Moth Pheromone Research Articles

Stimulus duration encoding occurs early in the moth olfactory pathway

Pheromones convey rich ethological information and guide insects’ search behavior. Insects navigating in turbulent environments are tasked with the challenge of coding the temporal structure of an odor plume, obliging recognition of the onset and offset of whiffs of odor. The coding mechanisms that shape odor offset recognition remain elusive. We designed a device to deliver sharp pheromone pulses and simultaneously measured the response dynamics from pheromone-tuned olfactory receptor neurons (ORNs) in male moths and Drosophila. We show that concentration-invariant stimulus duration encoding is implemented in moth ORNs by spike frequency adaptation at two time scales. A linear-nonlinear model fully captures the underlying neural computations and offers an insight into their biophysical mechanisms. Drosophila use pheromone cis-vaccenyl acetate (cVA) only for very short distance communication and are not faced with the need to encode the statistics of the cVA plume. Their cVA-sensitive ORNs are indeed unable to encode odor-off events. Expression of moth pheromone receptors in Drosophila cVA-sensitive ORNs indicates that stimulus-offset coding is receptor independent. In moth ORNs, stimulus-offset coding breaks down for short ( < 200 ms) whiffs. This physiological constraint matches the behavioral latency of switching from the upwind surge to crosswind cast flight upon losing contact with the pheromone.

Pheromonal variation and mating between two mitotypes of fall armyworm (Spodoptera frugiperda) in Africa

In the Americas, the fall armyworm (Spodoptera frugiperda) exists in two genetically distinct strains, the corn (C) and rice (R) strains. Despite their names, these strains are not associated with host plant preferences but have been shown to vary in pheromone composition and male responses. Recently, S. frugiperda was detected in Africa as an invasive species, but knowledge about variation in strain types, pheromone composition and inter-strain mating of populations of the pest in the continent has not been fully examined. Therefore, this study aimed to investigate variations, if any in the pheromone composition of female moths, male moth responses, and mating between C and R mitotypes of S. frugiperda populations in Kenya, as well as their geographic distribution. Strains (mitotypes) of S. frugiperda were identified using mitochondrial DNA (mtDNA) markers, and their pheromonal composition determined by coupled gas chromatography–mass spectrometric (GC–MS) analysis. Male moth responses to these compounds were evaluated using GC-electroantennographic detection (EAD), electroantennogram (EAG), and wind tunnel assays. Oviposition assays were used to determine whether R and C mitotype moths could mate and produce eggs. The results showed that both the R and C mitotypes were present, and there were no statistically significant differences in their distribution across all sampled locations. Five pheromone compounds including (Z)-7-dodecenyl acetate (Z7-12:OAc), (Z)-7-tetradecenyl acetate (Z7-14:OAc), (Z)-9-tetradecenyl acetate (Z9-14:OAc), (Z)-11-tetradecenyl acetate (Z11-14:OAc) and (Z)-11-hexadecenyl acetate (Z11-16:OAc), were detected in the pheromone glands of female moths of both mitotypes, with Z9-14:OAc being the most abundant. The relative percentage composition of Z9-14:OAc was similar in both mitotypes. However, the R mitotype had a 2.7 times higher relative percentage composition of Z7-12:OAc compared to the C mitotype moth, while the C mitotype moth had a 2.4 times higher relative percentage composition of Z11-16:OAc than the R mitotype moth. Male moths of both mitotypes exhibited similar responses to the pheromone compounds, showing the strongest responses to Z9-14:OAc and Z7-12:OAc in electrophysiological and behavioural assays. There was mating between R and C mitotypes with egg production comparable to mating within the same mitotype. Our results revealed that differences between the two S. frugiperda mitotypes are characterized by female moth pheromone composition rather than male moth responses to the pheromones, and that this does not prevent hybridisation between the mitotypes, which may have implications for their management.

Multiple amino acid changes are responsible for the shift of tuning breadth along the evolutionary trajectory of a moth pheromone receptor.

