Male-specific Compounds Research Articles

New Insights into the Composition of Aggregation Pheromones in Polygraphus poligraphus, Polygraphus punctifrons, Polygraphus subopacus and Polygraphus proximus

Four-eyed bark beetles of the genus Polygraphus have been involved in large bark beetle outbreaks in different parts of the world, resulting in major economic losses. A striking example is the invasive species Polygraphus proximus which is a pest on Abies sibirica in Russia. In Sweden, Polygraphus poligraphus has been involved in bark beetle outbreaks on Norway spruce, Picea abies, together with the European spruce bark beetle Ips typographus. Two related species, Polygraphus punctifrons and Polygraphus subopacus are also present in Sweden. Recently, aggregation pheromones or pheromone components have been identified for these four Polygraphus species. However, questions remain regarding the complete composition of their pheromones, particularly for P. subopacus and P. proximus, whose aggregation pheromones appear to be very similar. In an attempt to better understand the chemical communication of these species, additional studies were conducted on P. poligraphus, P. punctifrons and P. subopacus using solid phase microextraction coupled with gas chromatography and mass spectrometry (SPME–GC–MS), electroantennography (EAG) as well as SPME–GC–MS and GC–MS with electroantennographic detection (EAD). Field experiments were also conducted. In P. punctifrons, some male-specific compounds were found in addition to those previously identified. In EAG and SPME–GC–MS/EAD studies, all three Polygraphus species responded strongly to grandisol. Using a chiral column, GC–MS/EAD revealed that they were able to detect both enantiomers of grandisol. In summary, this work presents our current understanding of the aggregation pheromones in four Polygraphus species and the challenges we have met in identifying species-specific pheromone blends for some of these species.

Shared Pheromone Compounds in Neotropical Rice Stink Bugs: The Role of Zingiberenol and Sesquipiperitol.

The small black stem bug, Paratibraca (= Glyphepomis) spinosa (Campos and Grazia 1998), is a rice pest in Brazil and is part of a complex of stink bugs that includes Oebalus poecilus (Dallas) and Tibraca limbativentris Stål. Together, these pentatomid species pose a serious threat to rice crops throughout South America. In this study, we identified the sex pheromone of P. spinosa. Our findings revealed that male P. spinosa produces four male-specific compounds identical to those produced by T. limbativentris males. These include (3S,6S,7R)-1,10-bisaboladien-3-ol and (3R,6S,7R)-1,10-bisaboladien-3-ol (zingiberenols), both sharing the same absolute configuration as the 1,10-bisaboladien-3-ol from T. limbativentris; along with two minor male-specific components: the cis and trans isomers of 2,10-bisaboladien-1-ol (sesquipiperitol). Y-tube olfactometer bioassays demonstrated that P. spinosa females were attracted to the odours of live males, whereas males showed no attraction to either sex. Females were particularly drawn to male volatiles, especially the fraction containing the compounds 1,10-bisaboladien-3-ol and 2,10-bisaboladien-1-ol. When tested with synthetic compounds, females were attracted to a blend of the two isomers, (3S,6S,7R)-1,10-bisaboladien-3-ol and (3R,6S,7R)-1,10-bisaboladien-3-ol, in the same ratio as naturally produced by males. Given that the sex pheromone compounds identified in P. spinosa are identical to those of T. limbativentris, we conducted bioassays with live heterospecific insects. In olfactometer bioassays, the females of both species were attracted by the odours of heteroespecific males.

Identification and field verification of the aggregation pheromone components produced by male Holotrichia parallela Motschulsky (Coleoptera: Scarabaeidae).

