Sex Pheromone Composition Research Articles

The novel function of an orphan pheromone receptor reveals the sensory specializations of two potential distinct types of sex pheromones in noctuid moth

Sex pheromones play crucial role in mating behavior of moths, involving intricate recognition mechanisms. While insect chemical biology has extensively studied type I pheromones, type II pheromones remain largely unexplored. This study focused on Helicoverpa armigera, a representative species of noctuid moth, aiming to reassess its sex pheromone composition. Our research unveiled two previously unidentified candidate type II sex pheromones—3Z,6Z,9Z-21:H and 3Z,6Z,9Z-23:H—in H. armigera. Furthermore, we identified HarmOR11 as an orphan pheromone receptor of 3Z,6Z,9Z-21:H. Through AlphaFold2 structural prediction, molecular docking, and molecular dynamics simulations, we elucidated the structural basis and key residues governing the sensory nuances of both type I and type II pheromone receptors, particularly HarmOR11 and HarmOR13. This study not only reveals the presence and recognition of candidate type II pheromones in a noctuid moth, but also establishes a comprehensive structural framework for PRs, contributing to the understanding of connections between evolutionary adaptations and the emergence of new pheromone types.

Geographic variation in genetic composition, sexual communication and mating compatibility of the False Codling Moth, Thaumatotibia leucotreta for optimisation of area-wide control

Intraspecific variation in sex pheromones is a driver of reproductive isolation and speciation in insects. The False Codling Moth (FCM) Thaumatotibia leucotreta (Lepidoptera: Tortricidae) is a quarantine pest endemic to sub-Saharan Africa (SSA). The currently available precision control measures for FCM use female sex pheromone components to lure males into traps. However, the existing data on the composition of the female sex pheromone, especially the isomer ratios of the main pheromone component (E/Z)-8-dodecenyl acetate, are inconsistent for populations in SSA. This inconsistency led to speculation about possible reproductive isolation between geographically separated FCM populations and the potential need for local adjustment of pheromone-based FCM control tools. This, however, requires a comparative evaluation of geographic variation in FCM sexual communication and inter-population mating compatibility. We therefore investigated genetic isolation and mating compatibility between five geographically isolated FCM populations in South Africa and analysed the ratio of (E)- and (Z)-8-dodecenyl acetate in females from these populations. The five studied populations were found to form three genetically distinct groups with high genetic distances between each other. Mating compatibility tests showed that mating is possible across these groups, however, males preferred females of their own population when given choices; without a choice, males successfully mated with and transferred spermatophores to females from all other populations. The ratio of (E)- and (Z)-8-dodecenyl acetate was similar (c. 4:1) across the populations, indicating that this main female pheromone component does not cause the observed intra-population mating preferences. It remains to be investigated if qualitative/quantitative variation in other sex pheromone components influences intra-population recognition in South African FCM, providing a base for the development of regionally-specific lures for area-wide control programmes.

Identification of sex pheromone of red swamp crayfish Procambarus clarkii and exploration of the chemosensory mechanism of their antennae

Red swamp crayfish, Procambarus clarkii, is a globally invasive species, which has caused great damage to biodiversity, agriculture, and fishing. Therefore, the development of effective management methods, such as pheromone control, is necessary for biological control and biodiversity protection. However, the components of P. clarkii sex pheromones have not yet been explored, and the chemosensory mechanism of the P. clarkii antennae after stimulation by sex pheromone also remains unknown. In this study, we isolated and identified the candidate bioactive component of the female P. clarkii sex pheromone using ultrafiltration centrifugation, semi-preparative liquid phase separation and omics technologies and conducted bioassays to determine its attraction ability. Meanwhile, RNA-Seq technology was used to analyze the potential chemosensory mechanism of antennae. Our results indicated that the male P. clarkii were uniaxially attracted to the female crude conditioned water (FCW), medium fraction (MF, isolated by ultrafiltration centrifugation), and preparative fragment 6 of females (PFF6, isolated by semi-preparative liquid phase separation). Metabolomic analysis revealed the presence of 18 differential metabolites between the PFF6 and PFM6 samples, among which 15 were significantly upregulated in the PFF6 sample. Bioassay test also showed that mestranol, especially at concentrations of 10−5–10−2 mol∙l−1, could significantly attract P. clarkii males; therefore, mestranol was identified as the candidate sex pheromone component of P. clarkii females. Furthermore, RNA-Seq results showed that most differentially expressed genes (DEGs) enriched in lipid metabolism and signal transduction pathways were up-regulated in P. clarkii males. In addition, high expressions of Ca2+-binding protein and ion transporting ATPases may enhance the sensitivity of the antennae of P. clarkii males towards sex pheromones. Our study provides data on P. clarkii sex pheromone composition and reveals the molecular mechanism of sex pheromone response in P. clarkii. Moreover, our study provides a referable method for the isolation of candidate bioactive molecules from the P. clarkii sex pheromone.

