Reference Genes for Expression Analysis Using RT-qPCR in Cnaphalocrocis medinalis (Lepidoptera: Pyralidae)
2022
Xiaoyu Zhao | Jiawen Guo | Yanhui Lu | Tianyi Sun | Junce Tian | Jianlei Huang | Hongxing Xu | Zhengliang Wang | Zhongxian Lu
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 (EF1&alpha:, AK, EF1&beta:, 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 EF1&beta:, PGK, and RPL18, related to developmental stages: RPS3 and RPL18 in larval tissues: EF1&beta: and PGK in larvae feeding on different rice varieties: EF1&alpha:, EF1&beta:, and PGK in larvae temperature treatments: PGK and RPL13, related to different adult ages: PGK, EF1&alpha:, 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.
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