Marie Bengtsson

Host plant volatiles synergize response to sex pheromone in codling moth, Cydia pomonella

Plant volatile compounds synergize attraction of codling moth males Cydia pomonella to sex pheromone (E,E)-8,10-dodecadien-1-ol (codlemone). Several apple volatiles, known to elicit a strong antennal response, were tested in a wind tunnel. Two-component blends of 1 pg/min codlemone and 100 pg/min of either racemic linalool, (E)-β-farnesene, or (Z)-3-hexen-1-ol attracted significantly more males to the source than codlemone alone (60, 58, 56, and 37%, respectively). In comparison, a blend of codlemone and a known pheromone synergist, dodecanol, attracted 56% of the males tested. Blends of pheromone and plant volatiles in a 1:100 ratio attracted more males than 1:1 or 1:10,000 blends. Adding two or four of the most active plant compounds to codlemone did not enhance attraction over blends of codlemone plus single-plant compounds. Of the test compounds, only farnesol was attractive by itself; at a release rate of 10,000 pg/min, 16% of the males arrived at the source. However, attraction to a 1:10,000 blend of codlemone and farnesol (42%) was not significantly different from attraction to codlemone alone (37%). In contrast, a codlemone mimic, (E)-10-dodecadien-1-ol, which attracted 2% males by itself, had a strong antagonistic effect when blended in a 1:10,000 ratio with codlemone.

Attraction of codling moth males to apple volatiles

The attraction of the codling moth, Cydia pomonella, to apple volatile compounds known to elicit an antennal response was tested both in the field and in a wind tunnel. In the field, (E)‐β‐farnesene captured male moths. The addition of other apple volatiles, including (E,E)‐α‐farnesene, linalool, or (E,E)‐farnesol to (E)‐β‐farnesene did not significantly augment trap catch. Few females were caught in traps which also caught male moths, but female captures were not significantly different from blank traps. In the wind tunnel, males were attracted to (E,E)‐farnesol, but not to (E)‐β‐farnesene. The addition of (E,E)‐α‐farnesene to (E)‐β‐farnesene had a synergistic effect on male attraction. The male behavioural sequence elicited by plant volatiles, including upwind flight behaviour, was indistinguishable from the behaviour elicited by sex pheromone.

Flying the Fly: Long-range Flight Behavior of Drosophila melanogaster to Attractive Odors

The fruit fly, Drosophila melanogaster Meigen (Diptera: Drosophilidae), is a model for how animals sense, discriminate, and respond to chemical signals. However, with D. melanogaster our knowledge of the behavioral activity of olfactory receptor ligands has relied largely on close-range attraction, rather than on long-range orientation behavior. We developed a flight assay to relate chemosensory perception to behavior. Headspace volatiles from vinegar attracted 62% of assayed flies during a 15-min experimental period. Flies responded irrespective of age, sex, and mating state, provided they had been starved. To identify behaviorally relevant chemicals from vinegar, we compared the responses to vinegar and synthetic chemicals. Stimuli were applied by a piezoelectric sprayer at known and constant release rates. Re-vaporized methanol extracts of Super Q-trapped vinegar volatiles attracted as many flies as vinegar. The main volatile component of vinegar, acetic acid, elicited significant attraction as a single compound. Two other vinegar volatiles, 2-phenyl ethanol and acetoin, produced a synergistic effect when added to acetic acid. Geosmin, a microbiological off-flavor, diminished attraction to vinegar. This wind tunnel assay based on a conspicuous and unambiguous behavioral response provides the necessary resolution for the investigation of physiologically and ecologically relevant odors and will become an essential tool for the functional analysis of the D. melanogaster olfactory system.

Floral to green: mating switches moth olfactory coding and preference

Mating induces profound physiological changes in a wide range of insects, leading to behavioural adjustments to match the internal state of the animal. Here, we show for the first time, to our knowledge, that a noctuid moth switches its olfactory response from food to egg-laying cues following mating. Unmated females of the cotton leafworm (Spodoptera littoralis) are strongly attracted to lilac flowers (Syringa vulgaris). After mating, attraction to floral odour is abolished and the females fly instead to green-leaf odour of the larval host plant cotton, Gossypium hirsutum. This behavioural switch is owing to a marked change in the olfactory representation of floral and green odours in the primary olfactory centre, the antennal lobe (AL). Calcium imaging, using authentic and synthetic odours, shows that the ensemble of AL glomeruli dedicated to either lilac or cotton odour is selectively up- and downregulated in response to mating. A clear-cut behavioural modulation as a function of mating is a useful substrate for studies of the neural mechanisms underlying behavioural decisions. Modulation of odour-driven behaviour through concerted regulation of odour maps contributes to our understanding of state-dependent choice and host shifts in insect herbivores.

