Reverse-transcription quantitative polymerase chain reaction (RT-qPCR) is commonly used to quantify gene expression. For normalization, the expression of each gene is compared with a reference “housekeeping” gene that is stably expressed under relevant stress. Unfortunately, there have been no reports on the stability of such reference genes under various treatments of the Spodoptera frugiperda. In this study, we used five tools (RefFinder, GeNorm, NormFinder, BestKeeper, and ΔCt methods) to evaluate the stability of 12 candidate reference genes (RPS18, β-tubulin, GAPDH, RPS7, RPS15, RPL7, RPL32, Actin-5C, EF1-α, EF1-γ, RPL27, and ACE) in different instars, tissues, and treatments (high and low temperature, UV-A, and emamectin benzoate). Several ribosomal proteins (RPS7, RPS15, RPL32, RPS18, and RPL7), GAPDH, Actin-5C, and β-tubulin, were relatively stable, suggesting that they are ideal housekeeping genes for various treatments. ACE was extremely unstable under various experimental treatments, rendering it unsuitable as an internal reference. This study identified the reference housekeeping genes stably expressed by S. frugiperda under different treatments, thus setting a foundation for further exploration of the physiological and biochemical mechanisms.

中文翻译：

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草地夜蛾（鳞翅目：夜蛾科）中候选RT-qPCR管家基因的表达稳定性

逆转录定量聚合酶链反应 (RT-qPCR) 通常用于量化基因表达。为了标准化，将每个基因的表达与在相关压力下稳定表达的参考“看家”基因进行比较。不幸的是，在草地贪夜蛾的各种处理下，没有关于这些参考基因的稳定性的报道。. 在本研究中，我们使用了五种工具（RefFinder、GeNorm、NormFinder、BestKeeper 和 ΔCt 方法）来评估 12 个候选参考基因（RPS18、β-微管蛋白、GAPDH、RPS7、RPS15、RPL7、RPL32、Actin-5C）的稳定性、EF1-α、EF1-γ、RPL27 和 ACE）在不同龄期、组织和处理（高温和低温、UV-A 和甲维菌素苯甲酸酯）中。几种核糖体蛋白（RPS7、RPS15、RPL32、RPS18 和 RPL7）、GAPDH、肌动蛋白 5C 和 β-微管蛋白相对稳定，表明它们是各种治疗的理想看家基因。ACE 在各种实验处理下极不稳定，使其不适合作为内部参考。本研究鉴定了草地贪夜蛾稳定表达的参考看家基因 为进一步探索生理生化机制奠定基础。