Abstract
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.
Highlights
Eucryptorrhynchus scrobiculatus Motschulsky (Coleoptera: Curculionidae), an important wood-boring pest of the tree-of-heaven Ailanthus altissima Swingl [1,2,3] that is widely distributed in 21 provinces in China [4], is one of the most damaging invasive insects in this country
Considered an invasive species, A. altissima is a serious threat to ecosystems in North America; as E. scrobiculatus feeds on this tree, it has great promise as a potential biological control agent [1, 6]
To obtain an overview of transcript abundance, expression levels of the 18 candidate reference genes were investigated in different tissues of E. scrobiculatus
Summary
Eucryptorrhynchus scrobiculatus Motschulsky (Coleoptera: Curculionidae), an important wood-boring pest of the tree-of-heaven Ailanthus altissima Swingl [1,2,3] that is widely distributed in 21 provinces in China [4], is one of the most damaging invasive insects in this country. Considered an invasive species, A. altissima is a serious threat to ecosystems in North America; as E. scrobiculatus feeds on this tree, it has great promise as a potential biological control agent [1, 6]. To study the biology of E. scrobiculatus, it is important to identify plant cell wall-degrading enzymes and functional genes involved in host-parasite interactions; reference genes must first be selected. E. scrobiculatus transcriptomes are available and provide comprehensive information to facilitate such studies.
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