The chafer beetle, Holotrichia parallela, causes damage to numerous economically significant crops worldwide. Adult beetles exhibit aggregation behavior likely mediated by a male-produced pheromone. Advancements in biological research technology have facilitated the identification of insect aggregation pheromones and promoted their applications as bait for trapping and monitoring pests. Currently, only a few active components of aggregation pheromones from Holotrichia species have been identified. However, the specific components of aggregation pheromones produced by H. parallela remain unknown. In this study, we initially observed from Y-tube olfactometer assays that both male and female H. parallela were significantly attracted to volatiles emitted by males, but not to those from females. We then collected hindgut crude extracts of male adults and carried out gas chromatography-mass spectrometry analysis to identify potential aggregation pheromone components. Pentadecyl acetate, cis-13-docosenol, and behenic acid were identified as male-specific compounds in comparison to female extracts, serving as components of the aggregation pheromone in H. parallela. We further evaluated their attractiveness to H. parallea in both laboratory and field experiments. In laboratory settings, pentadecyl acetate, cis-13-docosenol, and behenic acid evoked significant responses to both males and females at specific concentrations, as evidenced by both electroantennography tests and behavioral bioassays. Under field conditions, traps baited with these three compounds captured significantly more H. parallela adults compared to control traps. In this study, we found that pentadecyl acetate, cis-13-docosenol, and behenic acid are specifically present in male H. parallela, serving as aggregation pheromones. Both laboratory and field-trapping experiments suggest their potential as monitoring and controlling tools against H. parallela adults. © 2024 Society of Chemical Industry.

Confirmation that Monochamol is a Male Produced Aggregation-Sex Pheromone for Monochamus maculosus Haldeman (Coleoptera: Cerambycidae).

The recognition of cerambycids as frequent and damaging invaders led to an increase in the interest in the chemical ecology of the group with the identification of pheromones and pheromone-like attractants for well over 100 species. Pheromone components of the Cerambycidae are often phylogenetically conserved, with a single compound serving as a pheromone component for several related species. In the subfamily Lamiinae, the compound 2-(undecyloxy)ethanol (monochamol) has been identified as an aggregation-sex pheromone for several species of the genus Monochamus. In other species, including Monochamus maculosus Haldeman, field trials have demonstrated that monochamol is a pheromone attractant, but at that point it was still unknown as to whether it was a pheromone for this species. Here we report the identification, and laboratory and field trials of a pheromone component produced by adult male M. maculosus. Chemical analyses of headspace volatile collections sampled from field collected beetles of both sexes revealed the presence of one male-specific compound that was identified as 2-(undecyloxy)ethanol. Electroantennography analyses showed that monochamol elicited responses from the antennae of female beetles. Traps baited with monochamol in the field captured M. maculosus adults of both sexes corroborating the identification of monochamol as the sex-aggregation pheromone of this species. The attractivity of monochamol to adult M. maculosus in our field trapping experiment was synergized by the addition of the host volatile α-pinene.

Identification of a Male-Produced Aggregation Sex Pheromone in Rosalia batesi, an Endemic Japanese Longhorn Beetle.

The longhorned beetle Rosalia batesi Harold (Coleoptera; Cerambycidae) is endemic to Japan, where its range extends from Hokkaido to Kyushu. The colorful adults are well-known to entomologists and collectors worldwide. It is a hardwood-boring species with larvae that develop in dead broad-leaf trees. In laboratory bioassays, females were attracted to males, which suggested that males produce a sex pheromone. The congeneric species R. alpina is native to Europe, and another congener, R. funebris, is distributed in North America. The pheromone components produced by males of these species had been previously identified as two compounds from different biosynthetic pathways. In the present study, volatiles were collected from beetles of both sexes, and the analyses of the resulting extracts revealed a single male-specific compound, which was identified as 3,5-dimethyl-6-(1-methylbutyl)-pyran-2-one; this is the same compound as the pheromone of the European R. alpina. This alkylated pyrone structure is, so far, unique among known cerambycid pheromones. In field bioassays with traps baited with the racemic synthetic pheromone, significant numbers of both sexes of R. batesi were attracted in an approximately equal ratio, indicating that the compound is an aggregation-sex pheromone rather than a sex pheromone.

Laboratory Evidence of 2-Isobutyl-3-methoxypyrazine as a Male-Released Aggregative Cue in Labidostomis lusitanica (Germar) (Coleoptera: Chrysomelidae).

In spite of its incidence on pistachio trees, the chemical ecology of Labidostomis lusitanica (Germar) (Coleoptera: Chrysomelidae) has been neglected so far. In this work, we provide the first evidence of a biologically active male-specific compound that may be promoting field aggregation. Headspace collections through solid-phase microextraction from feral males and females reported the presence of 2-isobutyl-3-methoxypyrazine exclusively in males. Electroantennographic recordings revealed that males and females responded in a dose-dependent manner to increasing stimuli of 2-isobutyl-3-methoxypyrazine, with females overall displaying a higher response than males. In dual-choice tests, both males and females showed a significant preference for the compound in comparison to a pure air stimulus. In light of these results, the possible role of 2-isobutyl-3-methoxypyrazine as an aggregation cue in L. lusitanica is discussed.

Identification of sex-specific compounds in the invasive four-eyed fir bark beetle Polygraphus proximus

Polygraphus proximus, a four-eyed fir bark beetle, is an invasive bark beetle species which has caused extensive damage to forests of Abies sibirica in southern and western Siberia and to Abies species in the European part of Russia. There is a high risk that the pest insect will spread to areas where it is currently not considered present, such as the European Union. In these areas, it threatens to attack conifer forests of various species which may result in major environmental and economic impact. The aim of this study was to identify pheromone components of P. proximus that can be used as pheromone baits. Males and females of P. proximus were allowed to bore into the bark of stem sections of Abies sibirica at the laboratory, and volatiles were collected with solid-phase microextraction (SPME). Analyses of these extracts with gas chromatography-mass spectrometry (GC–MS) revealed several sex-specific compounds. In total, twelve male-specific compounds and one female-specific compound were identified. The major male-specific compound determined by GC peak area was (Z)‐2‐(3,3‐dimethylcyclohexylidene)‐ethanol [(Z)-DMCHE] and the minor male-specific compounds were 3-methyl-3-buten-1-ol, 3-methyl-2-buten-1-ol, 3-methyl-2-butenal, benzyl alcohol, fragranol, 7-methyl-3-methylene-6-octen-1-ol, (Z)- and (E)-2-(3,3-dimethylcyclohexylidene)-acetaldehyde, geraniol, geranial and papayanol. The only female-specific compound was identified as 1-hexanol. Two of the male-specific compounds, (Z)‐DMCHE and 3-methyl-2-buten-1-ol were shown to attract males and females of P. proximus in field studies. Thus, we now for the first time can present the structures of two male-specific components that are biologically active parts of P. proximus aggregation pheromone. However, some chemical communication overlap between P. proximus and P. subopacus needs to be further investigated as (Z)‐DMCHE also attracted males and females of P. subopacus.

Identification of Components of the Aggregation Pheromone of the Guam Strain of Coconut Rhinoceros Beetle, Oryctes rhinoceros, and Determination of Stereochemistry

The coconut rhinoceros beetle, Oryctes rhinoceros (Linnaeus 1758) (Coleoptera: Scarabaeidae: Dynastinae) (CRB), is endemic to tropical Asia where it damages both coconut and oil palm. A new invasion by CRB occurred on Guam in 2007 and eradication attempts failed using commonly applied Oryctes rhinoceros nudivirus (OrNV) isolates. This and subsequent invasive outbreaks were found to have been caused by a previously unrecognized haplotype, CRB-G, which appeared to be tolerant to OrNV. The male-produced aggregation pheromone of the endemic, susceptible strain of O. rhinoceros (CRB-S) was previously identified as ethyl 4-methyloctanoate. Following reports from growers that commercial lures containing this compound were not attractive to CRB-G, the aim of this work was to identify the pheromone of CRB-G. Initial collections of volatiles from virgin male and female CRB-G adults from the Solomon Islands failed to show any male- or female-specific compounds as candidate pheromone components. Only after five months were significant quantities of ethyl 4-methyloctanoate and 4-methyloctanoic acid produced by males but not by females. No other male-specific compounds could be detected, in particular methyl 4-methyloctanoate, 4-methyl-1-octanol, or 4-methyl-1-octyl acetate, compounds identified in volatiles from some other species of Oryctes. Ethyl 4-methyloctanoate elicited a strong electroantennogram response from both male and female CRB-G, but these other compounds, including 4-methyloctanoic acid, did not. The enantiomers of ethyl 4-methyloctanoate and 4-methyloctanoic acid were conveniently prepared by enzymatic resolution of the commercially-available acid, and the enantiomers of the acid, but not the ester, could be separated by gas chromatography on an enantioselective cyclodextrin phase. Using this approach, both ethyl 4-methyloctanoate and 4-methyloctanoic acid produced by male CRB-G were shown to be exclusively the (R)-enantiomers whereas previous reports had suggested male O. rhinoceros produced the (S)-enantiomers. However, re-examination of the ester and acid produced by male CRB-S from Papua New Guinea showed that these were also the (R)-enantiomers. In field trapping experiments carried out in the Solomon Islands, both racemic and ethyl (R)-4-methyloctanoate were highly attractive to both male and female CRB-G beetles. The (S)-enantiomer and the corresponding acids were only weakly attractive. The addition of racemic 4-methyloctanoic acid to ethyl 4-methyloctanoate did significantly increase attractiveness, but the addition of (R)- or (S)-4-methyloctanoic acid to the corresponding ethyl esters did not. Possible reasons for the difference in assignment of configuration of the components of the CRB pheromone are discussed along with the practical implications of these results.

