사후 조건의 꿀벌 내역봉에서 qRT-PCR을 위한 안정적인 reference 유전자 선발

초록

The western honey bee (Apis mellifera) is essential for pollination, supporting both ecosystem stability and agricultural productivity. Recently, mass colony losses have been reported worldwide, including in Korea. Investigating gene expression in postmortem honey bee samples may help identify the causes of colony collapse. However, selecting stable reference genes under postmortem conditions is critical for accurate gene expression analysis using quantitative real-time PCR (qRT-PCR). In this study, we evaluated the expression stability of 10 candidate reference genes (RPS5, RPS18, GAPDH, ARF1, RAB1a, PPI, PGK, SDH, TBP, and EF1) in the head tissues of honey bees collected at postmortem intervals (PMIs) ranging from 1 to 7 days. We assessed gene stability using Cq value distribution and four algorithms: NormFinder, BestKeeper, geNorm, and RefFinder. In addition, expression levels of the target gene EcR were compared using both single and multiple reference genes. Among the candidates, RAB1a showed the highest overall stability across all PMIs and was most consistent with multi-gene normalization results, making it the most suitable single reference gene. Abdominal tissues exhibited greater variability due to decomposition, suggesting head tissue is more appropriate for postmortem gene expression analysis. These results provide a foundation for postmortem molecular studies in honey bees and may contribute to understanding colony loss and estimating PMI in apiculture.

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