title: Exploring valid reference genes for quantitative real-time PCR analysis in Plutella xylostella (Lepidoptera: Plutellidae).
abstract: Quantitative real-time PCR (qRT-PCR), a primary tool in gene expression analysis, requires an appropriate normalization strategy to control for variation among samples. The best option is to compare the mRNA level of a target gene with that of reference gene(s) whose expression level is stable across various experimental conditions. In this study, expression profiles of eight candidate reference genes from the diamondback moth, Plutella xylostella, were evaluated under diverse experimental conditions. RefFinder, a web-based analysis tool, integrates four major computational programs including geNorm, Normfinder, BestKeeper, and the comparative ΔCt method to comprehensively rank the tested candidate genes. Elongation factor 1 (EF1) was the most suited reference gene for the biotic factors (development stage, tissue, and strain). In contrast, although appropriate reference gene(s) do exist for several abiotic factors (temperature, photoperiod, insecticide, and mechanical injury), we were not able to identify a single universal reference gene. Nevertheless, a suite of candidate reference genes were specifically recommended for selected experimental conditions. Our finding is the first step toward establishing a standardized qRT-PCR analysis of this agriculturally important insect pest.
authors: Institute of Pesticide Science, Hunan Agricultural University, Changsha 410128, China.
authors: Wei
authors: W
authors: Fu
authors: Wen
authors: W
authors: Xie
authors: Zhuo
authors: Z
authors: Zhang
authors: Shaoli
authors: S
authors: Wang
authors: Qingjun
authors: Q
authors: Wu
authors: Yong
authors: Y
authors: Liu
authors: Xiaomao
authors: X
authors: Zhou
authors: Xuguo
authors: X
authors: Zhou
authors: Youjun
authors: Y
authors: Zhang
mesh_terms: D000818:Animals; D020869:Gene Expression Profiling; D005796:Genes / Q000235:genetics*; D009036:Moths / Q000235:genetics*; D020648:Peptide Elongation Factor 1 / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D012016:Reference Values
chemical_list: D020648:Peptide Elongation Factor 1
keywords: Plutella xylostella; abiotic factor.; biotic factor; qRT-PCR analysis; reference gene
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