Adverse effects of the Bordeaux mixture copper-based fungicide on the non-target vineyard pest Lobesia botrana.

BACKGROUND: Bordeaux mixture is a copper-based fungicide commonly used in vineyards to prevent fungal and bacterial infections in grapevines. However, this fungicide may adversely affect the entomological component, including insect pests. Understanding the impacts of Bordeaux mixture on the vineyard pest Lobesia botrana is an increasing concern in the viticultural production. RESULTS: Bordeaux mixture had detrimental effects on the development and reproductive performance of L. botrana. Several physiological traits were adversely affected by copper-based fungicide exposure, including a decrease in larval survival and a delayed larval development to moth emergence, as well as a reduced reproductive performance through a decrease in female fecundity and fertility and male sperm quality. However, we did not detect any effect of Bordeaux mixture on the measured reproductive behaviors (mating success, pre-mating latency and mating duration). CONCLUSION: Ingestion by larvae of food contaminated with Bordeaux mixture had a negative effect on the reproductive performance of the pest L. botrana, which could affect its population dynamics in vineyards. Although this study highlighted collateral damage of Bordeaux mixture on L. botrana, the potential impact of copper-based fungicides on vineyard diversity, including natural predators is discussed and needs to be taken in consideration in integrated pest management. © 2024 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Arthropods on grapes benefit more from fungicide reduction than from organic farming.

BACKGROUND: Pesticides are considered main contributors to global arthropod declines and therefore may decrease the provision of ecosystem services such as natural pest control. Organic farming and cultivating pest- and disease-resistant varieties can allow pesticide applications and their impacts on nontarget organisms and the environment to be reduced. We investigated the effects of organic versus conventional management and fungus-resistant versus susceptible wine grape varieties on arthropod biodiversity and pest control of grape berry moths in 32 vineyards in the Palatinate region, Germany. Hazard quotients of applied pesticides were calculated for each vineyard. RESULTS: The cultivation of fungus-resistant varieties led to significantly reduced hazard quotients and in turn enhanced abundances of natural enemies, particularly theridiid and philodromid spiders. Unexpectedly, organic management resulted in higher hazard quotients than conventional management and reduced numbers of natural enemies, particularly earwigs. Pest predation rates showed no significant differences between grape varieties or management types. CONCLUSION: Widespread benefits of organic management on arthropod biodiversity found in other crops were absent in our viticultural study region. This is likely due to the dominant role of fungal diseases in viticulture, which requires high numbers of fungicide treatments under both conventional and organic viticulture. Thus, fungicide reduction through the cultivation of fungus-resistant grape varieties is one key element to fostering the abundance of arthropods in general and beneficial arthropods in particular. Beyond vineyards, this is potentially relevant in numerous other crop types. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

The deterrent ability of Xenorhabdus nematophila and Photorhabdus laumondii compounds as a potential novel tool for Lobesia botrana (Lepidoptera: Tortricidae) management.

The grapevine moth, Lobesia botrana (Lepidoptera: Tortricidae), is a critical pest for vineyards and causes significant economic losses in wine-growing areas worldwide. Identifying and developing novel semiochemical cues (e.g. volatile bacterial compounds) which modify the ovipositional and trophic behaviour of L. botrana in vineyard fields could be a novel control alternative in viticulture. Xenorhabdus spp. and Photorhabdus spp. are becoming one of the best-studied bacterial species due to their potential interest in producing toxins and deterrent factors. In this study, we investigated the effect of the deterrent compounds produced by Xenorhabdus nematophila and Photorhabdus laumondii on the ovipositional moth behaviour and the larval feeding preference of L. botrana. Along with the in-vitro bioassays performed, we screened the potential use of 3 d cell-free bacterial supernatants and 3 and 5 d unfiltered bacterial ferments. In addition, we tested two application systems: (i) contact application of the bacterial compounds and (ii) volatile bacterial compounds application. Our findings indicate that the deterrent effectiveness varied with bacterial species, the use of bacterial cell-free supernatants or unfiltered fermentation product, and the culture times. Grapes soaked in the 3 d X. nematophila and P. laumondii ferments had âˆ¼ 55% and âˆ¼ 95% fewer eggs laid than the control, respectively. Likewise, the volatile compounds emitted by the 5 d P. laumondii fermentations resulted in âˆ¼ 100% avoidance of L. botrana ovipositional activity for three days. Furthermore, both bacterial fermentation products have larval feeding deterrent effects (∼65% of the larva chose the control grapes), and they significantly reduced the severity of damage caused by third instar larva in treated grapes. This study provides insightful information about a novel bacteria-based tool which can be used as an eco-friendly and economical alternative in both organic and integrated control of L. botrana in vineyard.

