Atrijuglans hetaohei Yang (Lepidoptera: Gelechioidea), is one of the major pests that seriously damage the walnut fruits. Although the morphology and physiology of A. hetaohei have been widely studied, suitable reference genes for normalizing target gene expression have not been identified. In this study, the expressions of eight candidate reference genes including GAPDH, β‐Tubulin, TATA‐binding protein (TBP), Histone, Ubiquitin‐conjugating enzyme 9 (UBC9), arginine kinase (AK), 28S and elongation factor 1 alpha (EF1α) in different developmental stages and various larval tissues of A. hetaohei were determined using reverse transcription‐quantitative polymerase chain reaction. Moreover, the expression stability of the selected reference genes was evaluated by ΔCt method, BestKeeper, NormFinder, geNorm and ReFinder, a comprehensive software platform. These results demonstrated that the best reference genes were GAPDH and 28S at different developmental stages of A. hetaohei; and the optimum references in various larval tissues of A. hetaohei were 28S, Histone and TBP. Our study should be useful to analyze the expression profiles of target genes and form a solid foundation for future research on the understanding of the genes' biological functions in A. hetaohei.

中文翻译：

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在 Atrijuglans hetaohei（鳞翅目：Gelechioidea）中筛选用于逆转录-定量聚合酶链反应归一化的潜在参考基因

Atrijuglans hetaohei Yang（鳞翅目：Gelechioidea），是严重危害核桃果实的主要害虫之一。尽管已经广泛研究了河套黑 A. 的形态和生理学，但尚未确定用于标准化靶基因表达的合适参考基因。本研究中，8个候选参考基因的表达包括GAPDH、β-微管蛋白、TATA结合蛋白（TBP）、组蛋白、泛素结合酶9（UBC9）、精氨酸激酶（AK）、28S和延伸因子1α（使用逆转录-定量聚合酶链反应测定了河套黑松鼠不同发育阶段和各种幼虫组织中的EF1α)。此外，所选参考基因的表达稳定性通过ΔCt法、BestKeeper、NormFinder、geNorm和ReFinder（一个综合软件平台）进行评估。这些结果表明，最佳内参基因为河套黑松不同发育阶段的GAPDH和28S；和桃黑猕猴桃各种幼虫组织中的最佳参照物为28S、组蛋白和TBP。我们的研究应该有助于分析目标基因的表达谱，并为未来了解河桃黑A.基因生物学功能的研究奠定坚实的基础。