Abstract
Bombyx mori bidensovirus (BmBDV) is a unique bipartite DNA virus that causes chronic disease in silkworms, and its interactions with the host give rise to variations of genes expression. Reverse transcription quantitative real-time PCR (RT-qPCR) has extensive applications in mRNA quantification. To obtain reproducible and meaningful quantification of transcripts, it is crucial to identify the best reference genes in preliminary work. In this work, we analyzed the expression of seven candidate reference genes in silkworm larvae infected or uninfected with BmBDV, and compared their expression stability with three statistical methods (geNorm, NormFinder, and BestKeeper). The results revealed that the RPL3 and 28S rRNA were the stable internal control, and UBQ was the least stable gene. Furthermore, the expression of Bombyx mori serine protease (BmSP142) was assessed using RPL3 as internal. The expression of BmSP142 was significantly different between BmBDV-resistant and BmBDV-susceptible silkworm after virus infection, suggesting that BmSP142 might be associated with the resistance to BmBDV in silkworm.
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