Limonene enhances rice plant resistance to a piercing-sucking herbivore and rice pathogens.

Terpene synthases (TPSs) are key enzymes in terpenoids synthesis of plants and play crucial roles in regulating plant defence against pests and diseases. Here, we report the functional characterization of OsTPS19 and OsTPS20, which were upregulated by the attack of brown planthopper (BPH). BPH female adults performed concentration-dependent behavioural responses to (S)-limonene showing preference behaviour at low concentrations and avoidance behaviour at high concentrations. Overexpression lines of OsTPS19 and OsTPS20, which emitted higher amounts of the monoterpene (S)-limonene, decreased the hatching rate of BPH eggs, reduced the lesion length of sheath blight caused by Rhizoctonia solani and bacterial blight caused by Xanthomonas oryzae. While knockout lines of OsTPS19 and OsTPS20, which emitted lower amounts of (S)-limonene, were more susceptible to these pathogens. Overexpression of OsTPS19 and OsTPS20 in rice plants had adverse effects on the incidence of BPH, rice blast, and sheath blight in the field and had no significant impacts on rice yield traits. OsTPS19 and OsTPS20 were found to be involved in fine-tuning the emission of (S)-limonene in rice plants and play an important role in defence against both BPH and rice pathogens.

A Highly Expressed Antennae Odorant-Binding Protein Involved in Recognition of Herbivore-Induced Plant Volatiles in Dastarcus helophoroides.

Natural enemies such as parasitoids and parasites depend on sensitive olfactory to search for their specific hosts. Herbivore-induced plant volatiles (HIPVs) are vital components in providing host information for many natural enemies of herbivores. However, the olfactory-related proteins involved in the recognition of HIPVs are rarely reported. In this study, we established an exhaustive tissue and developmental expression profile of odorant-binding proteins (OBPs) from Dastarcus helophoroides, an essential natural enemy in the forestry ecosystem. Twenty DhelOBPs displayed various expression patterns in different organs and adult physiological states, suggesting a potential involvement in olfactory perception. In silico AlphaFold2-based modeling and molecular docking showed similar binding energies between six DhelOBPs (DhelOBP4, 5, 6, 14, 18, and 20) and HIPVs from Pinus massoniana. While in vitro fluorescence competitive binding assays showed only recombinant DhelOBP4, the most highly expressed in the antennae of emerging adults could bind to HIPVs with high binding affinities. RNAi-mediated behavioral assays indicated that DhelOBP4 was an essential functional protein for D. helophoroides adults recognizing two behaviorally attractive substances: p-cymene and γ-terpinene. Further binding conformation analyses revealed that Phe 54, Val 56, and Phe 71 might be the key binding sites for DhelOBP4 interacting with HIPVs. In conclusion, our results provide an essential molecular basis for the olfactory perception of D. helophoroides and reliable evidence for recognizing the HIPVs of natural enemies from insect OBPs' perspective.

Silencing of an odorant binding protein (SaveOBP10) involved in the behavioural shift of the wheat aphid Sitobion avenae (Fabricius).

Insects are highly reliant on their active olfactory system in which odorant binding proteins play a role to selectivity and sensitivity during odour perception and processing. This study sets out to determine whether and to which extent the antennal loaded SaveOBP10 in English grain aphid Sitobion avenae, contributes in olfactory processing during host selection. To understand this possible relationship, we purified the SaveOBP10 recombinant protein and performed fluorescence ligand binding tests, molecular docking, RNA interference (RNAi) and behavioural trials. The results showed that SaveOBP10 had strong binding affinities (Ki ≤5 µM) with most of wheat plant volatiles at pH 5.0 as compared to pH 7.4. In Y-tube olfactometer bioassays, the S. avenae was attracted behaviourally towards pentadecane, butylated hydroxytoluene, tetradecane and ß-caryophyllene however repelled by naphthalene. After RNAi of SaveOBP10, the aphid showed nonattraction towards ß-caryophyllene and nonsignificant behavioural response to pentadecane, butylated hydroxytoluene and tetradecane. Furthermore, the three-dimensional structure modelling and molecular docking of SaveOBP10 were performed to the volatiles with high binding abilities. Together these findings indicate that SaveOBP10 can bind more strongly to the volatiles that involved in S. avenae behaviour regulation and possibly will contribute effectively in S. avenae integrated pest management.

