다양한 조건의 살충제에 노출된 꿀벌에서의 qRT-PCR을 위한 안정 발현 reference 유전자 선정

Abstract

Pesticides have been considered to be a major factor that leads to a significant decline in honey bee populations, including colony collapse disorder. To understand pesticide detoxification physiology in honey bees, it is essential to identify the gene expression in different pesticideexposed conditions. For the accurate determination of target gene quantification through quantitative real-time polymerase chain reaction (qRT-PCR), reference gene stably expressing across different samples should be selected. Therefore, in this study, to select the optimal reference genes, we analyzed the amplification stability of five candidate reference genes (RPS5, RPS18, GAPDH, ARF1, and RAB1A) from honey bees exposed to seven pesticides (acetamiprid, imidacloprid, flupyradifurone, fenitrothion, carbaryl, amitraz, and bifenthrin) across various condition of samples, including different tissues and exposure time and concentration of pesticides using four programs (geNorm, NormFinder, BestKeeper, and RefFinder). Although the stability values of genes varied depending on different analysis algorithms, our results suggest that RPS5 is the most appropriate reference gene for the identification of target gene expression levels in qRT-PCR assays for honeybees under various conditions of pesticide treatment.