Mattias C. Larsson

Green leaf volatile-detecting olfactory receptor neurones display very high sensitivity and specificity in a scarab beetle

In the Japanese scarab beetle, Phyllopertha diversa, olfactory receptor neurones specific for the detection of so‐called general green leaf volatiles (GLV) display a high specificity and sensitivity. Three main types of green‐leaf‐volatile‐detecting receptor neurones specific to (Z)‐3‐hexenyl acetate, (E)‐2‐hexenal and (Z)‐3‐hexenol, respectively, were identified. Each type responded at a very low stimulus concentration to the key stimulus, and required at least a thousand‐fold increase in concentration to respond to any of the other GLVs tested. Flower‐odour‐and pheromone‐detecting receptor neurones were also identified. Olfactory sensilla housing plant‐odour‐ or pheromone‐detecting receptor neurones displayed clear morphological differences, and were also separated into different antennal regions.

Characteristic Odor of Osmoderma eremita Identified as a Male-Released Pheromone

Osmoderma eremita (Scopoli) is an endangered scarab beetle living in hollow trees. It has mainly been known for its characteristic odor, typically described as a fruity, peachlike or plumlike aroma. The odor emanating from a single beetle can sometimes be perceived from a distance of several meters. In this paper, we show that the characteristic odor from O. eremita is caused by the compound (R)-(+)-γ-decalactone, released in large quantities mainly or exclusively by male beetles. Antennae from male and female beetles responded in a similar way to (R)-(+)-γ-decalactone in electroantennographic recordings. Field trapping experiments showed that (R)-(+)-γ-decalactone is a pheromone attracting female beetles. Lactones similar to (R)-(+)-γ-decalactone are frequently used as female-released sex pheromones by phytophagous scarabs. This is, however, the first evidence of a lactone used as a male-produced pheromone in scarab beetles. We propose that the strong signal from males is a sexually selected trait used to compete for females and matings. The signal could work within trees but also act as a guide to tree hollows, which are an essential resource for O. eremita. Males may, thus, attract females dispersing from their natal tree by advertising a suitable habitat. This signal could also be exploited by other males searching for tree hollows or for females, which would explain the catch of several males in our traps.

Olfactory receptor neurons detecting plant odours and male volatiles in Anomala cuprea beetles (Coleoptera: Scarabaeidae)

We have identified several types of olfactory receptor neurons in male and female Anomala cuprea beetles. The receptor neurons were sensitive to female sex pheromone components, flower volatiles, green leaf volatiles and unknown volatiles from males. Olfactory sensilla were located on three lamellae forming the antennal club. There was a clear spatial separation between some types of sensilla on each lamella. Receptor neurons for the two sex pheromone components were situated in sensilla placodea covering a specific area on each lamella in both males and females. All sex pheromone receptor neurons were found in these sensilla. Most other receptor neurons were located in a longitudinal, heterogeneous streak formed by various types of sensilla. Receptor neurons for plant-derived compounds appeared to be specialists with a high sensitivity to their respective key compound. The most remarkable among these are the green leaf volatile-specific receptor neurons, which were both sensitive and selective, with the key compound being at least 1000 times as effective as any other compound. These green leaf volatile detectors are apparently homologous to detectors recently found in the scarab Phyllopertha diversa. Our results emphasize the role of single-sensillum recordings as a tool in the identification of biologically active odours.

Candidate chemosensory Genes in Female Antennae of the Noctuid Moth Spodoptera littoralis

Chemical senses are crucial for all organisms to detect various environmental information. Different protein families, expressed in chemosensory organs, are involved in the detection of this information, such as odorant-binding proteins, olfactory and gustatory receptors, and ionotropic receptors. We recently reported an Expressed Sequence Tag (EST) approach on male antennae of the noctuid moth, Spodoptera littoralis, with which we could identify a large array of chemosensory genes in a species for which no genomic data are available. Here we describe a complementary EST project on female antennae in the same species. 18,342 ESTs were sequenced and their assembly with our previous male ESTs led to a total of 13,685 unigenes, greatly improving our description of the S. littoralis antennal transcriptome. Gene ontology comparison between male and female data suggested a similar complexity of antennae of both sexes. Focusing on chemosensation, we identified 26 odorant-binding proteins, 36 olfactory and 5 gustatory receptors, expressed in the antennae of S. littoralis. One of the newly identified gustatory receptors appeared as female-enriched. Together with its atypical tissue-distribution, this suggests a role in oviposition. The compilation of male and female antennal ESTs represents a valuable resource for exploring the mechanisms of olfaction in S. littoralis.

