Ylva Hillbur

Antennal transcriptome analysis of the chemosensory gene families in the tree killing bark beetles, Ips typographus and Dendroctonus ponderosae (Coleoptera: Curculionidae: Scolytinae)

BackgroundThe European spruce bark beetle, Ips typographus, and the North American mountain pine beetle, Dendroctonus ponderosae (Coleoptera: Curculionidae: Scolytinae), are severe pests of coniferous forests. Both bark beetle species utilize aggregation pheromones to coordinate mass-attacks on host trees, while odorants from host and non-host trees modulate the pheromone response. Thus, the bark beetle olfactory sense is of utmost importance for fitness. However, information on the genes underlying olfactory detection has been lacking in bark beetles and is limited in Coleoptera. We assembled antennal transcriptomes from next-generation sequencing of I. typographus and D. ponderosae to identify members of the major chemosensory multi-gene families.ResultsGene ontology (GO) annotation indicated that the relative abundance of transcripts associated with specific GO terms was highly similar in the two species. Transcripts with terms related to olfactory function were found in both species. Focusing on the chemosensory gene families, we identified 15 putative odorant binding proteins (OBP), 6 chemosensory proteins (CSP), 3 sensory neuron membrane proteins (SNMP), 43 odorant receptors (OR), 6 gustatory receptors (GR), and 7 ionotropic receptors (IR) in I. typographus; and 31 putative OBPs, 11 CSPs, 3 SNMPs, 49 ORs, 2 GRs, and 15 IRs in D. ponderosae. Predicted protein sequences were compared with counterparts in the flour beetle, Tribolium castaneum, the cerambycid beetle, Megacyllene caryae, and the fruit fly, Drosophila melanogaster. The most notable result was found among the ORs, for which large bark beetle-specific expansions were found. However, some clades contained receptors from all four beetle species, indicating a degree of conservation among some coleopteran OR lineages. Putative GRs for carbon dioxide and orthologues for the conserved antennal IRs were included in the identified receptor sets.ConclusionsThe protein families important for chemoreception have now been identified in three coleopteran species (four species for the ORs). Thus, this study allows for improved evolutionary analyses of coleopteran olfaction. Identification of these proteins in two of the most destructive forest pests, sharing many semiochemicals, is especially important as they might represent novel targets for population control.

Field attractants for Pachnoda interrupta selected by means of GC-EAD and single sensillum screening

The sorghum chafer, Pachnoda interrupta Olivier (Coleoptera: Scarabaeidae: Cetoniinae), is a key pest on sorghum, Sorghum bicolor (L.) Moench (Poaceae), in Ethiopia. At present there is a lack of efficient control methods. Trapping shows promise for reduction of the pest population, but would benefit from the development of attractive lures. To find attractants that could be used for control of P. interrupta, either by mass trapping or by monitoring as part of integrated pest management, we screened headspace collections of sorghum and the highly attractive weed Abutilon figarianum Webb (Malvaceae) for antennal activity using gas chromatograph-coupled electroantennographic detection (GC-EAD). Compounds active in GC-EAD were identified by combined gas chromatography and mass spectrometry (GC-MS). Field trapping suggested that attraction is governed by a few influential compounds, rather than specific odor blends. Synthetic sorghum and abutilon odor blends were attractive, but neither blend outperformed the previously tested attractants eugenol and methyl salicylate, of which the latter also was part of the abutilon blend. The strong influence of single compounds led us to search for novel attractive compounds, and to investigate the role of individual olfactory receptor neurons (ORNs) in the perception of kairomones. We screened the response characteristics of ORNs to 82 putative kairomones in single sensillum recordings (SSR), and found a number of key ligand candidates for specific classes of ORNs. Out of these key ligand candidates, six previously untested compounds were selected for field trapping trials: anethole, benzaldehyde, racemic 2,3-butanediol, isoamyl alcohol, methyl benzoate and methyl octanoate. The compounds were selected on the basis that they activated different classes of ORNs, thus allowing us to test potential kairomones that activate large non-overlapping populations of the peripheral olfactory system, while avoiding redundant multiple activations of the same ORN type. Field trapping results revealed that racemic 2,3-butanediol is a powerful novel attractant for P. interrupta.

Identification of sex pheromone components of the pea midge, Contarinia pisi (Diptera: Cecidomyiidae)

Abstract Three components in extract of pheromone glands of female pea midges, Contarinia pisi, were found to be active on male pea midge antennae by coupled gas chromatographic-electroantennographic detection. The EAD active components were identified as 2-acetoxytridecane, (2S,11S)-diacetoxytridecane, and (2S,12S)-diacetoxytridecane. A blend of these compounds proved to be highly attractive to males in windtunnel experiments.