Sex pheromone recognition is essential for mating in many insects and plays a major role in maintaining reproductive barriers. A previous study from our lab reported the evolutionary history of the pheromone receptor OR5 in Spodoptera moths. Using heterologous expression in Xenopus oocytes and site-directed mutagenesis, we found that eight amino acid substitutions were sufficient to recapitulate the evolution from an ancestral broadly-tuned to a highly specific receptor. Here, we confirmed this result using expression in Drosophila olfactory neurons. This further confirmed that multiple amino acid changes explain the shift in tuning breadth of Spodoptera OR5 during evolution.

Chemosensory Receptor Expression in the Abdomen Tip of the Female Codling Moth, Cydia pomonella L. (Lepidoptera: Tortricidae).

In insects, the chemical senses influence most vital behaviors, including mate seeking and egg laying; these sensory modalities are predominantly governed by odorant receptors (ORs), ionotropic receptors (IRs), and gustatory receptors (GRs). The codling moth, Cydia pomonella, is a global pest of apple, pear, and walnut, and semiochemically based management strategies limit the economic impacts of this species. The previous report of expression of a candidate pheromone-responsive OR in female codling moth ovipositor and pheromone glands raises further questions about the chemosensory capacity of these organs. With an RNA-sequencing approach, we examined chemoreceptors' expression in the female codling moth abdomen tip, sampling tissues from mated and unmated females and pupae. We report 37 ORs, 22 GRs, and 18 IRs expressed in our transcriptome showing overlap with receptors expressed in adult antennae as well as non-antennal candidate receptors. A quantitative PCR approach was also taken to assess the effect of mating on OR expression in adult female moths, revealing a few genes to be upregulated or downregulating after mating. These results provide a better understanding of the chemosensory role of codling moth female abdomen tip organs in female-specific behaviors. Future research will determine the function of specific receptors to augment current semiochemical-based strategies for codling moth management.

From citation metrics to citation ethics: Critical examination of a highly-cited 2017 moth pheromone paper

From citation metrics to citation ethics: Critical examination of a highly-cited 2017 moth pheromone paper

Lipases and carboxylesterases affect moth sex pheromone compounds involved in interspecific mate recognition

Moth sex pheromones are a classical model for studying sexual selection. Females typically produce a species-specific pheromone blend that attracts males. Revealing the enzymes involved in the interspecific variation in blend composition is key for understanding the evolution of these sexual communication systems. The nature of the enzymes involved in the variation of acetate esters, which are prominent compounds in moth pheromone blends, remains unclear. We identify enzymes involved in acetate degradation using two closely related moth species: Heliothis (Chloridea) subflexa and H. (C.) virescens, which have different quantities of acetate esters in their sex pheromone. Through comparative transcriptomic analyses and CRISPR/Cas9 knockouts, we show that two lipases and two esterases from H. virescens reduce the levels of pheromone acetate esters when expressed in H. subflexa females. Together, our results show that lipases and carboxylesterases are involved in tuning Lepidoptera pheromones composition.

A tale of two copies: Evolutionary trajectories of moth pheromone receptors

Pheromone communication is an essential component of reproductive isolation in animals. As such, evolution of pheromone signaling can be linked to speciation. For example, the evolution of sex pheromones is thought to have played a major role in the diversification of moths. In the crop pests Spodoptera littoralis and S. litura, the major component of the sex pheromone blend is (Z,E)-9,11-tetradecadienyl acetate, which is lacking in other Spodoptera species. It indicates that a major shift occurred in their common ancestor. It has been shown recently in S. littoralis that this compound is detected with high specificity by an atypical pheromone receptor, named SlitOR5. Here, we studied its evolutionary history through functional characterization of receptors from different Spodoptera species. SlitOR5 orthologs in S. exigua and S. frugiperda exhibited a broad tuning to several pheromone compounds. We evidenced a duplication of OR5 in a common ancestor of S. littoralis and S. litura and found that in these two species, one duplicate is also broadly tuned while the other is specific to (Z,E)-9,11-tetradecadienyl acetate. By using ancestral gene resurrection, we confirmed that this narrow tuning evolved only in one of the two copies issued from the OR5 duplication. Finally, we identified eight amino acid positions in the binding pocket of these receptors whose evolution has been responsible for narrowing the response spectrum to a single ligand. The evolution of OR5 is a clear case of subfunctionalization that could have had a determinant impact in the speciation process in Spodoptera species.

