AUTHOR=Xie Jiaoxin , Liu Tinghui , Khashaveh Adel , Yi Chaoqun , Liu Xiaoxu , Zhang Yongjun 

TITLE=Identification and Evaluation of Suitable Reference Genes for RT-qPCR Analysis in Hippodamia variegata (Coleoptera: Coccinellidae) Under Different Biotic and Abiotic Conditions

JOURNAL=Frontiers in Physiology

VOLUME=12

YEAR=2021

URL=https://www.frontiersin.org/journals/physiology/articles/10.3389/fphys.2021.669510

DOI=10.3389/fphys.2021.669510

ISSN=1664-042X

ABSTRACT=<p>Reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) is an accurate and convenient technique for quantifying expression levels of the target genes. Selection of the appropriate reference gene is of the vital importance for RT-qPCR analysis. <italic>Hippodamia variegata</italic> is one of the most important predatory natural enemies of aphids. Recently, transcriptome and genome sequencings of <italic>H. variegata</italic> facilitate the gene functional studies. However, there has been rare investigation on the detection of stably expressed reference genes in <italic>H. variegata</italic>. In the current study, by using five analytical tools (Delta Ct, geNorm, NormFinder, BestKeeper, and RefFinder), eight candidate reference genes, namely, <italic>Actin</italic>, <italic>EF1</italic>α, <italic>RPL7</italic>, <italic>RPL18</italic>, <italic>RPS23</italic>, <italic>Tubulin-</italic>α, <italic>Tubulin-</italic>β, and <italic>TufA</italic>, were evaluated under four experimental conditions including developmental stages, tissues, temperatures, and diets. As a result, a specific set of reference genes were recommended for each experimental condition. These findings will help to improve the accuracy and reliability of RT-qPCR data, and lay a foundation for further exploration on the gene function of <italic>H. variegata</italic>.</p>