agreement with the molecular data but in weak conflict with adult morphology. However, the larval data agree with adult morphology and conflict with the molecular data in supporting monophyly of the inquilines.     

Exocrine glands of the ant Myrmoteras iriodum

This paper describes the morphological characteristics of nine major exocrine glands in workers of the formicine ant Myrmoteras iriodum. The elongate mandibles reveal along their entire length a conspicuous intramandibular gland, which contains both class‐1 and class‐3 secretory cells. The secretory cells of the mandibular glands show a peculiar appearance, with a branched end apparatus, which is unusual for ants. The other major glands (pro‐ and postpharyngeal gland, infrabuccal cavity gland, labial gland, metapleural gland, venom gland and Dufour gland) show common features for formicine ants. The precise function of the glands could not yet be experimentally demonstrated, and to clarify this will depend on the availability of live material of these enigmatic ants in future.     

Venom and Dufour's glands of the emerald cockroach wasp Ampulex compressa (Insecta,Hymenoptera, Sphecidae): Structural and biochemical aspects

The digger wasp species Ampulex compressa produces its venom in two branched gland tubules. They terminate in a short common duct, which is bifurcated at its proximal end. One leg is linked with the venom reservoir, the other one extends to the ductus venatus. Each venom gland tubule possesses, over its entire length, a cuticle-lined central duct. Around this duct densely packed class 3 gland units each composed of a secretory cell and a canal cell are arranged. The position of their nuclei was demonstrated by DAPI staining. The brush border of the secretory cells surrounds the coiled end-apparatus. Venom is stored in a bladder like reservoir, which is surrounded by a thin reticulated layer of muscle fibres. The reservoir as a whole is lined with class 3 gland units. The tubiform Dufour's gland has a length of about 350 μm (∅ 125 μm) only and is surrounded by a network of pronounced striated muscle fibres. The glandular epithelium is mono-layered belonging to the class 1 type of insect epidermal glands. The gland cells are characterized by conspicuous lipid vesicles. Secretion of material via the gland cuticle into the gland lumen is apparent. Analysis of the polypeptide composition demonstrated that the free gland tubules and the venom reservoir contain numerous proteins ranging from 3.4 to 200 kDa. The polypeptide composition of the Dufour's gland is completely different and contains no lectin-binding glycoproteins, whereas a dominant component of the venom droplets is a glycoprotein of about 80 kDa. Comparison of the venom reservoir contents with the polypeptide pattern of venom droplets revealed that all of the major proteinaceous constituents are secreted. The secreted venom contains exclusively proteins present in the soluble contents of the venom gland. The most abundant compound class in the Dufour's gland consisted of n-alkanes followed by monomethyl-branched alkanes and alkadienes. Heptacosane was the most abundant n-alkane. Furthermore, a single volatile compound, 2-methylpentan-3-one, was identified in various concentrations in the lipid extract of the Dufour's gland.     

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.     

Chemical composition of the poison apparatus secretions of the African weaver ant, Oecophylla longinoda, and their role in behaviour

ABSTRACT. Major workers of Oecophylla longinoda emit venom from the tip of the abdomen, as it is brought immediately above the head. The sources of the venom are (a) the poison gland, which contains formic acid, and (b) Dufour's gland, which contains hydrocarbons, including n-undecane and other n-alkanes, 4-tridecene, 8-heptadecene and 4, 7-heptadecadiene. The venom elicits a 'mass attack' response in other major workers. Formic acid and n-undecane presented together experimentally also evoke this response, the mixture being considerably more effective than either compound tested separately. These compounds act in combination with the mandibular gland secretions to form a complex alarm/defence system.     

Effect of the venom of the gregarious parasitoid Apanteles glomeratus on its hemocytic encapsulation by the host, Pieris

When eggs from the lateral oviduct of the gregarious parasitoid Apanteles glomeratus were injected with calyx fluid and venom apparatus material into host larvae, Pieris rapae crucivora, most of the eggs were not encapsulated. Apanteles eggs deposited by the parasitoid from which the venom apparatus was removed were usually encapsulated by the host. These results indicate that the parasitoid venom apparatus material is an important factor in suppressing the encapsulation of 1- or 2-day-old eggs in the host. In order to clearly demonstrate that the venom suppresses egg encapsulation but not the encapsulation of other foreign objects, DEAE-Sephadex A-50 ion-exchange particles stained with 0.001% (w/v) Congo Red solution were injected into hosts together with venom apparatus material. The Sephadex particles were encapsulated by host hemocytes. The results suggest that the venom does not inhibit the encapsulation ability of the host.     

Biased parasitoid sex ratios: Wolbachia,functional traits,local and landscape effects

Adult sex ratio (ASR) is a key demographic parameter, being essential for the survival and dynamics of populations. Biased ASRs are adaptations to the environment on different scales, resulting from different mechanisms such as inbreeding, mating behaviour, resource limitations, endosymbionts such as Wolbachia, and changes in density or spatial distribution. Parasitoid ASRs are known to be strongly biased, but little information is available on how they are affected by large-scale variables such as landscape composition or fragmentation. We examined whether landscape scale variables affect the ASR of several parasitoid species belonging to the same tritrophic gall inducer community. We examined the effects of various explanatory variables on parasitoid ASR: the ovipositor length (a species level functional trait), resource amount (gall size) and density (local scale) as well as habitat amount, land use and landscape history (landscape scale). We controlled for the incidence and prevalence of Wolbachia infections. We found that parasitoid ASR is best explained by and positively correlated with ovipositor length and gall diameter. The interaction of functional traits with habitat availability also significantly explained parasitoid ASRs. Our results support the hypothesis that large-scale environmental characteristics affect parasitoid ASRs in addition to intrinsic and local characteristics.