Trehalose-6-Phosphate Synthase Contributes to Rapid Cold Hardening in the Invasive Insect Lissorhoptrus oryzophilus (Coleoptera: Curculionidae) by Regulating Trehalose Metabolism.

Rapid cold hardening (RCH) is known to rapidly enhance the cold tolerance of insects. Trehalose has been demonstrated to be a cryoprotectant in Lissorhoptrus oryzophilus, an important invasive pest of rice in China. Trehalose synthesis mainly occurs through the Trehalose-6-phosphate synthase (TPS)/trehalose-6-phosphate phosphatase (TPP) pathway in insects. In this study, the TPS gene from L. oryzophilus (LoTPS) was cloned and characterized for the first time. Its expression and trehalose content changes elicited by RCH were investigated. Our results revealed that RCH not only increased the survival rate of adults but also upregulated the expression level of LoTPS and increased the trehalose content under low temperature. We hypothesized that upregulated LoTPS promoted trehalose synthesis and accumulation to protect adults from low-temperature damage. To further verify the function of the LoTPS gene, we employed RNA interference (RNAi) technology. Our findings showed that RCH efficiency disappeared and the survival rate did not increase when the adults were fed dsRNA of LoTPS. Additionally, inhibiting LoTPS expression resulted in no significant difference in trehalose content between the RCH and non-RCH treatments. Furthermore, the expression patterns of trehalose transporter (TRET) and trehalase (TRE) were also affected. Collectively, these results indicate the critical role of LoTPS in L. oryzophilus cold resistance after RCH induction. LoTPS can enhance survival ability by regulating trehalose metabolism. These findings contribute to further understanding the role of TPS in insect cold resistance and the invasiveness of L. oryzophilus. Moreover, RNAi of LoTPS opens up possibilities for novel control strategies against L. oryzophilus in the future.

Synthesis of Imidazole-Based Molecules under Ultrasonic Irradiation Approaches.

Imidazole-based compounds are a series of heterocyclic compounds that exhibit a wide range of biological and pharmaceutical activities. However, those extant syntheses using conventional protocols can be time-costly, require harsh conditions, and result in low yields. As a novel and green technique, sonochemistry has emerged as a promising method for organic synthesis with several advantages over conventional methods, including enhancing reaction rates, improving yields, and reducing the use of hazardous solvents. Contemporarily, a growing body of ultrasound-assisted reactions have been applied in the preparation of imidazole derivatives, which demonstrated greater benefits and provided a new strategy. Herein, we introduce the brief history of sonochemistry and focus on the discussion of the multifarious approaches for the synthesis of imidazole-based compounds under ultrasonic irradiation and its advantages in comparison with conventional protocols, including typical name-reactions and various sorts of catalysts in those reactions.

Evaluation of Afidopyropen Toxicity at Environmentally Relevant Doses to the Asian Honeybee (Apis cerana) Using Physiological and Transcriptome Analysis.

Afidopyropen is a novel biogenic pesticide widely applied to control sap-feeding pests, and a few studies have evaluated the side effects of afidopyropen on pollinators, excluding the Asian honeybee. Thus, we estimated the physiological influences of afidopyropen in Apis cerana, which could cause significant death and nutritional deficiency in bees after continuous dietary intake (14 days) at the field recommended dose. Moreover, we found afidopyropen ingestion-induced changes in the activity of detoxification enzymes (AChE, GR, CarE) and expression of genes critical for the central nervous system and chemosensory function in the antennae, brain, midgut, and malpighian tubule of exposed bees. However, there was no evidence that there was a long-term impact on foraging activity when observing foragers treated with apfidopyropen as newly emerged workers. Overall, our study provides vital information to improve bee health, which will improve outcomes for beekeepers, increase pollination services, and strengthen pollinator communities.

Glutathione S-Transferase Highly Expressed in Holotrichia parallela Antennae Inactivates the Odorant Unsaturated Aldehyde Volatiles.

