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Reference Gene Selection for Expression Analyses by qRT-PCR in Dendroctonus valens
Chunchun Zheng1, Dongfang Zhao1, Yabei Xu1
1Key Laboratory of Beijing for the Control of Forest Pests, Beijing Forestry University, Beijing 100083, China.
Insects
|May 31, 2020
Summary
This study identifies stable reference genes for gene expression analysis in the pine pest, *Dendroctonus valens*. Recommended gene pairs, 18S rRNA with PRS18 for adults and 18S rRNA with TUB for larvae, ensure accurate research under varying temperatures.
Area of Science:
- Entomology
- Molecular Biology
- Genetics
Background:
- *Dendroctonus valens* is a significant pest impacting *Pinus* species.
- Accurate gene expression analysis is crucial for understanding pest biology and developing control strategies.
- Identifying reliable reference genes is essential for normalizing gene expression data in *D. valens*.
Purpose of the Study:
- To determine the most stable reference genes for quantitative gene expression analysis in adult and mature larval stages of *Dendroctonus valens*.
- To evaluate candidate reference genes under diverse temperature conditions relevant to the pest's environment.
- To provide validated reference gene recommendations for future molecular studies on *D. valens*.
Main Methods:
- Candidate reference gene sequences (*ACT*, *TUB*, *SHDA*, *PRS18*, *18S rRNA*, *CYP4G55*) were retrieved from transcriptome data.
- Real-time quantitative PCR (qPCR) was employed to analyze the expression levels of these candidate genes.
- Statistical algorithms (geNorm, NormFinder, BestKeeper) were utilized to assess the expression stability of the selected reference genes.
Main Results:
- In adult *D. valens*, *18S rRNA*, *PRS18*, and *TUB* exhibited stable expression, with *18S rRNA* being the most stable, followed by *PRS18* and then *TUB*.
- In mature larvae, *TUB*, *18S rRNA*, and *SDHA* showed stable expression, with *TUB* as the most stable, followed by *18S rRNA* and *SDHA*.
- The combination of *18S rRNA* and *PRS18* is recommended for adults, while *18S rRNA* and *TUB* are recommended for mature larvae.
Conclusions:
- Validated reference genes are essential for reliable gene expression studies in *Dendroctonus valens*.
- Specific reference gene combinations (*18S rRNA*/ *PRS18* for adults, *18S rRNA*/ *TUB* for larvae) are recommended for accurate normalization under varying temperatures.
- These findings will enhance the precision of molecular research on this economically important pine pest.
