Beijing Key Laboratory for Forest Pest Control, Beijing Forestry University, Beijing, P. R. China.
PLoS One. 2020 Feb 3;15(2):e0228308. doi: 10.1371/journal.pone.0228308. eCollection 2020.
Eucryptorrhynchus scrobiculatus is an important wood-boring pest of Ailanthus altissima in China, where it causes a large number of these trees to weaken or even die. To identify genes related to economic traits or specific cellular processes in E. scrobiculatus, gene expression in multiple tissue/organ samples is commonly surveyed, and reference genes are required in this process as a control for normalization. In the present study, 18 candidate reference genes from E. scrobiculatus were identified, and the expression levels of these reference genes were estimated through quantitative real-time PCR. Differences in expression levels were analyzed with four algorithms (geNorm, NormFinder, BestKeeper and delta Ct) and comprehensively with RefFinder. With the most stable levels of expression in different tissues, RPL13, RPS3 and RPL36 were determined to be suitable for use as candidate reference genes. Moreover, the expression profile of one target gene (glycoside hydrolase family 45, GH45) confirmed the reliability of the selected candidate reference genes. This study provides the first set of suitable candidate reference genes for gene expression studies in E. scrobiculatus, and the findings will facilitate subsequent transcriptomics and functional gene research on this pest.
星天牛是中国臭椿的重要蛀干害虫，大量危害导致臭椿衰弱甚至死亡。为了鉴定星天牛经济性状或特定细胞过程相关的基因，通常会调查多个组织/器官样本中的基因表达情况，在此过程中需要参考基因作为归一化的对照。本研究从星天牛中鉴定了 18 个候选参考基因，并通过定量实时 PCR 估计了这些参考基因的表达水平。使用 geNorm、NormFinder、BestKeeper 和 delta Ct 这四个算法以及 RefFinder 综合分析了表达水平的差异。根据不同组织中表达水平最稳定的基因，确定 RPL13、RPS3 和 RPL36 适合作为候选参考基因。此外，一个靶基因（糖苷水解酶家族 45，GH45）的表达谱证实了所选候选参考基因的可靠性。本研究为星天牛基因表达研究提供了第一组合适的候选参考基因，研究结果将有助于后续对该害虫的转录组学和功能基因研究。
