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Honey bees are exposed to various pesticides via both acute and chronic routes. Transcriptional profiling based on quantitative real-time PCR could be used as a representative method to assess the effects of pesticides on honey bee physiology. In this method, the identification of reference genes left unaffected by pesticide treatment is required for accurate analysis; however, there have been no reports regarding such reference genes. In this study, the expression level fluctuations of two putative reference genes ( arf1  and  rab1 ), along with three commonly used reference genes ( rpS18 ,  actin , and  gapdh ), were investigated in different stages and tissues following treatment with fluvalinate, coumaphos, imidacloprid, and flupyradifurone, and their performances were compared using geNorm, BestKeeper, NormFinder and RefFinder. The expression stability of  rab1a  was the highest, followed by that of  arf1 ,  rpS18 ,  actin , and  gapdh , suggesting that  rab1  and  arf1  can be employed as reliable reference genes in pesticide studies.

Selection of stable reference genes for real-time quantitative PCR in honey bee pesticide toxicity studies