Sublethal Exposure to Cadmium Induces Chemosensory Dysfunction in Fire Ants.

Ants easily accumulate cadmium (Cd) from the food web in terrestrial ecosystems. Cd contamination may cause olfactory dysfunction and consequently disorders in the social behavior of ants. To explore the molecular mechanism underlying the effect of Cd exposure on the chemosensory process of ants, we characterized the Cd-induced variations in the expression of genes involved in chemoreception and electrophysiological and behavioral sensitivity to semiochemicals by using the red imported fire ant, Solenopsis invicta, as a model system. As a result, Cd exposure increased Cd accumulation and decreased the survival rate of S. invicta. Cd exposure altered the expression profiles of odor binding protein genes of S. invicta (SiOBPs). Specifically, SiOBP15 protein expression was upregulated upon Cd exposure. Both SiOBP7 and SiOBP15 exhibited high binding affinities to limonene, nonanal, and 2,4,6-trimethylpyridine. S. invicta exposed to Cd showed less sensitive electrophysiological and behavioral response to the three chemicals but exhibited sensitive perception to undecane. Silencing of SiOBP7 and SiOBP15 abolished the behavioral response of S. invicta to nonanal and undecane, respectively, suggesting that SiOBP7 and SiOBP15 play essential roles in the chemoreception of S. invicta. In general, our results suggest that Cd contamination may interfere with olfactory signal transduction by altering the expression of SiOBPs, consequently evoking chemosensory dysfunction in fire ants.

Molecular Characterization and Expression Patterns of Two Pheromone-Binding Proteins from the Diurnal Moth Phauda flammans (Walker) (Lepidoptera: Zygaenoidea: Phaudidae).

Sex pheromone-binding proteins (PBPs) play an important role in sex pheromone recognition in Lepidoptera. However, the mechanisms of chemical communication mediating the response to sex pheromones remain unclear in the diurnal moths of the superfamily Zygaenoidea. In this study, Phauda flammans (Walker) (Lepidoptera: Zygaenoidea: Phaudidae) was used as a model insect to explore the molecular mechanism of sex pheromone perception in the superfamily Zygaenoidea. Two novel pheromone-binding proteins (PflaPBP1 and PflaPBP2) from P. flammans were identified. The two pheromone-binding proteins were predominantly expressed in the antennae of P. flammans male and female moths, in which PflaPBP1 had stronger binding affinity to the female sex pheromones Z-9-hexadecenal and (Z, Z, Z)-9, 12, 15-octadecatrienal, PflaPBP2 had stronger binding affinity only for (Z, Z, Z)-9, 12, 15-octadecatrienal, and no apparent binding affinity to Z-9-hexadecenal. The molecular docking results indicated that Ile 170 and Leu 169 are predicted to be important in the binding of the sex pheromone to PflaPBP1 and PflaPBP2. We concluded that PflaPBP1 and PflaPBP2 may be responsible for the recognition of two sex pheromone components and may function differently in female and male P. flammans. These results provide a foundation for the development of pest control by exploring sex pheromone blocking agents and the application of sex pheromones and their analogs for insect pests in the superfamily Zygaenoidea.

A Salivary Odorant-Binding Protein Mediates Nilaparvata lugens Feeding and Host Plant Phytohormone Suppression.

Odorant-binding proteins (OBPs) typically act as transporters of odor molecules and play an important role in insect host location. Here, we identified an OBP in brown planthopper (BPH) Nilaparvata lugens salivary glands via transcriptome sequencing. Real-time quantitative PCR and Western blotting analysis results showed that NlugOBP11 was highly expressed in salivary glands and secreted into rice plant during feeding, suggesting that it assists in BPH feeding on rice. Functional analysis in N. lugens saliva revealed that silencing this gene by RNA interference decreased the BPH stylet performance in the phloem of rice plants, reduced sap sucking, and ultimately led to insect death. Moreover, overexpression of NlugOBP11 in rice protoplasts or Nicotiana benthamiana leaves inhibited the production of defense-related signaling molecule salicylic acid in rice plant. The results demonstrate that NlugOBP11 is not only essential for BPH feeding, but also acts as an effector that inhibits plant defense.