Odorant-degrading enzymes in insects play a vital role in maintaining olfactory sensitivity. However, the role and molecular mechanism of glutathione S-transferases (GSTs) in odorant inactivation has been rarely studied. In the present study, 31 GSTs were identified from the antennal transcriptome of Holotrichia parallela. HpGSTd1 possesses the highest transcriptome expression level. Recombinant HpGSTd1 showed degradation activity toward various unsaturated aldehyde volatiles. Furthermore, the metabolite of cinnamaldehyde was identified by high-resolution mass spectrometry (HRMS). The molecular docking analysis and site-directed mutagenesis revealed the key residues of HpGSTd1 in degrading odorants. In addition, the unsaturated aldehyde volatiles elicited the behavioral and electrophysiological responses of H. parallela. Taken together, our findings suggest that HpGSTd1 may play an essential role in inactivating odorants in H. parallela, which provides new insights for identifying molecular targets and exploring effective olfactory regulators for this underground pest.

Response of Odorant Receptors with Phenylacetaldehyde and the Effects on the Behavior of the Rice Water Weevil (Lissorhoptrus oryzophilus).

The rice water weevil (RWW), Lissorhoptrus oryzophilus Kuschel (Coleoptera: Curculionidae), is a destructive rice pest that threatens the rice industry worldwide. Odorant receptors (ORs) and odorant receptor coreceptors (Orcos) play an important role in the process of insects' whole life activities; however, there are no related functional studies on RWW. On this basis, a heterologous study of LoryOR20/LoryOrco in Xenopus laevis oocytes was performed to detect the effects of certain natural compounds on RWWs and four active compounds were found. Electroantennogram (EAG) recordings and a behavior test showed that RWWs exhibited a significant response to phenylacetaldehyde (PAA) and an EAG measurement of dsRNA-LoryOR20-treated RWWs revealed a significant decrease in response to PAA. Our results revealed an olfactory molecular mechanism for the recognition of PAA by RWWs, thus providing a potential genetic target at the peripheral olfactory sensing level, contributing to the development of novel control strategies for pest management.

1-nonene plays an important role in the response of maize-aphid-ladybird tritrophic interactions to nitrogen.

Plant volatile organic compounds (VOCs) are the key distress signals involved in tritrophic interactions, by which plants recruit predators to protect themselves from herbivores. However, the effect of nitrogen fertilization on VOCs that mediate tritrophic interactions remains largely unidentified. In this study, a maize (Zea mays)-aphid (Rhopalosiphum padi)-ladybird (Harmonia axyridis) tritrophic interaction model was constructed under high-nitrogen (HN) and low-nitrogen (LN) regimens. H. axyridis had a stronger tendency to be attracted by aphid-infested maize under HN conditions. Then, volatiles were collected and identified from maize leaves on which aphids had fed. All of the HN-induced volatiles (HNIVs) elicited an electroantennogram (EAG) response from H. axyridis. Of these HNIVs, 1-nonene was attractive to H. axyridis under simulated natural volatilization. Furthermore, our regression showed that the release of 1-nonene was positively correlated with H. axyridis visitation rates. Supplying 1-nonene to maize on which aphids had fed under LN enhanced attractiveness to H. axyridis. These results supported the conclusion that 1-nonene was the active compound that mediated the response to nitrogen in the tritrophic interaction. In addition, the 1-nonene synthesis pathway was hypothesized, and we found that the release of 1-nonene might be related to the presence of salicylic acid (SA) and abscisic acid (ABA). This research contributes to the development of novel environmentally friendly strategies to optimize nitrogen fertilizer application and to improve pest control in maize crops.

Application of the Reuterin System as Food Preservative or Health-Promoting Agent: A Critical Review.

The reuterin system is a complex multi-component antimicrobial system produced by Limosilactobacillus reuteri by metabolizing glycerol. The system mainly includes 3-hydroxypropionaldehyde (3-HPA, reuterin), 3-HPA dimer, 3-HPA hydrate, acrolein and 3-hydroxypropionic acid, and has great potential to be applied in the food and medical industries due to its functional versatility. It has been reported that the reuterin system possesses regulation of intestinal flora and anti-infection, anti-inflammatory and anti-cancer activities. Typically, the reuterin system exerts strong broad-spectrum antimicrobial properties. However, the antimicrobial mechanism of the reuterin system remains unclear, and its toxicity is still controversial. This paper presents an updated review on the biosynthesis, composition, biological production, antimicrobial mechanisms, stability, toxicity and potential applications of the reuterin system. Challenges and opportunities of the use of the reuterin system as a food preservative or health-promoting agent are also discussed. The present work will allow researchers to accelerate their studies toward solving critical challenges obstructing industrial applications of the reuterin system.