Identification of male produced compounds in the bark beetle Polygraphus subopacus and establishment of (Z)-2-(3,3-dimethylcyclohexylidene)-ethanol as an aggregation pheromone component

Bark beetles of the genus Polygraphus have recently been involved in large bark beetle outbreaks in central Sweden, together with the European spruce bark beetle Ips typographus. Three species of Polygraphus can be found in this region; Polygraphus poligraphus, Polygraphus punctifrons and Polygraphus subopacus. Efficient pheromone traps would facilitate further investigations of these species and their role in bark beetle outbreaks. Pheromone compounds have previously been identified in P. poligraphus and P. punctifrons, but not in P. subopacus. Thus, we allowed males and females of P. subopacus to bore in the bark of stem sections of Norway spruce (Picea abies) in the laboratory. Volatile organic compounds from boring insects were sampled with SPME and analysed with GC–MS and several male-specific compounds were observed. The male specific compounds were 3-methyl-3-buten-1-ol, 3-methyl-2-buten-1-ol, 3-methyl-2-butenal, grandisol, fragranol, (Z)-2-(3,3-dimethylcyclohexylidene)-ethanol, (E)-2-(3,3-dimethylcyclohexylidene)-ethanol, (Z)-2-(3,3-dimethylcyclohexylidene)-acetaldehyde, (E)-2-(3,3-dimethylcyclohexylidene)-acetaldehyde, geranial and γ-isogeraniol. (Z)-2-(3,3-dimethylcyclohexylidene)-ethanol, [(Z)-DMCHE], was identified from GC–MS analysis to be the major male-specific compound while the (E)-isomer, [(E)-DMCHE], was found as a minor compound. These two compounds gave positive responses in EAG analyses with antennae from males and females of P. subopacus. Thus, (Z)- and (E)-DMCHE were used in a field experiment in central Sweden but only (Z)-DMCHE was found to be attractive to males and females of P. subopacus. Consequently, (Z)-DMCHE was established to be a component of P. subopacus aggregation pheromone.

Large-scale characterization of sex pheromone communication systems in Drosophila

Insects use sex pheromones as a reproductive isolating mechanism to attract conspecifics and repel heterospecifics. Despite the profound knowledge of sex pheromones, little is known about the coevolutionary mechanisms and constraints on their production and detection. Using whole-genome sequences to infer the kinship among 99 drosophilids, we investigate how phylogenetic and chemical traits have interacted at a wide evolutionary timescale. Through a series of chemical syntheses and electrophysiological recordings, we identify 52 sex-specific compounds, many of which are detected via olfaction. Behavioral analyses reveal that many of the 43 male-specific compounds are transferred to the female during copulation and mediate female receptivity and/or male courtship inhibition. Measurement of phylogenetic signals demonstrates that sex pheromones and their cognate olfactory channels evolve rapidly and independently over evolutionary time to guarantee efficient intra- and inter-specific communication systems. Our results show how sexual isolation barriers between species can be reinforced by species-specific olfactory signals.

Cervidins A-D: Novel Glycine Conjugated Fatty Acids from the Tarsal Gland of Male Whitetail Deer, Odocoileus virginianus.

Sexually mature male deer are known to rub-urinate, a process where urine is deposited on the tarsal gland. The resulting mixture of compounds from urine and secretions from the tarsal gland are used to signal sex, age, maturation status, and other information at close distance. We examined the difference in metabolites of tarsal gland extracts from male and female whitetail deer, Odocoileus virginianus, harvested during the mating season. Using NMR spectroscopy and high-pressure liquid chromatography linked to high resolution mass spectrometry (HPLC/HR-MS) we identified a homologous series of four male-specific compounds. The compounds are novel glycine conjugates of 10-hydroxy-6,9-oxido fatty acids, which we term cervidins A-D. Cervidins were deemed to possess the absolute configuration 6S,9R,10R through comparison of their spectroscopic data with those of known compounds. In addition, cholesterol 3-sulfate and 3-(3-hydroxyphenyl)-propanoic acid were found to be present in the extracts. Our results clearly demonstrate the diversity of potential semiochemicals contained in the mammalian integument.