Chemical ecology of triatomines: current knowledge and implications for Chagas disease vector management

AbstractTriatomines (Hemiptera: Heteroptera: Reduviidae), commonly known as conenose or kissing bugs, includes species of outstanding medical importance given their role as vectors of the protozoan parasite Trypanosoma cruzi Chagas (Kinetoplastida: Trypanosomatidae), the aetiological agent of Chagas disease. Herein, we present an updated review of the chemical ecology of triatomines, with special reference to molecules involved in the communication of these insects. Infochemicals, including hormones and semiochemicals (pheromones and allelochemicals), have a pivotal role in mediating information and shaping interactions within and between triatomines, during distinct behavioural contexts. We analyse the new findings about the chemical composition of alarm, aggregation and sex pheromones, as well as on cuticular hydrocarbons, host attractants and repellents. We also discuss how triatomine hormones may serve as communication signals and highlight the importance of the reverse chemical ecology approach in studying the complex chemoecology of these arthropod vectors. In the final section, we outline how knowledge about triatomine chemical ecology can be used for implementing real-world control operations with proven epidemiological impact.

Comparative expression profiles of carboxylesterase orthologous CXE14 in two closely related tea geometrid species, Ectropis obliqua Prout and Ectropis grisescens Warren.

Insect carboxylesterases (CXEs) can be expressed in multiple tissues and play crucial roles in detoxifying xenobiotic insecticides and degrading olfactory cues. Therefore, they have been considered as an important target for development of eco-friendly insect pest management strategies. Despite extensive investigation in most insect species, limited information on CXEs in sibling moth species is currently available. The Ectropis obliqua Prout and Ectropis grisescens Warren are two closely related tea geometrid species, which share the same host of tea plant but differ in geographical distribution, sex pheromone composition, and symbiotic bacteria abundance, providing an excellent mode species for studies of functional diversity of orthologous CXEs. In this study, we focused on EoblCXE14 due to its previously reported non-chemosensory organs-biased expression. First, the EoblCXE14 orthologous gene EgriCXE14 was cloned and sequence characteristics analysis showed that they share a conserved motif and phylogenetic relationship. Quantitative real-time polymerase chain reaction (qRT-PCR) was then used to compare the expression profiles between two Ectropis spp. The results showed that EoblCXE14 was predominately expressed in E. obliqua larvae, whereas EgriCXE14 was abundant in E. grisescens at multiple developmental stages. Interestingly, both orthologous CXEs were highly expressed in larval midgut, but the expression level of EoblCXE14 in E. obliqua midgut was significantly higher than that of EgriCXE14 in E. grisescens midgut. In addition, the potential effect of symbiotic bacteria Wolbachia on the CXE14 was examined. This study is the first to provide comparative expression profiles of orthologous CXE genes in two sibling geometrid moth species and the results will help further elucidate CXEs functions and identify a potential target for tea geometrid pest control.

Sex pheromone signal and stability covary with fitness.

If sexual signals are costly, covariance between signal expression and fitness is expected. Signal–fitness covariance is important, because it can contribute to the maintenance of genetic variation in signals that are under natural or sexual selection. Chemical signals, such as female sex pheromones in moths, have traditionally been assumed to be species-recognition signals, but their relationship with fitness is unclear. Here, we test whether chemical, conspecific mate finding signals covary with fitness in the moth Heliothis subflexa. Additionally, as moth signals are synthesized de novo every night, the maintenance of the signal can be costly. Therefore, we also hypothesized that fitness covaries with signal stability (i.e. lack of temporal intra-individual variation). We measured among- and within-individual variation in pheromone characteristics as well as fecundity, fertility and lifespan in two independent groups that differed in the time in between two pheromone samples. In both groups, we found fitness to be correlated with pheromone amount, composition and stability, supporting both our hypotheses. This study is, to our knowledge, the first to report a correlation between fitness and sex pheromone composition in moths, supporting evidence of condition-dependence and highlighting how signal–fitness covariance may contribute to heritable variation in chemical signals both among and within individuals.

