Fine structure and physicochemical analysis of the metathoracic scent glands of Lincus Malevolus (Rolston) and L. Spurcus (Rolston) (Heteroptera : Pentatomidae)

Bugs of the genus Lincus (Heteroptera : Pentatomidae) are attracted by volatile compounds emitted from the inflorescences of fertile palms. To define the basis of their chemical ecology, we have studied the metathoracic scent glands (MTG) of males and females of 2 species, L. spurcus and L. malevolus. The metathoracic scent gland system belongs to the diastomien type. The 2 glandular pores located between the mesothoracic and metathoracic coxae are associated with “crescent-like” evaporation areas. The large median reservoir, which is composed of one type of flattened pigmented epithelial cells, is flanked by multitubular lateral glands. These glands result from the apposition of 2 cell-type glandular units. The strip-like accessory gland is embedded in the reservoir wall. At its level, the thinner cuticular intima forms finger-like invaginations where a protein secretory product is secreted. Extracts of the volatile fraction of the metathoracic gland secretion were analyzed by capillary gas chromatography (GC) and by GC-mass spectrometry (GC-MS). These analyses exhibited a typical pentatomid MTG composition. The glands of L. spurcus and L. malevolus males and females contain 11 compounds: (E)-2-hexenal, 4-keto-(E)-2-hexenal, (E)-2-hexenoic acid, decane, (E)-2-octenal, undecene, undecane, (E)-2-octenly acetate, (E)-2-decenal, tridecane and (E)-2-decenyl acetate, including 3 major compounds, which represent 60–85% of the secretion in the 2 species: (E)-2-hexenal, (E)-2-octenal, and n-undecane. The 4-keto-(E)-2-hexenal is present only in the L. malevolus MTG, and represents 26% of its secretion. The female extracts of both species are characterized by the presence of (E)-2-hexenoic acid, which was detected in the male extracts as traces.     

Vitamin E supplementation increases the attractiveness of males' scent for female European green lizards

Background

In spite that chemoreception is important in sexual selection for many animals, such as reptiles, the mechanisms that confer reliability to chemical signals are relatively unknown. European green lizards (Lacerta viridis) have substantial amounts of α-tocopherol ( = vitamin E) in their femoral secretions. Because vitamin E is metabolically important and can only be attained from the diet, its secretion is assumed to be costly. However, its role in intraspecific communication is unknown.

Methodology/Principal Findings

Here, we experimentally show that male European green lizards that received a dietary supplement of vitamin E increased proportions of vitamin E in their femoral secretions. Furthermore, our experiments revealed that females preferred to use areas scent marked by males with experimentally increased vitamin E levels in their secretions. Finally, female preferences were stronger when vitamin E differences between a pair of males'' secretions were larger.

Conclusions/Significance

Our results demonstrate that female green lizards are able to discriminate between males based on the vitamin E content of the males'' femoral secretions. We suggest that the possible cost of allocating vitamin E to secretions, which might be dependent on male quality, may be a mechanism that confers reliability to scent marks of green lizards and allows their evolution as sexual signals.     

Isolation and Characteristic of Overlong-Chain Unsaturated Aldehydes from the Freshwater Sponge <Emphasis Type="Italic">Lubomirskia baicalensis</Emphasis>

Several unusual overlong-chain unsaturated aldehydes (22 : 1, 22 : 2, 23 : 1, 24 : 1, 24 : 2, and 25 : 2) were found in total lipids of the endemic sponge Lubomirskia baicalensis from Baikal Lake. Tetracos-17-enal was identified as the major aldehyde of the mixture using GC-MS and ¹H and ¹³C NMR spectroscopy. A procedure for the isolation of total overlong-chain aldehydes was suggested. We think that the overlong-chain aldehydes defend the sponge from fouling and predators.     

