A plant volatile alters the perception of sex pheromone blend ratios in a moth.

Mate finding in most moths is based on male perception of a female-emitted pheromone whose species specificity resides in component chemistry and proportions. Components are individually detected by specialized olfactory receptor neurons (ORNs) projecting into the macroglomerular complex (MGC) of the male brain. We asked how robust ratio recognition is when challenged by a plant volatile background. To test this, we investigated the perception of the pheromone blend in Agrotis ipsilon, a moth species whose females produce a blend of Z7-dodecenyl acetate (Z7-12:Ac), Z9-tetradecenyl acetate (Z9-14:Ac), and Z11-hexadecenyl acetate in a 4:1:4 ratio optimally attractive for males. First, we recorded the responses of specialist ORNs for Z7 and Z9 and showed that heptanal, a flower volatile, activated Z7 but not Z9 neurons. Then, we recorded intracellularly the responses of MGC neurons to various ratios and showed that heptanal altered ratio responses of pheromone-sensitive neurons. Finally, we analyzed the behavior of males in a wind tunnel and showed that their innate preference for the 4:1:4 blend was shifted in the presence of heptanal. Pheromone ratio recognition may thus be altered by background odorants. Therefore, the olfactory environment might be a selective force for the evolution of pheromone communication systems.

Identification of the Aggregation Pheromone of the Date Palm Root Borer Oryctes agamemnon.

Laboratory and field investigations aimed to characterize the chemical communication system of the date palm pest Oryctes agamemnon. Live males or extracts of male effluvia attracted conspecifics in an olfactometer, whereas female effluvia attracted only males. Volatile emissions from adults feeding on sugarcane were sampled and analysed by gas chromatography (GC) and GC/mass spectrometry (GC/MS). Males emitted a blend of 1) ethyl 4-methyloctanoate, 2) 4-methyloctanoic acid, 3) 4-methyloctanyl acetate, and 4) 4-methyloctanol in variable ratio. Single sensillum recordings demonstrated that compounds 1, 2, and 3 are detected by specific olfactory receptor neurons. Olfactometric experiments showed that compounds 1 and 3 attract both sexes of O. agamemnon, but females are more attracted by compound 1 and males by compound 3. Compound 2 was more attractive for females, especially virgin ones. Field experiments confirmed that compound 1 and compound 2 attracted O. agamemnon of both sexes and showed synergy with palm odors. No clear activity of compound 3 was observed. A mix of compounds 1 and 2 with date palm core odor was significantly the most attractive, and captured more females than males. The male aggregation pheromone of O. agamemnon appears therefore to be based on a mixture in contrast to previously identified Oryctes pheromones. Our results provide the basis for developing mass trapping to control this pest.

Pheromone modulates plant odor responses in the antennal lobe of a moth.

In nature, male moths are exposed to a complex plant odorant environment when they fly upwind to a sex pheromone source in their search for mates. Plant odors have been shown to affect responses to pheromone at various levels but how does pheromone affects plant odor perception? We recorded responses from neurons within the non-pheromonal "ordinary glome ruli" of the primary olfactory center, the antennal lobe (AL), to single and pulsed stimulations with the plant odorant heptanal, the pheromone, and their mixture in the male moth Agrotis ipsilon. We identified 3 physiological types of neurons according to their activity patterns combining excitatory and inhibitory phases. Both local and projection neurons were identified in each physiological type. Neurons with excitatory responses to heptanal responded also frequently to the pheromone and showed additive responses to the mixture. Moreover, the neuron's ability of resolving successive pulses generally improved with the mixture. Only some neurons with combined excitatory/inhibitory, or purely inhibitory responses to heptanal, also responded to the pheromone. Although individual mixture responses were not significantly different from heptanal responses in these neurons, pulse resolution was improved with the mixture as compared with heptanal alone. These results demonstrate that the pheromone and the general odorant subsystems interact more intensely in the moth AL than previously thought.

Is the evolution of insect odorscapes under anthropic pressures a risk for herbivorous insect invasions?

