State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
Department of Fruit Science, College of Horticulture, China Agricultural University, Beijing 100193, China.
Viruses. 2022 Jun 10;14(6):1269. doi: 10.3390/v14061269.
Several members of the genus infect fruit trees and are distributed worldwide. Prunus necrotic ringspot virus (PNRSV) is one of the most prevalent viruses, causing significant losses. can be infected by several ilarviruses, leading to obvious symptoms, including PNRSV, which suggests that cucumbers could be good hosts for the study of the pathogenesis of ilarviruses. Real-time quantitative PCR is an optimal choice for studying gene expression because of its simplicity and its fast and high sensitivity, while its accuracy is highly dependent on the stability of the reference genes. In this study, we assessed the stability of eleven reference genes with geNorm, NormFinder, ΔCt method, BestKeeper, and the ranking software, RefFinder. The results indicated that the combined use of and was the most accurate normalization method. In addition, the host genes , , and were selected to test the reliability of the reference genes. This study provides useful information for gene expression analysis during PNRSV infection and will facilitate gene expression studies associated with ilarvirus infection.
几种属侵染果树，分布于世界各地。李坏死环斑病毒（PNRSV）是最常见的病毒之一，造成重大损失。可以被几种 ilarviruses 感染，导致明显的症状，包括 PNRSV，这表明黄瓜可能是研究 ilarviruses 发病机制的良好宿主。实时定量 PCR 是研究基因表达的最佳选择，因为它简单、快速、高灵敏度，但其准确性高度依赖于参考基因的稳定性。在这项研究中，我们使用 geNorm、NormFinder、ΔCt 方法、BestKeeper 和排名软件 RefFinder 评估了 11 个参考基因的稳定性。结果表明，和的组合使用是最准确的归一化方法。此外，选择宿主基因、和来测试参考基因的可靠性。本研究为 PNRSV 感染过程中的基因表达分析提供了有用的信息，并将有助于与 ilarvirus 感染相关的基因表达研究。