Purification and Functional Characterization of a Soluble Trehalase in Lissorhoptrus oryzophilus (Coleoptera: Curculionidae).

Trehalase is the only enzyme known for the irreversible splitting of trehalose and plays a major role in insect growth and development. In this report, we describe a basic study of the trehalase gene fragment encoding a soluble trehalase from Lissorhoptrus oryzophilus (LoTRE1). Sequence alignment and phylogenetic analysis suggested that LoTRE1 was similar to some known insect trehalases and belongs to the Coleoptera trehalase group. Additionally, LoTRE1 was expressed mainly in the fat body. Purified protein was obtained using heterologous expression of LoTRE1 in Escherichia coli, and the recombinant protein exhibited the ability to decompose trehalose. Enzyme-substrate docking indicated the potential involvement of other residues in the catalytic activity, in addition to Asp 333. Moreover, feeding of adults on LoTRE1 dsRNA silenced the transcription of LoTRE1 and thereby reduced the activity of trehalase and increased the trehalose content; it also led to a 12% death rate. This study reveals essential molecular features of trehalase and offers insights into the structural aspects of this enzyme, which might be related to its function. Taken together, the findings demonstrate that LoTRE1 is indispensable for adults of this pest and provide a new target for the control of L. oryzophilus.

Impact of repeated freeze-thaw cycles environment on the allelopathic effect to Secale cereale L. seedlings.

Allelopathy, as environmental stress, plays a prominent role in stress ecotoxicity, and global warming directly increases freeze-thaw cycles (FTCs) frequency in the winter. Yet, the effect between FTCs environment and allelopathy stress is rarely known, and the interaction of allelopathy stresses lacks consideration. Here, we addressed interactions between artemisinin stress (AS) and A. trifida extract stress (AES) under Non-FTCs and FTCs environments. The results found that AS and AES had an antagonistic relation under Non-FTCs environment, while a strong synergism and cooperation under FTCs environment affect the growth and physiology in S. cereale seedlings. Besides, AS and AES under FTCs environment had more inhibition on the growth of roots and shoots, chlorophylls, photosynthetic parameters, and relative water content; while more promotion on malondialdehyde, soluble sugar, and soluble protein. Moreover, the antioxidant enzyme activities of superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), and ascorbate peroxidase (APX) were increased by AS and AES, showing a good resistance of S. cereale seedlings to allelopathy stress, but FTCs environment significantly weakened this resistance. Thus, the allelopathic effect of AS and AES on S. cereale seedlings was significantly emphasized by FTCs environment.

Engineered chimeric insecticidal crystalline protein improves resistance to lepidopteran insects in rice (Oryza sativa L.) and maize (Zea mays L.).

The insecticidal crystalline proteins (Crys) are a family of insect endotoxin functioning in crop protection. As insects keep evolving into tolerance to the existing Crys, it is necessary to discover new Cry proteins to overcome potential threatens. Crys possess three functional domains at their N-termini, and the most active region throughout evolution was found at the domain-III. We swapped domain-IIIs from various Cry proteins and generated seven chimeric proteins. All recombinants were expressed in Escherichia coli and their toxicity was assessed by dietary exposure assays. Three of the seven Crys exhibited a high toxicity to Asian corn borer over the controls. One of them, Cry1Ab-Gc, a chimeric Cry1Ab being replaced with the domain-III of Cry1Gc, showed the highest toxicity to rice stem borer when it was over-expressed in Oryza sativa. Furthermore, it was also transformed into maize, backcrossed into commercial maize inbred lines and then produced hybrid to evaluate their commercial value. Transgenic maize performed significant resistance to the Asian corn borer without affecting the yield. We further showed that this new protein did not have adverse effects on the environment. Our results indicated that domain III swapped of Crys could be used as an efficient method for developing new engineered insecticidal protein.

Response characteristics of highland barley (Hordeum vulgare) seedlings to the stress of salinity and artemisinin under freeze­thaw environment.

In Qinghai-Tibet Plateau, crops are commonly subjected to freeze-thaw and salt stress factors simultaneously, and allelopathy is common, which affects the growth of highland barley (Hordeum vulgare L.), the largest food crop in Tibet. In order to explore the effects of artemisinin, salt and freeze-thaw (FAS) stress on physiological characteristics of highland barley seedlings, hydroponic experiment was carried out with the addition of 20mg/L artemisinin and 150mMNaCl as well as the simulation of freeze-thaw environment. The results suggested that under combined stress, the soluble protein content in combined stresses of artemisinin, FAS increased by 97.8%, the variation of relative conductivity in FAS group was lower than that in combined salt and freeze-thaw stress (FS), the relative water content decreased significantly (P <0.05), the malondialdehyde (MDA), H2 O2 and soluble sugar content in FAS group accumulated but less than those in FS group, and the superoxide dismutase (SOD) activity in combined artemisinin and freeze-thaw stress (FA) and FAS groups decreased. In addition, after freeze-thaw treatment, photosynthesis was weakened, and internal CO2 conentration (C i ) in FAS group significantly decreased (P <0.05). This study proved that appropriate amount of artemisinin can alleviate the damage of salt and freeze-thaw stress on barley seedlings.

Physiological response of barley seedlings to salinity and artemisinin combined stresses under freeze-thaw environment.

In the Qinghai-Tibet Plateau, both the large daily temperature difference and soil salinization make plants susceptible to abiotic stresses such as freeze-thaw and salinity. Meanwhile, crops in this area can be affected by artemisinin, an antimalarial secondary metabolite produced in Artemisia. Under freeze-thaw and salinity stresses, artemisinin was induced as an allelopathy stress factor to explore the physiological response of highland barley, including the relative electrical conductivity (RC), soluble protein (SP) content, malondialdehyde (MDA) content, antioxidant enzyme activity, and water use efficiency (WUE). Compared with the control group, the contents of RC and MDA in seedling leaves under stress were significantly increased by 24.74-402.37% and 20.18-77.95%, indicating that cell membrane permeability was greatly damaged, and WUE was significantly decreased by 15.77-238.59%. The activity of enzymes increased under single stress and decreased under combined stress. Salinity, artemisinin, and freeze-thaw stress show a synergistic relationship; that is, compound stresses were more serious than single stress. In summary, the results of this study revealed the physiological and ecological responses of barley seedlings under different habitat stresses and the interactions among different stress factors.

Response characteristics of highland barley under freeze-thaw, drought and artemisinin stresses.

The freeze-thaw of early spring in China's Qinghai-Tibet Plateau is often accompanied by severe droughts. Artemisia annua, widely distributed in China, releases allelopathic substances, mainly artemisinin, to the environment and exerts a wide range of effects on crops. This paper studied the physiological effects of highland barley under freeze-thaw, drought, and artemisinin stress through indoor simulation experiments. The physiological response characteristics of superoxide dismutase (SOD) activity, catalase (POD) activity, net photosynthetic rate, relative water content (RWC), relative electrical conductivity, malondialdehyde (MDA) content, and soluble protein content in highland barley were analyzed. The results showed that artemisinin and drought contributed to the increase of SOD activity and the decrease of POD activity. Under the freeze-thaw stress, the SOD and POD activities both decreased firstly and then increased, but the effect of compound stress on POD was more complicated. Either artemisinin, drought, or low temperature could reduce the net photosynthetic rate of highland barley. Low temperature had more significant impacts on photosynthesis, and compound stress would show a single stress superimposed effect. Artemisinin, drought, and low temperature could reduce the RWC of highland barley, and increase the relative electrical conductivity and the concentration of soluble protein (except for low temperature stress above zero, which reduces the concentration of soluble protein). However, the effect of compound stress on soluble protein is more complex. The single stress of artemisinin and drought had no obvious effect on MDA content, while the MDA content was increased significantly under the freeze-thaw stress and the compound stress of artemisinin and drought, and the MDA content reached its peak at T1. The results are helpful to explore the effects of freeze-thaw, drought and artemisinin stress on the growth of highland barley under the background of the aridification of the Qinghai-Tibet Plateau, and provide ideas for rational agricultural management.

The herbivore-induced plant volatile tetradecane enhances plant resistance to Holotrichia parallela larvae in maize roots.