Olfaction is directly involved in the insect capacity to exploit new habitats by guiding foraging behaviors. We searched in the literature whether some traits of olfactory systems and behaviors are associated with invasiveness and the impact of anthropogenic activities thereof. Human activities dramatically modify habitats and alter insect odorscapes. Air pollution, for instance, decreases lifetime and active range of semiochemicals. Plasticity and behavioral adaptability of invasive species are decisive by allowing host shifts and adaptative responses to new habitats. Changes in biophysical environments also impact on the use of semiochemicals in biocontrol. Although no evidence for a unique ensemble of olfactory traits associated with invasiveness was found, a growing number of case studies reveal characteristics with risk-predicting value, opening the paths to better invasion-control strategies.

Effects of Multi-Component Backgrounds of Volatile Plant Compounds on Moth Pheromone Perception.

The volatile plant compounds (VPC) alter pheromone perception by insects but mixture effects inside insect olfactory landscapes are poorly understood. We measured the activity of receptor neurons tuned to Z7-12Ac (Z7-ORN), a pheromone component, in the antenna and central neurons in male Agrotis ipsilon while exposed to simple or composite backgrounds of a panel of VPCs representative of the odorant variety encountered by a moth. Maps of activities were built using calcium imaging to visualize which areas in antennal lobes (AL) were affected by VPCs. We compared the VPC activity and their impact as backgrounds at antenna and AL levels, individually or in blends. At periphery, VPCs showed differences in their capacity to elicit Z7-ORN firing response that cannot be explained by differences in stimulus intensities because we adjusted concentrations according to vapor pressures. The AL neuronal network, which reformats the ORN input, did not improve pheromone salience. We postulate that the AL network evolved to increase sensitivity and to encode for fast changes of pheromone at some cost for signal extraction. Comparing blends to single compounds indicated that a blend shows the activity of its most active component. VPC salience seems to be more important than background complexity.

Insect olfactory communication in a complex and changing world.

Insect intraspecific olfactory communication occurs in a complex sensory environment. Here we present recent results on how the olfactory system extracts specific information from a sensory background, and integrates it with complementary information to improve odor source localization. Recent advances on mechanisms of olfactory mixture processing, multi-modal integration, as well as plasticity of sensory processing are reviewed. Significant progress in the understanding of neural coding and molecular bases of olfaction reinforce our perception of the tremendous adaptability of insects to a changing environment. However several reports demonstrate that anthropogenic environmental perturbations interfere with insect olfactory communication and might as a consequence significantly alter the functioning of ecosystems and agroecosystems.

Modulatory effects of pheromone on olfactory learning and memory in moths.

Pheromones are chemical communication signals known to elicit stereotyped behaviours and/or physiological processes in individuals of the same species, generally in relation to a specific function (e.g. mate finding in moths). However, recent research suggests that pheromones can modulate behaviours, which are not directly related to their usual function and thus potentially affect behavioural plasticity. To test this hypothesis, we studied the possible modulatory effects of pheromones on olfactory learning and memory in Agrotis ipsilon moths, which are well-established models to study sex-pheromones. To achieve this, sexually mature male moths were trained to associate an odour with either a reward (appetitive learning) or punishment (aversive learning) and olfactory memory was tested at medium- and long-term (1 h or 1.5 h, and 24 h). Our results show that male moths can learn to associate an odour with a sucrose reward, as well as a mild electric shock, and that olfactory memory persists over medium- and long-term range. Pheromones facilitated both appetitive and aversive olfactory learning: exposure to the conspecific sex-pheromone before conditioning enhanced appetitive but not aversive learning, while exposure to a sex-pheromone component of a heterospecific species (repellent) facilitated aversive but not appetitive learning. However, this effect was short-term, as medium- and long-term memory were not improved. Thus, in moths, pheromones can modulate olfactory learning and memory, indicating that they contribute to behavioural plasticity allowing optimization of the animal's behaviour under natural conditions. This might occur through an alteration of sensitization.

Plant terpenes affect intensity and temporal parameters of pheromone detection in a moth.

