Identification and Validation of Reference Genes for Quantitative Gene Expression Analysis in Ophraella communa

文献类型： 外文期刊

第一作者： Zhang, Yan

作者： Zhang, Yan;Chen, Jiqiang;Chen, Guangmei;Ma, Chao;Chen, Hongsong;Gao, Xuyuan;Tian, Zhenqi;Cui, Shaowei;Tian, Zhenya;Guo, Jianying;Wan, Fanghao;Zhou, Zhongshi;Chen, Hongsong;Gao, Xuyuan

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关键词： Ophraella communa; RT-qPCR; reference gene; normalization; gene expression

期刊名称：FRONTIERS IN PHYSIOLOGY （ 影响因子：4.566； 五年影响因子：4.804 ）

ISSN： 1664-042X

年卷期： 2020 年 11 卷

页码：

收录情况： SCI

摘要： Ophraella communa is an effective bio-control agent of the invasive common weed. By now, the reference genes in O. communa have not yet been screened and validated. The aim of this study was to screen for the most stable reference genes in different backgrounds, such as different developmental stages, sexes, tissues, and male reproductive system with different body sizes. We selected 12 common housekeeping genes involved in different biological processes, including GAPDH, ACT1, ACT2, ARF1, ARF4, SDH, beta TUBC, RPL4, RPL19, RPS18, EF1 alpha, and COX as the candidate reference genes. To analyze the stability of the candidate reference genes, we first used three dedicated algorithms, GeNorm, NormFinder, and BestKeeper, and further comprehensive ranking was provided by ReFinder. The results showed that RPL19 and RPL4 exhibited the least variation in different developmental stages/sexes and in male reproductive systems with different body sizes. COX proved to be most suitable for normalizing the gene expression levels in different tissues, and coincidentally, RPL19 was also found to be second in terms of stability in this study. To the best of our knowledge, this is the first study to identify suitable reference genes for analyzing gene expression in O. communa; thus, this study would lay the foundation for future research on the molecular physiology and biochemistry of O. communa and other insects.

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