The Role of TcCYP6K1 and TcCYP9F2 Influences Trehalose Metabolism under High-CO[sub.2] Stress in Tribolium castaneum

Due to issues with pesticide residues and resistance caused by traditional pesticides, the use of modified atmosphere storage technology has become increasingly popular. However, research has shown that prolonged exposure to high concentrations of CO[sub.2] for pest control in this technology can lead to insect resistance to hypoxia. Therefore, it is necessary to find out the resistance mechanism. This study identifies TcCYP6K1 and TcCYP9F2 as key factors in the response to high CO[sub.2] in Tribolium castaneum , suggesting that these genes may affect the resistance to high CO[sub.2] by influencing in the synthesis or breakdown of the carbohydrate metabolism pathways. These findings provide a theoretical basis for the combined use of novel nucleic acid pesticides and modified atmosphere treatment. Cytochrome P450 monooxygenases (CYP), crucial detoxification enzymes in insects, are involved in the metabolism of endogenous substances as well as the activation and degradation of exogenous compounds. In this study, T. castaneum was utilized to investigate the roles of TcCYP6K1 and TcCYP9F2 genes influencing in the trehalose metabolism pathway under high-CO[sub.2] stress. By predicting the functional sequences of TcCYP6K1 and TcCYP9F2 genes and analyzing their spatiotemporal expression patterns, it was discovered that both genes belong to the CYP3 group and exhibit high expression levels during the larval stage, decreasing during the pupal stage, while showing high expression in the fatty body, intestine, and malpighian tubules. Furthermore, following the knockdown of TcCYP6K1 and TcCYP9F2 genes in combination with treating larvae with 75% CO[sub.2] , it was observed that larval mortality increased, and glycogen content significantly decreased, while trehalose content increased significantly. Additionally, membrane-bound trehalase enzyme activity declined, TPS gene expression was significantly upregulated, GS gene expression was significantly downregulated, and ATP content showed a marked decrease. In conclusion, CYP genes are critical responsive genes of T. castaneum to high CO[sub.2] levels, potentially impacting the insect’s resistance to carbon dioxide through their involvement in the synthesis or breakdown of the carbohydrate metabolism pathway. These findings could serve as a theoretical basis for the utilization of novel pesticides in low-oxygen grain storage techniques and offer new insights for environmentally friendly pest control strategies in grain