In moths, the components of the female pheromone blend are detected in the male antennae by pheromone olfactory receptor neurons (Ph-ORNs) expressing narrowly tuned olfactory receptors. Responses to sex pheromones have generally been thought to be independent from the odorant background. However, interactions between pheromone components and plant volatiles have been reported at behavioral and detection levels. To document the mechanisms of such interactions, we analyzed Ph-ORN responses of Spodoptera littoralis to the main pheromone component, Z9E11-14:Ac, in the presence of 4 monoterpenes. To mimic natural contexts in which plant odors and pheromone emanate from different sources, the 2 stimuli were presented with different temporal patterns and from independent sources. Linalool reversibly reduced the firing response to Z9E11-14:Ac and produced an off effect. Geraniol and geranyl and linalyl acetates reduced the responses to Z9E11-14:Ac with a longer time course. Pulses of linalool over prolonged pheromone stimulation resulted in a discontinuous firing activity. Pulses of pheromone were better separated over a background of linalool, compared with odorless air. The data confirm that plant compounds may modulate the intensity and the temporal coding by Ph-ORNs of pheromone information. This modulation might positively affect mate location at high pheromone density especially nearby a pheromone source.

Thymol Affects Congruency Between Olfactory and Gustatory Stimuli in Bees.

Honey bees learn to associate sugars with odorants in controlled laboratory conditions and during foraging. The memory of these associations can be impaired after exposure to contaminants such as pesticides. The sub-lethal effects of acaricides such as 5-methyl-2-(propan-2-yl)-phenol (thymol) introduced into colonies to control varroa mites are of particular concern to beekeeping, due to detrimental effects of some acaricides on bees. Here we assess whether various odorant/sugar pairs are identically memorized in a differential appetitive olfactory conditioning experiment and whether this learning is affected by thymol exposure. Responses to odorants in retrieval tests varied according to the sugar they were paired with, a property called congruency. Interestingly, congruency was altered by pre-exposure to some thymol concentrations during retrieval tests, although electroantennography recordings showed it left odorant detection intact. This highlights the importance of taking into account subtle effects such as odor/sugar congruency in the study of the effect of pesticides on non-target insects, in addition to the simpler question of memory impairment.

Insect Odorscapes: From Plant Volatiles to Natural Olfactory Scenes.

Olfaction is an essential sensory modality for insects and their olfactory environment is mostly made up of plant-emitted volatiles. The terrestrial vegetation produces an amazing diversity of volatile compounds, which are then transported, mixed, and degraded in the atmosphere. Each insect species expresses a set of olfactory receptors that bind part of the volatile compounds present in its habitat. Insect odorscapes are thus defined as species-specific olfactory spaces, dependent on the local habitat, and dynamic in time. Manipulations of pest-insect odorscapes are a promising approach to answer the strong demand for pesticide-free plant-protection strategies. Moreover, understanding their olfactory environment becomes a major concern in the context of global change and environmental stresses to insect populations. A considerable amount of information is available on the identity of volatiles mediating biotic interactions that involve insects. However, in the large body of research devoted to understanding how insects use olfaction to locate resources, an integrative vision of the olfactory environment has rarely been reached. This article aims to better apprehend the nature of the insect odorscape and its importance to insect behavioral ecology by reviewing the literature specific to different disciplines from plant ecophysiology to insect neuroethology. First, we discuss the determinants of odorscape composition, from the production of volatiles by plants (section "Plant Metabolism and Volatile Emissions") to their filtering during detection by the olfactory system of insects (section "Insect Olfaction: How Volatile Plant Compounds Are Encoded and Integrated by the Olfactory System"). We then summarize the physical and chemical processes by which volatile chemicals distribute in space (section "Transportation of Volatile Plant Compounds and Spatial Aspects of the Odorscape") and time (section "Temporal Aspects: The Dynamics of the Odorscape") in the atmosphere. The following sections consider the ecological importance of background odors in odorscapes and how insects adapt to their olfactory environment. Habitat provides an odor background and a sensory context that modulate the responses of insects to pheromones and other olfactory signals (section "Ecological Importance of Odorscapes"). In addition, insects do not respond inflexibly to single elements in their odorscape but integrate several components of their environment (section "Plasticity and Adaptation to Complex and Variable Odorscapes"). We finally discuss existing methods of odorscape manipulation for sustainable pest insect control and potential future developments in the context of agroecology (section "Odorscapes in Plant Protection and Agroecology").

A Background of a Volatile Plant Compound Alters Neural and Behavioral Responses to the Sex Pheromone Blend in a Moth.

