The parasitic mite Varroa destructor devastates honey bee (Apis mellifera) colonies around the world. Entering a brood cell shortly before capping, the Varroa mother feeds on the honey bee larvae. The hormones 20-hydroxyecdysone (20E) and juvenile hormone (JH), acquired from the host, have been considered to play a key role in initiating Varroa’s reproductive cycle. This study focuses on differential expression of the genes involved in the biosynthesis of JH and ecdysone at 6 time points during the first 30 hours after cell capping in both drone and worker larvae of A. mellifera. This time frame, covering the conclusion of the honey bee brood cell invasion and the start of Varroa’s ovogenesis, is critical to the successful initiation of a reproductive cycle. Our findings support a later activation of the ecdysteroid cascade in honey bee drones compared to worker larvae, which could account for the increased egg production of Varroa in A. mellifera drones. The JH pathway was generally downregulated confirming its activity is antagonistic to the ecdysteroid pathway during the larva development. Nevertheless, the genes involved in JH synthesis revealed an increased expression in drones. The upregulation of jhamt gene involved in methyl farnesoate (MF) synthesis came into attention since the MF is not only a precursor of JH but it is also an insect pheromone in its own right as well as JH-like hormone in Acari. This could indicate a possible kairomone effect of MF for attracting the mites into the drone brood cells, along with its potential involvement in ovogenesis after the cell capping, stimulating Varroa’s initiation of egg laying.

Sheets 1-3: Moulting hormone pathway: differential gene expressions between time points in Drones (Sheet 1), Workers (Sheet 2); differential gene expressions between Drones and Workers at each time point (Sheeet 3); Gene IDs, log2-fold-changes (logFC) and statistical significance (P values).

Sheet 4-6: Juvenile hormone pathway: differential gene expressions between time points in Drones (Sheet 4) and Workers (Sheet 5); differential gene expressions between Drones and Workers at each time point (Sheet 6); Gene IDs, log2-fold-changes (logFC) and statistical signifficance (P values).

Sheet 7-9: Fibroins: differential gene expressions between time points in Drones (Sheet 7) and Workers (Sheet 8); differential gene expressions between Drones and Workers at each time point (Sheet 9); Gene IDs, log2-fold-changes (logFC) and statistical significance (P values).

Sheet 10: The results of statistical analysis for jhamt relative expression during temporal and drone versus worker analysis. The significant p values for ANOVA and Bonferroni post hoc are highlighted in yellow.

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