BACKGROUND: Many herbivore-induced volatiles have been proven to act as signaling compounds to regulate nearby plant defense responses. However, the precise roles of key volatiles produced by maize roots after Holotrichia parallela larva feeding remain largely unknown. RESULTS: We investigated changes in phytohormones and volatiles in maize roots after H. parallela larval infestation. Marked increases in the phytohormone jasmonic acid (JA) and the volatiles jasmone and tetradecane were induced by herbivores, whereas the salicylic acid content decreased. In addition, pre-exposure to tetradecane markedly increased the levels of the stress hormone JA, its precursors and derivatives, and related gene expression. In addition, pre-exposure altered the production of defensive benzoxazinoid secondary metabolites, resulting in increased plant resistance to H. parallela larvae. Plants pre-exposed to jasmone did not differ from control plants. In addition, bioassays showed that H. parallela larval growth was suppressed by feeding maize roots after pre-exposure to tetradecane. CONCLUSION: These results demonstrate that tetradecane may function as a potent defense induction signal that prepares neighboring plants for incoming attacks. © 2021 Society of Chemical Industry.

Physiological effects of different stubble height and freeze-thaw stress on Secale cereale L. seedlings.

BACKGROUND: As a biennial plant, Secale cereale L is usually harvested in the autumn in the northern part of China where the temperature difference between day and night is of great disparity Through the pot experiment, the seedlings were cut to 2, 6 and 10 cm stubble height, and the simulated freeze-thaw (FT) stress (10/- 5 °C) was carried out after 6 days regrowth. The physiological effects of FT with different stubble height were revealed by analyzing the relative water content (RWC), osmotic adjustment substance concentration (soluble sugar and protein), membrane peroxidation (MDA) and catalase (CAT) activity. RESULTS: The results demonstrated that under freeze stress (- 5 °C), the content of soluble protein and MDA decreased and the seedlings of 2 cm treatment kept higher level of soluble protein and MDA, while the seedlings of 6 and 10 cm treatments kept higher level of the RWC, soluble sugar content, and CAT activity. After FT stress, the content of soluble sugar and protein, RWC in the 6 cm treatment were higher than those in 2 cm and 10 cm treatments, and the CAT activity in 10 cm treatment was the highest while the MDA content is lower. CONCLUSION: These data suggest that keeping high stubble height is more adaptive for short-term FT stress.

Identification and analysis of chemosensory genes encoding odorant-binding proteins, chemosensory proteins and sensory neuron membrane proteins in the antennae of Lissorhoptrus oryzophilus.

The rice water weevil, Lissorhoptrus oryzophilus Kuschel (Coleoptera: Curculionidae), is a destructive pest that causes damage to rice crops worldwide. The olfactory system is critical for host or mate location by weevils, but only limited information about the molecular mechanism of olfaction-related behaviour has been reported in this insect. In this study, we conducted SMRT-seq transcriptome analysis and obtained 54,378 transcripts, 38,706 of which were annotated. Based on these annotations, we identified 40 candidate chemosensory genes, including 31 odorant-binding proteins (OBPs), six chemosensory proteins (CSPs) and three sensory neuron membrane proteins (SNMPs). Phylogenetic analysis showed that LoryOBPs, LoryCSPs and LorySNMPs were distributed in various clades. The results of tissue expression patterns indicated that LoryOBPs were highly abundant in the antennae, whereas LoryCSPs were highly abundant not only in the antennae but also in the abdomen, head and wings. Our findings substantially expand the gene database of L. oryzophilus and may serve as a basis for identifying novel targets to disrupt key olfactory genes, potentially providing an eco-friendly strategy to control this pest in the future.

Identification of Candidate Carboxylesterases Associated With Odorant Degradation in Holotrichia parallela Antennae Based on Transcriptome Analysis.

Insects rely on their olfactory systems in antennae to recognize sex pheromones and plant volatiles in surrounding environments. Some carboxylesterases (CXEs) are odorant-degrading enzymes (ODEs), degrading odorant signals to protect the olfactory neurons against continuous excitation. However, there is no report about CXEs in Holotrichia parallela, one of the most major agricultural underground pests in China. In the present study, 20 candidate CXEs were identified based on transcriptome analysis of female and male antennae. Sequence alignments and phylogenetic analysis were performed to investigate the characterization of these candidate CXEs. The expression profiles of CXEs were compared by RT-qPCR analysis between olfactory and non-olfactory tissues of both genders. HparCXE4, 11, 16, 17, 18, 19, and 20 were antenna-biased expressed genes, suggesting their possible roles as ODEs. HparCXE6, 10, 11, 13, and 16 showed significantly higher expression profiles in male antennae, whereas HparCXE18 was expressed more in female antennae. This study highlighted candidate CXE genes linked to odorant degradation in antennae, and provided a useful resource for further work on the H. parallela olfactory mechanism and selection of target genes for integrative control of H. parallela.

