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

• Published in: Insects (Basel, Switzerland) Vol. 13; no. 11; p. 1046
• Main Authors: Zhao, Xiaoyu, Guo, Jiawen, Lu, Yanhui, Sun, Tianyi, Tian, Junce, Huang, Jianlei, Xu, Hongxing, Wang, Zhengliang, Lu, Zhongxian
• Format: Journal Article
• Language: English
• Published: Basel MDPI AG 13.11.2022; MDPI
• Subjects: Algorithms; Behavior; Butterflies & moths; Cnaphalocrocis medinalis; Cotton; Dehydrogenases; Developmental stages; Environmental changes; Experiments; expression stability; Gene expression; Genes; Glyceraldehyde-3-phosphate dehydrogenase; Humidity; Insects; Kinases; Larvae; Mating; migratory insect; Molecular modelling; Pests; Physiological effects; Physiological responses; Proteins; reference genes; Reliability analysis; Reliability aspects; Rice; rRNA 18S; RT-qPCR; Seeds; Takeoff; Trypsin; China
• ISSN: 2075-4450; 2075-4450
• DOI: 10.3390/insects13111046

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.