Construction, petro-collecting/dispersing capacities, antimicrobial activity, and molecular docking study of new cationic surfactant-sulfonamide conjugates.

Surfactants with their diverse activities have been recently involved in controlling the spread of new coronavirus (COVID-19) pandemic as they are capable of disrupting the membrane surrounding the virus. Using hybrids approach, we constructed a novel series of cationic surfactant-sulfonamide conjugates (3a-g) through quaternization of the as-prepared sulfonamide derivatives (2a-g) with n-hexadecyl iodide followed by structural characterization by spectroscopy (IR and NMR). Being collective properties required in petroleum-processing environment, the petro-collecting/dispersing capacities on the surface of waters with different degrees of mineralization, and the antimicrobial performance against microbes and sulfate-reducing bacteria (SRB) that mitigate microbiological corrosion were investigated for the synthesized conjugates. Among these conjugates, 3g (2.5% aq. solution) exhibited the strongest ability to disperse the thin petroleum film on the seawater surface, whereas KD is 95.33% after 96 h. In diluted form, 3f collected the petroleum layer on distilled water surface (Kmax = 32.01) for duration exceeds 4 days. Additionally, almost all compounds revealed high potency and comparable action with standard antimicrobials, especially 3b and 3f, which emphasize their role as potential biocides. Regarding biocidal activity against SRB, 3g causes a significant reduction in the bacterial count from 2.8 × 106 cells/mL to Nil. Moreover, the conducted molecular docking study confirms the strong correlation between RNA polymerase binding with bioactivity against microbes over other studied proteins (threonine synthase and cyclooxygenase-2).

Expression Profiles and Biochemical Analysis of Chemosensory Protein 3 from Nilaparvata lugens (Hemiptera: Delphacidae).

Insects have evolved highly sensitive olfactory sensory systems to detect plant hosts and mates, with plant volatiles playing an important role in informing insect behavior. Chemosensory proteins (CSPs) are thought to play a key role in this process, but in this respect, there is limited information on brown planthopper Nilaparvata lugens, one of the most destructive pests of rice. To expand our understanding of CSP function in N. lugens we explored expression profiles and binding characteristics of NlugCSP3. The ligands with higher binding affinity were also validated by molecular docking and behavioral assays. NlugCSP3 mRNA was expressed at relatively higher levels in antennae and abdomen of 3-day-old unmated macropterous males as well as in antennae of 3-day mated macropterous and brachypterous females. Fluorescence competitive binding assays revealed that 5 out of 25 candidate volatiles are strong binders (Ki < 10 µM). Behavioral assays revealed that nonadecane and 2-tridecanone, which have high binding affinities in fluorescence competition-binding assays, displayed strong attractiveness to N. lugens. Pursuing this further, molecular docking analysis identified key amino acid residues involved in binding volatile compounds. Overall, our data provide a base for further investigation of the potential physiological functions of CSP3 in Nilaparvata lugens, and extend the function of NlugCSP3 in chemoreception of N. lugens.

The larvicidal activity of natural inspired piperine-based dienehydrazides against Culex pipiens.

A series of piperine-based dienehydrazide derivatives were designed and synthesized to be used as insecticides against Culex pipiens. The chemical structure of compound 5n was confirmed by single-crystal x-ray diffraction. Their insecticidal activities of synthesized compounds were tested against third-instar larval of Cx. pipiens at concentrations ranging from 0.1 to 1.2 mg/mL. Among all derivatives, compounds 5a, 5b, 5f, 5g, 5m, 5n, 5o, 5p, and 5u displayed good activities. The final mortality rates at the concentration of 0.75 mg/mL after 48 h treatment, were found to be in the range from 80.00 to 83.33% and with LC50 values ranging from 0.221 to 0.094 mg/mL. These compounds demonstrated higher insecticidal activities than piperine and Deltamethrin (a commercial positive control). Molecular modelling reveals several molecular interactions between synthesized compounds and the substrate binding sits of acetylcholinesterase (AChE) that are predicted to be responsible for its binding and inhibition activity. .

An Odorant Binding Protein (SaveOBP9) Involved in Chemoreception of the Wheat Aphid Sitobion avenae.

Odorant binding proteins play a key role in the olfactory system and are involved in the odor perception and discrimination of insects. To investigate the potential physiological functions of SaveOBP9 in Sitobion avenae, fluorescence ligand binding experiments, molecular docking, RNA interference, and behavioral tests were performed. Fluorescence binding assay results showed that SaveOBP9 had broad and high (Ki < 10 µM) binding abilities with most of the wheat volatiles, but was more obvious at pH 7.4 than pH 5.0. The binding sites of SaveOBP9 to the volatiles were predicted well by three-dimensional docking structure modeling and molecular docking. Moreover, S. avenae showed a strong behavioral response with the four compounds of wheat. The reduction in mRNA transcript levels after the RNA interference significantly reduced the expression level of SaveOBP9 and induced the non-significant response of S. avenae to the tetradecane, octanal, decanal, and hexadecane. This study provides evidence that SaveOBP9 might be involved in the chemoreception of wheat volatile organic compounds and can successfully contribute in the integrated management programs of S. avenae.

Rice defense responses are induced upon leaf rolling by an insect herbivore.

BACKGROUND: Plant defense against herbivores begins with perception. The earlier plant detects the harm, the greater plant will benefit in its arm race with the herbivore. Before feeding, the larvae of the rice pest Cnaphalocrocis medinalis, initially spin silk and fold up a leaf. Rice can detect and protect itself against C. medinalis feeding. However, whether rice could perceive C. medinalis leaf rolling behavior is currently unknown. Here, we evaluated the role of leaf rolling by C. medinalis and artificial leaf rolling in rice plant defense and its indirect effect on two important C. medinalis parasitoids (Itoplectis naranyae and Apanteles sp.) through a combination of volatile profiling, gene-transcriptional and phytohormonal profiling. RESULTS: Natural leaf rolling by C. medinalis resulted in an increased attraction of I. naranyae when compared to the undamaged plant after 12 h. Volatile analysis revealed that six out of a total 22 components significantly increased in the headspace of C. medinalis rolled plant when compared to undamaged plant. Principal component analysis of these components revealed similarities in the headspace of undamaged plant and artificially rolled plant while the headspace volatiles of C. medinalis rolled plant deferred significantly. Leaf rolling and feeding by C. medinalis up-regulated the plant transcriptome and a series of jasmonic acid (JA) and salicylic acid (SA) related genes. While feeding significantly increased JA level after 12 to 36 h, rolling significantly increased SA level after 2 to 12 h. Compared to artificial rolling, natural rolling significantly increased JA level after 36 h and SA level after 2 and 12 h. CONCLUSIONS: Our findings suggest that natural leaf rolling by C. medinalis can be perceived by rice plant. The detection of this behavior may serve as an early warning signal in favor of the rice plant defenses against C. medinalis.

Molecular Screening of Behaviorally Active Compounds with CmedOBP14 from the Rice Leaf Folder Cnaphalocrocis medinalis.

Odorant binding proteins (OBPs) play a key role in chemoreception in insects. In an earlier study, we identified CmedOBP14 from the rice leaf folder, Cnaphalocrocis medinalis, with potential physiological functions in olfaction. Here, we performed a competitive binding assay under different pH conditions as well as knockdown via RNA interference to determine the specific role of CmedOBP14 in C. medinalis. CmedOBP14 displayed broad binding affinities to many host-related compounds, with higher affinities at pH 7.4 compared with pH 5.0. After treatment with CmedOBP14-dsRNA, the transcript level of OBP14 was significantly decreased at 72 h compared with controls, and the electroantennogram response evoked by nerolidol, L-limonene and beta-ionone was reduced. Furthermore, behavioral assays revealed consistent patterns among these compounds, especially for nerolidol, with adults could no longer able to differentiate 0.1% nerolidol from controls. RNAi experiments suggest that at least in part, CmedOBP14 mediates the ability to smell nerolidol and beta-ionone.