Quantitative proteomics suggests changes in the carbohydrate metabolism of maize in response to larvae of the belowground herbivore Holotrichia parallela.

The larvae of Holotrichia parallela, a destructive belowground herbivore, may cause yield losses of up to 20% in maize in a typical year. To understand the protein-level mechanisms governing the response of maize to this herbivore, tandem mass tag (TMT) quantitative proteomics was used for the comparative analysis of protein abundance in the maize roots after H. parallela larval attack. A total of 351 upregulated proteins and 303 downregulated proteins were identified. Pathway enrichment analysis revealed that the differentially abundant proteins (DAPs) were most strongly associated with carbohydrate and energy metabolism pathways, such as glycolysis, pentose phosphate pathway and fructose and mannose metabolism. Most glycolysis-related proteins were significantly induced. In addition, H. parallela larval attack decreased the glucose concentrations in the roots. This study demonstrates that maize can manipulate carbohydrate metabolism by modifying glycolysis and pentose phosphate pathway response to root-feeding herbivorous attackers. The results of this study may help to establish a foundation for further functional studies of key protein-mediated responses to H. parallela larvae in maize.

Odorant binding proteins promote flight activity in the migratory insect, Helicoverpa armigera.

Migratory insects are capable of actively sustaining powered flight for several hours. This extraordinary phenomenon requires a highly efficient transport system to cope with the energetic demands placed on the flight muscles. Here, we provide evidence that the role of the hydrophobic ligand binding of odorant binding proteins (OBPs) extends beyond their typical function in the olfactory system to support insect flight activity via lipid interactions. Transcriptomic and candidate gene analyses show that two phylogenetically clustered OBPs (OBP3/OBP6) are consistently over-expressed in adult moths of the migrant Old-World bollworm, Helicoverpa armigera, displaying sustained flight performance in flight activity bioassays. Tissue-specific over-expression of OBP6 was observed in the antennae, wings and thorax in long-fliers of H. armigera. Transgenic Drosophila flies over-expressing an H. armigera transcript of OBP6 (HarmOBP6) in the flight muscle attained higher flight speeds on a modified tethered flight system. Quantification of lipid molecules using mass spectrometry showed a depletion of triacylglyerol and phospholipids in flown moths. Protein homology models built from the crystal structure of a fatty acid carrier protein identified the binding site of OBP3 and OBP6 for hydrophobic ligand binding with both proteins exhibiting a stronger average binding affinity with triacylglycerols and phospholipids compared with other groups of ligands. We propose that HarmOBP3 and HarmOBP6 contribute to the flight capacity of a globally invasive and highly migratory noctuid moth, and in doing so, extend the function of this group of proteins beyond their typical role as chemosensory proteins in insects.

Three Host Plant Volatiles, Hexanal, Lauric Acid, and Tetradecane, are Detected by an Antenna-Biased Expressed Odorant Receptor 27 in the Dark Black Chafer Holotrichia parallela.

Insects rely on olfaction to locate their host plants by antennae in complex chemical environments. Odorant receptor (OR) genes are thought to play a crucial role in the process. ORs function together with odorant coreceptors to determine the specificity and sensitivity of olfactory reception. The dark black chafer, Holotrichia parallela Motschulsky (Coleoptera: Scarabaeidae), is a destructive underground pest. To understand the molecular basis of H. parallela olfactory reception, an olfactory-biased expressed odorant receptor HparOR27 and HparOrco (HparOR40) were identified from antennal transcriptome analysis and prediction of the sequence structure. Tissue expression analysis showed that HparOR27 was mainly expressed in adult antennae throughout developmental stages. The functions of HparOR27 were analyzed using the Xenopus laevis oocyte expression system. HparOR27 was broadly responsive to three host plant volatiles, including hexanal, lauric acid, and tetradecane. Electroantennogram tests confirmed that three ligands were electrophysiologically active in antennae of female adults. A Y-tube olfactometer test indicated that hexanal was a repellent for adults of both sexes. Taken together, our data support the identification of odorant receptors and provide a molecular basis for eco-friendly pest control.