The accessory gland and metathoracic scent gland function in Oncopeltus fasciatus

The paper provides further clues as to the physiological function and biological significance of the ‘accessory gland’ in the metathoracic scent apparatus of the lygaeid Oncopeltus fasciatus. From various lines of evidence (gas chromatographic, cytochemical) it is concluded as probable that the accessory gland secretes small quantities of a mucopolysaccharide secretory product together with water. The difficulty of reconciling these data with other data indicating that the accessory gland is involved in the biosynthesis of the scent aldehydes is discussed. It is suggested that the water secreted into the median scent reservoir by the accessory gland provides O. fasciatus with a means of volumetric compensation for a diminished output of scent repellent.     

External thoracic scent efferent system of Scutelleridae (Hemiptera: Heteroptera)

The metathoracic scent glands in the Heteroptera produce defence secretions which are spread outside the body through and by using the thoracic external scent efferent system. That complex system was studied in 18 species from 11 genera of four subfamilies, Elvisurinae, Eurygastrinae, Hoteinae and Scutellerinae of the family Scutelleridae (Pentatomoidea). The results have been compared with published data. The pattern of that system is more consistent at the level of genus, mostly very similar in the congeneric species, but mostly variable within higher taxonomic levels, tribes and subfamilies. Five types of the external scent efferent system are recognized within the family Scutelleridae, basic two of them in studied species: (i) peritreme well developed, covering large part of metapleuron, evaporatorium small, developed only on metapleuron, (ii) evaporatorium large, more conspicuous than moderate-sized to small peritreme, extending to mesopleuron as large structure. The results do not support a hypothesis that the system of structures associated externally with metathoracic scent glands is in correlation with type of a habitat. However, these structures are well usable as diagnostic characters for scutellerid genera (e.g. Cantao, Hyperonchus, Scutellera and Solenosthedium).     

Morphology and Volatile Compounds of Metathoracic Scent Gland in Tessaratoma papillosa (Drury) (Hemiptera: Tessaratomidae)

Tessaratoma papillosa (Drury) (Hemiptera: Tessaratomidae) is a serious insect pest of litchi and longan in South China. When disturbed, this insect could release large quantities of disagreeable odorous volatiles from its scent gland. Knowledge on the scent gland and its secretion is crucial for developing the semiochemical methods to manage this pest. Morphology and ultrastructure of the metathoracic scent glands (MTGs) were studied under stereo and scanning electron microscopy, and the volatile compounds of MTGs from both male and female T. papillosa were analyzed with coupled gas chromatography?Cmass spectrometry (GC?CMS). The MTG complex is located between the metathorax and the first abdominal segment at the ventral surface of the insect, which has a well-developed single double valve cystic-shaped orange median reservoir, paired colorless lateral glands in both sides, and a long and wavy tubular accessory gland that inlays tightly into the ventral edge around the median reservoir. The MTG opens to the body surface through paired ostioles located between the meso- and metacoxae of the evaporatorium with mushroom bodies. The GC?CMS analyses showed that female and male adults have nine major volatile components in common. Tridecane is the most abundant in both females and males, reaching up to 47.1% and 51.8% of relative amount, respectively. The minor component is benzophenone with only 0.28% and 0.14%. Furthermore, undecane, tetradecane, 3-methyl-tridecane, and cyclopentadecane were found only in males. The possible function of volatile compounds of MTG contents in T. papillosa is addressed.     

Observations on the metathoracic scent gland system of the backswimmer,Notonecta glauca L. (Heteroptera: Notonectidae)

The closing apparatus in the metathoracic scent gland system of Notonecta glauca is described, and notes on the distribution of metathoracic scent glands in the family Notonectidae are included.     

