| Summary: | <i>Dendroctonus valens</i> is the main pest of the genus <i>Pinus</i>. To facilitate gene expression analyses, suitable reference genes for adults and mature larvae of <i>D. valens</i> under different temperature conditions were determined. In particular, we obtained the sequences of candidate reference genes, <i>ACT</i>, <i>TUB</i>, <i>SHDA</i>, <i>PRS18</i>, <i>18S rRNA</i>, and <i>CYP4G55</i>, from transcriptome data. Real-time quantitative PCR was used to analyze gene expression, and geNorm, NormFinder, and BestKeeper were used to evaluate expression stability. Under different temperature conditions, the expression levels of <i>18S rRNA</i>, <i>PRS18</i>, and <i>TUB</i> were stable in adults, in which <i>18S rRNA</i> > <i>PRS18</i> > <i>TUB</i>. In mature larvae, the expression levels of <i>TUB</i>, <i>18S rRNA</i>, and <i>SDHA</i> were stable, in which <i>TUB</i> > <i>18S rRNA</i> > <i>SDHA</i>. The combination of <i>18S rRNA</i> and <i>PRS18</i> is recommended for studies of gene expression in adults and the combination of <i>18S rRNA</i> and <i>TUB</i> is effective for studies of gene expression in mature larvae of <i>D. valens</i> under different temperature conditions.
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