Selection of reference genes for tissue/organ samples of adults of Eucryptorrhynchus scrobiculatus
Selection of reference genes for tissue/organ samples of adults of Eucryptorrhynchus scrobiculatus
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DOI：
10.1371/journal.pone.0228308
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发表时间：
2020-02-03
期刊：
影响因子：
3.7
通讯作者：
Wen, Junbao
中科院分区：
文献类型：
--
作者：
Gao, Peng;Wang, Jingzhen;Wen, Junbao
臭椿是我国重要的蛀木害虫，它会导致臭椿树大量衰弱甚至死亡。为了鉴定与 E. scrobiculatus 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，并且在此过程中需要参考基因作为标准化的对照。在本研究中，鉴定了 18 个来自 E. scrobiculatus 的候选内参基因，并通过定量实时 PCR 估计了这些内参基因的表达水平。使用四种算法（geNorm、NormFinder、BestKeeper 和 delta Ct）并使用 RefFinder 综合分析表达水平的差异。 RPL13、RPS3和RPL36在不同组织中的表达水平最为稳定，被确定适合用作候选内参基因。此外，一种靶基因（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。本研究为瘰疬基因表达研究提供了第一套合适的候选内参基因，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。
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.