Sex Pheromone of the Alfalfa Plant Bug, Adelphocoris lineolatus: Pheromone Composition and Antagonistic Effect of 1-Hexanol (Hemiptera: Miridae)

The sex pheromone composition of alfalfa plant bugs, Adelphocoris lineolatus (Goeze), from Central Europe was investigated to test the hypothesis that insect species across a wide geographical area can vary in pheromone composition. Potential interactions between the pheromone and a known attractant, (E)-cinnamaldehyde, were also assessed. Coupled gas chromatography-electroantennography (GC-EAG) using male antennae and volatile extracts collected from females, previously shown to attract males in field experiments, revealed the presence of three physiologically active compounds. These were identified by coupled GC/mass spectrometry (GC/MS) and peak enhancement as hexyl butyrate, (E)-2-hexenyl butyrate and (E)-4-oxo-2-hexenal. A ternary blend of these compounds in a 5.4:9.0:1.0 ratio attracted male A. lineolatus in field trials in Hungary. Omission of either (E)-2-hexenyl-butyrate or (E)-4-oxo-2-hexenal from the ternary blend or substitution of (E)-4-oxo-2-hexenal by (E)-2-hexenal resulted in loss of activity. These results indicate that this Central European population is similar in pheromone composition to that previously reported for an East Asian population. Interestingly, another EAG-active compound, 1-hexanol, was also present in female extract. When 1-hexanol was tested in combination with the ternary pheromone blend, male catches were reduced. This compound showed a dose-response effect with small doses showing a strong behavioral effect, suggesting that 1-hexanol may act as a sex pheromone antagonist in A. lineolatus. Furthermore, when (E)-cinnamaldehyde was field tested in combination with the sex pheromone, there was no increase in male catch, but the combination attracted both males and females. Prospects for practical application are discussed.

Dispersal From Natal Patch Correlates With the Volatility of Female Sex Pheromones in Parasitoid Wasps

Chemical communication via pheromones is considered the oldest and most widespread form of communication in nature. However, the way that the enormous diversity of species-specific pheromones evolved is still of debate. One possible process driving pheromone evolution is the mate-finding and dispersal behavior, as long-distance mate-finding requires highly volatile compounds. In contrast, less volatile compounds might be sufficient attractants in species that search for mates within proximity. In the parasitoid wasp genus Leptopilina, the composition of species-specific sex pheromones ranges from highly volatile iridoid compounds through combinations of iridoids with low volatile cuticular hydrocarbons (CHCs) to only CHCs. To study the selective forces shaping the composition of sex pheromones in Leptopilina, we examined the dispersal behavior, i.e. the proportion of male and female wasps dispersing after emergence, in four species with known sex pheromone compositions. If males and females disperse immediately, long-range mate attraction might become necessary, favoring volatile iridoids over CHCs. If mating occurs directly on the host patch, short-range mate attraction by low volatile CHCs might suffice. Our analyses have revealed that the dispersal behavior of Leptopilina males and females after emergence does indeed differ between species with differently volatile sex pheromones. Specifically, males of species with iridoid sex pheromones start to disperse immediately before their females’ emergence, whereas males of species with CHC sex pheromones delay dispersal until their conspecific females emerge. While the differences in female dispersal behavior turned out to be species-specific, differences in male dispersal correlated with the volatility of female-produced sex pheromones of each species. This study significantly contributes to our understanding of the evolution of sex pheromones by differences in dispersal behavior.

Polyphenism of visual and chemical secondary sexually-selected wing traits in the butterfly Bicyclus anynana: How different is the intermediate phenotype?

