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Exploring valid reference genes for quantitative real-time PCR analysis in Plutella xylostella (Lepidoptera: Plutellidae)

文献类型： 外文期刊

作者： Fu, Wei 2 ; Xie, Wen 2 ; Zhang, Zhuo 3 ; Wang, Shaoli 2 ; Wu, Qingjun 2 ; Liu, Yong 3 ; Zhou, Xiaomao 1 ; Zhou, Xuguo; Zh 1 ;

作者机构： 1.Hunan Agr Univ, Inst Pesticide Sci, Changsha 410128, Hunan, Peoples R China

2.Chinese Acad Agr Sci, Dept Plant Protect, Inst Vegetables & Flowers, Beijing 100081, Peoples R China

3.Hunan Acad Agr Sci, Hunan Plant Protect Inst, Changsha 410125, Hunan, Peoples R China

4.Key Lab Integrated Management Pests & Dis Hort Cr, Changsha 410125, Hunan, Peoples R China

5.Key Lab

关键词： Abiotic factor;Biotic factor;Plutella xylostella;qRT-PCR analysis;Reference gene

期刊名称：INTERNATIONAL JOURNAL OF BIOLOGICAL SCIENCES （ 影响因子：6.58； 五年影响因子：6.478 ）

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收录情况： SCI

摘要： 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.

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