Octadecanal as the Male-Produced Aggregation Pheromone of the Coconut Weevil, Amerrhinus ynca (Coleoptera: Curculionidae)

The beetle Amerrhinus ynca Sahlberg, 1823 (Coleoptera: Curculionidae) causes damage to several species of the Arecaceae family. The larvae of this palm weevil damage palm trees by causing perforations in the leaf rachis, obstructing the flow of nutrients to the plant. To date, are no registered insecticides for the control of A. ynca. An efficient control strategy would be the use of synthetic pheromones of this beetle for integrated pest management tactics. In this work, the aggregation pheromone of A. ynca was identified from adults collected as larvae and pupae from the rachis of coconut palm. The volatile compounds produced by A. ynca were collected via headspace aeration of adult beetles, and the identification of the pheromone was performed with gas chromatography coupled to mass spectrometry (GC-MS), GC-Fourier transform infrared spectroscopy (GC-FTIR) and GC-electroantenographic detector (GC-EAD). Dual-choice olfactometry experiments were performed to evaluate the bioactivity of the synthetic pheromone. GC analyses showed the presence of a male-specific compound that was identified as octadecanal. In GC-EAD analyses, synthetic octadecanal elicited responses of A. ynca female antennae. Behavioral bioassays showed attractiveness of synthetic compound for males and females, indicating octadecanal as the aggregation pheromone of A. ynca.

Identification of Zingiberenol and Murgantiol as Components of the Aggregation-Sex Pheromone of the Rice Stink Bug, Mormidea v-luteum (Heteroptera: Pentatomidae).

Mormidea v-luteum (Lichtenstein, 1796) feeds on commercial crops, such as rice and ryegrass, causing damage that slows growth and reduces productivity. With the aim of developing an eco-friendly control technique, we investigated the compounds involved in chemical communication in this species. The volatiles produced by a group of seven males or females allocated to different aerated glass chambers were collected for 24h and analyzed by gas chromatograph/mass spectrometry and gas chromatography/Fourier transform infrared spectroscopy. The analyses showed six male-specific compounds, identified as two isomers of zingiberenol (compounds 1 and 2), three isomers of murgantiol (3, 5 and 6) and sesquipiperitol (4). Compounds 1 and 5 were the major components and were produced in a ratio of 6:4. The absolute stereochemistry of the two isomers of zingiberenol was established as (1S,4R,1'S)-1 and (1R,4R,1'S)-2 by chiral gas chromatography. Stereochemistry was not determined for all the other molecules. To confirm the attractiveness of these chemicals, bioassays were performed in a Y-tube olfactometer, first using crude extracts and, subsequently, synthetic compounds. Male volatiles were attractive to both sexes, demonstrating an aggregation pheromone. In bioassays with synthetic compounds, (1'S)-zingiberenol was highly attractive to both males and females. However, when (1'S)-murgantiol was tested, only females were attracted. Interestingly, when a mixture of zingiberenol and murgantiol isomers was tested, it was attractive to both sexes, with females more attracted to the mixture than to zingiberenol alone; males did not distinguish between treatments. Thus, the bioassay data suggest that the molecules have different functions in chemical communication of this species: zingiberenol acts primarily as an aggregation pheromone, while murgantiol plays a role as a sex pheromone.

Camphor and Borneol as the Male-Produced Sex Pheromone of the Shield Bug, Orsilochides leucoptera (Hemiptera: Scutelleridae).

The ability of stink bugs to release high amounts of strong-smelling and irritating defensive compounds is related to their metathoracic gland (MTG), which is an exocrine gland with defensive, sexual, alarm and aggregation signal functions. Orsilochides leucoptera (Scutelleridae) is a widespread species in the Neotropical region that feeds on plants of the families Malvaceae, Poaceae and Euphorbiaceae. A series of compounds (ketones, alcohols and esters) have been identified in the MTGs among the three species of Scutelleridae whose MTG secretions have been investigated thus far; however, no sex pheromone compounds have been described for any scutellerid species. The aim of this work was to study sex pheromone communication within this family of stink bugs, and identify the compounds present in the MTG of O. leucoptera. Analysis by gas chromatography/mass spectrometry (GC/MS) revealed two male specific compounds identified as (R)-camphor (1) and (R)-borneol (2), which were attractive to females in Y-tube olfactometer bioassays. Also, GC/MS analysis of secretions from MTG of males and females of O. leucoptera, identified eight additional compounds: 2-(E)-hexenal (3), (E)-4-oxo-2-hexenal (4), (E)-hex-2-enyl acetate (5), (R)-linalool (6), (R)-α-terpineol (7), dodecane (8), 1-tridecene (9) and n-tridecane (10). From these, (R)-α-terpineol was detected only in the gland of males, and is probably a biosynthetic intermediate of the pheromone components. Most of the MTG compounds identified in O. leucoptera have been identified in other heteropteran species. Camphor is often a toxic and repellent compound for insects. However, we report it, for the first time, as a sex pheromone component of an insect.

Morphological, chemical and electrophysiological investigations of Telchin licus (Lepidoptera: Castniidae).

The giant sugarcane borer Telchin licus (Drury, 1773) (Lepidoptera: Castniidae) is a day-flying moth pest of sugarcane, pineapples and bananas. To better understand the chemical communication in this species, we examined the morphology of its olfactory system and the chemical composition of its body parts. The ventral surface of the clubbed antennae of T. licus has six morphological types of sensilla: sensilla trichodea, basiconica, chaetica, squamiforma, coeloconica, and auricillica. The telescopic ovipositor shows no evidence of a sexual gland, or female-specific compounds. On the other hand, the midleg basitarsus of males releases (E,Z)-2,13-octadecadienol and (Z,E)-2,13-octadecadienol, which are electroantennographically active in both sexes. These compounds are known female sex pheromones in the Sesiidae family and are male-specific compounds in another castniid moth, although further investigations are necessary to elucidate their ecological role in the Castniidae family.

A male-produced aggregation-sex pheromone of the beetle Arhopalus rusticus (Coleoptera: Cerambycidae, Spondylinae) may be useful in managing this invasive species

The longhorned beetle Arhopalus rusticus (Coleoptera: Cerambycidae, Spondylinae) is a common species in conifer forests of the Northern Hemisphere, but with global trade, it has invaded and become established in New Zealand, Australia, and South America. Arhopalus rusticus is a suspected vector of the phytopathogenic nematode, Bursaphelenchus xylophilus, the causative agent of pine wilt disease, which is a major threat to pine forests worldwide. Here, we report the identification of a volatile, male-produced aggregation-sex pheromone for this species. Headspace odours from males contained a major male-specific compound, identified as (2 S, 5E)-6,10-dimethyl-5,9-undecadien-2-ol (common name (S)-fuscumol), and a minor component (E)-6,10-dimethyl-5,9-undecadien-2-one (geranylacetone). Both compounds are known pheromone components for species in the same subfamily. In field trials in its native range in Slovenia, (S)-fuscumol was significantly more attractive to beetles of both sexes, than racemic fuscumol and a blend of host plant volatiles commonly used as an attractant for this species. Fuscumol-baited traps also caught significant numbers of another spondylidine species, Spondylis buprestoides (L.), and a rare click beetle, Stenagostus rufus (De Geer). The pheromone can be exploited as a cost-effective and environmentally safe tool for detection and monitoring of this invasive species at ports of entry, and for monitoring the beetle’s distribution and population trends in both endemic and invasive populations.

Identification of Aggregation-Sex Pheromone Components for a "Living Fossil", the False Click Beetle, Palaeoxenus dohrni Horn (Coleoptera: Eucnemidae).

Insect pheromones have rarely been exploited in surveys or studies of rare and endangered species, despite their potential as effective and highly selective attractants for target species. Here, we report the identification, synthesis, and field bioassays of a male-produced aggregation-sex pheromone blend of a rare false click beetle species endemic to southern California, Dohrn's elegant eucnemid beetle, Palaeoxenus dohrni Horn (Coleoptera: Eucnemidae). This species is the only extant species in its genus and subfamily. Analyses of extracts of headspace volatiles collected from adult beetles revealed several male-specific compounds. Two of these compounds, identified as (E)-2-nonen-4-one and (R)-2-nonanol, elicited electroantennographic responses from antennae of beetles of both sexes. In field bioassays, a blend of the two compounds attracted both sexes, whereas the individual compounds were not attractive. The identification of an attractant pheromone should provide a useful tool for bioconservation and ecological studies of this iconic species.