Specificity and redundancy in the olfactory system of the bark beetle Ips typographus: single-cell responses to ecologically relevant odors.

We screened 150 olfactory sensilla in single-sensillum recordings to unravel the mechanisms underlying host selection in the spruce bark beetle, Ips typographus (Coleoptera: Curculionidae: Scolytinae). Odor stimuli comprised of pheromone (various bark beetle spp.), host, and non-host compounds elicited strong and selective responses from 106 olfactory receptor neurons (ORNs), 45 of which were tuned to pheromone compounds, 37 to host compounds, and 24 to non-host volatiles (NHV). In addition, 26 ORNs responded only weakly to any odor stimulus. Strongly responding ORNs were classified into 17 classes. Seven classes responded primarily to the Ips pheromone compounds: cis-verbenol, ipsenol, ipsdienol (two classes), 2-methyl-3-buten-2-ol, amitinol, or verbenone, respectively. Six classes responded to the host compounds: alpha-pinene, myrcene, p-cymene, myrcene and p-cymene, 1,8-cineole, or Delta3-carene, respectively. Four classes responded to NHV: 3-octanol, 1-octen-3-ol, trans-conophthorin, or indiscriminately to the repellent green leaf volatiles (GLVs) 1-hexanol, Z3-hexen-1-ol and E2-hexen-1-ol, respectively. Indiscriminate responses from GLV neurons might explain a behavioral redundancy among these GLVs. This is the first description of individual bark beetle ORNs dedicated to NHV perception. These comprise almost 25% of the strongly responding neurons, demonstrating that a large proportion of the olfactory system is devoted to signals from plants that the insect avoids.

Detection of fruit- and flower-emitted volatiles by olfactory receptor neurons in the polyphagous fruit chafer Pachnoda marginata (Coleoptera: Cetoniinae)

Abstract. Olfactory receptor neurons on the antennae of the African fruit chafer species Pachnoda marginata (Coleoptera: Scarabaeidae) were examined through extensive use of gas chromatography linked with electrophysiological recordings from single olfactory receptor neurons. Contacted neurons were stimulated with a large number of extracted volatiles from 22 different fruits and with 64 synthetic plant compounds. Extracted fruit volatiles were identified using linked gas chromatography-mass spectrometry. In total, 48 different odor compounds were found to elicit responses. Analysis of the response spectra of the contacted neurons (n=232) revealed the presence of 28 classes of receptor neurons. The neurons exhibited strong selectivity as well as high sensitivity. Eleven of the identified classes were selectively activated by single compounds, while the remaining were activated by 2–6 compounds. Several receptor neurons that were activated by more than one compound responded to compounds sharing basic structural similarities. The results support the growing hypothesis that a significant proportion of plant-odor receptor neurons in insects are highly sensitive and selective for single odors.

Functional characterization of a sex pheromone receptor in the pest moth Spodoptera littoralis by heterologous expression in Drosophila

Moth sex pheromone communication is recognised as a long‐standing model for insect olfaction studies, and a widespread knowledge has been accumulated on this subject thanks to numerous chemical, electrophysiological and behavioural studies. A key step has been the identification of candidate sex pheromone receptors, opening new routes to understanding the specificity and sensitivity of this communication system, but only few of these receptors have as yet been functionally characterised. In this context, we aim at unravelling the molecular bases of pheromone reception in the noctuid moth Spodoptera littoralis. Taking advantage of a collection of antennal‐expressed sequence tags, we previously identified three fragments of candidate pheromone receptors in this species. Here, we report full‐length cloning of one of these receptors, named SlitOR6. Both sequence and expression pattern analyses were consistent with its annotation as a pheromone receptor, which we further confirmed by functional characterization. Using Drosophila antennae as a heterologous expression system, we identified a single component of the pheromone blend of S. littoralis, (Z,E)‐9,12‐tetradecadienyl acetate, as the ligand of SlitOR6. Two strategies were employed: (i) expressing SlitOR6 in the majority of Drosophila olfactory neurons, in addition to endogenous receptors, and monitoring the responses to pheromone stimuli by electroantennography; (ii) replacing the Drosophila pheromone receptor OR67d with SlitOR6 and monitoring the response by single sensillum recordings. Results were fully congruent and responses to (Z,E)‐9,12‐tetradecadienyl acetate were highly specific in both heterologous systems. This approach appears to be efficient and reliable for studying moth pheromone receptors in an in vivo context.