Recognition of intra-specific olfactory signals within a complex environment of plant-related volatiles is crucial for reproduction in male moths. Sex pheromone information is detected by specific olfactory receptor neurons (Phe-ORNs), highly abundant on the male antenna. The information is then transmitted to the pheromone processing macroglomerular complex (MGC) within the primary olfactory center, the antennal lobe, where it is processed by local interneurons and projection neurons. Ultimately a behavioral response, orientation toward the pheromone source, is elicited. Volatile plant compounds (VPCs) are detected by other functional types of olfactory receptor neurons (ORNs) projecting in another area of the antennal lobe. However, Phe-ORNs also respond to some VPCs. Female-produced sex pheromones are emitted within a rich environment of VPCs, some of which have been shown to interfere with the detection and processing of sex pheromone information. As interference between the different odor sources might depend on the spatial and temporal features of the two types of stimuli, we investigated here behavioral and neuronal responses to a brief sex pheromone blend pulse in a VPC background as compared to a control background in the male noctuid moth Agrotis ipsilon. We observed male orientation behavior in a wind tunnel and recorded responses of Phe-ORNs and MGC neurons to a brief sex pheromone pulse within a background of individual VPCs. We also recorded the global input signal to the MGC using in vivo calcium imaging with the same stimulation protocol. We found that VPCs eliciting a response in Phe-ORNs and MGC neurons masked responses to the pheromone and decreased the contrast between background odor and the sex pheromone at both levels, whereas α-pinene did not interfere with first order processing. The calcium signal produced in response to a VPC background was tonic, lasting longer than the VPC stimulus duration, and masked entirely the pheromone response. One percent heptanal and linalool, in addition to the masking effect, caused a clear delay in responses of MGC neurons to the sex pheromone. Upwind flight toward the pheromone in a wind tunnel was also delayed but otherwise not altered by different doses of heptanal.

Perception of cuticular hydrocarbons by the olfactory organs in Periplaneta americana (L.) (Insecta: Dictyoptera).

Despite the importance of cuticular hydrocarbons (CHs) in insect chemical communication, direct proof that they are detected and recognized by insects by contact or by olfactory receptors are rare. In Periplaneta americana, CHs induce aggregation. The aim of our study was to investigate how CHs are detected by P. americana antennae. Using solid phase microextraction and gas chromatography, the three main CHs of the species profile were identified in the volatiles emitted by these insects. Gas chromatography coupled to electroantennography recordings demonstrated that the antennae responded to these three CHs. Furthermore, CHs had an attraction effect in Y-olfactometer bioassays when presented at high concentrations. As CHs can be perceived by P. americana, at least from a short distance, they could play a role in attracting conspecifics during aggregation processes, in addition to inducing aggregation when direct contact is possible.

Olfactory signal coding in an odor background.

Insects communicating with pheromones are confronted with an olfactory environment featuring a diversity of volatile organic compounds from plant origin. These volatiles constitute a rich and fluctuant background from which the information carried by the pheromone signal must be extracted. Thus, the pheromone receptor neurons must encode into spike trains the quality, intensity and temporal characteristics of the signal that are determinant to the recognition and localization of a conspecific female. We recorded and analyzed the responses of the pheromone olfactory receptor neurons of male moths to sex pheromone in different odor background conditions. We show that in spite of the narrow chemical tuning of the pheromone receptor neurons, the sensory input can be altered by odorant background.

Unexpected plant odor responses in a moth pheromone system.

Male moths rely on olfactory cues to find females for reproduction. Males also use volatile plant compounds (VPCs) to find food sources and might use host-plant odor cues to identify the habitat of calling females. Both the sex pheromone released by conspecific females and VPCs trigger well-described oriented flight behavior toward the odor source. Whereas detection and central processing of pheromones and VPCs have been thought for a long time to be highly separated from each other, recent studies have shown that interactions of both types of odors occur already early at the periphery of the olfactory pathway. Here we show that detection and early processing of VPCs and pheromone can overlap between the two sub-systems. Using complementary approaches, i.e., single-sensillum recording of olfactory receptor neurons, in vivo calcium imaging in the antennal lobe, intracellular recordings of neurons in the macroglomerular complex (MGC) and flight tracking in a wind tunnel, we show that some plant odorants alone, such as heptanal, activate the pheromone-specific pathway in male Agrotis ipsilon at peripheral and central levels. To our knowledge, this is the first report of a plant odorant with no chemical similarity to the molecular structure of the pheromone, acting as a partial agonist of a moth sex pheromone.