ANTENNAL AND BEHAVIORAL RESPONSES OF GRAPEVINE MOTH Lobesia botrana FEMALES TO VOLATILES FROM GRAPEVINE

Grapevine moth Lobesia botrana is the economically most important insect of grapevine Vitis vinifera in Europe. Flower buds, flowers, and green berries of Chardonnay grapevine are known to attract L. botrana for oviposition. The volatile compounds collected from these phenological stages were studied by gas chromatography—mass spectrometry, and the antennal response of L. botrana females to these headspace collections was recorded by gas chromatography—electroantennography. The compounds found in all phenological stages, which consistently elicited a strong antennal response, were pentadecane, nonanal, and α -farnesene. In a wind tunnel, gravid L. botrana females flew upwind to green grapes, as well as to headspace collections from these berries released by a piezoelectric sprayer release device. However, no females landed at the source of headspace volatiles, possibly due to inappropriate concentrations or biased ratios of compounds in the headspace extracts.

Antennal and behavioural response of codling moth Cydia pomonella to plant volatiles

Identification of host volatile compounds attractive to codling moth Cydia pomonella, a most important insect of apple, will contribute to the development of safe control techniques. Synthetic apple volatiles in two doses were tested for antennal and behavioural activity in codling moth. Female antennae strongly responded to (Z)3‐hexenol, (Z)3‐hexenyl benzoate, (Z)3‐hexenyl hexanoate, (±)‐linalool and E,E‐α‐farnesene. Two other compounds eliciting a strong antennal response were the pear ester, ethyl (E,Z)‐2,4‐decadienoate, and its corresponding aldehyde, E,E‐2,4‐decadienal, which is a component of the larval defence secretion of the European apple sawfly. Attraction of codling moth to compounds eliciting a strong antennal response was tested in a wind tunnel. Male moths were best attracted to a blend of (E,E)‐α‐farnesene, (E)‐beta‐farnesene and ethyl (E,Z)‐2,4‐decadienoate. The aldehyde E,E‐2,4‐decadienal had an antagonistic effect when added to the above mixture.

A predicted sex pheromone receptor of codling moth Cydia pomonella detects the plant volatile pear ester

Plant volatiles mediate host discrimination and host finding in phytophagous insects. Understanding how insects recognize these signals is a current challenge in chemical ecology research. Pear ester, ethyl (E,Z)-2,4-decadienoate, is a powerful, bisexual attractant of codling moth Cydia pomonella (Lepidoptera, Tortricidae) and strongly synergizes the male response to female-produced sex pheromone. We show here that the codling moth odorant receptor (OR) CpomOR3 is dedicated to detecting this plant volatile. Heterologous expression of CpomOR3 in Drosophila T1 trichoid and ab3A basiconic sensilla, followed by a screening with codling moth pheromone compounds and known plant volatile attractants, confirms that CpomOR3 binds to pear ester. Although CpomOR3 does not respond to any of the pheromone components tested, a phylogenetic analysis of lepidopteran chemosensory receptor genes reveals a close relationship of CpomOR3 with pheromone receptors (PRs) in moths. This corroborates the interaction of ecological and social chemosensory cues during premating communication. The finding that a plant volatile compound, pear ester, is a specific ligand for a PR-like lepidopteran receptor adds to our understanding of insect-plant interactions and emphasizes the interaction of natural and sexual selection during the phylogenetic divergence of insect herbivores.