Attraction of the larval predator Elater ferrugineus to the sex pheromone of its prey, Osmoderma eremita and its implication for conservation biology

Elater ferrugineus is a threatened click beetle inhabiting old hollow trees. Its larvae consume larvae of other saproxylic insects including the threatened scarab beetle Osmoderma eremita. Recently, (R)-(+)-γ-decalactone was identified as a male-produced sex pheromone of O. eremita. Here we present evidence that E. ferrugineus adults use this odor as a kairomone for location of their prey. In field trapping experiments, significantly more trapping events of E. ferrugineus beetles were observed in Lindgren funnel traps baited with (R)-(+)-γ-decalactone than in control traps (20 vs. 1, respectively). Analyses of headspace collections from E. ferrugineus beetles indicate that the predator itself does not produce the substance. Both sexes were attracted to the prey pheromone, suggesting that E. ferrugineus males use the odor as an indirect cue for location of mates or of the tree hollows, which make up their habitat. When compared to pitfall traps, the Lindgren system was significantly more effective in trapping E. ferrugineus, and no difference could be established for O.eremita, showing the high potential to use odor-based systems to catch both species. We suggest that (R)-(+)-γ-decalactone could be used as a master signal in monitoring programs for these vulnerable beetle species, which are both regarded as indicators of the associated insect fauna of the threatened habitat of old hollow trees.

Natural odor ligands for olfactory receptor neurons of the female mosquito Aedes aegypti: use of gas chromatography-linked single sensillum recordings

SUMMARY Female Aedes aegypti are vectors of dengue and yellow fever. Odor volatiles are the predominant cues that drive the host-seeking behavior of Ae. aegypti. Odorant molecules are detected and discriminated by olfactory receptor neurons (ORNs) housed in sensory hairs, sensilla, located on the antennae and maxillary palps. In a previous study, we used odor volatiles that are behaviorally and/or electrophysiologically active for Ae. aegypti and other mosquito species to show that antennal ORNs of female Ae. aegypti are divided into functionally different classes. In the present study, we have, for the first time, conducted gas chromatography-coupled single sensillum recordings (GC–SSR) from antennal trichoid and intermediate sensilla of female Ae. aegypti in order to screen for additional putative host attractants and repellents. We used headspace collections from biologically relevant sources, such as different human body parts (including feet, trunk regions and armpit), as well as a plant species used as a mosquito repellent, Nepeta faassenii. We found that a number of ORN types strongly responded to one or more of the biological extracts. GC–SSR recordings revealed several active components, which were subsequently identified through GC-linked mass spectrometry (GC–MS). Electrophysiologically active volatiles from human skin included heptanal, octanal, nonanal and decanal.

Genetic Variability and Robustness of Host Odor Preference in Drosophila melanogaster

Chemosensory stimuli play a crucial role for host selection in insects, including the fruit fly Drosophila melanogaster. Drosophila has been instrumental in unraveling the neurological basis of olfactory processing in insects. Basic knowledge regarding chemical ecology and thorough studies of olfactory preferences are still lacking to a great extent in D. melanogaster, however. We have characterized repeatable variation in olfactory preference between five classical D. melanogaster wild-type strains toward a large array of natural host odors and synthetic compounds. By recording the rate of attraction over up to 24 hr, we could compare stimuli varying in attractiveness and characterize phenotypic parameters on the basis of individual stimuli and the whole stimulus array. Behavioral differences between strains were predominantly due to variation in a single phenotypic parameter: their overall responsiveness toward optimal and suboptimal olfactory stimuli. These differences were not explained by variation in olfactory sensitivity, locomotory activity, or general vigor monitored by survival. Comparisons with three recently established wild-type strains indicated that a high behavioral threshold against accepting suboptimal olfactory stimuli is the characteristic phenotype of wild D. melanogaster.