Big Data and Machine Learning to Improve European Grapevine Moth (Lobesia botrana) Predictions.

Machine Learning (ML) techniques can be used to convert Big Data into valuable information for agri-environmental applications, such as predictive pest modeling. Lobesia botrana (Denis & Schiffermüller) 1775 (Lepidoptera: Tortricidae) is one of the main pests of grapevine, causing high productivity losses in some vineyards worldwide. This work focuses on the optimization of the Touzeau model, a classical correlation model between temperature and L. botrana development using data-driven models. Data collected from field observations were combined with 30 GB of registered weather data updated every 30 min to train the ML models and make predictions on this pest's flights, as well as to assess the accuracy of both Touzeau and ML models. The results obtained highlight a much higher F1 score of the ML models in comparison with the Touzeau model. The best-performing model was an artificial neural network of four layers, which considered several variables together and not only the temperature, taking advantage of the ability of ML models to find relationships in nonlinear systems. Despite the room for improvement of artificial intelligence-based models, the process and results presented herein highlight the benefits of ML applied to agricultural pest management strategies.

Identification of Exoenzymes Secreted by Entomopathogenic Fungus Beauveria pseudobassiana RGM 2184 and Their Effect on the Degradation of Cocoons and Pupae of Quarantine Pest Lobesia botrana.

Beauveria pseudobassiana RGM 2184 has shown 80% maximum efficacy against the pest Lobesia botrana in the autumn and winter seasons. This suggests that the strain possesses an interesting battery of enzymes that are cold-adapted to penetrate the thick and hydrophobic cocoon of L. botrana. In this study, screening of the proteolytic, lipolytic, and chitinolytic activity of enzyme extracts secreted by the RGM 2184 strain was carried out in various culture media. The enzyme extracts with the highest activity were subjected to zymography and mass spectrometry. These analyses allowed the identification of two proteases, two lipases, and three chitinases. Comparative analysis indicated that the degree of similarity between these enzymes was substantially reduced when the highest degree of taxonomic relatedness between RGM 2184 and the entomopathogenic fungus strain was at the family level. These results suggest that there is a wide variety of exoenzymes in entomopathogenic fungi species belonging to the order Hypocreales. On the other hand, exoenzyme extract exposure of cocoons and pupae of L. botrana provoked damage at 10 °C. Additionally, an analysis of the amino acid composition of the RGM 2184 exoenzyme grouped them close to the cold-adapted protein cluster. These results support the use of this strain to control pests in autumn and winter. Additionally, these antecedents can form a scaffold for the future characterization of these exoenzymes along with the optimization of the strain's biocontrol ability by overexpressing them.

The causes and consequences of pest population variability in agricultural landscapes.

Variability in population densities is key to the ecology of natural systems but also has great implications for agriculture. Farmers' decisions are heavily influenced by their risk aversion to pest outbreaks that result in major yield losses. However, the need for long-term pest population data across many farms has prevented researchers from exploring the drivers and implications of pest population variability (PV). Here, we demonstrate the critical importance of PV for sustainable farming by analyzing 13 years of pest densities across >1300 Spanish olive groves and vineyards. Variable populations were more likely to cause major yield losses, but also occasionally created temporal windows when densities fell below insecticide spray thresholds. Importantly, environmental factors regulating pest variability were very distinct from factors regulating mean density, suggesting variability needs to be uniquely managed. Finally, we found diversifying landscapes may be a win-win situation for conservation and farmers, as diversified landscapes promote less abundant and less variable pest populations. Therefore, we encourage agricultural stakeholders to increase the complexity of the landscapes surrounding their farms through conserving/restoring natural habitat and/or diversifying crops.

Sexual audience affects male's reproduction investment without consequences on reproductive outputs.

Males evolved plastic strategies to respond to male-male competition and exhibit adaptive traits and behaviors maximizing their access to the females and limiting sperm competition. Mating behaviors allow males to express quick responses to current sexual audience, that is, the number of nearby conspecifics prone to mate. In contrast, physiological responses are frequently delayed because they are constrained by the time and resources having to be mobilized to produce and export sperm and associated products. This is especially critical in species for which males produce spermatophores. Here we investigated in what extend moth males (the tortricid moth Lobesia botrana) producing spermatophores exhibit plastic behavioral and physiological responses to different sexual audiences before and during mating and the consequences for their reproductive output. We found that males adjusted their mating behaviors and spermatophore size to a potentially elevated risk of sperm competition perceived before mating. In addition, males responded to the closed presence of females during mating by reducing their mating duration. Surprisingly, the various behavioral and physiological responses we highlighted here were not fully reflected in their reproductive performance as we did not reveal any effect on fecundity and fertility of their mate. The selective pressure exerted on males experiencing male-male competition could thus be sufficient to trigger adjustment in male mating behaviors but constrains physiological responses according to the perception of competition.

Dual effects of entomopathogenic fungi on control of the pest Lobesia botrana and the pathogenic fungus Eutypella microtheca on grapevine.

BACKGROUND: Entomopathogenic fungi (EPF) are the natural enemies of insect pests. Nevertheless, research on the use of EPF for simultaneous prevention of pest and disease agents on the same crop is limited. In this study, we explored the potential dual effects of three strains of the EPF Metarhizium anisopliae on the control of detrimental agents of Vitis vinifera L., including different developmental stages (larvae, pupae, and adult) of the insect pest Lobesia botrana and the phytopathogenic fungus Eutypella microtheca. METHODS: Laboratory pathogenicity trials were performed to examine the effects of the three M. anisopliae strains on the mortality rate of L. botrana. In addition, field trials were conducted to assess the biocontrol potential of one selected M. anisopliae strain on the larval stage of L. botrana. Moreover, inhibitory effects of the three EPF strains on E. microtheca growth were examined in vitro. RESULTS: All the M. anisopliae strains were highly effective, killing all stages of L. botrana as well as inhibiting the growth of E. microtheca. The in vitro mortality of larvae treated with the strains was over 75%, whereas that of treated pupae and adults was over 85%. The three EPF strains showed similar efficacy against larvae and adult stages; nevertheless, pupal mortality was observed to be strain dependent. Mortality of L. botrana larvae ranged from 64 to 91% at field conditions. Inhibition of E. microtheca growth reached 50% in comparison to the control. CONCLUSIONS: Our study showed that M. anisopliae strains were highly effective in ensuring control of two different detrimental agents of V. vinifera L., providing new evidence to support the dual effects of entomopathogenic fungi.

Feeding Volatiles of Larval Sparganothis pilleriana (Lepidoptera: Tortricidae) Attract Heterospecific Adults of the European Grapevine Moth.

Plants release volatiles in response to caterpillar feeding. These herbivore-induced plant volatiles (HIPVs) attract natural enemies of the herbivores and repel or attract conspecific adult herbivores in a tri-trophic interaction which has been considered to be an indirect plant defense against herbivores. Recently, we demonstrated the attraction of male and female European grapevine moth, Lobesia botrana (Denis & Schiffermüller) (Lepidoptera: Tortricidae) to a blend of phenylacetonitrile and acetic acid, two compounds identified as HIPVs in heterospecific apple-leafroller interactions. The ecological basis of our findings is not clearly understood. Thus, this work was undertaken to investigate HIPVs in the grapevine-leafroller interaction and study the response of heterospecific adults L. botrana, to these volatiles. We collected headspace volatiles emitted from uninfested grapevines and grapevines infested with larvae of a generalist herbivore, the grapevine leafroller moth, Sparganothis pilleriana (Denis & Schiffermüller), and analyzed them using gas chromatography/mass spectrometry. Infested grape leaves released three compounds (phenylacetonitrile, indole, and 2-phenylethanol) not found from uninfested leaves. Nine different blends, comprising a full factorial set of the three compounds with each blend containing acetic acid, were tested in a field-cage trial. Only lures containing phenylacetonitrile caused a significant increase in trap catches compared to the other lures and blank traps. Electroantennographic tests show that L. botrana can detect the compounds. The results confirm our hypothesis that phenylacetonitrile is released during grapevines infestation with herbivores, and attracts adult L. botrana.

Organic Farming and Cover-Crop Management Reduce Pest Predation in Austrian Vineyards.

Habitat simplification and intensive use of pesticides are main drivers of global arthropod declines and are, thus, decreasing natural pest control. Organic farming, complex landscapes, and local vineyard management practices such as implementation of flower-rich cover-crop mixtures may be a promising approach to enhance predator abundance and, therefore, natural pest control. We examined the effect of organic versus integrated management, cover-crop diversity in the vineyard inter-rows, and landscape composition on the natural pest control of Lobesia botrana eggs and pupae. Predation of L. botrana pupae was reduced by organic farming and species-poor cover-crops by about 10%. Predation rates of L. botrana eggs did not differ significantly in any of the studied management options. Dominant predators were earwigs (Forficulidae), bush crickets (Tettigoniidae), and ants (Formicidae). Negative effects of organic viticulture are most likely related to the negative nontarget effects on arthropods related to the frequent sulfur and copper applications in combination with the avoidance of strongly damaging insecticides by integrated winegrowers. While a 10% difference in predation rates on a single pest stage is unlikely to have strong practical implications, our results show that the assumed effectiveness of environmentally friendly agriculture needs to be evaluated for specific crops and regions.

Models Applied to Grapevine Pests: A Review.

This paper reviews the existing predictive models concerning insects and mites harmful to grapevine. A brief conceptual description is given on the definition of a model and about different types of models: deterministic vs. stochastics, continuous vs. discrete, analytical vs. computer-based, and descriptive vs. data-driven. The main biological aspects of grapevine pests covered by different types of models are phenology, population growth and dynamics, species distribution, and invasion risk. A particular emphasis is put on forecasting epidemics of plant disease agents transmitted by insects with sucking-piercing mouthparts. The most investigated species or groups are the glassy-winged sharpshooter Homalodisca vitripennis (Germar) and other vectors of Xylella fastidiosa subsp. fastidiosa, a bacterium agent of Pierce's disease; the European grape berry moth, Lobesia botrana (Denis and Schiffermuller); and the leafhopper Scaphoideus titanus Ball, the main vector of phytoplasmas agents of Flavescence dorée. Finally, the present and future of decision-support systems (DSS) in viticulture is discussed.

Lobesia botrana: A Biological Control Approach with a Biopesticide Based on Entomopathogenic Fungi in the Winter Season in Chile.

Lobesia botrana (Denis and Shiffermüller) (Lepidoptera: Tortricidae) is one of the main pests that affect the production and export of table grapes in Chile. Because this pest has quarantine status, the fruit must be fumigated with methyl bromide, which reduces the fruit's export competitiveness in the destination market. In the present study, to help resolve this issue, six native entomopathogenic fungi were identified through multilocus analysis, including three Beauveria pseudobassiana and three Metarhizium robertsii. These fungi were evaluated in the laboratory to control L. botrana in its pupal stage in a silk cocoon and compared against a biological control product. Formulations with additional carbon sources improved the performance of the fungi. The treatments with outstanding performance contained the fungal strains B. pseudobassiana RGM 2184 and M. robertsii RGM 678. These strains were evaluated in the field during the winter season in two different regions of the country; the strains reached maximum efficacies of 80% and 88%, respectively, at 21 days post first application. Therefore, entomopathogenic fungi can contribute to reducing pupal populations in winter, thereby decreasing the moth population in spring-summer.

Landscape simplification increases vineyard pest outbreaks and insecticide use.

Diversifying agricultural landscapes may mitigate biodiversity declines and improve pest management. Yet landscapes are rarely managed to suppress pests, in part because researchers seldom measure key variables related to pest outbreaks and insecticides that drive management decisions. We used a 13-year government database to analyse landscape effects on European grapevine moth (Lobesia botrana) outbreaks and insecticides across c. 400 Spanish vineyards. At harvest, we found pest outbreaks increased four-fold in simplified, vineyard-dominated landscapes compared to complex landscapes in which vineyards are surrounded by semi-natural habitats. Similarly, insecticide applications doubled in vineyard-dominated landscapes but declined in vineyards surrounded by shrubland. Importantly, pest population stochasticity would have masked these large effects if numbers of study sites and years were reduced to typical levels in landscape pest-control studies. Our results suggest increasing landscape complexity may mitigate pest populations and insecticide applications. Habitat conservation represents an economically and environmentally sound approach for achieving sustainable grape production.

