Reference Genes for Expression Analysis Using RT-qPCR in Cnaphalocrocis medinalis (Lepidoptera: Pyralidae).

Cnaphalocrocis medinalis is a destructive migratory rice pest. Although many studies have investigated its behavioral and physiological responses to environmental changes and migration-inducing factors, little is known about its molecular mechanisms. This study was conducted to select suitable RT-qPCR reference genes to facilitate future gene expression studies. Here, thirteen candidate housekeeping genes ( EF1α , AK , EF1β , GAPDH , PGK , RPL13 , RPL18 , RPS3 , 18S rRNA , TBP1 , TBP2 , ACT , and UCCR ) were selected to evaluate their stabilities under different conditions using the ∆CT method; the geNorm, NormFinder, BestKeeper algorithms; and the online tool RefFinder. The results showed that the most stable reference genes were EF1β , PGK , and RPL18 , related to developmental stages; RPS3 and RPL18 in larval tissues; EF1β and PGK in larvae feeding on different rice varieties; EF1α , EF1β , and PGK in larvae temperature treatments; PGK and RPL13 , related to different adult ages; PGK , EF1α , and ACT , related to adult nutritional conditions; RPL18 and PGK , related to adult mating status; and, RPS3 and PGK , related to different adult take-off characteristics. Our results reveal reference genes that apply to various experimental conditions and will greatly improve the reliability of RT-qPCR analysis for the further study of gene function in this pest.