Functional differentiation of two general odorant-binding proteins to sex pheromones in Spodoptera frugiperda

A precise chemosensory system can help insects complete various important behavioral responses by accurately identifying different external odorants. Therefore, deeply understanding the mechanism of insect recognition of important odorants will help us develop efficient and environmentally-friendly behavioral inhibitors. Spodoptera frugiperda is a polyphagous pest that feeds on >350 different host plants worldwide and also harms maize production in China. However, the molecular mechanism of the first step for males to use odorant-binding proteins (OBPs) to recognize sex pheromones remains unclear. Here, we obtained 50 OBPs from the S. frugiperda genome, and the expression level of SfruGOBP1 in females was significantly higher than that in males, whereas SfruGOBP2 displayed male-biased expression. Fluorescence competitive binding assays showed that only SfruGOBP2 showed binding affinities for the four sex pheromones of female S. frugiperda. Subsequently, we identified some key amino acid residues that can participate in the interaction between SfruGOBP2 and sex pheromones using molecular docking and site-directed mutagenesis methods. These findings will help us explore the interaction mechanism between GOBPs and sex pheromones in moths, and provide important target genes for developing new mating inhibitors of S. frugiperda in the future.

Controlling with Mass Trapping and Determination of Damage Rates of Codling Moth, Cydia pomonella L. (Lepidoptera: Tortricidae) at Good Agricultural Practices of Walnut Orchards in Hatay Province

The current study was carried out in 2018-2019 at good agricultural practices of walnut orchards to control with mass-trapping and determination of damage rates of codling moth, Cydia pomonella L. (Lepidoptera: Tortricidae) in Hatay province of Turkey. The study was conducted at 312.43 decares good agricultural practices of walnut orchards containing 3,928 number of Chandler varieties tree, located at the Kışlak village of the Yayladağ district of Hatay province. Delta traps with codling moth pheromones were used. Traps were hanged 1.5 m high, pheromone capsules were replaced with new ones in every forty days. In the first year, a total of 235 codling moth adults were caught by the 50 delta traps during the sampling period. The average population density of adults caught by traps was determined to be 4.61. In the second year, a total of 70 codling moth adults were caught by the 50 delta traps during the sampling period. The average population density of adults caught by traps was determined to be 1.4. In the first year, a total of 38,400 walnut fruits were harvested and a 100 walnut fruit was harmed by the codling moth larvae. Thus, the damage rates of the codling moth were estimated as 0.26 percent. In the second year, a total of 443,100 walnut fruits were harvested and a 9,065 walnut fruit was harmed by the codling moth larvae. Thus, the damage rates of the codling moth were estimated as 2.04 percent.

Insect Pest Pheromone Lures May Enhance the Activity of Insectivorous Bats in Mediterranean Vineyards and Apple Orchards