Structural Transformation Detection Contributes to Screening of Behaviorally Active Compounds: Dynamic Binding Process Analysis of DhelOBP21 from Dastarcus helophoroides.

In light of reverse chemical ecology, the fluorescence competitive binding assays of functional odorant binding proteins (OBPs) is a recent advanced approach for screening behaviorally active compounds of insects. Previous research on Dastareus helophoroides identified a minus-C OBP, DhelOBP21, which preferably binds to several ligands. In this study, only (+)-ß-pinene proved attractive to unmated adult beetles. To obtain a more in-depth explanation of the lack of behavioral activity of other ligands we selected compounds with high (camphor) and low (ß-caryophyllene) binding affinities. The structural transformation of OBPs was investigated using well-established approaches for studying binding processes, such as fluorescent quenching assays, circular dichroism, and molecular dynamics. The dynamic binding process revealed that the flexibility of DhelOBP21 seems conducive to binding specific ligands, as opposed to broad substrate binding. The compound (+)-ß-pinene and DhelOBP21 formed a stable complex through a secondary structural transformation of DhelOBP21, in which its amino-terminus transformed from random coil to an α-helix to cover the binding pocket. On the other hand, camphor could not efficiently induce a stable structural transformation, and its high binding affinities were due to strong hydrogen-bonding, compromising the structure of the protein. The other compound, ß-caryophyllene, only collided with DhelOBP21 and could not be positioned in the binding pocket. Studying structural transformation of these proteins through examining the dynamic binding process rather than using approaches that just measure binding affinities such as fluorescence competitive binding assays can provide a more efficient and reliable approach for screening behaviorally active compounds.

Transgenic cry1C(⁎) gene rough rice line T1C-19 does not change the host preferences of the non-target stored product pest, Rhyzopertha dominica (Fabricius) (Coleoptera: Bostrichidae), and its parasitoid wasp, Anisopteromalus calandrae (Howard) (Hymenoptera: Pteromalidae).

Rough rice grains are often stored for extended periods before they are used or consumed. However, during storage, the rough rice is vulnerable to insect infestation, resulting in significant economic loss. Previous studies have shown that volatiles cues, physical characteristics, and taste chemicals on the grains could be the important key behavior factors for storage insect pests to locate the hosts and select oviposition sites. It is also well known that the transgenic Bt rough rice line T1C-19, which expresses a cry1C(⁎) gene has a high resistance to Lepidoptera pests. However, there were no evidences to show the consequences of host preference for non-target insect pests after growing Bt transgenic rice. In this study, the potential key factors of Bt rough rice were investigated for their impacts on the behaviors of non-target pest lesser grain borer Rhyzopertha dominica, the main weevil pest of grain and its parasitic wasps Anisopteromalus calandrae, the natural enemy of the beetle. Both electronic nose and electronic tongue analyses showed that the parameters of Bt rough rice were analogous to those of the non-Bt rough rice. The volatile profiles of Bt and non-Bt rough rice examined by gas chromatographic mass spectrometry (GC-MS) were similar. For most volatile compounds, there were no significantly quantitative differences in compound quantities between Bt and non-Bt rough rice. The densities of sclereids and trichomes on the rough rice husk surface were statistically equal in Bt and non-Bt rough rice. The non-target pest, R. dominica, and its parasitoid wasp, A. calandrae, were attracted to both rough rice and could not distinguish the transgenic T1C-19 from the isogenic rough rice. These results demonstrated that Bt rough rice has no negative impacts on the host preference behaviors of non-target stored product pest R. dominica and its parasitoid A. calandrae.

Insight Into the Ultrastructure of Antennal Sensilla of Mythimna separata (Lepidoptera: Noctuidae).

