Selection of Bactrocera tau (Walker) Reference Genes for Quantitative Real-Time PCR

The selection of appropriate reference genes is critical for standardizing quantitative real-time reverse transcription polymerase chain reaction (RT-qPCR) data, thereby ensuring accurate and reliable results of gene expression analysis. In this study, we identified 10 candidate reference genes (encoding <i>α-tubulin</i>, <i>G6PDH</i>, <i>Rab1</i>, <i>RT</i>, <i>RPS13</i>, <i>β-tubulin</i>, <i>DPH1</i>, <i>HSP90</i>, <i>GAPDH</i>, and <i>CP</i>) and evaluated their suitability for use as reference genes in the pest insect, <i>Bactrocera tau</i>. Analysis was conducted using three software-based methods-Delta CT, NormFinder, and BestKeeper-alongside the online tool RefFinder. Expression levels of these genes were analyzed across various <i>B. tau</i> developmental stages and body parts. The overall ranking of reference gene stability scores was as follows: <i>α-tubulin</i> > <i>G6PDH</i> > <i>CP</i> > <i>β-tubulin</i> > <i>RT</i> > <i>HSP90</i> > <i>GAPDH</i> > <i>DPH1</i> > <i>RPS13</i> > <i>Rab1</i>. Ultimately, <i>α-tubulin</i> and <i>G6PDH</i> were identified as the most stable reference genes for <i>B. tau</i>.