University of Oulu

Fu, X., & Meyer-Rochow, V. B. (2021). Selection and Validation of Suitable Reference Genes for RT-qPCR Analysis in the Rare Aquatic Firefly Aquatica leii (Coleoptera: Lampyridae). Insects, 12(4), 359. doi:10.3390/insects12040359

Selection and validation of suitable reference genes for RT-qPCR analysis in the rare aquatic firefly Aquatica leii (Coleoptera: Lampyridae)

Saved in:
Author: Fu, Xinhua1,2; Meyer-Rochow, Victor Benno3,4
Organizations: 1College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, China
2Firefly Conservation Research Centre, Wuhan 430070, China
3Department of Ecology and Genetics, Oulu University, SF-90140 Oulu, Finland
4Agricultural Science and Technology Research Institute, Andong National University, Andong 36729, Korea
Format: article
Version: published version
Access: open
Online Access: PDF Full Text (PDF, 3.2 MB)
Persistent link:
Language: English
Published: Multidisciplinary Digital Publishing Institute, 2021
Publish Date: 2021-05-10


Aquatica leii Fu and Ballantyne is a species of rare aquatic firefly and endemic in China. It is considered good material to study the molecular mechanism of sexual flash communication systems. To improve conservation and behavioral research strategies, large-scale genetic studies involving gene-expression analysis are required and reverse transcription-quantitative polymerase chain reaction (RT-qPCR) is the most commonly used method. However, there have been very few reports on appropriate reference genes in any species of firefly. Here, we evaluated eight widely utilized reference genes including 18S, Actin, Reep5, Odc1, Tub, Gapdh, Ef1a and S27Ae for their expression stabilities in A. leii under three different conditions, i.e., life stage, tissue and dsRNA injection. Based on the gene stability ranking calculated by RefFinder, which integrates four algorithms (geNorm, delta Ct method, NormFinder, and BestKeeper), we recommend S27Ae and Reep5 as the most appropriate reference genes for molecular studies in different life stages; Ef1a and Odc1 for different tissues; Tub and Odc1 for RNAi studies. The most appropriate reference genes in all treatments are S27Ae and Tub. The results of this study will help improve accuracy and reliability to normalize RT-qPCR data in A. leii for further molecular analysis.

see all

Series: Insects
ISSN: 2075-4450
ISSN-E: 2075-4450
ISSN-L: 2075-4450
Volume: 12
Issue: 4
Article number: 359
DOI: 10.3390/insects12040359
Type of Publication: A1 Journal article – refereed
Field of Science: 1184 Genetics, developmental biology, physiology
1181 Ecology, evolutionary biology
Funding: This work was funded by National Natural Science Foundation grant of China, Nr. 31872281 to X.F.
Copyright information: © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (