Ophraella communa is an efficient biocontrol agent used against the invasive weed Ambrosia artemisiifolia. It is an herbivorous insect that feeds on specific plants; the olfactory functions of this insects plays an important role in their search for host plants. There are no reports on O. communa sensilla types, morphology, or chemosensory protein (CSP) genes. In this study, we observed the external structure and distribution of antennal sensilla in adult O. communa antennae by scanning electron microscopy; moreover, we cloned 11 CSPs (CSP1–CSP11) and elucidated their tissue-expression profiles using quantitative real-time polymerase chain reaction. Six types of sensilla were identified: sensilla trichodea (including two subtypes), sensilla chaetica, sensilla basiconica (including two subtypes), sensilla styloconica, sensilla coeloconica, and Böhm bristles. Both male and female antennae had all six types of sensilla, and no sexual dimorphism was noted in sensillar types or distribution. We also found that the expression levels of CSP2, CSP3, CSP4, CSP6, and CSP7 in male and female antennae were higher than those in other tissues, which suggests that these five CSPs may be related to olfactory function in O. communa. Ultimately, our results lay the foundation for interpreting the olfactory functions of adult O. communa.

Chemosensory systems play a pivotal role in insect survival and reproduction by mediating the detection of volatile organic compounds in the environment. Protaetia brevitarsis (Coleoptera: Scarabaeidae), a phytophagous pest widely distributed across East Asia, poses a significant threat to agro-horticultural systems through crop damage. We conducted antennal transcriptome sequencing of adult beetles and identified 117 chemosensory-related genes, including 66 odorant receptors (ORs), 20 ionotropic receptors, 10 gustatory receptors, 13 odorant-binding proteins (OBPs), four chemosensory proteins, and four sensory neuron membrane proteins. Tissue-specific expression profiling revealed the antennal enrichment of five PbreOBP genes and twenty-three ORs. Notably, sexual dimorphism was observed in OR expression patterns. PbreOR1/6/17/18/21/22/30/32 exhibited male-biased antennal expression, whereas PbreOR25/26/29/38/41/44/61 demonstrated female-biased antennal expression, indicating their potential involvement in sex-specific behaviors, such as pheromone detection and oviposition site selection. A comprehensive description of the antenna chemosensory-related genes of P. brevitarsis has deepened our understanding of the olfactory mechanisms in coleopteran insects. This study also provides a basis for understanding the molecular mechanisms underlying olfaction in P. brevitarsis.

The olfactory system plays a key role in various insect behaviors, and odorant-binding proteins participate in the first step of the olfactory process. Ophraella communa Lesage is an oligophagous phytophagous insect that is a specific biological control agent for Ambrosia artemisiifolia L. The leaf beetle must identify and locate A. artemisiifolia through olfaction; however, its odorant-binding protein (OBP) function has not yet been reported. In this study, OcomOBP7 was cloned, and its tissue expression profile and binding ability were analyzed using RT-qPCR and fluorescence binding assays, respectively. Sequence analysis demonstrated that OcomOBP7 belongs to the classical OBP family. The RT-qPCR results showed that OcomOBP7 was specifically expressed in the antennae, indicating that OcomOBP7 may be involved in chemical communication. The fluorescence binding assay showed that OcomOBP7 has an extensive binding ability to alkenes. The electroantennography experiments showed that O. communa antennal response to α-pinene and ocimene decreased significantly after interference because the two odors specifically bound to OcomOBP7. In summary, α-pinene and ocimene are odorant ligands corresponding to OcomOBP7, indicating that OcomOBP7 is involved in the chemical recognition of A. artemisiifolia. Our study lays a theoretical foundation for research into O. communa attractants, which is helpful for the better biological control of A. artemisiifolia by O. communa.

The polyphagous pest Oxycetonia jucunda Faldermann can cause substantial damage to a range of economically important crops, with the adult beetles feeding directly on floral tissues and young leaves. RT-qPCR is widely used to analyze gene expression, for which the selection of stable reference genes is essential for enabling an accurate normalization of expression. However, no systematic evaluations of suitable reference genes for RT-qPCR analysis using different tissues of O. jucunda have been conducted. To assess their applicability as reliable normalization controls, we used five computational methods to examine the stability of seven potential reference genes (GAPDH, EF1α, RPS3, RPS18, RPL18, RPS31, and UBC5A) across six adult tissues, with three biological replicates per tissue. The findings revealed RPS3 and RPS31 to be the most stably expressed. This pair of reference genes was further validated by normalizing the expression of the odorant-binding protein 3 (OBP3) target gene. Our findings will provide important foundational data for the accurate analysis of functional gene expression in O. jucunda.

The melon fly, Zeugodacus cucurbitae (Coquillett), is a globally significant agricultural pest causing substantial economic losses. Odorant-binding proteins (OBPs) are critical of the insect olfactory system, yet their specific physiological functions in Z. cucurbitae remain largely uncharacterized. In this study, we functionally characterized ZcucOBP14 and investigated its putative involvement in host chemoreception and insecticide tolerance. Sequence alignment and phylogenetic analysis indicated that ZcucOBP14 belongs to the Minus-C OBP subfamily, and quantitative reverse transcription PCR (RT-qPCR) showed that it was predominantly expressed in both the head and abdomen. Fluorescence binding assays revealed that ZcucOBP14 exhibited broad binding affinity to 11 host plant volatiles, three sex pheromones, and two insecticides. Subsequent electroantennography (EAG) and behavioral bioassays identified isopulegol, 1-hexanol, linalool, and α-pinene as key ligands regulating the behavioral responses of Z. cucurbitae. RNA interference (RNAi)-mediated knockdown of ZcucOBP14 significantly reduced EAG responses to key ligands, eliminated behavioral preference, and increased insecticide-induced mortality by 20%. Molecular docking further identified that Tyr71, Ile67, Trp50, Val107, Phe116 and Leu70 were critical residues involved in ligand interactions. Collectively, these findings highlight the indispensable role of ZcucOBP14 in olfactory perception and its contribution to insecticide tolerance, laying a solid theoretical foundation for the development of novel behavior-modifying agents, attractants, and optimized integrated pest management (IPM) strategies against this pest.