The Asian grass lizard (Takydromus sexlineatus) does not respond to the scent of a native mammalian predator

Lacertid lizards use chemical cues emitted by saurophagous snakes to evade predation. Whether these lizards can detect and respond to the chemical cues of predatory mammals has not been studied. As many mammals carry distinct body odours and/or use chemical cues for intraspecific communication, lizards can be expected to use these chemicals as early warning cues. To test this idea, we observed the behaviour of Asian grass lizards (Takydromus sexlineatus) that had been transferred to an unfamiliar test arena containing one of four scent treatments. No particular scent was applied to the arena in the control situation. Diluted aftershave served as a pungency control. In the snake treatment, scent of the Oriental whip snake (Ahaetulla prasina) was applied. We included this treatment to learn how Asian grass lizards react to predator chemical cues. Finally, in the mongoose treatment, the lizards were confronted with scent cues of several small Indian mongooses (Herpestes auropunctatus). Snake scent elicited foot shakes, startles and tail vibrations. These are behaviours that in lacertid lizards are associated with stressful situations such as predatory encounters. Surprisingly, lizards confronted with mongoose scent exhibited none of these stress-indicating behaviours. In fact, their behaviour did not differ from that of lizards subjected to an odourless control treatment. These results raise concern. Mongooses are rapidly invading ecosystems worldwide. If lizards that have co-evolved with mongooses are unable to detect these predators’ presence through chemical cues, it seems highly unlikely that evolutionary naïve lizards will develop this ability rapidly.     

High congruence of intraspecific variability in floral scent and genetic patterns in Gentianella bohemica Skalický (Gentianaceae)

Gentianella bohemica Skalický (Gentianaceae) is a critically endangered species endemic to the Bohemian Massif in the border region of Germany, Czechia and Austria. It consists of a restricted number of extremely scattered populations which are known to form distinct genetic groups. The objective of this work was to test for differences in the floral scent between Gentianella bohemica and Gentianella germanica and within these two species among populations, and to test for a correlation of scent and genetic similarity among the populations of G. bohemica. Floral scent was collected from the inflorescences/plants of eight flowering populations of G. bohemica and three populations of G. germanica using dynamic headspace methods, followed by GC/MS analyses. Both species emitted several aromatic and terpenoid compounds and multivariate analyses revealed differences in scent between the two species and within species among G. bohemica populations. Volatile components overlapped as expected for closely related species but floral scent was taxon-specific. Floral scent differentiation among G. bohemica populations was in high congruence with the genetic differentiation suggesting that scent differences among populations have a genetic basis and showing that scent is a suitable chemotaxonomic marker in this species.     

The metathoracic scent gland system in Hydrocorisae (Heteroptera: Nepomorpha)

Abstract. The metathoracic scent gland system is a basic feature of the order Heteroptera. It occurs widely not only in the terrestrial forms (Geocorisae) and water-surface bugs (Amphibicorisae) but also in the totally aquatic bugs (Hydrocorisae) and their littoral relatives (Ochteridae, Gelastocoridae). In Hydrocorisae the metathoracic scent gland conforms to Carayon's (1971) omphalian type (orifice median and undivided or, if divided, orifices close together towards the mid-ventral line) but shows marked differences in structure and physiological function between species from different families. There is taxonomic interest in the distribution of the three distinctly different types of metathoracic occlusion apparatus. Naucoridae and Belostomatidae have a median lip-valve, Notonectidae and Gelastocoridae a pair of stop-valves, Corixidae a median flap-valve. The valve opener muscles are usually dorso-ventral but are ventral in Corixidae; the cuticular microsculpture in the metathoracic efferent system specifically in Hydrocorisae with stop-valves (Notonectidae, Gelastocoridae) is similar to that widely present in Geocorisae. Groupings of Hydrocorisae from variations in the metathoracic occlusion apparatus differ from others based upon variations in the chemical constitution of the secretions. It is considered that the facts now known on metathoracic scent gland structure and function accord well with the hypothesis that Hydrocorisae constitute a polyphyletic assemblage.     

