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1 June 2013 Selection of Endogenous Reference Genes for Gene Expression Analysis in the Mediterranean Species of the Bemisia tabaci (Hemiptera: Aleyrodidae) Complex
Yun-Lin Su, Wen-Bo He, Jia Wang, Jun-Min Li, Shu-Sheng Liu, Xiao-Wei Wang
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Abstract

Quantitative real-time reverse transcription polymerase chain reaction is widely used for gene expression analysis, and robust normalization against stably expressed endogenous reference genes (ERGs) is necessary to obtain accurate results. In this study, the stability of nine housekeeping genes of the sweetpotato whitefly, Bemisia tabaci (Hemiptera: Aleyrodidae) Mediterranean were evaluated in various conditions by quantitative real-time reverse transcription polymerase chain reaction using geNorm and Normfinder programs. Both programs suggested α-tubulin/ubiquitin and 18S small subunit ribosomal RNA the most stable genes for bacterium- and insecticide-treated whiteflies, respectively. For developmental stages, organs, and the samples including salivary glands and the whole body, transcription initiation factor TFIID subunit was calculated as the most stably expressed gene by both programs. In addition, we compared the RNA-seq data with the results of geNorm and Normfinder and found that the stable genes revealed by RNA-seq analysis were also the ERGs recommended by geNorm and Normfinder. Furthermore, the use of the most stable gene suggested by RNA-seq analysis as an ERG produced similar gene expression patterns compared with results generated from the normalization against the most stable gene selected by geNorm and Normfinder and multiple genes recommended by geNorm. It indicates that RNA-seq data are reliable and provide a great source for ERG candidate exploration. Our results benefit future research on gene expression profiles of whiteflies and possibly other organisms.

© 2013 Entomological Society of America
Yun-Lin Su, Wen-Bo He, Jia Wang, Jun-Min Li, Shu-Sheng Liu, and Xiao-Wei Wang "Selection of Endogenous Reference Genes for Gene Expression Analysis in the Mediterranean Species of the Bemisia tabaci (Hemiptera: Aleyrodidae) Complex," Journal of Economic Entomology 106(3), 1446-1455, (1 June 2013). https://doi.org/10.1603/EC12459
Received: 9 November 2012; Accepted: 19 April 2013; Published: 1 June 2013
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KEYWORDS
Bemisia tabaci
endogenous reference gene
gene expression
quantitative PCR
RNA-seq
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