Coordinated neural, metabolic and muscular transcriptomic signatures associated with mite-biting behavior in honeybees (Apis mellifera L.)

Biting behavior is an important natural defense mechanism in honeybees (Apis mellifera) against Varroa destructor. Significant variation in this behavior exists across genetic lines of honeybees, with certain colonies exhibiting higher mite-biting activity than others. Selective breeding for enhanced biting behavior provides a promising strategy for sustainable mite control and colony resilience. However, successful implementation of such breeding programs requires a comprehensive understanding of the genomic mechanism underlying this trait. In this study, RNA-seq analysis of mandible transcriptomes of 1-day and 8-day old worker honeybees from high mite biting (HB) and low mite biting (LB) colonies were performed. A total of 9,345 genes (97.30%) showed a significant differential expression between LB and HB honeybees across different ages (one-way ANOVA, FDR < 0.05). Comparison of LB vs. HB workers collected on day 1 detected 166 down-regulated and 403 up-regulated genes, whereas workers collected on day 8 identified 82 down-regulated and 46 up-regulated genes. Furthermore, the Weighted Gene Coexpression Network Analysis (WGCNA) exhibited the brown and yellow modules with significantly higher expression in HB compared with LB on both day 1 (FC = 1.52, FDR = 0.0019 and FC = 1.47, FDR = 2.7 x 10-4, respectively) and day 8 (FC = 1.27, FDR = 0.056 and FC = 1.16, FDR = 0.073, respectively). Gene Ontology (GO) enrichment analysis identified over-representation of biological processes involved in muscle contraction, chitin binding, neural signaling, oxidative phosphorylation, neuron development, electron transport chain, mitochondrial ATP synthesis, stress response, sensory perception and metabolic processes. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways analysis identified significant enrichment of various pathways including oxidative phosphorylation, cytoskeleton-related pathways, carbon metabolism, motor proteins, ribosome-associated pathways, and citrate cycle (TCA cycle). The present findings demonstrate mite-biting behavior is associated with coordinated activation of neural, energetic and muscular system rather than a single molecular mechanism. These findings provide a basis to improve honeybee health, enhance resistance to Varroa and other ectoparasite, and eventually support sustainable beekeeping and agricultural pollination systems.