Comparative morphology of the odoriferous system in three predatory stink bugs (Heteroptera: Asopinae)

The metathoracic scent system in Heteroptera produces and releases defensive volatile compounds. The odor produced by predatory stink bugs differs from phytophagous bugs, suggesting a variation between the structure and function of the metathoracic scent system. The anatomy and ultrastructure of the external thoracic efferent system, scent gland, and reservoir in the stink bug predators Brontocoris tabidus, Podisus nigrispinus, and Supputius cincticeps (Heteroptera: Pentatomidae: Asopinae) were studied. External thoracic efferent systems of B. tabidus, P. nigrispinus, and S. cincticeps have anatomical differences in ostiole, peritreme, and evaporatorium. Scent glands have a secretory portion and a reservoir. The reservoir has irregular projections, and in B. tabidus, it is enlarged and heart shaped, whereas in P. nigrispinus and S. cincticeps it is flattened and semicircular. The secretory tissue of the scent gland has well-developed globular secretory cells that produce odorous compounds, and the reservoir has a single layer of cubical cells lined by a cuticular intima. Secretory cells are type III with an intracellular end apparatus, well-developed nucleus with decondensed chromatin, and cytoplasm rich in mitochondria, lysosomes, granules, and smooth endoplasmic reticulum. These findings suggest that there are differences in physiological function of the odoriferous system and the volatile compounds produced by the secretory cells, which may indicate variation in defensive behavior of these species.     

Change in protective coloration in the striated shieldbug <Emphasis Type="Italic">Graphosoma lineatum</Emphasis> (Heteroptera: Pentatomidae): predator avoidance and generalization among different life stages

There are two major forms of protective coloration, camouflage and warning coloration, which often entail different colour pattern characteristics. Some species change strategy between or within life stages and one such example is the striated shieldbug, Graphosoma lineatum. The larvae and the pale brownish-and-black striated pre-diapause adults are more cryptic in the late summer environment than is the red-and black striation that the adults change to after diapause in spring. Here we investigate if the more cryptic pre-diapause adult and larval coloration may affect the aposematic function of the coloration as compared to the red adult form. In a series of trials we presented fifth instar larvae, pale or red adults to shieldbug-naïve domestic chicks, Gallus gallus domesticus, to investigate the birds’ initial wariness, avoidance learning, and generalization between the three prey types. The naïve chicks found the red adults most aversive followed by pale adults, and they found the larvae the least aversive. The birds did not find the larvae unpalatable and did not learn to avoid them, while they learned to avoid the two adult forms and then to a similar degree. Birds generalized asymmetrically between life stages, positively from larvae to adults and negatively from adults to larvae. We conclude that the lower conspicuousness in the pale forms of G. lineatum may entail a reduced aposematic function, namely a reduced initial wariness in inexperienced birds. The maintenance of the colour polymorphism is discussed.     

Diel fragrance pattern correlates with olfactory preferences of diurnal and nocturnal flower visitors in Salix caprea (Salicaceae)

Floral scent, often a complex mixture of several volatile organic compounds (VOCs), has generally been interpreted as an adaptation to attract pollinators. However, not many studies have analysed which VOCs are functionally relevant for the reproductive success of a plant. Here, we show that, in Salix caprea (Salicaceae), temporal changes in floral scent emission during the day and night attract two different types of flower visitor: bees during the day and moths during the evening and night. We analysed the contribution of the two flower visitor groups to the reproductive success of the plant. The differences in scent emitted during the peak activity times of flower visitors (day versus night) were quantified and the response of 13 diurnal/nocturnal pollinator taxa to the floral scents was tested using gas chromatographic and electroantennographic techniques. Many of the c. 40 identified scent compounds were physiologically active, and bees and moths responded to nearly identical sets of compounds, although the response strengths differed. In bioassays, bees preferred the most abundant 1,4‐dimethoxybenzene over lilac aldehyde, a compound with increased emission at night, whereas moths preferred lilac aldehyde over 1,4‐dimethoxybenzene. Pollination by wind plus nocturnal pollinators (mainly moths) or by wind alone contributed less to seed set than pollination by wind plus diurnal pollinators (mainly bees). This suggests that the emission of scent during the night and attracting moths have no significant effect on reproductive success. It is possible that the emission of lilac aldehydes and other compounds at night is s result of phylogenetic constraints. Future studies should investigate whether moths may produce a marginal fitness gain in some years and/or some populations. © 2014 The Linnean Society of London, Botanical Journal of the Linnean Society, 2014, 175 , 624–640.     

