Identification of reference genes for gene expression analysis at different developmental stages of the bumblebee Bombus terrestris (Hymenoptera: Apidae)

Bumblebees are important pollinators for many wild plants and crops. To explore gene expression characteristics in depth, stable reference genes will be essential across different developmental stages. In this context, we evaluated the stability of eight common reference genes at ten developmental stages in the bumblebee Bombus terrestris (B. terrestris) with four algorithms and two rank approaches. The results showed that all the primers for the reference genes exhibited high specificity, linear correlation (R2-value 0.9917~0.9978) and a good PCR amplification efficiency (E-value 95.4~108.6%). The combined use of RPL13 and RP49 was suggested to be the most suitable reference genes. Our candidate reference genes will help to improve the precision of the detection of gene expression at different developmental stages of bumblebee species.