LABORATORY AND FIELD STUDY OF THE ATTRACTION OF MALE PEA MIDGES, Contarinia pisi, TO SYNTHETIC SEX PHEROMONE COMPONENTS

Behavioral activity of the recently identified sex pheromone components of the pea midge, Contarinia pisi, (2S,11S)-diacetoxytridecane, (2S,12S)-diacetoxytridecane, and 2-acetoxytridecane, was tested in wind tunnel and field-trapping experiments. In the wind tunnel, the attractancy of the three-component blend in a 7 : 10 : 0.1 ratio (following the above order, mimicking the ratios found in gland extract) did not differ significantly from female gland extract, whereas a mixture of the two major components (7 : 10) only attracted 2% of the males to the source. In the field, traps baited with the three-component blend caught by far the largest number of males. Traps baited with the two major components only caught slightly more than the blank traps, and catches in traps baited with 2-acetoxytridecane alone did not differ from catches in the blank traps. Traps baited with the racemate of all three components did not catch more than the blank traps, indicating that some of the enantiomers are inhibitory.

A chiral sex pheromone system in the pea midge, Contarinia pisi.

The sex pheromone of the pea midge consists of 2-acetoxytridecane, (2S,11S)-diacetoxytridecane and (2S,12S)-diacetoxytridecane. The responses of male pea midges to the corresponding stereoisomers of (2S,11S)-diacetoxytridecane and (2S,12S)-diacetoxytridecane were tested in field trapping experiments and by electroantennographic recordings. When added at 20% of the pheromone component to the sex pheromone blend, the (2S,11R)- and (2R,11S)-stereoisomers of (2S,11S)-diacetoxytridecane, were shown to have a strong inhibitory effect on male attraction in the field. At the same dose, (2R,11R)-diacetoxytridecane, (2R,12R)-diacetoxytridecane, and meso-2,12-diacetoxytridecane, did not have a significant effect on male behavior. It was also shown that substitution of either (2S,11S)-diacetoxytridecane or (2S,12S)-diacetoxytridecane with the related stereoisomers reduced trap catches to the level of blank traps. The electroantennographic recordings showed similar dose–response curves for the pheromone components and the stereoisomers shown to have an inhibitory effect. It seems likely that male antennae have receptors for both pheromone components and for inhibitory stereoisomers. Scanning electron microscopy and transmission electron microscopy of the antennae revealed three types of sensilla involved in chemoreception: sensilla circumfila, sensilla trichodea, and sensilla coeloconica. The sensilla circumfila and trichodea are both innervated by two sensory cells, whereas the sensilla coeloconica are innervated by four to five cells.

Comparative Study of Antennal and Maxillary Palp Olfactory Sensilla of Female Biting Midges (Diptera: Ceratopogonidae: Culicoides) in the Context of Host Preference and Phylogeny

ABSTRACT Culicoides biting midges (Diptera: Ceratopogonidae) are vectors of disease, including bluetongue and African horse sickness. Host preference of these insects is primarily regulated by olfactory cues, detected by olfactory sensilla on the antennae and maxillary palps. In this study, we analyzed the sensillum repertoire of biting midge species with known host preferences. Five different morphological sensillum types, sensilla trichodea, s. chaetica, s. ampullacea, s. coeloconica, and grooved peg sensilla, were present on the antennae of all species. In addition sensilla basiconica were present on the maxillary palps. We found that the numbers of short blunt-tipped s. trichodea, s. coeloconica, and s. basiconica are significantly higher in the ornithophilic Culicoides festivipennis (Kieffer) compared with the mammalophilic Culicoides obsoletus (Meigen) and Culicoides chiopterus (Meigen). In contrast, we found that the mammalophilic Culicoides pulicaris (L.) and the opportunistic Culicoides punctatus (Meigen) have intermediate numbers of these sensillum types. Comparison with available data from other species strongly suggests that these differences in the number of specific sensillum types, in general, are a reflection of host preference and not of phylogeny. We discuss the putative function of the individual sensillum types in relation to host volatile detection.

