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

• Published in: Genes Vol. 12; no. 8; p. 1253
• Main Authors: Yang, An-Pei, Wang, Yu-Sheng, Huang, Cong, Lv, Zhi-Chuang, Liu, Wan-Xue, Bi, Si-Yan, Wan, Fang-Hao, Wu, Qiang, Zhang, Gui-Fen
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 17.08.2021; MDPI
• Subjects: Abamectin; Actin; Adaptability; Animals; Antennae; Chlorpyrifos; Developmental stages; Gene expression; Genes, Insect - genetics; Insecticides; Insecticides - pharmacology; Insects; Kinases; Larva - genetics; Larva - pathogenicity; Lepidoptera - genetics; Lepidoptera - pathogenicity; Lycopersicon esculentum - genetics; Lycopersicon esculentum - growth & development; Lycopersicon esculentum - parasitology; Molecular modelling; Moths - genetics; Moths - pathogenicity; Pest Control, Biological; Pests; Proteins; Real-Time Polymerase Chain Reaction - standards; Reference Standards; Tomatoes; Tuta absoluta; Beijing China; China; Germany; reference genes; South American tomato leafminer; expression stability; environmental factors; RT-qPCR; biological factors
• ISSN: 2073-4425; 2073-4425
• DOI: 10.3390/genes12081253

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 under specific experimental conditions have not yet been clarified. In this study, seven candidate reference genes ( , , , , , , and ) 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 ( , , and ) for different developmental stages (egg, larva, pupa, unmated adult), two genes ( and ) for adult tissues (antenna, head, thorax, abdomen, leg), two genes ( and ) for insecticides ( , chlorpyrifos, abamectin-aminomethyl, and chlorantraniliprole), two genes ( and ) for temperature-induced stresses (0, 25, and 40 °C), and two genes ( and ) for VOC-induced stresses (nonanal, α-phellandrene, and tomato leaves). Our results provide a reference for selecting appropriate reference genes for further study of the functional genes of under different experimental conditions.