Polyphenism is a type of phenotypic plasticity supposedly adaptive to drastic and recurrent changes in the environment such as seasonal alternation in temperate and tropical regions. The butterfly Bicyclus anynana shows polyphenism with well-described wet and dry seasonal forms in sub-Saharan Africa, displaying striking morphological, physiological and behavioural differences in response to higher or lower developmental temperatures. During the seasonal transition in the wild, the intermediate phenotype co-occurs with wet and dry phenotypes. In this study, we aimed to characterize the secondary sexually-selected wing traits of the intermediate form to infer its potential fitness compared to wet and dry phenotypes. Among the previously described wing morphological traits, we first showed that the area of the fifth eyespot on the ventral hindwing is the most discriminant trait to identify wet, dry and intermediate phenotypes in both sexes. Second, we characterized the intermediate form for two secondary sexually-selected wing traits: the area and UV reflectance of the dorsal forewing pupil and the composition of the male sex pheromone. We showed that values of these two traits are often between those of the wet and dry phenotypes. Third, we observed increasing male sex pheromone production in ageing dry and wet phenotypes. Our results contrast with previous reports of values for sexually-selected traits in wet and dry seasonal forms, which might be explained by differences in rearing conditions or sample size effects among studies. Wet, dry and intermediate phenotypes display redundant sexually dimorphic traits, including sexually-selected traits that can inform about their developmental temperature in sexual interactions.

Comparison of the Sex Pheromone Composition of Harmonia axyridis Originating from Native and Invaded Areas.

The multicolored Asian lady beetle, Harmonia axyridis (Coleoptera: Coccinellidae), originates from South-East Asia and is now considered as an invasive species at a worldwide scale, with populations encountered in North and South America, Africa, and Europe. Several previous studies suggested that invasive populations display different behavioral and physiological traits, leading to a better fitness than native individuals. H. axyridis sex pheromone was identified recently, but only from individuals established in Europe. In this study, we compare the composition of the female sex pheromone of H. axyridis from two populations: (i) an invasive population in North America, and (ii) a native population in South-East China. We found the females originating from both populations to release in similar proportions the same five pheromonal compounds, namely β-caryophyllene, β-elemene, methyl-eugenol, α-humulene, and α-bulnesene. However, females from the North American strain release all five compounds in larger amount than the Chinese ones. Whether invasive individuals were selected during the process of invasion through their capacity to better call and find sexual partners remains to be confirmed.

Screening some Common Molecular Markers and a Desaturase Marker, Linked to Sex Pheromone Biosynthesis, in Three Z-Strain and an E-Strain Populations of the European Corn Borer, Ostrinia nubilalis, Occurring in Central Europe

The European corn borer moth, (ECB), Ostrinia nubilalis (Lepidoptera: Crambidae, Pyraustinae) is one of the most destructive pests of maize worldwide. ECB has two pheromone-strains, separated by specific ratios of isomers of E- and Z11-tetradecenyl acetates (E11- and Z11-14Ac), but appearing morphologically identical. Accordingly, E- and Z-ECB pheromone traps are available for the respective populations for practical monitoring of the flight, however, traps for Z-strain are unreliable for practical usage in some parts of Central-Europe. E- and Z-ECB populations occur in sympatry in some areas, while in allelopatry in other areas. Determining the strains before the flight of adults, when difference in the composition of their respective pheromones is manifested, would be of practical interest for early warning. In addition to the known fatty-acyl-reductase (FAR) marker, further markers would allow more comprehensive studies. We screened the following common markers for mitochondrial and nuclear DNA regions: partial cytochrome c oxidase I (COI), cytochrome B (CytB), the second spacer of the internal transcribed spacer (ITS2), Elongation factor 1a (EF1a) and actin gene (Act). In addition, a marker of the Δ11-desaturase gene (11desat), linked to biosynthesis of female-produced sex pheromone, was also included, because we reported earlier a differential expression for this site. Three Z-ECB populations locating at distant sites within Hungary, an area where only Z-strain occurs, and an E-ECB population in Slovenia, known as the closest-occurring E-strain, were included into the study. Separate laboratory colonies were established from each population, and F1 generations were sampled to verify the identity of pheromone strains, by analysing the composition of sex pheromone by gas chromatography linked to an electroantennographic detector (GC-EAD). Molecular studies were conducted using specimens taken from the F2 generations. Results of genetic studies showed that there were no differences between the Z and E populations for the common markers. In contrast to this, several nucleic acid changes (11 nt in 4 positions) were found between the three Z-populations (Hungary) and the E-population (Slovenia) in the desaturase marker. Further study is required to reveal whether the differences found in this study are consistent across E-populations, thus making these markers suitable for diagnostic purposes.