Dual effects of entomopathogenic fungi on control of the pest Lobesia botrana and the pathogenic fungus Eutypella microtheca on grapevine

BACKGROUND: Entomopathogenic fungi (EPF) are the natural enemies of insect pests. Nevertheless, research on the use of EPF for simultaneous prevention of pest and disease agents on the same crop is limited. In this study, we explored the potential dual effects of three strains of the EPF Metarhizium anisopliae on the control of detrimental agents of Vitis vinifera L., including different developmental stages (larvae, pupae, and adult) of the insect pest Lobesia botrana and the phytopathogenic fungus Eutypella microtheca. METHODS: Laboratory pathogenicity trials were performed to examine the effects of the three M. anisopliae strains on the mortality rate of L. botrana. In addition, field trials were conducted to assess the biocontrol potential of one selected M. anisopliae strain on the larval stage of L. botrana. Moreover, inhibitory effects of the three EPF strains on E. microtheca growth were examined in vitro. RESULTS: All the M. anisopliae strains were highly effective, killing all stages of L. botrana as well as inhibiting the growth of E. microtheca. The in vitro mortality of larvae treated with the strains was over 75%, whereas that of treated pupae and adults was over 85%. The three EPF strains showed similar efficacy against larvae and adult stages; never-theless, pupal mortality was observed to be strain dependent. Mortality of L. botrana larvae ranged from 64 to 91% at field conditions. Inhibition of E. microtheca growth reached 50% in comparison to the control. CONCLUSIONS: Our study showed that M. anisopliae strains were highly effective in ensuring control of two different detrimental agents of V. vinifera L., providing new evidence to support the dual effects of entomopathogenic fungi.

Pest Management and Ochratoxin A Contamination in Grapes: A Review.

Ochratoxin A (OTA) is the most toxic member of ochratoxins, a group of toxic secondary metabolites produced by fungi. The most relevant species involved in OTA production in grapes is Aspergillus carbonarius. Berry infection by A. carbonarius is enhanced by damage to the skin caused by abiotic and biotic factors. Insect pests play a major role in European vineyards, and Lepidopteran species such as the European grapevine moth Lobesia botrana are undoubtedly crucial. New scenarios are also emerging due to the introduction and spread of allochthonous pests as well as climate change. Such pests may be involved in the dissemination of OTA producing fungi even if confirmation is still lacking and further studies are needed. An OTA predicting model is available, but it should be integrated with models aimed at forecasting L. botrana phenology and demography in order to improve model reliability.

Study of the Bacillus thuringiensis Cry1Ia Protein Oligomerization Promoted by Midgut Brush Border Membrane Vesicles of Lepidopteran and Coleopteran Insects, or Cultured Insect Cells.

Bacillus thuringiensis (Bt) produces insecticidal proteins that are either secreted during the vegetative growth phase or accumulated in the crystal inclusions (Cry proteins) in the stationary phase. Cry1I proteins share the three domain (3D) structure typical of crystal proteins but are secreted to the media early in the stationary growth phase. In the generally accepted mode of action of 3D Cry proteins (sequential binding model), the formation of an oligomer (tetramer) has been described as a major step, necessary for pore formation and subsequent toxicity. To know if this could be extended to Cry1I proteins, the formation of Cry1Ia oligomers was studied by Western blot, after the incubation of trypsin activated Cry1Ia with insect brush border membrane vesicles (BBMV) or insect cultured cells, using Cry1Ab as control. Our results showed that Cry1Ia oligomers were observed only after incubation with susceptible coleopteran BBMV, but not following incubation with susceptible lepidopteran BBMV or non-susceptible Sf21 insect cells, while Cry1Ab oligomers were persistently detected after incubation with all insect tissues tested, regardless of its host susceptibility. The data suggested oligomerization may not necessarily be a requirement for the toxicity of Cry1I proteins.

Potential biological control of the pupal stage of the European grapevine moth Lobesia botrana by the entomopathogenic fungus Beauveria pseudobassiana in the winter season in Chile.

OBJECTIVE: Lobesia botrana, the European grapevine moth, affects Vitis vinifera L. and other species of economic importance in a number of countries through damage caused by its larvae in berries and associated secondary diseases such as Botrytis cinerea. Control of the moth in urban areas is difficult due to poor chemical management of infested plants in houses. Additionally, in winter, L. botrana is in its pupal stage covered with a cocoon that prevents the penetration of chemical pesticides. For this reason, the objective of this work was to control the pupal stage with a formulation based on the entomopathogenic fungus Beauveria pseudobassiana in urban areas. RESULTS: The strain RGM 1747 was identified as B. pseudobassiana by multilocus sequence analysis. The biocontrol activity of this formulated fungus against the infestation of vines with breeding pupae without cocoons showed 100% infection 21 days after inoculation under winter conditions. Finally, the biocontrol activity of the formulated fungus against natural infestations of L. botrana in winter in urban areas reached an efficacy of 51%. This result suggests that the B. pseudobassiana formulation is able to penetrate the cocoon and contributes to the integrated pest management of L. botrana.