Species Specificity of Aldehyde and Fatty Acid Profiles of Four Family Group Representatives within the Insect Infraorder Pentatomomorpha (Hemiptera: Heteroptera)

Reactive α,β‐unsaturated aldehydes, including 4‐oxoalk‐2‐enals, are known to be present in volatile secretions of numerous heteropteran insect species. Because the aldehydes are likely to originate from metabolism of fatty acids (FAs), the present study aimed to examine and compare the aldehyde and FA profiles of four model heteropteran species. The model species consisted of adult family group representatives within the infraorder Pentatomomorpha (Hemiptera: Heteroptera): seed bug (Lygaeus equestris (Lygaeoidea)), dock leaf bug (Coreus marginatus (Coreoidea)), red firebug (Pyrrhocoris apterus (Pyrrhocoroidea)), and European stink bug (Graphosoma lineatum (Pentatomoidea)). Solid‐phase microextraction combined with two‐dimensional gas‐chromatography/time‐of‐flight mass spectrometry was used to establish the profiles of volatile secretions in stressed living insects. The FA profiles of acylglyceride and phospholipid fractions deposited in fat body and/or hemolymph were obtained by liquid chromatography/mass spectrometry and gas chromatography with flame ionization detection techniques. Our results based on multivariate statistical analyses of the data imply that volatile secretion blends as well as fat body and/or hemolymph lipid profiles are species specific but the differences in volatile blends between different species do not mirror the changes in corresponding fat body and/or hemolymph lipid profiles of stressed and non‐stressed individuals.     

Scent gland apparatus in the Western conifer seed bug Leptoglossus occidentalis Heidemann (Heteroptera: Coreidae)

The coreid Leptoglossus occidentalis is a Nearctic bug responsible for severe seed losses to pine orchards. When disturbed, adults and nymphs emit a defensive secretion deemed an allomone. Here we describe the gross morphology of the scent gland apparatus and the related evaporatory structures in nymphs and adults of L. occidentalis, through light microscopy and scanning electron microscopy. Adults of both sexes possess a metathoracic scent gland complex (MTG) including a central orange‐yellow reservoir and a pair of white lateral glands, connected by ducts to the reservoir. The MTG belongs to the diastomian type, with two ostioles located on the metathorax associated with a microsculptured cuticular accumulation area, i.e. evaporatory area, which can prevent the spread of the secretion on to non‐evaporative cuticle and increase scent fluid evaporation. A high number of male‐specific sternal gland pores were observed. These pores and associated glands are likely the source of an attractant pheromone, which could be extremely useful in monitoring and combating this invasive pest. In nymphs, MTG is replaced by two dorsal abdominal scent glands (DAGs) located between the 4th and the 6th urotergites. DAGs are reddish cuticle‐lined sacs with gland cells forming the gland wall; the scent substances are released through two orifices lying on the mid‐dorsal abdominal line between urotergites IV–V and V–VI. Also in nymphs, peculiar cuticular evaporatory areas surround both orifices.     

The comparative morphology of epidermal glands in Pentatomoidea (Heteroptera)

The Heteroptera show a diversity of glands associated with the epidermis. They have multiple roles including the production of noxious scents. Here, we examine the cellular arrangement and cytoskeletal components of the scent glands of pentatomoid Heteroptera in three families, Pentatomidae (stink bugs), Tessaratomidae, and Scutelleridae (shield-backed bugs or jewel bugs). The glands are; (1) the dorsal abdominal glands, (2) the tubular glands of the composite metathoracic gland, and (3) the accessory gland component of the composite metathoracic gland. The dorsal abdominal glands are at their largest in nymphs and decrease in size in adults. The metathoracic gland is an adult-specific gland unit with a reservoir and multiple types of gland cells. The accessory gland is composed of many unicellular glands concentrated in a sinuous line across the reservoir wall. The lateral tubular gland is composed of two-cell units. The dorsal abdominal glands of nymphs are composed of three-cell units with a prominent cuticular component derived from the saccule cell sitting between the duct and receiving canal. The cuticular components that channel secretion from the microvilli of the secretory cell to the exterior differ in the three gland types. The significance of the numbers of cells comprising gland units is related to the role of cells in regenerating the cuticular components of the glands at moulting in nymphs.     

