Fu Xinhua, Meyer-Rochow Victor Benno
College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, China.
Firefly Conservation Research Centre, Wuhan 430070, China.
Insects. 2021 Apr 16;12(4):359. doi: 10.3390/insects12040359.
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 , , , , , , and for their expression stabilities in 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 and as the most appropriate reference genes for molecular studies in different life stages; and for different tissues; and for RNAi studies. The most appropriate reference genes in all treatments are and The results of this study will help improve accuracy and reliability to normalize RT-qPCR data in for further molecular analysis.
傅氏环萤是一种珍稀的水生萤火虫，为中国特有物种。它被认为是研究性闪光通讯系统分子机制的良好材料。为了改进保护和行为研究策略，需要开展涉及基因表达分析的大规模遗传学研究，而逆转录定量聚合酶链反应（RT-qPCR）是最常用的方法。然而，关于萤火虫任何物种中合适的内参基因的报道非常少。在此，我们评估了八个广泛使用的内参基因，包括……（此处原文未完整列出基因名称），研究它们在三种不同条件下，即生命阶段、组织和双链RNA注射处理下，在傅氏环萤中的表达稳定性。基于RefFinder计算的基因稳定性排名，RefFinder整合了四种算法（geNorm、delta Ct法、NormFinder和BestKeeper），我们推荐……（此处原文未完整列出基因名称）作为不同生命阶段分子研究的最合适内参基因；……（此处原文未完整列出基因名称）作为不同组织的内参基因；……（此处原文未完整列出基因名称）作为RNA干扰研究的内参基因。所有处理中最合适的内参基因是……（此处原文未完整列出基因名称）。本研究结果将有助于提高傅氏环萤RT-qPCR数据标准化的准确性和可靠性，以进行进一步的分子分析。
