Screening Potential Reference Genes in Tuta absoluta with Real-Time Quantitative PCR Analysis under Different Experimental Conditions

文献类型： 外文期刊

第一作者： Yang, An-Pei

作者： Yang, An-Pei;Wang, Yu-Sheng;Lv, Zhi-Chuang;Liu, Wan-Xue;Bi, Si-Yan;Wan, Fang-Hao;Wu, Qiang;Zhang, Gui-Fen;Yang, An-Pei;Huang, Cong

作者机构：

关键词： biological factors; environmental factors; expression stability; reference genes; RT-qPCR; South American tomato leafminer

期刊名称：GENES （ 影响因子：4.096； 五年影响因子：4.339 ）

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年卷期： 2021 年 12 卷 8 期

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收录情况： SCI

摘要： Tuta absoluta is one of the most significant invasive pests affecting tomato plants worldwide. RT-qPCR has emerged as one of the most sensitive and accurate methods for detecting gene expression data. The screening of stable internal reference genes is the most critical step for studying the molecular mechanisms of environmental adaptability. The stable reference genes expressed in T. absoluta under specific experimental conditions have not yet been clarified. In this study, seven candidate reference genes (RPL27, RPS13, RPS15, EF1-alpha, TUB, TBP, and beta-actin) and their optimal numbers were evaluated under biotic (developmental stages and adult tissues) and abiotic (insecticide, temperature, and plant VOC) conditions using four software programs. Our results identified the following reference genes and numbers as optimal: three genes (EF1-alpha, RPS13, and RPL27) for different developmental stages (egg, larva, pupa, unmated adult), two genes (RPS13 and TBP) for adult tissues (antenna, head, thorax, abdomen, leg), two genes (TBP and RPS13) for insecticides (Bacillus thuringiensis, chlorpyrifos, abamectin-aminomethyl, and chlorantraniliprole), two genes (RPL27 and TUB) for temperature-induced stresses (0, 25, and 40 degrees C), and two genes (RPS13 and TUB) for VOC-induced stresses (nonanal, alpha-phellandrene, and tomato leaves). Our results provide a reference for selecting appropriate reference genes for further study of the functional genes of T. absoluta under different experimental conditions.

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