Behavioral and electrophysiological response of sorghum chafer Pachnoda interrupta (Coleoptera: Scarabaeidae) to plant compounds

Behavioural and antennal responses of the sorghum chafer Pachnoda interrupta (Olivier) were tested to the synthetic compounds, such as eugenol, methyl salicylate, methyl anthranilate, isoamyl acetate and butyl butyrate. In the field, all odour-baited traps, except isoamyl acetate applied on cotton dispensers, were significantly more attractive than blank (unbaited) traps and all compounds, except isoamyl acetate, elicited dose-dependent responses in both male and female antennae. Dispenser type (cotton wick or rubber septum), trap location (inside or outside sorghum fields) and season (mating/July or feeding/September) affected the performance of the different compounds as lures. In July, methyl salicylate applied on cotton was the most attractive lure, whereas the most attractive treatments in September were eugenol and isoamyl acetate on rubber septa. Possibly due to odour and visual competition from the sorghum plants, traps placed outside the sorghum fields caught significantly more beetles than traps placed inside the fields. The trapping efficiency of a locally produced trap was found not to be competitive with that of the commercially available Japanese beetle trap. Overall, the results suggest that trapping with semiochemicals has a potential in sorghum chafer management, either for mass trapping and/or as part of an integrated pest management programme.

Optimisation of the pheromone blend of the swede midge, Contarinia nasturtii, for monitoring

BACKGROUND The swede midge, Contarinia nasturtii Kieffer, is a serious pest in crucifers. Its pheromone is a blend of (2S,9S)-diacetoxyundecane, (2S,10S)-diacetoxyundecane and (2S)-acetoxyundecane. The pheromone is used in monitoring traps, and this study examines possible ways to optimise the traps. RESULTS Two dispenser types were compared: polyethylene dispensers and cotton dispensers. Polyethylene dispensers attracted male C. nasturtii for more than 6 weeks, whereas cotton dispensers were attractive for only 2 weeks. All three pheromone components were important for attraction of male midges in the field. The importance of the stereoisomeric compositions of the pheromone compounds was also tested-both in the wind tunnel and in the field. In the case of 2,9-diacetoxyundecane and 2-acetoxyundecane, the non-natural stereoisomers did not inhibit male C. nasturtii attraction, whereas one or both of the stereoisomers of 2,10-diacetoxyundecane did. CONCLUSION Pheromone traps with the synthetic pheromone in a 1:2:0.02 ratio emitted from PE dispensers were highly effective and long lasting. As the mixture of stereoisomers of 2,10-diacetoxyundecane strongly inhibited attraction of male C. nasturtii while those of 2,9-diacetoxyundecane and 2-acetoxyundecane did not have any inhibitory effect, it is possible to produce traps that are effective and long lasting but cheaper to produce and maintain.

Conserved, Highly Specialized Olfactory Receptor Neurons for Food Compounds in 2 Congeneric Scarab Beetles, Pachnoda interrupta and Pachnoda marginata

Few studies have systematically addressed evolutionary changes in olfactory neuron assemblies, either by genetic drift or as an adaptation to specific odor environments. We have studied the sense of olfaction in 2 congeneric scarab beetles, Pachnoda interrupta Olivier and Pachnoda marginata Drury (Coleoptera: Scarabaeidae: Cetoniinae), which are both opportunistic polyphages, feeding mainly on fruit and flowers. The 2 species occur in dissimilar habitats: P. interrupta is found in dry savannah, and P. marginata in tropical parts of equatorial Africa. To study how these species may have adapted their sense of olfaction to their odor environments, we utilized single-unit electrophysiology on olfactory sensilla with a wide selection of food-related compounds. Despite the differences in habitat, we found that the species shared most of the physiological types of olfactory receptor neurons (ORNs) encountered, although their proportions frequently varied between the species. The high degree of conservation in olfaction between the species implies that a similar sensory strategy is efficient for food search in both habitats. However, shifts in proportions of receptor neuron classes, and slight shifts in response profiles and/or presence of some ORN classes unique to either species, may reflect adaptation to a different set of hosts.

Gall midge olfaction: Pheromone sensitive olfactory neurons in Contarinia nasturtii and Mayetiola destructor

This study describes the morphology and function of the antennal sensilla in two gall midge species, Contarinia nasturtii and Mayetiola destructor, where multi-component sex pheromones have been identified. Both species possess sensilla trichodea, s. coeloconica, s. chaetica and s. circumfila. Sensilla circumfila, which consist of several sensilla that bifurcate and fuse into one structure, are unique for the gall midges. In C. nasturtii s. circumfila are sexually dimorphic. In males, they form elongated loops suspended on cuticular spines, whereas in females they run like worm-like structures directly on the antennal surface. Single sensillum recordings demonstrated that olfactory sensory neurons housed in male s. circumfila in C. nasturtii responded to the female sex pheromone. In M. destructor, s. circumfila were attached to the antennal surface in both sexes, and displayed no response to sex pheromone components. A sexual dimorphism was also found in the number of s. trichodea per antennal segment in both C. nasturtii (male 1 vs. female 7) and M. destructor (male 13 vs. female 10). OSNs located in male M. destructor s. trichodea responded to the sex pheromone. This is the first gall midge single sensillum study, and the first demonstration of the functional significance of s. circumfila.