Plant volatiles mediate attraction to host and non-host plant in apple fruit moth, Argyresthia conjugella

Plant volatiles mediate host finding in insect herbivores and lead to host fidelity and habitat‐specific mating, generating premating reproductive isolation and facilitating sympatric divergence. The apple fruit moth, Argyresthia conjugella Zeller (Lepidoptera: Argyresthiidae), is a particularly suitable species to study the cues and behavioural mechanisms leading to colonization of a new host: it recurrently oviposits on the non‐host plant, apple Malus domestica Borkh. (Rosaceae), where the larvae cannot complete their development. The larval host of the apple fruit moth (Lepidoptera, Argyresthiidae), is rowan Sorbus aucuparia L. (Rosaceae). Fruit setting in rowan, however, fluctuates strongly over large areas in Scandinavia. Every 2–4 years, when too few rowanberries are available for egg laying in forests, apple fruit moth females oviposit instead on apple in nearby orchards, but not on other fruits, such as pear or plum. This poses the question of which cues mediate attraction to rowan and apple, and how apple fruit moth discriminates rowan from apple. Chemical analysis and antennal recordings showed that 11 out of 15 rowan volatiles eliciting an antennal response in A. conjugella females co‐occur in rowan and apple headspace, in a different proportion. In the field, A. conjugella was attracted to several of these plant volatiles, especially to 2‐phenyl ethanol, methyl salicylate, and decanal. Addition of anethole to 2‐phenyl ethanol had a strong synergistic effect, the 1 : 1 blend is a powerful attractant for A. conjugella males and females. These results confirm that volatiles common to both plants may account for a host switch in A. conjugella from rowan to apple. Some of the most attractive compounds, including 2‐phenyl ethanol, anethole, and decanal, which have been found in several apple cultivars, were not present in the headspace of the apple cultivar, Aroma, which is also susceptible to attack by A. conjugella. This supports the idea that the odour signal from apple is suboptimal for attraction of A. conjugella, but is nonetheless sufficient for attraction, during times when rowan is not available for egg laying.

Behavioral observations of codling moth, Cydia pomonella, in orchards permeated with synthetic pheromone

Mating disruption of codling moth, Cydia pomonella, was studied in apple orchards treated with the main pheromone compound codlemone, (E,E)-8,10-dodecadienol, and a blend of codlemone and codlemone acetate, (E,E)-8,10-dodecadienyl acetate, a strong pheromone antagonist. Codlemone alone and the pheromone/antagonist-blend had a similar effect on the behavior of males emerging into air-permeated orchards. Male flights within tree canopy and upwind orientation along tree rows were strongly enhanced by both formulations, compared to untreated plots. However, the codlemone/codlemone acetate-blend increased the rate of cross-wind and downwind flights within the orchard, compared to codlemone alone. The major difference between these two formulations was that males from nearby, untreated orchards were attracted towards orchards treated with codlemone, but not towards treatments with codlemone/codlemone acetate. Additional tests were done with an equilibrium blend of codlemone and its geometric isomers. Aerial pheromone concentrations in the orchards were recorded by the field electroantennogram technique. Decreasing pheromone concentrations towards the upper part of the tree canopy, together with the stimulation of male flight activity by synthetic pheromone explains failures to control codling moth at high population densities with current dispenser formulations.

Coding and interaction of sex pheromone and plant volatile signals in the antennal lobe of the codling moth Cydia pomonella.

SUMMARY In the codling moth Cydia pomonella (Lepidoptera: Tortricidae) plant volatiles attract males and females by upwind flight and synergise the male response to the female-produced sex pheromone, indicating a close relationship between the perception of social and environmental olfactory signals. We have studied the anatomical and functional organisation of the antennal lobe (AL), the primary olfactory centre, of C. pomonella with respect to the integration of sex pheromone and host-plant volatile information. A three-dimensional reconstruction of the glomerular structure of the AL revealed 50±2 and 49±2 glomeruli in males and females, respectively. These glomeruli are functional units involved in the coding of odour quality. The glomerular map of the AL was then integrated with electrophysiological recordings of the response of individual neurons in the AL of males and females to sex pheromone components and behaviourally active plant volatiles. By means of intracellular recordings and stainings, we physiologically characterised ca. 50 neurons in each sex, revealing complex patterns of activation and a wide variation in response dynamics to these test compounds. Stimulation with single chemicals and their two-component blends produced both synergistic and inhibitory interactions in projection neurons innervating ordinary glomeruli and the macroglomerular complex. Our results show that the sex pheromone and plant odours are processed in an across-fibre coding pattern. The lack of a clear segregation between the pheromone and general odour subsystems in the AL of the codling moth suggests a level of interaction that has not been reported from other insects.