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- Title
Screening potential reference genes for quantitative real-time PCR analysis in the oriental armyworm, Mythimna separata.
- Authors
Li, Hong-Bo; Dai, Chang-Geng; Zhang, Chang-Rong; He, Yong-Fu; Ran, Hai-Yan; Chen, Shi-Hong
- Abstract
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-α, GAPDH, RPS7, RPS13, RPL32 and TUB) in M. separata using the comparative ΔCt 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-α and TUB were optimal for different developmental stages; TUB, RPS13 and EF1-α 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-α, RPL32 and GAPDH were 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.
- Subjects
MYTHIMNA separata; INSECT pests; GENE targeting; GENE expression; MEDICAL screening; QUANTITATIVE research; POLYMERASE chain reaction
- Publication
PLoS ONE, 2018, Vol 13, Issue 4, p1
- ISSN
1932-6203
- Publication type
Article
- DOI
10.1371/journal.pone.0195096