Integrated Transcriptomic and Metabolomic Insights into “I See You” (ISY) Defensive Behavior in Apis cerana Against Vespa velutina

<i>Vespa velutina</i> (<i>V. velutina</i>) is a globally widespread predator of honeybees, posing a serious threat to them. <i>Apis cerana</i> (<i>A. cerana</i>) has acquired the ability to partially thwart the predation of the wasps, but the molecular mechanism by which it responds to <i>V. velutina</i> attacks is unknown. Here, both transcriptomic and metabolomic analyses were integrated to decipher what happened in the brain tissues of <i>A. cerana</i> exposed to <i>V. velutina</i> attacks. Transcriptomic analysis revealed 296 differentially expressed genes (DEGs) that were significantly enriched in neural signaling pathways (particularly serotonin and dopamine transmission), the cAMP signaling pathway, and energy metabolism. Metabolomic results showed that <i>V. velutina</i> attacks affected 38 metabolic pathways involving 86 differentially expressed metabolites (DEMs), primarily including amino acids, nucleotides, and carbohydrates. Several metabolites, such as isocitrate, xanthine, and adenosine, were upregulated in response to the hornet presence. Combined pathway analysis indicated an upregulation of the purine and tyrosine metabolism pathways, highlighting their critical role in <i>A. cerana</i> against invasive hornets. The results of this study will help to understand the molecular mechanisms of <i>A. cerana</i> in response to <i>V. velutina</i> attacks and provide valuable insights for enhancing collective defensive behavior against <i>V. velutina</i> in honeybees.