The apple buprestid beetle, Agrilus mali Matsumura (Coleoptera: Buprestidae), can respond to various volatiles, but the underlying mechanism of odorant perception for this insect is poorly understood. Here, we cloned A. mali’s odorant-binding proteins 3 (AmalOBP3) and 8 (AmalOBP8) and characterized their expression patterns and binding profiles. Sequence and phylogenetic analyses showed that AmalOBP3 and AmalOBP8 were distributed in the classic and minus-C OBP subfamily, respectively. AmalOBP3 was specifically and abundantly expressed in antennae of both sexes. AmalOBP8 displayed high transcript levels in antennae of both sexes, abdomens of males, and wings of both sexes. Both AmalOBPs exhibited much higher expression in male antennae than in female antennae, suggesting that they could be important in perception of male-specific olfactory cues (e.g., some sex pheromones). Out of the 40 odorant ligands tested, AmalOBP3 and AmalOBP8 bound to 15 and 21 different odorants, respectively, indicating a distinct and selective binding profile for them. Both AmalOBPs seemed to have very strong binding affinity to aliphatic alcohols and aldehydes with 12 to 15 carbon atoms. Alcohols, esters, and terpenoids were more likely to be good ligands for both AmalOBPs than aldehydes and alkanes. Together with its broad expression in different tissues, strong binding with higher numbers of putative ligands for AmalOPB8 means that this protein can have more extensive functional roles in chemosensation of A. mali. Our results provide insights into the molecular basis of chemosensation in A. mali, as well as a basis for developing detection, monitoring, and management tools for this serious pest.

The apple buprestid beetle, Agrilus mali Matsumura, is an invasive pest causing significant damages to rare wild apple forests of Xinjiang. The morphology, abundance and distribution of antennal sensilla in both sexes of this pest were examined. We found that the antennae of A. mali females were longer than those of males. Five types of antennal sensilla were characterized, including trichodea (subtypes Tr.1, Tr.2, and Tr.3), chaetica (subtypes Sc.1, Sc.2, Sc.3, and Sc.4), basiconica (subtypes Ba. 1, Ba. 2, Ba. 3 and Ba.4), Böhm bristles (subtypes BB. 1, and BB. 2), and multiporous grooved sensilla. The most abundant sensilla of Ba.2 tended to occur mainly on flagellomeres 5–8 in both sexes. The last three flagellomeres tended to have the most abundant Tr.1 in both sexes. Overall, the abundance and distribution of these sensilla appeared to be highly conserved in both sexes, and their olfactory organs seemed to cluster on flagellomeres 6–8. However, some sex dimorphisms were also observed. Tr.3 and BB.2 were found only in females. Sensilla of Sc.2 were found on the pedicel and first two flagellomeres only in males. When compared with males, females showed a higher number of Sc.3, but a lower number of Sc.4 on the pedicel. These results indicate that contact cues could be important in intersexual communication in A. mali. The functional roles of these sensilla and their implications in A. mali behaviors are discussed, and further studies of identified chemosensitive sensilla can provide a foundation for developing semiochemical-based management strategies.

Insects may develop different behavioral phenotypes in response to heterogeneous environments (e.g., host plants), but the plasticity of their feeding behaviors has been rarely explored. In order to address the issue, clones of the English grain aphid, Sitobion avenae (Fabricius), were collected from wheat, and their probing behaviors were recorded on three plants. Our results demonstrated that S. avenae individuals on the alternative plants (i.e., barley and oat) tended to have higher frequency of non-probing (Np), increased duration of the pathway phase, increased phloem salivation, and decreased phloem ingestion (E2), compared to those on the source plant (i.e., wheat), showing the resistance of barley and oat to this aphid’s feeding. This aphid showed apparently high extents of plasticity for all test probing behaviors on barley or oat. Positive selection for higher extents of plasticity in E2 duration was identified on barley and oat. The factor ‘clone’ alone explained 30.6% to 70.1% of the total variance for each behavioral plasticity, suggesting that the divergence of probing behavior plasticity in S. avenae had a genetic basis. This aphid’s fitness correlated positively with the plasticity of Np frequency and E2 frequency. Some behaviors and their corresponding plasticities (e.g., the frequency of xylem ingestion and its plasticity) were found to be correlated characters, probably reflecting the limits for the evolution of higher extents of behavioral plasticity in this aphid. The differential probing behaviors and their plasticity in S. avenae can have significant implications for the adaptation and management of aphids on different plants.

The grassland caterpillar Gynaephora qinghaiensis (Lepidoptera: Lymantriidae) is a dominant pest species in the alpine meadows of the Tibetan Plateau. Elucidating changes in key gene expression patterns will provide molecular insights into the adaptive evolutionary mechanisms of insects. Quantitative real-time PCR (qRT-PCR) is currently the predominant analytical methodology for assessing gene expression levels. However, variability among samples can compromise result reliability. Thus, selecting stably expressed reference genes for target gene normalization under diverse scenarios is critical. To date, suitable reference genes for G. qinghaiensis under varying experimental conditions have remained unidentified. In this study, the transcriptome data of G. qinghaiensis were obtained using the RNA-seq technique, and 13 candidate reference genes were selected. Four independent algorithms—ΔCt, geNorm, NormFinder, and BestKeeper—as well as a comprehensive online platform, RefFinder, were employed to evaluate the stability under six experimental conditions (tissues, developmental stages, sexes, temperatures, starvation, and insecticide treatments). Our findings identified the following optimal reference gene combinations for each experimental condition: RPS18, RPS15, and RPL19 for tissue samples; RPL19, RPS15, and RPL17 across developmental stages; RPS18 and RPS15 for different sexes; RPS8 and EF1-α under varying temperature conditions; RPL17 and RPL15 during starvation; and RPL19 and RPL17 following insecticide treatments. To validate the feasibility of the reference genes, we examined the expression of the target gene HSP60 in different tissues and under different temperatures. Our results established essential reference standards for qRT-PCR with G. qinghaiensis samples, laying the foundation for precise gene expression quantification in the future.

Diorhabda rybakowi Weise is one of the dominant pests feeding on Nitraria spp., a pioneer plant used for windbreaking and sand fixation purposes, and poses a threat to local livestock and ecosystems. To clarify the key olfactory genes of D. rybakowi and provide a theoretical basis for attractant and repellent development, the optimal reference genes under two different conditions (tissue and sex) were identified, and the bioinformatics and characterization of the tissue expression profiles of two categories of soluble olfactory proteins (OBPs and CSPs) were investigated. The results showed that the best reference genes were RPL13a and RPS18 for comparison among tissues, and RPL19 and RPS18 for comparison between sexes. Strong expressions of DrybOBP3, DrybOBP6, DrybOBP7, DrybOBP10, DrybOBP11, DrybCSP2, and DrybCSP5 were found in antennae, the most important olfactory organ for D. rybakowi. These findings not only provide a basis for further in-depth research on the olfactory molecular mechanisms of host-specialized pests but also provide a theoretical basis for the future development of new chemical attractants or repellents using volatiles to control D. rybakowi.