The oriental armyworm, Mythimna separata (Walk), is one of the most serious pests of cereals in Asia and Australasia. The structure and distribution of the antennal sensilla of M. separata were studied by scanning electron microscopy and transmission electron microscopy. The results showed that antennae of both female and male M. separata are filiform in shape. Three groups and seven morphological sensillum types were recorded in both sexes, including uniporous sensilla (sensilla chaetica), multiporous sensilla (sensilla trichodea, basiconica, coeloconica, and styloconica), and aporous sensilla (sensilla squamiformia and Böhm bristles). S. trichodea, which were the most abundant sensilla, was made of three subtypes (ST I, ST II, and ST III) according to external features and two subtypes of s. basiconica (SB I and SB II) and s. coeloconica (SCo I and SCo II) were identified, respectively. Sexual dimorphisms in sensilla of M. separata were mainly perceived as the variations in the numbers of several sensilla subtypes. Also, the possible functions of the antennal sensilla were discussed. These results contribute to our understanding of the function of antennae in the behavior of M. separata.

Antennal sensilla in the parasitoid Sclerodermus sp. (Hymenoptera: Bethylidae).

Parasitoid wasps of the genus Sclerodermus (Hymenoptera: Bethylidae) are an important natural enemy of the Japanese pine sawyer beetle Monochamus alternatus Hope (Coleoptera: Cerambycidae). In this study, we used scanning electron microscopy to examine the external morphology of the antennal sensilla of Sclerodermus sp. Antennae of females and males comprised the scape, pedicel, and 11 flagellomere segments. Based on the morphology of the sensilla in each sex, seven types of sensillum were identified: sensilla trichodea (Tr.1, Tr.2 and Tr.3), sensilla basiconica (Ba.1, Ba.2, and Ba.3), sensilla styloconica (St.1 and St.2), sensilla placodea, sensilla coeloconica, sensilla squamiforma, and Bohm's bristles. Tr.2, Ba.1, and St.1 were only found in females, whereas Ba.2, Ba.3, and St.2 were only observed in males. Sensilla placodea were the most common, given that they occur on the antennae of many parasitoid Hymenoptera, whereas sensilla Tr were the most abundant, being distributed over the entire antennal surface. These sensilla are likely to have roles in the host locating and habitat searching behavior of adult Sclerodermus wasps. Therefore, our findings provide a basis for further studies of the host location behavior of this and other species of parasitic wasp.

Construction and analysis of cDNA libraries from the antennae of Batocera horsfieldi and expression pattern of putative odorant binding proteins.

A high-quality cDNA library was constructed from female and male antenna of the longhorned beetle, Batocera horsfieldi (Hope) (Coleoptera: Cerambycidae), a serious pest of Populus (Salicales: Salicaceae). The titer was approximately 2.37 × 106 pfu/mL, and this complies with the test requirement. From the libraries, 692 clones were selected randomly, sequenced, and further analyzed, and the recombinational efficiency reached 93.85%. By alignment and cluster analysis, we identified four odorant binding proteins, two pheromone-binding proteins (have the characteristic six conserved cysteine residues), four Minus-C odorant binding proteins (lost two conserved cysteines), and three chemosensory proteins. In this study, we describe the identification and characterization of four new cDNAs that encode Minus-C odorant binding proteins (Minus-C OBPs) from B. horsfieldi antennal cDNA libraries. Our investigation focused on the expression pattern of the Minus-C OBP genes in various tissues in both sexes at different developmental stages, using reverse transcription PCR (RT-PCR) and realtime PCR (qPCR) strategies. Minus-C OBP1, 2, and 3 were expressed in all tested tissues, with the exception of the head (without antenna, labial palps, and maxillary palps). Minus-C OBP4 was expressed in the antenna, legs, and abdomen, but not in the labial palps, maxillary palps, or head. The qPCR results revealed MinusC OBPs were expressed in the antenna throughout the adult life, and that the transcript levels of these genes depended on the sex, age, and mating status of adults.

Electrophysiological responses of the rice leaffolder, Cnaphalocrocis medinalis, to rice plant volatiles.

