Pan Ying-Na, Zhao Ru-Na, Fu Di, Yu Chun, Pan Chun-Ni, Zhou Wei, Chen Wen-Long
Guizhou Provincial Key Laboratory for Agricultural Pest Management of Mountainous Regions, Institute of Entomology, Guizhou University, Guiyang 550025, China.
Insects. 2022 Aug 26;13(9):773. doi: 10.3390/insects13090773.
Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) is a widely used tool for measuring gene expression; however, its accuracy relies on normalizing the data to one or more stable reference genes. (Wolff) is a polyphagous predatory natural enemy insect that preferentially feeds on more than 40 types of agricultural and forestry pests, such as those belonging to the orders Lepidoptera, Coleoptera, and Hymenoptera. However, to our knowledge, the selection of stable reference genes has not been reported in detail thus far. In this study, nine candidate reference genes (, , , , , , , , and ) were selected based on transcriptome sequencing results. The expression of these genes in various samples was examined at different developmental stages, in the tissues of male and female adults, and after temperature and starvation treatments. Five algorithms were used, including ΔCt, geNorm, NormFinder, BestKeeper, and RefFinder, to evaluate reference gene expression stability. The results revealed that the most stable reference genes were and at different developmental stages; , , and for female adult tissue samples; and for male adult tissue samples; and for various temperature treatments of nymphs; and for nymph samples under starvation stress; and and for all samples. Overall, we obtained a stable expression of reference genes under different conditions in , which provides a basis for future molecular studies on this organism.
定量逆转录-聚合酶链反应（qRT-PCR）是一种广泛用于测量基因表达的工具；然而，其准确性依赖于将数据归一化到一个或多个稳定的参考基因。（沃尔夫）是一种多食性捕食性天敌昆虫，优先取食40多种农林害虫，如鳞翅目、鞘翅目和膜翅目的害虫。然而，据我们所知，迄今为止尚未详细报道稳定参考基因的选择。在本研究中，基于转录组测序结果选择了9个候选参考基因（、、、、、、、和）。检测了这些基因在不同发育阶段、雌雄成虫组织以及温度和饥饿处理后的各种样本中的表达。使用了5种算法，包括ΔCt、geNorm、NormFinder、BestKeeper和RefFinder，来评估参考基因表达的稳定性。结果表明，在不同发育阶段最稳定的参考基因是和；在雌性成虫组织样本中是、和；在雄性成虫组织样本中是和；在若虫的各种温度处理中是和；在饥饿胁迫下的若虫样本中是和；在所有样本中是和。总体而言，我们在中获得了不同条件下参考基因的稳定表达，这为该生物未来的分子研究提供了基础。
