ArticleJournal of insect science (Online)2016
Identification and Validation of Reference Genes for the Normalization of Gene Expression Data in qRT-PCR Analysis in Aphis gossypii (Hemiptera: Aphididae).
Article in Journal of insect science (Online), 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 38 papers.
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38 citing papers in PubMed, 97 citations in OpenAlex.
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- Identification of candidate host-manipulating effector genes in Aphis gossypii (Hemiptera: Aphididae) using a combination of transcriptome, genome, and differential gene expression data.Journal of insect science (Online) · 2025Article
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- Evaluation and Validation of Reference Genes for Gene Expression Analysis Using qRT-PCR in the Sugarcane Stem BorerInsects · 2024Article
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- Selection and Validation of Reference Genes for Quantitative Real-Time PCR Normalization inFrontiers in physiology · 2022Article
- Early-Stage Defense Mechanism of the Cotton AphidFrontiers in immunology · 2022Article
- Identification of suitable reference genes for expression profiling studies using qRT-PCR in an important insect pest, Maruca vitrata.Molecular biology reports · 2021Article
- UDP-Glycosyltransferases from the UGT344 Family Are Involved in Sulfoxaflor Resistance inInsects · 2021Article
- Reference Gene Selection for Normalizing Gene Expression inFrontiers in physiology · 2021Article
- Selection and Validation of Reference Genes For qRT-PCR Analysis ofFrontiers in physiology · 2021Article
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Authors and funding
6 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
To obtain accurate and reliable results from quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) analysis, it is necessary to select suitable reference genes as standards for normalizing target gene expression data. QRT-PCR is a popular analytical methodology for studying gene expression and it has been used widely in studies of Aphis gossypii Glover in recent years. However, there is absence of study on the stability of the expression of reference genes in A. gossypii. In this study, eight commonly used candidate reference genes, including 18S, 28S, β-ACT, GAPDH, EF1α, RPL7, α-TUB, and TBP, were evaluated under various experimental conditions to assess their suitability for use in the normalization of qRT-PCR data. The optimal number of reference genes was determined using the geNorm program, and the suitability of particular reference genes was empirically validated by performing normalizations of expression data for the HSP70 gene. The results showed the most suitable combinations of reference genes for the different experimental conditions. For experiments based on divergent developmental stages, EF1α, β-ACT, and RPL7 are the optimal reference gene combination, both EF1α and β-ACT are the optimal combination used in the experiments of different geographical populations, whereas for experiments of the temperature changes, the combination of GAPDH and RPL7 is optimal, both 18S and β-ACT are an optimal combination for feeding assay experiments. These research results should be useful for the selection of the suitable reference genes to obtain reliable qRT-PCR data in the gene expression study of A. gossypii.
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