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Expression profiling in Bemisia tabaci under insecticide treatment: indicating the necessity for custom reference
Pei Liang1, Yajie Guo1, Xuguo Zhou2
1Department of Entomology, China Agricultural University, Beijing, P. R. China.
Plos One
|February 6, 2014
Summary
Selecting stable reference genes for quantitative PCR (qRT-PCR) is crucial for accurate results in pest research. This study identified elongation factor 1α (EF1α), α-tubulin (TUB1α), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as reliable reference genes for insecticide studies in Bemisia tabaci.
Area of Science:
- Molecular Biology
- Entomology
- Pest Management
Background:
- Accurate gene expression analysis using quantitative reverse transcription PCR (qRT-PCR) relies on stable reference genes.
- Existing reference genes lack universal applicability across diverse experimental conditions, including various biotic and abiotic stresses.
- The sweetpotato whitefly (Bemisia tabaci) is a globally significant invasive pest, necessitating reliable molecular tools for its study.
Purpose of the Study:
- To evaluate the stability of eight candidate reference genes in Bemisia tabaci under insecticide treatment.
- To identify the most stable reference genes for qRT-PCR analysis in this pest species when exposed to different insecticide classes.
- To validate the suitability of selected reference genes using target gene expression analysis.
Main Methods:
- Quantitative reverse transcription PCR (qRT-PCR) was employed to analyze the expression profiles of eight housekeeping genes.
- Bemisia tabaci were exposed to eight commercial insecticides from five different classes.
- Reference gene stability was assessed using statistical algorithms, and validated with target genes (Cyp6cm1 and GST).
Main Results:
- Elongation factor 1α (EF1α), α-tubulin (TUB1α), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were identified as the most stable reference genes under insecticide exposure.
- The ranking of reference gene stability varied significantly between different insecticide classes and even within the same class.
- Validation using target genes confirmed the differential stability of reference genes across various treatments.
Conclusions:
- Custom selection of reference genes is essential for each specific experimental condition, even when investigating the same abiotic or biotic factor.
- EF1α, TUB1α, and GAPDH are recommended as stable reference genes for Bemisia tabaci under general insecticide treatment.
- The findings underscore the need for rigorous validation of reference genes to ensure the accuracy of qRT-PCR data in insecticide resistance studies.
