Cerambycid beetles (Cerambycidae) are major forest pests, posing a serious threat to the security of forest resources worldwide. Extensive research has focused on the control of cerambycid beetles from physiological and biochemical perspectives. Despite the important roles of insect haemolymph in physiological processes, efficient collection methods for Cerambycidae are lacking. For the efficient and easy collection of large amounts of pure haemolymph from adult cerambycid beetles, a new method, named net centrifugation, was developed. Three species of cerambycid beetles with large differences in size, Anoplophora chinensis, Monochamus saltuarius and Saperda populnea, were selected for the study. Haemolymph was collected by the newly developed net centrifugation method—in which an inner nylon net is used during centrifugation under optimised conditions, and a relatively small wound is generated on the insect—as well as the traditional tearing method and double centrifugation method. Among the three methods evaluated, the net centrifugation method caused the least damage to cerambycid beetles during the whole operation. This method resulted in the most haemolymph from a single beetle, with the lowest turbidity, mostly pure haemocytes in the precipitate, the clearest haemolymph smears by microscopy and the highest quality of RNA extracted from haemocytes. The net centrifugation method has a high collection efficiency, providing important technical support for haemolymph extraction and entomological research.

Insects use a powerful and complex olfactory recognition system to sense odor molecules in the external environment to guide behavior. A large family of odorant receptors (ORs) mediates the detection of pheromone compounds. Anoplophora glabripennis is a destructive pest that harms broad-leaved tree species. Although olfactory sensation is an important factor affecting the information exchange of A. glabripennis, little is known about the key ORs involved. Here, we identified ninety-eight AglaORs in the Agla2.0 genome and found that the AglaOR gene family had expanded with structural and functional diversity. RT-qPCR was used to analyze the expression of AglaORs in sex tissues and in adults at different developmental stages. Twenty-three AglaORs with antennal-biased expression were identified. Among these, eleven were male-biased and two were female-biased and were more significantly expressed in the sexual maturation stage than in the post-mating stage, suggesting that these genes play a role in sexual communication. Relatively, two female-biased AglaORs were overexpressed in females seeking spawning grounds after mating, indicating that these genes might be involved in the recognition of host plant volatiles that may regulate the selection of spawning grounds. Our study provides a theoretical basis for further studies into the molecular mechanism of A. glabripennis olfaction.

Eogystia hippophaecolus (Hua, Chou, Fang et Chen) is a drilling pest that damages sea buckthorn, leading to substantial ecological and economic losses in areas where sea buckthorn is cultivated in China. Pheromone lures are widely used to monitor and trap male moths; however, the olfactory recognition mechanisms in male moths require further investigation. In this study, we characterized odorant receptor genes using phylogenetics, gene expression, sequence analyses and molecular docking. A phylogenetic analysis using the maximum likelihood method revealed that odorant receptor co-receptor (EhipOrco) genes clustered with other lepidopteran Orco genes, while pheromone receptor (EhipPR) genes clustered with other lepidopteran PR genes, suggesting that these genes have similar functions. The expression levels of 27 odorant receptor (EhipOR) genes in five tissues of male adults were analyzed using qPCR. Nine EhipOR genes were specifically expressed in the antennae, while several EhipOR genes were highly expressed in the head, feet, and external genitalia. As determined using molecular docking, EhipPR1–3 bound strongly to the primary sex pheromone components of E. hippophaecolus. Additionally, EhipPR1–3 demonstrated a strong binding affinity for longifolene among sea buckthorn volatiles. This study provides a foundation for future functional research on EhipORs and the olfactory recognition mechanisms of E. hippophaecolus.