Reference Gene Selection for Quantitative Gene Expression Analysis in Argynnis hyperbius

文献类型： 外文期刊

第一作者： Xin, Hong-Juan

作者： Xin, Hong-Juan;Liu, Chen-Yang;Yan, Feng;Wang, Lu-Dan;Zhai, Qing;Zhang, Huan-Huan;Shen, Chen-Hui

作者机构：

关键词： Argynnis hyperbius; reference gene; RT-qPCR; selection; validation

期刊名称：INSECTS （ 影响因子：2.9； 五年影响因子：3.3 ）

ISSN：

年卷期： 2025 年 16 卷 10 期

页码：

收录情况： SCI

摘要： Argynnis hyperbius (Lepidoptera: Nymphalidae), as an important environmental indicator species, has shown a gradual decline in its species richness amid intensifying climate change and increasing environmental pressures. Screening for optimal reference genes is fundamental to studying their physiological and adaptive changes using RT-qPCR technology. In this study, 10 commonly used reference genes (ACT, alpha-TUB, AK, GAPDH, EF1 alpha, BTF3, RPS3, RPL10, RPL32, and RPL27) were selected, and their expression stability under different developmental stages, genders, and temperature treatments was evaluated using the RefFinder website. The results indicated that selecting a pair of reference genes could achieve the most reliable normalization analysis under all experimental conditions. Specifically, AK and EF1 alpha were the most stably expressed reference genes across different developmental stages; ACT and RPL32 showed the most stable expression in different adult sexes; and EF1 alpha and RPL27 exhibited stable expression under different temperature treatments. Additionally, this study used EF1 alpha and RPL32, the most stable reference genes from all results, to normalize and analyze the relative transcription levels of HSP90 under different temperatures, revealing significant differences between the 4 degrees C group and both the 26 degrees C and 37 degrees C groups. The findings of this study will significantly improve the reliability of RT-qPCR detection and lay a foundation for in-depth research on the gene expression, physiology, and biochemical mechanisms of A. hyperbius.

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