Homeotic Protein Distal-Less Regulates NlObp8 and NlCsp10 to Impact the Recognition of Linalool in the Brown Planthopper Nilaparvata lugens.

Odorant-binding proteins (OBPs) and chemosensory proteins (CSPs), which are thought to play key roles in the olfactory recognition of insects, can be induced by the odorants they recognize, but little is known about the underlying regulatory mechanisms. Here, we found that NlOBP8 and NlCSP10 play coordinative roles in the chemoreception of brown planthoppers (BPHs) to the volatile component linalool. Also, the relative mRNA levels of NlObp8 and NlCp10 decreased upon exposure to linalool. Further, homeotic protein distal-less (Dll), which was also highly expressed in the antennae, was found to positively regulate the transcription of NlObp8 and NlCsp10 directly. Knocking down NlDll expression downregulated the expression of many additional olfactory functional genes and impaired the repellent behavior of BPHs to linalool. Our findings elucidate the direct regulatory role of Dll in BPHs' olfactory plasticity to linalool through modulating the olfactory functional gene expression and could provide guidance to sustainably control BPHs in the field.

NlugOBP8 in Nilaparvata lugens Involved in the Perception of Two Terpenoid Compounds from Rice Plant.

Odorant binding proteins (OBPs) play an important role in insect peripheral olfactory systems and exploring the physiological function of OBPs could facilitate the understanding of insects' chemical communication. Here, the functional analysis of an antenna-based NlugOBP8 from brown planthopper (BPH) Nilaparvata lugens (Stål) was performed both in vitro and in vivo. Recombinant NlugOBP8 exhibited strong binding affinity to 13 out of 26 rice plant volatiles and could form a stable complex with 9 of them according to the fluorescence binding and fluorescence quenching experiments. Circular dichroism spectra demonstrated that six volatiles could give rise to significant conformational change of recombinant NlugOBP8. H-tube olfactometer bioassay confirmed that BPHs were significantly attracted by nerolidol and significantly repelled by linalool, caryophyllene oxide, and terpinolene, respectively. Antennae of dsNlugOBP8-injected BPHs exhibited significantly lower electrophysiological response to linalool and caryophyllene oxide. Moreover, the repellent responses of BPHs to these two volatiles were also impaired upon silencing NlugOBP8. These data suggest that NlugOBP8 is involved in recognizing linalool and caryophyllene oxide and provide additional target for the sustainable control of BPHs.

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.

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.

BarH1 regulates odorant-binding proteins expression and olfactory perception of Monochamus alternatus Hope.

Insect odorant-binding proteins (OBPs) are a class of small soluble proteins that can be found in various tissues wherein binding and transport of small molecules are required. Thus, OBPs are not only involved in typical olfactory function by specific activities with odorants but also participate in other physiological processes in non-chemosensory tissues. To better understand the complex biological functions of OBPs, it is necessary to study the transcriptional regulation of their expression patterns. In this paper, an apparent gradient expression pattern of Obp19, that was highly and specifically expressed in antennae and played an essential role in the detection of camphene, was defined in the antennae of the Japanese pine sawyer. Further, the transcription factor BarH1, that also presented gradient expression pattern in antennae, was found to regulate expression of Obp19 directly through binding to its upstream DNA sequence. The condition of BarH1 gene silence, the gene expression levels of Obp19 significantly decreased. At the same time, additional olfactory genes also were regulated and thus influence camphene reception. These findings provide us an opportunity to incorporate Obps in the gene regulatory networks of insects, which contribute to a better understanding of the multiplicity and diversity of OBPs and the olfactory mediated behaviors.

Discovery of behaviorally active semiochemicals in Aenasius bambawalei using a reverse chemical ecology approach.

BACKGROUND: The invasive mealybug, Phenacoccus solenopsis, has caused serious damage to cotton crops throughout the world. Aenasius bambawalei is a dominant endoparasitoid of P. solenopsis. Exploration of behaviorally active semiochemicals may promote the efficacy of parasitoids used in biological control. Reverse chemical ecology, based on the physiological function of odorant-binding proteins (OBPs), provides an effective approach to screen behaviorally active compounds to target insect pests. Determination of the binding mechanisms and specificity towards different odorants in A. bambawalei may facilitate the development of more-efficient biological control strategies. RESULTS: We characterized the expression profile and analyzed the binding affinity of OBP28 in A. bambawalei. AbamOBP28 showed high expression in the wings and antennae of both male and female A. bambawalei. A fluorescence competitive binding assay indicated that AbamOBP28 displayed strong binding affinity to most candidate ligands. Circular dichroism spectra demonstrated that 1-octen-3-one, myrcene, dodecane, 2,4,4-trimethyl-2-pentene, nonanal, and limonene elicited conformational changes in AbamOBP28. Electrophysiological and behavioral bioassays revealed that diethyl sebacate, 2,4,4-trimethyl-2-pentene, and 1-octen-3-one evoked significant electroantennography responses and functioned as attractants in A. bambawalei at specific concentrations. Furthermore, three-dimensional structure modeling and molecular docking showed that hydrogen bonds were formed by Glu1 and Ser75 of AbamOBP28 with diethyl sebacate, respectively. CONCLUSION: These results demonstrate that AbamOBP28 is involved in the chemoreception of A. bambawalei. The identified protein provides a potential target for efficient enemy utilization and pest control, and the overall results may help develop protocols for more effective screening of behaviorally active semiochemicals. © 2021 Society of Chemical Industry.

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.

The role of chemosensory protein 10 in the detection of behaviorally active compounds in brown planthopper, Nilaparvata lugens.

Chemosensory proteins (CSPs) play important roles in insects' chemoreception, although their specific functional roles have not been fully elucidated. In this study, we conducted the developmental expression patterns and competitive binding assay as well as knock-down assay by RNA interference both in vitro and in vivo to reveal the function of NlugCSP10 from the brown planthopper (BPH), Nilaparvata lugens (Stål), a major pest in rice plants. The results showed that NlugCSP10 messenger RNA was significantly higher in males than in females and correlated to gender, development and wing forms. The fluorescence binding assays revealed that NlugCSP10 exhibited the highest binding affinity with cis-3-hexenyl acetate, eicosane, and (+)-ß-pinene. Behavioral assay revealed that eicosane displayed attractant activity, while cis-3-hexenyl acetate, similar to (+)-ß-pinene significantly repelled N. lugens adults. Silencing of NlugCSP10, which is responsible for cis-3-hexenyl acetate binding, significantly disrupted cis-3-hexenyl acetate communication. Overall, findings of the present study showed that NlugCSP10 could selectively interrelate with numerous volatiles emitted from host plants and these ligands could be designated to develop slow-release mediators that attract/repel N. lugens and subsequently improve the exploration of plans to control this insect pest.