Insectivorous bats may play a significant role in regulating populations of agricultural pests. Currently, few methods are available to enhance the activity of bats in agroecosystems. We asked whether synthetic sex pheromones, used in integrated pest management (IPM) to impede the mating success of major moth pests in vineyards and apple orchards, could also enhance the activity and richness of insectivorous bats, their natural enemies. We hypothesized that applying concentrated sex pheromones of pest moths will alter the movement patterns of male moths, indirectly affecting bat richness and activity. We compared the effect of sex pheromones on bats under two different agricultural management systems: conventional farming and IPM. We used synthetic sex pheromones of Lobesia botrana or Cydia pomonella; both are among the most destructive moth pests in vineyards and apple orchards, respectively. Using passive acoustic monitoring, we compared species richness and bat activity in plots without and with additional pheromones. In both IPM vineyards and IPM apple orchards, total bat activity and species richness significantly increased after applying the pheromone treatment, with a positive correlation between total bat activity and the numbers of moth pests in the vineyards. In conventional vineyards, bat species richness increased significantly, but not total bat activity. IPM vineyards had significantly higher species richness than conventional vineyards, both before and after the pheromone treatment. Our study shows that moth pheromone lures, commonly used as a pest control method, may also attract insectivorous bats, which in turn may further suppress the pests. These findings highlight the potential of insectivorous bats as pest control agents and call for further research directed at integrating them in IPM practices.

A novel domain-duplicated SlitFAR3 gene involved in sex pheromone biosynthesis in Spodoptera litura.

Fatty acyl reductases (FARs) are key enzymes that participate in sex pheromone biosynthesis by reducing fatty acids to fatty alcohols. Lepidoptera typically harbor numerous FAR gene family members. Although FAR genes are involved in the biosynthesis of sex pheromones in moths, the key FAR gene of Spodoptera litura remains unclear. In this work, we predicted 30 FAR genes from the S.litura genome and identified a domain duplication within gene SlitFAR3, which exhibited high and preferential expression in the sexually mature female pheromone glands (PGs) and a rhythmic expression pattern during the scotophase of sex pheromone production. The molecular docking of SlitFAR3, as predicted using a 3D model, revealed a co-factor NADPH binding cavity and 2 substrate binding cavities. Functional expression in yeast cells combined with comprehensive gas chromatography indicated that the SlitFAR3 gene could produce fatty alcohol products. This study is the first to focus on the special phenomenon of FAR domain duplication, which will advance our understanding of biosynthesis-related genes from the perspective of evolutionary biology.

Manufacturing specialized wax esters in plants

Biologically produced wax esters can fulfil different industrial purposes. These functionalities almost drove the sperm whale to extinction from hunting. After the ban on hunting, there is a niche in the global market for biolubricants with properties similar to spermaceti. Wax esters can also serve as a mechanism for producing insect sex pheromone fatty alcohols. Pheromone-based mating disruption strategies are in high demand to replace the toxic pesticides in agriculture and manage insect plagues threatening our food and fiber reserves. In this study we set out to investigate the possibilities of in planta assembly of wax esters, for specific applications, through transient expression of various mix-and-match combinations of genes in Nicotiana benthamiana leaves. Our synthetic biology designs were outlined in order to pivot plant lipid metabolism into producing wax esters with targeted fatty acyl and fatty alcohols moieties. Through this approach we managed to obtain industrially important spermaceti-like wax esters enriched in medium-chain fatty acyl and/or fatty alcohol moieties of wax esters. Via employment of plant codon-optimized moth acyl-CoA desaturases we also managed to capture unusual, unsaturated fatty alcohol and fatty acyl moieties, structurally similar to moth pheromone compounds, in plant-accumulated wax esters. Comparison between outcomes of different experimental designs identified targets for stable transformation to accumulate specialized wax esters and helped us to recognize possible bottlenecks of such accumulation.

Optimization of a pheromone lure by analyzing the peripheral coding of sex pheromones of Spodoptera frugiperda in China.

