Reference Genes for Expression Analysis Using RT-qPCR in <i>Cnaphalocrocis medinalis</i> (Lepidoptera: Pyralidae)
Xiaoyu Zhao,
Jiawen Guo,
Yanhui Lu,
Tianyi Sun,
Junce Tian,
Jianlei Huang,
Hongxing Xu,
Zhengliang Wang,
Zhongxian Lu
Affiliations
Xiaoyu Zhao
College of Life Sciences, China Jiliang University, Hangzhou 310018, China
Jiawen Guo
State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products, Institute of Plant Protection and Microbiology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China
Yanhui Lu
State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products, Institute of Plant Protection and Microbiology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China
Tianyi Sun
College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China
Junce Tian
State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products, Institute of Plant Protection and Microbiology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China
Jianlei Huang
College of Agriculture and Forestry, Hebei North University, Zhangjiakou 075000, China
Hongxing Xu
State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products, Institute of Plant Protection and Microbiology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China
Zhengliang Wang
College of Life Sciences, China Jiliang University, Hangzhou 310018, China
Zhongxian Lu
College of Life Sciences, China Jiliang University, Hangzhou 310018, China
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.