Geographic variation in pheromone component ratio and antennal responses, but not in attraction, to sex pheromones among fall armyworm populations infesting corn in Mexico

The fall armyworm, Spodoptera frugiperda (Lepidoptera: Noctuidae), is one of the most important pests of corn in Latin America. This insect presents two strains with behavioural and genetic differences. In Mexico, both strains are present, and at least two different FAW populations have been reported within the corn-strain. The objective of this study was to gather evidence of whether pheromonal communication varies among different S. frugiperda populations infesting corn in Mexico. First, we investigated any qualitative or quantitative difference in the composition of sex pheromones among populations; second, we studied whether male antennal responses to pheromone components vary among populations; and, finally, we investigated whether males from a region can discriminate between a synthetic pheromone blend characteristic of their region and blends formulated with the ratio of pheromone compounds emitted by females from other populations. Sex pheromone components were sampled by solid-phase microextraction and identified by gas chromatography coupled to mass spectrometry. Females from all populations consistently released three compounds: (Z)-9-tetradecenyl acetate (Z9-14:OAc), (Z)-11-hexadecenyl acetate (Z11-16:OAc) and (Z)-7-dodecenyl acetate (Z7-12:OAc). Our results showed that Z9-14:OAc and Z7-12:OAc evoked the highest and most variable antennal responses among the populations studied compared to Z11-16:OAc. However, males did not discriminate between local pheromone blends and those formulated from other populations in a field test. These results show that although there is geographic variation in the ratio of pheromone components and in the peripheral reception of them, males were not differentially attracted to different pheromone blends in the field.

Conservation and lineage-specific rearrangements in the GOBP/PBP gene complex of distantly related ditrysian Lepidoptera

General odorant binding proteins (GOBPs) and pheromone binding proteins (PBPs) form a monophyletic subfamily of insect odorant binding proteins (OBPs) specific for Lepidoptera, butterflies and moths. The GOBP/PBP genes include six subgroups (GOBP1–2, PBP-A–D) previously reported to form a complex arrayed in a conserved order in representative moths (superfamily Bombycoidea) and butterflies (Nymphalidae). Although our knowledge of lepidopteran genomes has increased greatly recently, the structure of the GOBP/PBP complex has been studied only for species that represent limited lineages of the highly diverged Ditrysia. To understand the evolution of this functionally important gene complex, we determined 69–149 kb genomic sequences that include GOBP2 and five PBP genes in three Ostrinia moths (Pyraloidea), O. nubilalis, O. furnacalis, and O. latipennis, using bacterial artificial chromosome (BAC) and fosmid clones. The structure of the GOBP2/PBP gene cluster was well conserved despite the different sex pheromone composition utilized by the three moths. Five expressed PBP genes in Ostrinia moths were the result of two duplications of PBP-A genes. Surprisingly, an allele containing a fusion gene between tandemly arrayed PBP-A genes was observed in O. nubilalis. We also revealed duplication and intra-chromosomal translocation of the GOBP1 gene in P. xylostella by fluorescence in situ hybridization (FISH) analysis. Additionally, we compared the structure of the GOBP/PBP gene complex of seventeen species covering six superfamilies and twelve families of the lepidopteran clade, Ditrysia, and found the gene order was basically conserved despite the frequent occurrence of lineage-specific gains, losses, inversions and translocations of these genes, compared with their neighboring genes. Our findings support the hypothesis that the structure of the GOBP/PBP gene complex was already established in the common ancestor of Ditrysia.

