Supplementary Tables

We performed an RNA-seq based comparison of gene expression levels in the antennae of honey bee drones and time-trained foragers (workers) collected at different times of the day and different activity states. Interestingly, olfaction-related genes (i.e. odorant receptor (Ors), odorant binding proteins (Obps), carboxyl esterases (CEst) etc.) showed stable gene expression differences between drone and worker antennae. Drone antennae showed higher expression of 24 Ors, of which 21 belong to the clade X which comprises the receptor for the major queen pheromone compound 9-ODA. This high number of drone-biased Ors suggests that more Ors than previously expected play a role in sex-pheromone communication. In addition, we found higher expression levels for many non-olfaction related genes including nitric oxide synthase (NOS), and the potassium channel Shaw. In contrast, workers showed higher expression of 67 Ors, which belong to different Or clades that are involved in pheromone communication as well as the perception of cuticular hydrocarbons and floral scents. Further, drone antennae showed higher expression of genes involved in energy metabolism, whereas worker antennae showed higher expression of genes involved in neuronal communication consistent with earlier reports on peripheral olfactory plasticity. Finally, drones that perform mating flight in the afternoon (innate) and foragers that are trained to forage in the afternoon (adapted) showed similar daily changes in the expression of two major clock genes, period and cryptochrome2. Most of the other genes showing changes with time or onset of daily flight activity were specific to drones and foragers.

本研究基于RNA测序（RNA-seq）技术，对不同时段采集、处于不同活动状态的蜜蜂雄蜂与经时段训练的采集工蜂的触角基因表达水平开展对比分析。值得注意的是，嗅觉相关基因（即嗅觉受体（Odorant Receptors, Ors）、嗅觉结合蛋白（Odorant Binding Proteins, Obps）、羧酸酯酶（Carboxyl Esterases, CEst）等）在雄蜂与工蜂触角中呈现出稳定的表达差异。雄蜂触角中24种嗅觉受体的表达量更高，其中21种属于X进化支，该进化支包含了主要蜂王信息素成分9-ODA的受体。这类雄蜂偏好型嗅觉受体的高占比提示，相较于此前认知，有更多嗅觉受体参与性信息素通讯过程。此外，本研究还发现多种非嗅觉相关基因的表达量上调，包括一氧化氮合酶（Nitric Oxide Synthase, NOS）以及钾通道Shaw。与之相对，工蜂触角中67种嗅觉受体的表达量更高，这些受体分属不同的嗅觉受体进化支，参与信息素通讯以及表皮碳氢化合物与花香的感知。进一步而言，雄蜂触角中参与能量代谢的基因表达量更高，而工蜂触角中参与神经元通讯的基因表达量更高，这一结果与此前关于外周嗅觉可塑性的研究报道相符。最后，具有午后婚飞先天行为的雄蜂，与经过午后采集训练的采集工蜂，在两种核心节律基因（period即周期基因与cryptochrome2即隐花色素2）的表达上呈现出相似的昼夜变化模式。其余多数随昼夜时段或日常飞行活动起始而表达变化的基因，则仅在雄蜂或工蜂中特异性存在。