BACKGOUND Sex pheromones of the fall armyworm, Spodoptera frugiperda, show differences in composition and proportions in different geographical populations, but always contain Z9-14:OAc as the major component. Odorant receptor neurons (ORNs) housed in the long trichoid sensilla (TS) of male antennae are essential to detect female-released sex pheromones in moths. RESULTS: In this study, we identified seven components from pheromone gland extracts of female S. frugiperda in the Yunnan population from China, including (Z)-7-dodecen-1-yl acetate (Z7-12:OAc), (Z)-9-tetradecenal (Z9-14:Ald), (Z)-9-dodecen-1-yl acetate (Z9-12:OAc), (Z)-9-tetradecen-1-yl acetate (Z9-14:OAc), (E)-11-tetradecen-1-yl acetate (E11-14:OAc), (Z)-11-tetradecen-1-yl acetate (Z11-14:OAc) and (Z)-11-hexadecen-1-yl acetate (Z11-16:OAc) at a ratio of 1.2:4:0.8:79.1:1.6:1.6:11.7 by gas chromatography coupled with mass spectrometry. Gas chromatography-electroantennographic detection showed that Z9-14:OAc, Z7-12:OAc and Z11-16:OAc are the male antennal active components. Peripheral coding of pheromones in males was investigated by single sensillum recording. Five functional neurons housed in three types of TS were identified based on profiles of neuronal responses, which are responsible for attractive component Z9-14:OAc, synergistic components Z7-12:OAc, Z11-16:OAc, interspecific pheromones (Z)-9-tetradecen-1-ol (Z9-14:OH) and (Z,E)-9,12-tetradecadien-1-yl acetate (Z9,E12-14:OAc), respectively. Wind tunnel and field tests demonstrated that a ternary combination of Z9-14:OAc, Z7-12:OAc and Z11-16:OAc at a ratio of 88:1:11 shows the strongest attractiveness to males. CONCLUSION: An optimized pheromone blend of Z9-14:OAc, Z7-12:OAc and Z11-16:OAc in an 88:1:11 ratio was identified for monitoring the invasive pest S. frugiperda in China. Five functional ORNs encoding intra- and interspecific pheromones were identified in male antennae, of which three neurons encode attractive component Z9-14:OAc, synergistic components Z7-12:OAc and Z11-16:OAc, respectively, and the other two neurons encode interspecific pheromones Z9-14:OH and Z9,E12-14:OAc, separately. © 2022 Society of Chemical Industry.

Release of moth pheromone compounds from Nicotiana benthamiana upon transient expression of heterologous biosynthetic genes

BackgroundUsing genetically modified plants as natural dispensers of insect pheromones may eventually become part of a novel strategy for integrated pest management.ResultsIn the present study, we first characterized essential functional genes for sex pheromone biosynthesis in the rice stem borer Chilo suppressalis (Walker) by heterologous expression in Saccharomyces cerevisiae and Nicotiana benthamiana, including two desaturase genes CsupYPAQ and CsupKPSE and a reductase gene CsupFAR2. Subsequently, we co-expressed CsupYPAQ and CsupFAR2 together with the previously characterized moth desaturase Atr∆11 in N. benthamiana. This resulted in the production of (Z)-11-hexadecenol together with (Z)-11-hexadecenal, the major pheromone component of C. suppressalis. Both compounds were collected from the transformed N. benthamiana headspace volatiles using solid-phase microextraction. We finally added the expression of a yeast acetyltransferase gene ATF1 and could then confirm also (Z)-11-hexadecenyl acetate release from the plant.ConclusionsOur results pave the way for stable transformation of plants to be used as biological pheromone sources in different pest control strategies.

Pheromone Receptor Knock-Out Affects Pheromone Detection and Brain Structure in a Moth.

Sex pheromone receptors are crucial in insects for mate finding and contribute to species premating isolation. Many pheromone receptors have been functionally characterized, especially in moths, but loss of function studies are rare. Notably, the potential role of pheromone receptors in the development of the macroglomeruli in the antennal lobe (the brain structures processing pheromone signals) is not known. Here, we used CRISPR-Cas9 to knock-out the receptor for the major component of the sex pheromone of the noctuid moth Spodoptera littoralis, and investigated the resulting effects on electrophysiological responses of peripheral pheromone-sensitive neurons and on the structure of the macroglomeruli. We show that the inactivation of the receptor specifically affected the responses of the corresponding antennal neurons did not impact the number of macroglomeruli in the antennal lobe but reduced the size of the macroglomerulus processing input from neurons tuned to the main pheromone component. We suggest that this mutant neuroanatomical phenotype results from a lack of neuronal activity due to the absence of the pheromone receptor and potentially reduced neural connectivity between peripheral and antennal lobe neurons. This is the first evidence of the role of a moth pheromone receptor in macroglomerulus development and extends our knowledge of the different functions odorant receptors can have in insect neurodevelopment.