Isophorone derivatives as a new structural motif of aggregation pheromones in Curculionidae

The beetle Homalinotus depressus (Coleoptera: Curculionidae) is a major pest of coconuts in the Northern region of Brazil, for which environmentally friendly methods of control are desired. Behavioral responses of H. depressus to airborne volatile extracts from conspecifics suggested the presence of a male-produced aggregation pheromone. GC analyses of these extracts showed the presence of four male-specific compounds. Analytical data in combination with the synthesis of standards led to the identification of the male-released semiochemicals as epoxyisophorone (1), isophorone (2), homalinol (3), and 2-hydroxyisophorone (4), of which (3) was the major constituent. The configuration of homalinol was determined to be cis on the basis of retention times of synthetic cis and trans synthetic standards. Enantiomers of cis-homalinol were obtained in high enantiomeric excess by using biocatalysis. Their separation on a GC enantioselective column (β-Dex325®), allowed us to unambiguously determine that the absolute configuration of natural homalinol was (1R,2R,6S). Field bioassays demonstrated that both the synthetic major compound per se and mixtures of all four male-specific compounds were attractive to H. depressus.

Aggregation Pheromone of the Bearded Weevil, Rhinostomus barbirostris (Coleoptera: Curculionidae): Identification, Synthesis, Absolute Configuration and Bioactivity.

The bearded weevil, Rhinostomus barbirostris (Coleoptera: Curculionidae: Dryophthorinae), attacks coconut trees, oil palms and other species of Arecaceae. Besides direct damage, R. barbirostris may be a vector of diseases in coconut and oil palms, such as stem bleeding (resinosis) and red ring disease. Currently, the only method to control this weevil is by visual observation of damage and removal of infected plants. Semiochemical-based trapping could improve the effectiveness of monitoring and management of R. barbirostris. In comparisons of volatiles released by R. barbirostris males and females by gas chromatography (GC) two male-specific compounds were observed. GC-mass spectrometry (MS) and GC-Fourier transform-infrared (FTIR) analyses of the natural compounds suggested these were diastereoisomers of 5-hydroxy-4-methylheptan-3-one, also known as sitophilure, a pheromone component of other dryophthorine species. Synthesis of the mixture of all four stereoisomers of sitophilure was performed in two steps, and the chemical structures were confirmed by comparing GC retention times and MS and FTIR spectra of natural and synthetic compounds. The absolute configurations of the two male-specific compounds were elucidated by enantioselective GC; the major component was the (4S,5R)-isomer, and the minor component (4S,5S)-sitophilure. In analyses by GC-electroantennography (EAG) the antennae of male and female R. barbirostris only responded to the (4S,5R)-isomer of the synthetic sitophilure. The stereoisomeric mixture of sitophilure was attractive to both sexes of R. barbirostris in laboratory experiments in the presence of sugar cane volatiles, and a similar result was obtained in a preliminary field trapping test.

Two Sex-Specific Volatile Compounds Have Sex-Specific Repulsion Effects on Adult Locusts Locusta migratoria manilensis (Meyen) (Orthoptera: Acrididae)

Gregarious locusts use volatile semiochemicals released from their bodies and feces to regulate their biological characteristics, and many of these compounds are sex-specific. The locust Locusta migratoria manilensis (Meyen) (Orthoptera: Acrididae) is a frequent plague species in China, but very little is known of its olfactory behavioral ecology. In our previous study, one malespecific compound (2-heptanone) was identified from body volatiles of gregarious adult L. m. manilensis. In the present study, six compounds were identified from fecal volatiles of gregarious adult L. m. manilensis using a gas chromatograph coupled to a mass spectrometer: 3-Methyl-1-butanol, cyclo hexanol, 2,5-dimethylpyrazine, 6-methyl-5-hepten-2-one and 2,6,6-trimethyl-2-cyclohexene-1,4-dione were present in the fecal volatiles of both sexes, and 2,2,6-trimethylcyclohexanone only in that of females. Olfactometric bioassays indicated that the male-specific 2-heptanone repelled the adult males, but had no effect on females. On the other hand, female-specific 2,2,6-trimethylcyclohexanone was found to repel the adult females, but had no effect on males. The two sex-specific compounds may help to improve reproductive efficiency of the locusts.