Screening potential reference genes for quantitative real-time PCR analysis in the oriental armyworm, Mythimna separata

文献类型： 外文期刊

第一作者： Li, Hong-Bo

作者： Li, Hong-Bo;Dai, Chang-Geng;Zhang, Chang-Rong;He, Yong-Fu;Ran, Hai-Yan;Chen, Shi-Hong

作者机构：

期刊名称：PLOS ONE （ 影响因子：3.24； 五年影响因子：3.788 ）

ISSN： 1932-6203

年卷期： 2018 年 13 卷 4 期

页码：

收录情况： SCI

摘要： The oriental armyworm, Mythimna separata, is a major insect pest in China and other Asian countries. Unfortunately, suitable reference genes for quantitative real-time PCR (qRT-PCR) have not been previously identified in M. separata for normalizing target gene expression. In this study, we evaluated the expression stability of eight candidate genes (18S, ACT, EF1-alpha, GAPDH, RPS7, RPS13, RPL32 and TUB) in M. separata using the comparative Delta Ct method, BestKeeper, Normfinder geNorm and ReFinder, a comprehensive software platform. The results indicated that the appropriate reference gene varied depending on the experimental conditions. We found that ACTIN, EF1-alpha and TUB were optimal for different developmental stages; TUB, RPS13 and EF1-alpha showed the most stable expresssion in different tissues; RPS13 and 18S were the best reference genes for monitoring expression under high temperature conditions; TUB, RPS13 and RPS7 exhibited the most stable expression under larval-crowding conditions; RPS7, EF1-alpha, RPL32 and GAPDHwere the best for pesticide exposure experiments. This study provides tools for reliable normalization of qRT-PCR data and forms a foundation for functional studies of target gene expression in M. separata.

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