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

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-?, GAPDH, RPS7, RPS13, RPL32 and TUB) in M. separata using the comparative ?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-? and TUB were optimal for different developmental stages; TUB, RPS13 and EF1-? 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-?, 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.

SUBMITTER: Li HB 

PROVIDER: S-EPMC5884543 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature