Reliable reference genes are quite important in calculating gene transcript levels by using real-time quantitative reverse transcription-PCR (RT-qPCR). Eotetranychus sexmaculatus is known as a dangerous mite causing significant yield reduction of rubber tree latex; however, selection of appropriate reference genes for validation of target gene expression in E. sexmaculatus has not been conducted yet. In the present study, nine candidate reference genes were analyzed for their expression stability in different life stages of E. sexmaculatus by using common algorithms including comparative ΔCq method, geNorm, NormFinder and BestKeeper. In addition, a comprehensive analysis software (RefFinder) was used to assign an overall final rank for each candidate gene. The results showed that β-actin and β-TUB were the best two reference genes and were subjected to evaluate expression of two protective enzyme genes (EsCu/ZnSOD and EsCAT1) in E. sexmaculatus. We found that the expression of EsCu/ZnSOD and EsCAT1 in E. sexmaculatus feeding on mite-resistant rubber tree germplasm was significantly lower compared with those feeding on mite-susceptible germplasm. These results will facilitate research in revealing molecular mechanisms underlying rubber tree resistance to the spider mite.

通过使用实时定量逆转录PCR（RT-qPCR），可靠的参考基因在计算基因转录水平方面非常重要。臭螨（Eotetranychus sexmaculatus）是一种危险的螨虫，可引起橡胶树胶乳的大量减产。然而，还没有选择合适的参考基因来验证性大肠埃希菌中靶基因的表达。在本研究中，分析了九个候选参考基因在大肠埃希菌不同生命阶段的表达稳定性。通过使用常见的算法，包括比较ΔCq方法，geNorm，NormFinder和BestKeeper。另外，使用综合分析软件（RefFinder）为每个候选基因分配总体最终排名。结果表明，β-肌动蛋白和β-TUB是最好的两个参考基因，并评估了两个保护酶基因（EsCu / ZnSOD和EsCAT1）在性大肠杆菌中的表达。我们发现EsCu / ZnSOD和EsCAT1在E. sexmaculatus中的表达以耐螨虫的橡胶树种质为食比以螨虫敏感的种质为食。这些结果将有助于研究揭示橡胶树抵抗红蜘蛛的分子机制。