Engineering Research Center of Biological Control, Ministry of Education, South China Agricultural University, Guangzhou 510640, China.
Engineering Research Center of Biotechnology for Active Substances, Ministry of Education, Chongqing Normal University, Chongqing 401331, China.
Genes (Basel). 2024 Feb 28;15(3):318. doi: 10.3390/genes15030318.
The citrus whitefly, , is a destructive pest that infests citrus plants. It is a major vector in transmitting plant viruses such as citrus yellow vein clearing virus (CYVCV), which has caused severe economic losses worldwide, and therefore efficient control of this pest is economically important. However, the scope of genetic studies primarily focused on is restricted, something that has potentially limited further study of efficient control options. To explore the functionalities of target genes, screening for specific reference genes using RT-qPCR under different experimental conditions is essential for the furtherance of biological studies concerning . The eight candidate reference genes were evaluated by dedicated algorithms (, , and Δ method) under five specific experimental conditions (developmental stage, sex, tissue, population and temperature). In addition, the software, a comprehensive evaluation platform integrating all of the above algorithms, ranked the expression stability of eight candidate reference genes. The results showed that the best reference genes under different experimental settings were and at different developmental stages; and in both sexes; and in different tissues; and under different populations; and and in different temperatures. The validation of selected reference genes was further identified using heat shock protein () 70 as a reporter gene. Our study, for the first time, provides a detailed compilation of internal reference genes for that are suitable for RT-qPCR analysis, which is robust groundwork for comprehensive investigation of the functional target genes of
橘潜叶蛾，是一种侵害柑橘植物的破坏性害虫。它是传播植物病毒的主要媒介，如柑橘黄龙脉褪绿病毒（CYVCV），该病毒已在全球范围内造成严重的经济损失，因此，有效控制这种害虫在经济上具有重要意义。然而，遗传研究的范围主要集中在 上，这可能限制了对有效控制选项的进一步研究。为了探索 目标基因的功能，在不同的实验条件下使用 RT-qPCR 筛选特定的参考基因，对于进一步研究 生物学至关重要。使用 RT-qPCR 在五种特定的实验条件（发育阶段、性别、组织、种群和温度）下，通过专用算法（geNorm、NormFinder、BestKeeper 和 ΔCt 方法）对 8 个候选参考基因进行了评估。此外，使用 软件，该软件是一个集成了所有上述算法的综合评估平台，对 8 个候选参考基因的表达稳定性进行了排名。结果表明，在不同的实验设置下，最佳的参考基因分别是不同发育阶段的 和 ；雌雄两性的 和 ；不同组织的 和 ；不同种群的 和 ；不同温度的 和 。通过热休克蛋白（HSP70）70 作为报告基因对选定的参考基因进行了进一步验证。我们的研究首次为 RT-qPCR 分析提供了适合的 详细参考基因清单，为全面研究 的功能目标基因奠定了坚实的基础。