Hexokinase Is Required for Sex Pheromone Biosynthesis in Helicoverpa armigera.

Simple SummarySex pheromone is an essential chemical signal for insect reproduction. Acetyl-CoA, which is generated from glycolysis, is the precursor of sex pheromone biosynthesis in Helicoverpa armigera. As the first speed-limited enzyme in glycolysis, the function of Hexokinase (HK) in sex pheromone biosynthesis remains elusive. This article employed H. armigera as a model. Results exhibited that the transcription profile of HaHK in female moth PGs was consistent with the release trend of sex pheromone. Knockdown of HaHK in female PGs caused significant decreases in acetyl-CoA content, sex pheromone production, and mating behaviors. Sugar feeding increased the transcription and enzymatic activity of HK. PBAN signal phospho-activated HaHK in PGs and Sf9 cells via PKA. In general, our study confirmed that PBAN/cAMP/PKA signal activated HaHK activity, and HaHK is required for sex pheromone biosynthesis in H. armigera.Acetyl-CoA, the precursor of sex pheromone biosynthesis in Helicoverpa armigera, is generated from glycolysis. As the first speed-limited enzyme in glycolysis, Hexokinase (HK) plays an important role in acetyl-CoA production. However, the function of HK in sex pheromone production remains unclear. This study employed H. armigera as material to explore the role of HK in sex pheromone production. Results demonstrated that the transcription profile of HaHK in female moth pheromone glands (PGs) was consistent with the release fluctuation of sex pheromone. Interference of HaHK prevented the increase of acetyl-CoA content induced by PBAN. Therefore, knockdown of HaHK in female PGs caused significant decreases in (Z)-11-hexadecenal (Z11-16:Ald) production, female capability to attract males, and mating rate. Furthermore, sugar feeding (5% sugar) increased the transcription and enzymatic activity of HK. Pheromone biosynthesis activating neuropeptide (PBAN) signal phospho-activated HaHK in PGs and Sf9 cells via protein kinase A (PKA), as shown by pharmacological inhibitor analysis. In general, our study confirmed that PBAN/cAMP/PKA signal activated HaHK, in turn promoted glycolysis to ensure the supply of acetyl-CoA, and finally facilitated sex pheromone biosynthesis and subsequent mating behavior.

Sex pheromone signal and stability covary with fitness.

If sexual signals are costly, covariance between signal expression and fitness is expected. Signal–fitness covariance is important, because it can contribute to the maintenance of genetic variation in signals that are under natural or sexual selection. Chemical signals, such as female sex pheromones in moths, have traditionally been assumed to be species-recognition signals, but their relationship with fitness is unclear. Here, we test whether chemical, conspecific mate finding signals covary with fitness in the moth Heliothis subflexa. Additionally, as moth signals are synthesized de novo every night, the maintenance of the signal can be costly. Therefore, we also hypothesized that fitness covaries with signal stability (i.e. lack of temporal intra-individual variation). We measured among- and within-individual variation in pheromone characteristics as well as fecundity, fertility and lifespan in two independent groups that differed in the time in between two pheromone samples. In both groups, we found fitness to be correlated with pheromone amount, composition and stability, supporting both our hypotheses. This study is, to our knowledge, the first to report a correlation between fitness and sex pheromone composition in moths, supporting evidence of condition-dependence and highlighting how signal–fitness covariance may contribute to heritable variation in chemical signals both among and within individuals.

