Differential energy pathways are required for rapid long-term memory formation in the oriental fruit fly, Bactrocera dorsalis

• Bactrocera dorsalis can rapidly form long-term memory. • Long-term memory formation is an energy-consuming process. • Early long-term memory formation requires energy from the TCA cycle, while late long-term memory formation requires energy from oxidative phosphorylation. To ensure the reliability of learned information, most insects require multiple intervals of experience before storing the information as Long-term memory (LTM), and this requirement has been validated in insects from the behavioral to the molecular level. Recent studies have shown that some insects can form LTM after a single experience, although the mechanisms underlying one-trial LTM formation are not well understood. Therefore, understanding the mechanisms underlying rapid learning and subsequent preference formation in insects is crucial. Here we show that the agricultural pest Bactrocera dorsalis can rapidly form LTM, which is dependent on protein synthesis, and that the formation of LTM requires high energy support at the cost of reduced survival. Furthermore, based on a liquid chromatography-mass spectrometry (LC-MS) metabolomics approach, we found that LTM-related processes are sequentially coupled to two processes for energy generation, the TCA cycle and oxidative phosphorylation. This was further confirmed by blocking these energy generation processes. Our results provide a theoretical basis for the development of behavioral modulators in oriental fruit flies that target energy generation intermediate metabolites, as well as a new perspective on the rapid formation of LTM in insects.