Cytochrome P450 monooxygenase (CYP) is one of the three detoxification metabolic enzymes in insects, and is involved in the metabolism and transformation of endogenous substances as well as the activation and degradation of exogenous compounds. This study aims to reveal the molecular mechanism of CYP9E2 in Tribolium castaneum in adapting to high-CO₂ stress. By predicting the sequence function of CYP9E2, analyzing the temporal and spatial expression profile of TcCYP9E2, and using RNAi to silence TcCYP9E2 combined with a high-CO₂ stress treatment, we measured the carbohydrate content, trehalase activity, and gene expression levels in trehalose metabolism of T. castaneum. A bioinformatics analysis showed that the predicted molecular weight of the protein encoded by TcCYP9E2 is 60.15, the theoretical isoelectric point is 8.63, there is no signal peptide, and the protein is hydrophilic. An evolutionary tree analysis showed that TcCYP9E2 belongs to the CYP6 family and belongs to the CYP3 group; and the spatiotemporal expression profile results showed that TcCYP9E2 was highly expressed in the larvae midgut 48 h after injection of dsCYP9E2, with survival rates decreasing with the increase in CO₂ concentration. Under the condition of 75% CO₂, the contents of glycogen, glucose, ATP, and membrane-bound trehalase decreased significantly after the injection of dsCYP9E2. The expression of TRE-1, TRE-2, and GP in trehalose metabolism and energy pathways was significantly downregulated.

High carbon dioxide concentrations can effectively control most storage pests. To estimate the toxicity effect of high concentrations of CO₂, four different concentrations of CO₂ (25% CO₂, 50% CO₂, 75% CO₂, and 95% CO₂) were used to treat Tribolium castaneum, and the biochemical (carbohydrate content and gene expression level) and physiological (mortality, pupation, eclosion rate, and weight) features of insects submitted to different treatments with CO₂ were evaluated. The T. castaneum mortality rate was 50% in approximately 2 days when exposed to a treatment with 95% CO₂. When the CO₂ concentration exceeded 75%, the pupation rate and eclosion rate of T. castaneum seriously declined. Higher than 25% CO₂ concentrations resulted in a lower weight and shrunken body size of T. castaneum. It was further found that different CO₂ concentration treatments all influenced the levels of the three carbohydrate contents in T. castaneum. In addition, according to the detection of trehalose metabolism pathway-related genes, T. castaneum mainly responds to stress factors via high expression of TPS, TRE1-2, and TRE1-3. Our results enrich the evaluation of the toxicity effect of CO₂ treatment on grain storage pests, providing a basis for further improving the method of regulating grain storage to control insect pests.

Heavy metal pollution, particularly cadmium (Cd) contamination in water and farmland, might accumulate in natural insect enemies through the food chain. In response to this heavy metal stress, natural enemy insects adapt by altering their metabolism and behaviors. As a result, this investigation aimed to elucidate how the development, reproduction, and feeding of Harmonia axyridis Pallas (Coleoptera: Coccinellidae) are affected under Cd contamination. Compared to the control group, the developmental period of H. axyridis was prolonged, with decreased survival, predation, and body weights. Notably, adult insects exhibited deformation, including molting difficulties and wing deformities, which indicated reduced fitness. The ovarian development of female insects was delayed with reduced size, and the pre-oviposition period was prolonged under Cd contamination. Additionally, the hatching rate of offspring was significantly reduced. The Vitellogenin 1 (Vg1) and Vitellogenin 2 (Vg2) exhibited considerable changes throughout their developmental stages. Our results confirmed that the accumulation of Cd has a significant impact on the growth, development, and normal molting of H. axyridis, affecting the reproduction of H. axyridis. The aforementioned results provide valuable insights into the potential ecological effects of Cd accumulation on the food chain, which can inform strategies for pest control, ecosystem stabilization in rice fields, and potentially novel bioremediation approaches. Thereby establishing a theoretical foundation for pest control and ecosystem stabilization in rice fields under Cd contamination while simultaneously providing novel insights for bioremediation strategies.