EFFECT OF MATING DISRUPTION ON THE PHEROMONE TRAP CATCHES OF CODLING MOTH (CYDIA POMONELLA L.) AND FRUIT DAMAGE IN WALNUT (JUGLANS REGIA L.)ORCHARDS UNDER ORGANIC FARMING

ABSTRACT The codling moth (Cydia pomonella L.), a key pest in walnut orchards in Bulgaria, is managed mainly with broad spectrum insecticides. Insecticide application often resulted in destruction of beneficial species and environmental pollution. The present study was conducted during 2018 - 2019 in Pazardzhik region of Bulgaria in an isolated orchard. The pheromone trap catches of C. pomonella in the organically maintained walnut orchard in the presence of mating disruption (MD) dispensers was compared with the pheromone trap catches in the conventional control orchard. The Pherocon VI sticky traps, type Delta were baited with standard PHEROCON CM L2 – codlemone. CIDETRAK® CMDA COMBO™ MESO™ mating disruption dispensers containing a unique combination of codling moth pheromone and a patented male and female behavior modifying kairomone were used at the rate of 20 pieces per/ha in the organic orchard. There was reduction in fruit damage in the organic orchards, despite the lack of treatment with pesticides. In spite of 9 - 10 treatments with insecticides, fruit damage was higher in theconventional control orchard compared to the organic orchard. The present results brought out that the mating disruption using CIDETRAK® CMDA COMBO™ MESO™ dispensers against codling moth can provide more effective control against C. pomonella, than the pesticide treatments.

Development of methods for application of pheromonic materials for monitoring and managing the number of apple phytophages

Synthetic sex pheromones have found wide application in plant protection as a mean of early detection of pests that allows observation of the phenology of insects to optimise protective measures. Insect sex pheromones can be applied for different purposes in protecting crops such as monitoring, determining the species composition and combating harmful species (by using sex pheromones for disorientation). The present work studies the species composition, dynamics of the number of basic pests of an apple tree, synchronisation of the seasonal and circadian activity of phytophages of an apple tree in the central zone of the Krasnodar Territory. The results of the field assessment of the male complex disorientation method of apple and eastern moths are presented. It was shown that the species-specificity of sex pheromones in the apple orchard depends on the faunistic diversity of Lepidoptera species with similar pheromone systems that develop at a given point in space and time. It was revealed that the behaviour of this complex changes during the season and over years, depending on the climate and natural dynamics of insect populations. The quantitative ratio and species-specificity of pheromones will probably be different in ecosystems with various species composition and different geographic zones. A novelty of this research is the division of the studied Lepidoptera phytophage species into three groups according to a decrease in the absolute species-specificity in the forest biotope in comparison with the garden one. The most widespread and coinciding in terms of summer synchronicity are apple (Cydia pomonella L.), plum (Grapholitha funebrana Tr.), eastern (Grapholitha molesta Tr.) and pomegranate moth (Euzophera bigella Zell.). The disorientation method, applied using a complex system of dispensers with apple and eastern moth pheromones, showed that installing 500 dispensers/ha allowed 99.3 % efficiency to be achieved. Fruit damage amounted to 1.2 % and 2.7 % in the experimental and control groups, respectively. The duration of the disorienting effect of the pheromone formulations lasted for over 4 months.

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

Simple SummaryIt is well acknowledged that some of the volatile plant compounds (VPC) naturally present in insect natural habitats alter the perception of their own pheromone when presented individually as a background to pheromone. However, the effects of mixing VPCs as they appear to insects in natural olfactory landscapes are poorly understood. We measured the activity of brain neurons and neurons that detect a sex pheromone component in a moth antenna, while exposed to simple or composite backgrounds of VPCs representative of the odorant variety encountered by this moth. Maps of activities were built using calcium imaging to visualize which brain areas were most affected by VPCs. In the antenna, we observed differences in VPC capacity to elicit firing response that cannot be explained by differences in stimulus intensities because we adjusted concentrations according to volatility. The neuronal network, which reformats the input from antenna neurons in the brain, did not improve pheromone salience. We postulate that moth olfactory system evolved to increase sensitivity and encode fast changes of concentration 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 more important than background complexity.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.