Evaluation of reference genes for quantitative real-time PCR normalization in the scarab beetleHolotrichia oblita

文献类型： 外文期刊

第一作者： Xie, Minghui

作者： Xie, Minghui;Zhong, Yongzhi;Lin, Lulu;Zhang, Guangling;Su, Weihua;Chen, Haoliang;Ni, Wanli;Qu, Mingjing

作者机构：

期刊名称：PLOS ONE （ 影响因子：3.24； 五年影响因子：3.788 ）

ISSN： 1932-6203

年卷期： 2020 年 15 卷 10 期

页码：

收录情况： SCI

摘要： Quantitative real-time polymerase chain reaction (qPT-PCR) is commonly used to analyze gene expression, however, the accuracy of the normalized results is affected by the expression stability of reference genes.Holotrichia oblita(Coleoptera: Scarabaeidae) causes serious damage to crops. Reliable reference genes inH.oblitaare needed for qRT-PCR analysis. Therefore, we evaluated 13 reference genes under biotic and abiotic conditions. RefFinder provided a comprehensive stability ranking, and geNorm suggested the optimal number of reference genes for normalization.RPL13aandRPL18were the most suitable reference genes for developmental stages, tissues, and temperature treatments;RPL13aandRPS3were the most suitable for pesticide and photoperiod treatments;RPS18andRPL18were the most suitable for the two sexes. We validated the normalized results using odorant-binding protein genes as target genes in different tissues. Compared with the selected suitable reference genes, the expression ofOBP1in antennae, abdomen, and wings, andOBP2in antennae and wings were overestimated due to the instability ofACTb. These results identified several reliable reference genes inH.oblitafor normalization, and are valuable for future molecular studies.

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