Identification and field evaluation of a new blend of the sex pheromone of Hypsipyla grandella

Abstract: The objective of this work was to identify and carry out a field testing of the sex pheromone composition of the mahogany shoot borer, Hypsipyla grandella, from a population in Southern Brazil. Abdominal glands of H. grandella females were excised and extracted using n-hexane; the gland extracts were analysed by gas chromatography (GC-FID and GC-MS). Wind-tunnel and electrophysiology experiments were conducted to evaluate the role of gland compounds in the mating behavior of H. grandella males. In the field, pheromone traps containing the identified pheromone were tested. In addition to the two previously identified compounds - (9Z,12E)-tetradecadien-1-ol and (9Z,12E)-tetradecadienyl acetate -, in insects from Central America populations, two additional compounds were identified in the blend: (9Z)-tetradecen-1-ol and (9Z)-tetradecenyl acetate. Gas chromatography-electroantennographic analyses showed that these four components elicited antennal responses from conspecific males. Wind tunnel bioassays using different amounts of the components Z9-14:OH, Z9,E12-14:OH, Z9-14:OAc, and Z9,E12-14:OAc, elicited male responses similar to the response to conspecific calling females. When the binary and quaternary mixtures were tested in field conditions, males were attracted to the traps containing the quaternary mixture. The new pheromone blend identified in H. grandella males has a great potential to be exploited as a monitoring tool or control measure in the field.

Female sex pheromone of a nettle caterpillar,Monema flavescens, in China

Monema flavescens Walker (Lepidoptera: Limacodidae) is a multivoltine, generalist moth whose larvae cause serious damage to many types of trees. Pheromone lures prepared according to a study of a Japanese population were found to be ineffective at attracting M. flavescens nettle caterpillars in China, and some studies have shown intraspecific geographical differences in the composition of sex pheromones. We therefore reexamined the sex pheromone composition of M. flavescens in a Chinese population. In this study, the electroantennographically (EAG) active compounds in an extract from Chinese virgin females of M. flavescens were identified as (E)-8-decen-1-ol (E8-10:OH), (Z)-7,9-decadien-1-ol (Z7,9-10:OH), (Z)-9,11-dodecadien-1-ol (Z9,11-12:OH), and (Z)-9,11-dodecadienal (Z9,11-12:Ald) via coupled gas chromatographic-electroantennographic detection (GC-EAD) and coupled GC-mass spectrometry (MS). Pheromone dimorphism might occur in this species, as this mixture of compounds in Chinese females was different from that of E8-10:OH and E7,9-10:OH extracted from Japanese females in previous research. In wind tunnel and field tests, the males were significantly attracted to a blend of the pheromone components E8-10:OH, Z7,9-10:OH, and Z9,11-12:OH in a 100:5:4 ratio. The addition of Z9,11-12:Ald did not change the male response. The optimized three-component lure blend may provide a useful tool for monitoring and controlling Chinese populations of M. flavescens.

Disposable Polydimethylsiloxane (PDMS)-Coated Fused Silica Optical Fibers for Sampling Pheromones of Moths.

In the past decades, the sex pheromone composition in female moths has been analyzed by different methods, ranging from volatile collections to gland extractions, which all have some disadvantage: volatile collections can generally only be conducted on (small) groups of females to detect the minor pheromone compounds, whereas gland extractions are destructive. Direct-contact SPME overcomes some of these disadvantages, but is expensive, the SPME fiber coating can be damaged due to repeated usage, and samples need to be analyzed relatively quickly after sampling. In this study, we assessed the suitability of cheap and disposable fused silica optical fibers coated with 100 μm polydimethylsiloxane (PDMS) by sampling the pheromone of two noctuid moths, Heliothis virescens and Heliothis subflexa. By rubbing the disposable PDMS fibers over the pheromone glands of females that had called for at least 15 minutes and subsequently extracting the PDMS fibers in hexane, we collected all known pheromone compounds, and we found a strong positive correlation for most pheromone compounds between the disposable PDMS fiber rubs and the corresponding gland extracts of the same females. When comparing this method to volatile collections and the corresponding gland extracts, we generally found comparable percentages between the three techniques, with some differences that likely stem from the chemical properties of the individual pheromone compounds. Hexane extraction of cheap, disposable, PDMS coated fused silica optical fibers allows for sampling large quantities of individual females in a short time, eliminates the need for immediate sample analysis, and enables to use the same sample for multiple chemical analyses.

Differentiation in putative male sex pheromone components across and within populations of the African butterfly Bicyclus anynana as a potential driver of reproductive isolation.