Spodoptera frugiperda (J. E. Smith, 1797), an omnivorous crop pest worldwide, reproduces prolifically. Validamycin, a competitive natural inhibitor of trehalase, is regarded as an effective and safe insecticide. Pupae were injected with a validamycin gradient (0.5–10 µg/µL) to block trehalase; enzyme activity and the Vitellogenin gene (Vg)/its receptor gene (VgR) expression (rpL10 reference) were subsequently quantified to determine the compound’s impact on S. frugiperda ontogeny and fecundity. Validamycin directly inhibited pupal membrane-bound trehalase, sharply lowering glycogen. Both pupal and adult mortality rose with dose, yielding marked abnormalities versus the Control Check (CK) group. At 0.5 μg/μL validamycin, eggs blackened and clumped in the lateral oviduct, blocking release; treated females produced far fewer eggs by day 4, exhibited ovarian atrophy, shorter lifespan, and low hatchability. The expression levels of Vg and VgR in the ovaries of the fall armyworm were consistent with the changes in the ovarian developmental phenotype. Validamycin significantly inhibited the activity of trehalase in S. frugiperda, severely hindering their normal eclosion and lowering the potential reproductive capacity of S. frugiperda. Simultaneously, it directly affects ovarian development and the lifespan of female moths. The results provide data to support the development of new methods for controlling S. frugiperda.

Nilaparvata lugens is a migratory pest with high fecundity and outstanding drug resistance, which poses a devastating danger to rice production. This study investigated the reproductive regulation mechanism of N. lugens, specifically silencing the trehalose-6-phosphate synthase gene (TPS) via RNAi to elucidate how TPS governs the trehalose metabolic network through modulation of trehalose biosynthesis. Insect fecundity hinges on the synchronized progression of oogenesis and the tightly controlled expression of vitellogenin (Vg). In N. lugens, this coordination is orchestrated by an integrated molecular network that converges juvenile hormone signaling (JH), 20-hydroxyecdysone pathways (20E), insulin/IGF signaling (IIS), and the target of rapamycin cascade (TOR), collectively dictating the reproductive output of the species. Using TPS knockdown as the entry point, this study dissects the lipid-metabolic circuitry of N. lugens and uncovers how hormonal signaling cascades orchestrate reproduction by precisely modulating vitellogenin (Vg) and its cognate receptor VgR. Synthesized double-stranded terpene synthase genes (dsTPSs) can degrade mRNA, inhibit protein translation, and ultimately lead to the silencing of TPS genes, simultaneously crippling energy provision and hormonal signaling to orchestrate a multi-pronged suppression of reproduction. This dual-action intervention offers a promising molecular target for environmentally friendly management of N. lugens.

The mass rearing of natural enemy insects is a cornerstone of effective green pest control, making the development of optimized artificial diets critical for sustaining large populations. However, existing artificial diets often have limitations, including inhibitory effects on the reproductive development of natural enemies. In this study, we innovatively incorporated 20% catalpol into the artificial diets for Harmonia axyridis, aiming to comprehensively assess the effects of such dietary regimens on larval development, reproductive performance, and the regulation of energy metabolism in H. axyridis. Our results demonstrated that catalpol supplementation significantly shortened the duration of the 4th instar larval and pupal stages and increased pupal weight. Furthermore, catalpol treatment enhanced reproductive capacity, as evidenced by increased fecundity, improved hatching rate stability, and enhanced ovarian development. This effect was directly correlated with the significant upregulation of the vitellogenin genes Vg1 and Vg2 and their receptor gene VgR at 3 days post-emergence. Regarding energy metabolism, catalpol supplementation led to increased glycogen reserves and glucose content. The expression of the membrane-bound trehalase enzyme gene TRE2 was significantly higher than that in the control group, and the expression level of the soluble trehalase enzyme gene TRE1 also showed a significant increase. These findings indicate that catalpol effectively optimized energy metabolism, thereby accelerating developmental processes and enhancing reproductive capacity. In this study, we elucidated the mechanism by which catalpol regulates physiological metabolism in predatory natural enemy insects. For the first time, catalpol was incorporated into the artificial diet of H. axyridis as a synergistic component, significantly enhancing its growth, reproductive capacity, and energy metabolism. These findings provide a scientific basis for developing efficient artificial feeding systems and improving the field adaptability of natural enemy insects.