Leafroller-induced phenylacetonitrile and acetic acid attract adult Lobesia botrana in European vineyards.

We recently identified unique caterpillar-induced plant volatile compounds emitted from apple leaves infested with the larvae of various leafroller species. In subsequent field tests, binary blends of phenylacetonitrile+acetic acid and 2-phenylethanol+acetic acid were found to be attractive to a range of tortricid leafroller species (Tortricidae: Tortricinae) in both the Southern and Northern Hemispheres. In this work, the caterpillar-induced plant volatiles from the apple-leafroller system were tested in two vineyards in Spain and Hungary for their attractiveness to the grape frugivore Lobesia botrana (Tortricidae: Olethreutinae). As seen for Tortricinae species, a binary blend of phenylacetonitrile+acetic acid attracted significantly more male and female L. botrana to traps than acetic acid or blank lures. Traps baited with other caterpillar-induced plant volatile compounds (benzyl alcohol, 2-phenylethanol, indole, and (E)-nerolidol, each as a binary blend with acetic acid) did not catch significantly more moths than traps containing acetic acid alone. The catches of male and female moths support an optimistic future for new products in female tortricid surveillance and control that are based on combinations of kairomone compounds released from larval-damaged foliage.

Predicting Larval Stage Distribution of Lobesia botrana (Lepidoptera: Tortricidae) at Three Constant Temperatures.

European grapevine moth, Lobesia botrana (Denis and Schiffermüller), is an invasive pest species subject to regulatory control under the International Plant Protection Convention. An extensive infestation of this insect detected in California vineyards in October 2009 resulted in the initiation of postharvest research intended to ensure continuing trade of California table grapes by preventing the unintentional introduction of L. botrana to uninfested export destinations. Having the ability to accurately predict and confirm the particular stadia of larval development of insects obscured from direct observations, such as those being mass-reared in diet for use in phytosanitary control experiments, is necessary to be able to ensure that the most tolerant life stage is accurately targeted. Lobesia botrana were reared from egg to eclosion at three different temperatures: 16, 20, and 24°C (60% RH; 16:8 [L:D] h). Daily samples (≥15) were taken of larvae and pupae to determine the chronological distribution of life stages until daily samples comprised 100% pupae, after which only adults were counted. Head capsule measurements of all sampled larvae were taken (n = 2,820) and analyzed using nonlinear least squares (NLLS) fitting to Gaussian curves to describe stadia-specific head capsule demarcations and the probability of misclassifying instars due to overlapping life stage distributions.

Analysis of the grapevine moth Lobesia botrana antennal transcriptome and expression of odorant-binding and chemosensory proteins.

The grapevine moth, Lobesia botrana, is considered a harmful pest for vineyards in Chile as well as in North America and Europe. Currently, monitoring and control methods of L. botrana are based on its main sex pheromone component, being effective for low population densities. In order to improve control methods, antennal olfactory proteins in moths, such as odorant-binding proteins (OBPs) and odorant receptors (ORs) have been studied as promising targets for the discovery of new potent semiochemicals, which have not been reported for L. botrana. Therefore, the objective of this study was to identify the repertoire of proteins related to chemoreception in L. botrana by antennal transcriptome and analyze the relative expression of OBPs and CSPs in male and female antennae. Through next-generation sequencing of the antennal transcriptome by Ilumina HiSeq2500 we identified a total of 118 chemoreceptors, from which 61, 42 and 15 transcripts are related to ORs, ionotropic receptors (IRs) and gustatory receptors (GRs), respectively. Furthermore, RNA-Seq data revealed 35 transcripts for OBPs and 18 for chemosensory proteins (CSPs). Analysis by qRT-PCR showed 20 OBPs significantly expressed in female antennae, while 5 were more expressed in males. Similarly, most of the CSPs were significantly expressed in female than male antennae. All the olfactory-related sequences were compared with homologs and their phylogenetic relationships elucidated. Finally, our findings in relation to the improvement of L. botrana management are discussed.