Systematic Synthesis of Multifluorinated alpha,alpha-Difluoro-gamma-lactones through Intramolecular Radical Cyclization.

Carbon radicals from allyl O-(trimethylsilyl)-alpha-bromo-alpha,alpha-difluoroacetal can cyclize onto the olefinic part regiospecifically to give gamma-lactols in good yield. The lactols are then converted to the corresponding alpha,alpha-difluoro-gamma-lactones. Systematic synthesis of multifluorinated-alpha,alpha-difluoro-gamma-lactones has thus been accomplished through intramolecular radical cyclization as a key reaction. Semiempirical MO calculation study suggested a unique nature of alpha,alpha-difluoroacetate in that complete delocalization of the electrons in the SOMO orbital of alpha,alpha-difluoroacetyl radical occurred; this caused unsuccessful cyclization. To apply the present radical reaction, the first synthesis of both enantiomers of difluoroeldanolide, analogues of the sex pheromone of the male African sugarcane borer, has been demonstrated. Electrophysiological tests revealed that the difluorinated analogues were as active as the natural eldanolide on the olfactory receptors.

Structure and function of the antennal sensilla of the palm weevil Rhynchophorus palmarum (Coleoptera, Curculionidae).

The distribution, fine structure and function of the sensilla present on the antennal club of Rhynchophorus palmarum were studied. No sex dimorphism was observed. Scanning and transmission electron microscopy showed five types of hair-like structures, four of which were evenly distributed on the antennal club. Two types of hair (IV and V) showed wall pores, a characteristic of olfactory sensilla. The antenna numbers 11,190 +/- 3040 type IV and 7360 +/- 1500 type V hairs. Using single sensillum recording, we identified 17 types of olfactory receptor neurons (ORNs) on the basis of their responses to pheromone and host plant odors, triggering synergic behavioral responses. We characterized highly specific and sensitive ORNs tuned to the aggregation pheromone (18% ORNs; 0.01-1 ng response threshold) and to host plant odors such as ethyl acetate, ethanol, acetoin and guaiacol (10% ORNs; 1-10 ng response threshold). Eleven percent of the ORNs were more generalist, responding to several odors with low sensitivity. Nine percent of the ORNs showed a complex pattern of responses, being co-activated by the pheromone and plant odors. This suggests an interaction at the sensory neuron level between pheromone and plant odors, triggering synergic behavioral responses.

Electrophysiological responses to salts from antennal chaetoid taste sensilla of the ground beetle Pterostichus aethiops.

Antennal gustatory sensilla of the ground beetle Pterostichus aethiops (Pz., 1797) (Coleoptera, Carabidae) respond to salts, the three sensory cells, A-, B- and C-cells, producing action potentials that are distinguished by differences in their shape, amplitude, duration and polarity of spikes. The B-cell (salt cell) was highly sensitive to both ionic composition and concentration of the tested nine salt solutions showing phasic-tonic type of reaction with a pronounced phasic component. The stimulating effect was dominated by the cations involved, and in most cases, monovalent cations were more effective stimuli than divalent cations. Salt concentration/response relations were tested with NaCl at 1, 10, 100 and 1000 mmol l(-1): mean firing rates increased from 0.8 to 44 spikes per first second of the response, respectively. The pH value of the stimulating solutions also influenced the B-cell rate of firing. By contrast, the pH level of stimulus solutions influenced the A-cells' phasic-tonic response more than the ionic composition or concentration of these solutions. Compared to a standard 100 mmol l(-1) salt (NaCl) solution (pH 6.3), alkaline solutions of the salts NaCH3COO, Na2HPO4 and Na2B4O7 (pH 7.9, 8.5 and 9.3, respectively, all 100 mmol l(-1)) induced remarkably stronger responses in the A-cell. On the other hand, the reaction to an acid solution of NaH2PO4 (pH 4.5, 100 mmol l(-1)) was minimal. A-cell responses to neutral salts like NaCl, KCl, CaCl2, MgCl2 and C5H14NOCl (pH 6.1-6.5) varied largely in strength. Very low or no responses were observed with chlorides of divalent cations, CaCl2 and MgCl2, and choline chloride (C5H14NOCl), indicating that the ionic composition of the solutions also affected A-cell responses. Neural activity of the C-cell was not influenced by the salt solutions tested.

