Gong Zhongjun, Zhang Jing, Chen Qi, Li Huiling, Zhang Ziqi, Duan Yun, Jiang Yueli, Li Tong, Miao Jin, Wu Yuqing
Henan Key Laboratory of Crop Pest Control, Key Laboratory of Integrated Pest Management on Crops in Southern Region of North China, Institute of Plant Protection, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China.
Luohe Academy of Agricultural Sciences, Luohe 462300, China.
Insects. 2024 Aug 30;15(9):661. doi: 10.3390/insects15090661.
is among the world's most destructive pests. For accurate qPCR and gene expression studies, the selection of stable and appropriate reference genes is crucial. However, a thorough evaluation of potential reference genes for use in research is lacking. In this study, 11 reference genes (, , , , , , , , , ,and ) were evaluated under different biological conditions and environmental stresses. The stability of 11 potential reference gene transcripts was evaluated through various computational tools, including geNorm, BestKeeper, NormFinder, theΔCt method, and the RefFinder program. Under various developmental stages and RNAi conditions, and exhibited the greatest stability. , , and were the most stable genes in both male and female adults. Under differing tissue conditions, and stood out as the most reliable. Moreover, under varying photoperiod conditions, , and were the most stable genes. Lastly, and were the most stable genes across different temperatures. These findings offer essential criteria for selecting suitable reference genes across diverse experimental settings, thereby establishing a solid basis for accurate gene expression studies in using RT-qPCR.
是世界上最具破坏性的害虫之一。对于准确的定量聚合酶链反应（qPCR）和基因表达研究而言，选择稳定且合适的内参基因至关重要。然而，目前缺乏对用于该研究的潜在内参基因的全面评估。在本研究中，对11个内参基因（、、、、、、、、、和）在不同生物学条件和环境胁迫下进行了评估。通过包括geNorm、BestKeeper、NormFinder、ΔCt法和RefFinder程序在内的各种计算工具，评估了11个潜在内参基因转录本的稳定性。在不同发育阶段和RNA干扰（RNAi）条件下，和表现出最大的稳定性。在雌雄成虫中，、和是最稳定的基因。在不同组织条件下，和最为可靠。此外，在不同光周期条件下，、和是最稳定的基因。最后，在不同温度下，和是最稳定的基因。这些发现为在不同实验环境中选择合适的内参基因提供了重要标准，从而为利用逆转录定量聚合酶链反应（RT-qPCR）在中进行准确的基因表达研究奠定了坚实基础。