Sexual traits are often the most divergent characters among closely related species, suggesting an important role of sexual traits in speciation. However, to prove this, we need to show that sexual trait differences accumulate before or during the speciation process, rather than being a consequence of it. Here, we contrast patterns of divergence among putative male sex pheromone (pMSP) composition and the genetic structure inferred from variation in the mitochondrial cytochrome oxidase 1 and nuclear CAD loci in the African butterfly Bicyclus anynana (Butler, 1879) to determine whether the evolution of “pheromonal dialects” occurs before or after the differentiation process. We observed differences in abundance of some shared pMSP components as well as differences in the composition of the pMSP among B. anynana populations. In addition, B. anynana individuals from Kenya displayed differences in the pMSP composition within a single population that appeared not associated with genetic differences. These differences in pMSP composition both between and within B. anynana populations were as large as those found between different Bicyclus species. Our results suggest that “pheromonal dialects” evolved within and among populations of B. anynana and may therefore act as precursors of an ongoing speciation process.

Molecular identification of closely related mirine plant bugs, Apolygus spinolae and A. lucorum (Heteroptera: Miridae) by PCR-RFLP markers

The plant bugs Apolygus spinolae (Meyer-Dür) and Apolygus lucorum (Meyer-Dür) (Heteroptera: Miridae) are well-known as economically important pests of various agricultural crops in quite a few parts of the Palearctic Region. These two species are difficult to distinguish from one another due to their similar appearances. Earlier studies revealed that the two species can be easily separated by species-specific sex pheromone composition. Segments of two mitochondrial genes, large ribosomal RNA subunit (16S) and cytochrome c oxidase subunit I (COI), were compared between A. spinolae and A. lucorum female adults that had been identified by analyzing the pheromone composition. Based on the DNA sequence differences between the two species, molecular diagnostic markers using PCR and restriction fragment length polymorphism (RFLP) were developed. This rapid and accurate method was successfully applied to verify the species identification of male adults caught by pheromone traps and of nymphs collected from various host plants. These molecular markers and diagnostic methods could provide a useful tool for the identification of these sibling species and further contribute to the development of rational pest management strategies.

Sex Pheromone of the Rare Click Beetle Betarmon bisbimaculatus.

The click beetle Betarmon bisbimaculatus (Fabricius, 1803) (Coleoptera: Elateridae) has a scattered distribution throughout a large area in Europe and the near East. Due to its scarcity, little is known about the ecology, biology, and development of this peculiar species. Here, we studied the composition of the female-released sex pheromone of B. bisbimaculatus. Neryl hexanoate, neryl octanoate, and neryl decanoate, in a ratio of approximately 3:1:6, were the only volatile compounds present in the extracts of pheromone glands. A synthetic mixture of all three compounds in the natural ratio was highly attractive to males in field traps. When the compounds were tested individually, only traps baited with neryl hexanoate were attractive, but they caught only a sixth of the males compared to the mixture. Based on the similarity of their sex pheromones, we propose that the tribe Pomachiliini with B. bisbimaculatus is closely related to the tribe Agriotini. This study shows the potential of sex pheromone studies for monitoring of rare and threatened insects as well as for elucidating phylogenetic relationships.

Young male mating success is associated with sperm number but not with male sex pheromone titres.

BackgroundIntraspecific communication is of crucial importance throughout the animal kingdom and may involve a combination of visual, gustatory, olfactory and acoustic cues. Variation in male sex pheromone amount and composition may convey important information to female conspecifics, for instance on species identity or age. However, whether increased male pheromone titres are associated with fitness benefits for the female, thus indicating a role as an honest signal, is under debate.ResultsAgainst this background, we tested in the butterfly Bicyclus anynana (1) whether young males being successful or unsuccessful in gaining a mating differed in sex pheromone titres and (2) for associations between male pheromone titres and spermatophore mass, eupyrene sperm number, and a variety of female and offspring life-history traits. Successful and unsuccessful males did not differ in pheromone titres, however eupyrene sperm number was much higher in successful males. Pheromone titres were not associated with any fitness-related female or offspring trait measured in our study, though correlation analyses yielded evidence for trade-offs among specific traits. Patterns did not differ among control and olfaction-blocked females.ConclusionTherefore, we suggest that in young B. anynana pheromone titres do not indicate male quality.