Disruption of responses to pheromone by (Z)-11-hexadecenyl trifluoromethyl ketone, an analogue of the pheromone, in the cabbage armyworm Mamestra brassicae.

The effects of (Z)-11-hexadecenyl trifluoromethyl ketone (Z11-16:TFMK) a fluorinated pheromone analogue, on the responses to sex pheromone of the male cabbage armyworm, Mamestra brassicae, have been investigated in an actograph and by electroantennography (EAG). In spite of its structural proximity with the natural pheromone, Z11-16:TFMK was poorly active in EAG, and not active on male behaviour. When permeated in the air, Z11-16:TFMK reversibly inhibited the electroantennographic responses to (Z)-11-hexadecenyl acetate (Z11-16:Ac), the main component of the sex pheromone. In the actograph, the latency of the activation was increased and the intensity of the behavioural activity of males in response to Z11-16:Ac was significantly reduced in the presence of Z11-16:TFMK. These results, along with others previously reported by us, provide new pointers to the possible use of Z11-16:TFMK in pest-control strategies.

Responses of the olfactory receptor neurons of the corn stalk borer Sesamia nonagrioides to components of the pheromone blend and their inhibition by a trifluoromethyl ketone analogue of the main component.

Two types of olfactory hairs and three types of olfactory receptor neurons (ORN) have been characterized on the antennae of male Sesamia nonagrioides Lef for the first time. Type A sensilla housed a cell which fired large spikes in response to (Z)-11-hexadecenyl acetate (Z11-16:Ac), the major component of the sex pheromone, and a second cell firing smaller spikes in response to (Z)-11-hexadecenal (Z11-16:Ald), a minor component of the pheromone blend. Type B sensilla housed one cell firing large spikes to Z11-16:Ac and a cell firing smaller spikes to another minor component of the pheromone blend, (Z)-11-hexadecenyl alcohol (Z11-16:OH). No cell responding to dodecyl acetate, another minor component of the natural extract, was found. Fluorinated ketones were tested as inhibitors of the cell responses to pheromone compounds. The fluorinated derivatives tested, (Z)-11-hexadecenyl trifluoromethyl ketone (Z11-16:TFMK), n-hexadecyl trifluoromethyl ketone (16:TFMK), (Z,E)-9,11-tetradecadienyl trifluoromethyl ketone (Z9,E11-14:TFMK), 3-octylthio-1,1,1-trifluoropropan-2-one (OTFP), (Z)-11-tetradecenyl trifluoromethyl ketone (Z11-11:TFMK) and 1,1-difluoro-(Z)-11-hexadecenyl methyl ketone (Z11-16:DFMK), had no or only weak excitatory effects. However, the neuron responses to the pheromone compounds were significantly decreased in the presence of a constant stimulation with Z11-16:TFMK and the effect was reversible. The latencies of the responses to the acetate and aldehyde cells were significantly increased. The effects were not specific, since Z11-16:TFMK also inhibited the responses of the ORNs of Spodoptera littoralis Boisd. Correspondingly, Z9,E11-14:TFMK, an analogue of the main component of the pheromone of this latter insect, inhibited responses of S nonagrioides ORNs. Implications of these results on the utilization of Z11-16:TFMK as a communication disruptant are discussed.

Responses to Pheromones in a Complex Odor World: Sensory Processing and Behavior.

Insects communicating with pheromones, be it sex- or aggregation pheromones, are confronted with an olfactory environment rich in a diversity of volatile organic compounds of which plants are the main releaser. Certain of these volatiles can represent behaviorally relevant information, such as indications about host- or non-host plants; others will provide essentially a rich odor background out of which the behaviorally relevant information needs to be extracted. In an attempt to disentangle mechanisms of pheromone communication in a rich olfactory environment, which might underlie interactions between intraspecific signals and a background, we will summarize recent literature on pheromone/plant volatile interactions. Starting from molecular mechanisms, describing the peripheral detection and central nervous integration of pheromone-plant volatile mixtures, we will end with behavioral output in response to such mixtures and its plasticity.