Abstract
Dendroctonus valens is the main pest of the genus Pinus. To facilitate gene expression analyses, suitable reference genes for adults and mature larvae of D. valens under different temperature conditions were determined. In particular, we obtained the sequences of candidate reference genes, ACT, TUB, SHDA, PRS18, 18S rRNA, and CYP4G55, 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 18S rRNA, PRS18, and TUB were stable in adults, in which 18S rRNA > PRS18 > TUB. In mature larvae, the expression levels of TUB, 18S rRNA, and SDHA were stable, in which TUB > 18S rRNA > SDHA. The combination of 18S rRNA and PRS18 is recommended for studies of gene expression in adults and the combination of 18S rRNA and TUB is effective for studies of gene expression in mature larvae of D. valens under different temperature conditions.
Highlights
Dendroctonus valens (Coleoptera: Scolytidae) is a major pest of the genus Pinus, native to Canada and North America
Actin tissues (ACT), SDHA, 18S rRNA, RPS18, CYP4G55, and TUB were selected as candidate reference genes based on transcriptome sequencing data
Six candidate reference genes were screened from transcriptome data for D. valens
Summary
Dendroctonus valens (Coleoptera: Scolytidae) is a major pest of the genus Pinus, native to Canada and North America. The pest has caused serious harm to Pinus tabulaeformis and P. Sylvestris and has become one of the main invasive pests in China. In the implementation of qRT-PCR, RNA extraction, polymerase amplification, and cDNA synthesis can all lead to systematic errors [2,4]. To eliminate these sources of error, genes with constant expression levels across conditions are usually selected as references for normalization. The expression level of an ideal reference gene remains constant [5]. Common reference genes for insect studies include beta-actin, beta-tubulin, alpha-tubulin, ribosomal protein S18 (PRS18), 18S ribosomal
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