The rice leaffolder, Cnaphalocrocis medinalis Guenée (Lepidoptera: Pyralidae), is one of the most destructive pests of rice. Electrophysiological responses of this species to 38 synthetic volatiles known to be released from rice plants (Poaceae: Oryza spp.) were studied using the electroantennogram (EAG) method. Compounds that elicited the strongest EAG responses for each physiological condition were selected for EAG dose-response tests at five concentrations. These compounds included: methyl salicylate, heptanol, linalool, cyclohexanol, and 2-heptanone for one-day-old male moths; heptanol, hexanal, (Z)-2-hexen-1-ol, and nonadecane for one-day- old females; methyl salicylate, heptanol, (E)-2-hexen-1-ol, and (Z)-2-hexen-1-ol for three-day- old males; linalool, heptanol, (E)-2-hexen-1-ol, 2-heptanone, and hexanal for three-day-old females; 2-heptanone, cyclohexanol, linalool, heptanol, and methyl salicylate for five-day-old virgin females; and methyl benzoate, (Z)-2-hexen-1-ol, heptanol, linalool, and hexanal for five- day-old mated females. Female and male C. medinalis exhibited broad overlap in their EAG responses, and there was no clear difference between male and female EAG responses to different compounds. Statistical analyses revealed that both volatile compound chemical structure and C. medinalis physiological condition (age, sex, and mating condition) had an effect on EAG response.

Binding characterization of odorant-binding protein BhorOBP29 in Batocera horsfieldi (Hope) with host-plant volatiles.

Odorant binding proteins (OBPs) are involved in odorant discrimination and act as the first filter in the peripheral olfactory system. Previous studies have shown that BhorOBP29 is potentially involved in olfactory perception in an important wood-boring pest Batocera horsfieldi (Hope) (Coleoptera: Cerambycidae), however, its function remains unclear. Here, we investigated the ligand-binding profiles of recombinant BhorOBP29 with 22 compounds from its host plant using fluorescence competitive binding assays and fluorescence quenching assays. The results showed that BhorOBP29 could bind to five ligands relying mainly on hydrophobic interactions. Molecular docking analysis indicated that residues Ile48, Leu51, Met52, Trp57, Asn105, and Val119 were extensively involved in the interactions between BhorOBP29 and the five ligands. Furthermore, the site-directed mutagenesis analysis revealed that Leu51 and Met52 residues were indispensable for BhorOBP29-ligands binding. Finally, electroantennogram (EAG) assays confirmed that hexanal, (-)-limonene, and 2-methylbutyraldehyde elicited a concentration-dependent EAG response with a maximum at the concentration of 1/10 v/v. These findings suggest that BhorOBP29 may play a significant role in the perception of host plant volatiles by B. horsfieldi. This study may help to discover novel behavioral regulation and environmentally friendly strategies for controlling B. horsfieldi in the future.

Identification of odorant-binding proteins and functional analysis of antenna-specific BhorOBP28 in Batocera horsfieldi (Hope).

BACKGROUND: The important wood-boring pest Batocera horsfieldi has evolved a sensitive olfactory system to locate host plants. Odorant-binding proteins (OBPs) are thought to play key roles in olfactory recognition. Therefore, exploring the physiological function of OBPs could facilitate a better understanding of insect chemical communications. RESULTS: In this research, 36 BhorOBPs genes were identified via transcriptome sequencing of adults' antennae from B. horsfieldi, and most BhorOBPs were predominantly expressed in chemosensory body parts. Through fluorescence competitive binding and fluorescence quenching assays, the antenna-specific BhorOBP28 was investigated and displayed strong binding affinities forming stable complexes with five volatiles, including (+)-α-Pinene, (+)-Limonene, ß-Pinene, (-)-Limonene, and (+)-Longifolene, which could also elicit conformation changes when they were interacting with BhorOBP28. Batocera horsfieldi females exhibited a preference for (-)-Limonene, and a repellent response to (+)-Longifolene. Feeding dsOBP19 produced by a bacteria-expressed system with a newly constructed vector could lead to the knockdown of BhorOBP28, and could further impair B. horsfieldi attraction to (-)-Limonene and repellent activity of (+)-Longifolene. The analysis of site-directed mutagenesis revealed that Leu7, Leu72, and Phe121 play a vital role in selectively binding properties of BhorOBP28. CONCLUSION: By modeling the molecular mechanism of olfactory recognition, these results demonstrate that BhorOBP28 is involved in the chemoreception of B. horsfieldi. The bacterial-expressed dsRNA delivery system gains new insights into potential population management strategies. Through the olfactory process concluded that discovering novel behavioral regulation and environmentally friendly control options for B. horsfieldi in the future. © 2024 Society of Chemical Industry.