Han Shipeng, Qin Qiuju, Wang Da, Zhou Yayuan, He Yunzhuan
College of Plant Protection, Hebei Agricultural University, Baoding 071000, China.
Insects. 2021 Oct 3;12(10):902. doi: 10.3390/insects12100902.
As an accurate and convenient technique, the qRT-PCR is always used in the quantitative expression analysis of functional genes. Normalization of the data relies on stable reference genes. The fall armyworm (J. E. Smith) is an important invasive and migratory pest that seriously threatens corn production around the world. In this paper, we selected 10 candidate reference genes (, , , , , , , , , and ) and determined their expression levels under different conditions (different developmental stages, various tissues, mating status, hormones, diets, and temperatures). Subsequently, the stability of reference genes was evaluated by four algorithms (Delta Ct method, geNorm, NormFinder, BestKeeper). The optimal combination of reference genes for each treatment was obtained by geNorm. Finally, the comprehensive ranks were determined by the online tool RefFinder. Results showed that the most stable reference genes were , , and for developmental stages, , , and for different tissues, , , and for mating status, and under hormone treatment, , , and under diet treatment, , , and under temperature treatment. This study confirmed recent data on a few reference genes and provided an evaluation of a number of additional reference genes of under various conditions.
作为一种准确且便捷的技术，qRT-PCR 常用于功能基因的定量表达分析。数据的标准化依赖于稳定的内参基因。草地贪夜蛾（J. E. 史密斯）是一种重要的入侵性迁飞害虫，严重威胁着全球玉米生产。在本文中，我们选取了10个候选内参基因（，，，，，，，，，和），并测定了它们在不同条件下（不同发育阶段、各种组织、交配状态、激素、食物和温度）的表达水平。随后，通过四种算法（ΔCt 法、geNorm、NormFinder、BestKeeper）评估内参基因的稳定性。通过 geNorm 获得每种处理的内参基因最佳组合。最后，由在线工具 RefFinder 确定综合排名。结果表明，发育阶段最稳定的内参基因是，，和；不同组织是，，和；交配状态是，，和；激素处理下是和；食物处理下是，，和；温度处理下是，，和。本研究证实了关于一些内参基因的最新数据，并对草地贪夜蛾在各种条件下的多个其他内参基因进行了评估。