Lipophilic compounds in femoral secretions of males and females of the El Hierro giant lizard Gallotia simonyi (Lacertidae)

Many lizards use femoral gland secretions in reliable intraspecific communication. Based on mass spectra, obtained by GC–MS, we found 57 lipophilic compounds in femoral secretions of males and females of El Hierro giant lizards, Gallotia simonyi (fam. Lacertidae). Compounds included steroids (mainly cholesterol) and fatty acids ranging between n-C₁₆ and n-C₂₂ (mainly hexadecanoic and octadecanoic acids), followed by aldehydes, alcohols, ketones, squalene and waxy esters. There were important intersexual differences in the presence, abundance and number of compounds (more numerous in males). Males had higher proportions of the most odoriferous compounds (fatty acids and aldehydes), while females had higher proportions of more stable compounds (steroids, waxy alcohols, waxy esters and terpenoids). This suggests sexual differences in function of femoral secretions. In addition, some compounds could reflect the physiological state, allowing monitoring health of lizards from secretions samples, which is especially important given the critical conservation status of this lizard.     

A key role for floral scent in a wasp-pollination system in Eucomis (Hyacinthaceae)

Background and Aims

Floral scent may play a key role as a selective attractant in plants with specialized pollination systems, particularly in cases where floral morphology does not function as a filter of flower visitors. The pollination systems of two African Eucomis species (E. autumnalis and E. comosa) were investigated and a test was made of the importance of scent and visual cues as floral attractants.

Methods and Key Results

Visitor observations showed that E. autumnalis and E. comosa are visited primarily by pompilid wasps belonging to the genus Hemipepsis. These wasps carry considerably more Eucomis pollen and are more active on flowers than other visiting insects. Furthermore, experiments involving virgin flowers showed that these insects are capable of depositing pollen on the stigmas of E. autumnalis, and, in the case of E. comosa, pollen deposited during a single visit is sufficient to result in seed set. Experimental hand-pollinations showed that both species are genetically self-incompatible and thus reliant on pollinators for seed set. Choice experiments conducted in the field and laboratory with E. autumnalis demonstrated that pompilid wasps are attracted to flowers primarily by scent and not visual cues. Measurement of spectral reflectance by flower petals showed that flowers are cryptically coloured and are similar to the background vegetation. Analysis of headspace scent samples using coupled gas chromatography–mass spectrometry revealed that E. autumnalis and E. comosa scents are dominated by aromatic and monoterpene compounds. One hundred and four volatile compounds were identified in the floral scent of E. autumnalis and 83 in the floral scent of E. comosa, of which 57 were common to the scents of both species.

Conclusions

This study showed that E. autumnalis and E. comosa are specialized for pollination by pompilid wasps in the genus Hemipepsis and achieve specialization through cryptic colouring and the use of scent as a selective floral attractant.Key words: Eucomis, Pompilidae, wasp pollination, breeding system, pollination syndrome, pollinator shift, floral volatile, floral filter     

Isolation and characteristic of super long-chain unsaturated aldehydes from the freshwater sponge Lubomirskia baicalensis

Several unusual overlong-chain unsaturated aldehydes (22:1, 22:2, 23:1, 24:1, 24:2, and 25:2) were found in total lipids of the endemic sponge Lubomirskia baicalensis from Baikal Lake. Tetracos-17-enal was identified as the major aldehyde of the mixture using GC-MS and 1H and 13C NMR spectroscopy. A procedure for the isolation of total overlong-chain aldehydes was suggested. We think that the overlong-chain aldehydes defend the sponge from fouling and predators. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2005, vol. 31, no. 6; see also http://www.maik.ru.