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International audience; AbstractMandibular gland (MG) secretions of honey bee are synthesized through a three-step bifurcation pathway, and the initiate hydroxylation step is carried out by cytochrome P450 enzymes (P450). We have previously revealed that CYP6AS8 and CYP6AS11 are possibly participating in this process. In this study, we evaluated the function of CYP6AS8 in honey bee worker’s MG using RNA interference technology, and we investigated the expression pattern of CYP6AS8 in MGs from workers of different ages. We found that the expression of CYP6AS8 was detectable in MGs across the whole life cycle of Apis mellifera except for newly emerged workers and predominately expressed at day 15. RNAi knockdown revealed that the production of 10-HDA was synchronized with the expression level of CYP6AS8 in MG of honey bee workers; the downregulation of CYP6AS8 caused a significant reduction of 10-HDA content, demonstrating that CYP6AS8 is indispensable for 10-HDA biosynthesis. Our current results confirm that CYP6AS8 modulates 10-HDA biosynthesis in MGs of western honey bee worker and provided the first evidence to identify the certain P450 participate in the MG secretion biosynthesis of honey bee workers.