Quantitative real-time RT-PCR is considered to valuable method for the analysis of gene expression. But for the data accuracy of gene expression, the normalization of data using reference genes is essential. In this study, we focus on the analysis of the relative expression of nine commonly used candidate reference genes in the different developmental stages and female and male pupae/adults of Chilo suppressalis, and expression stabilities were analyzed by the ΔCt method, geNorm, NormFinder, and RefFinder. The results showed that H3 was the most stable reference genes in the different developmental stages while EF1 was the optimal genes in pupae/adult of different sexes of C. suppressalis. To validate, the expression profile of the gene encoding heat shock protein 60 (Hsp60) was investigated. The results showed that the transcription of target gene Hsp60 varied depending on the reference gene selected for normalization in different developmental stages, but the trend of expression of Hsp60 between the most stable reference gene combination and the least stable reference gene in pupae/adults of different sexes was the same. Therefore, it is imperative to select the reference genes to estimate target gene expression accurately in the specific experiment. This study provides scientific basis on functional analyses of the important genes contributed to the development and sex differentiation of C. suppressalis.

定量实时 RT-PCR 被认为是分析基因表达的有价值的方法。但是对于基因表达的数据准确性，使用参考基因​​对数据进行归一化是必不可少的。在这项研究中，我们侧重于九个常用的候选参考基因的相对表达的在不同发育阶段和雌雄蛹/成人的分析二化螟，和表达稳定性通过ΔCT法进行了分析，geNorm，NormFinder，和 RefFinder。结果表明，H3是不同发育阶段最稳定的参考基因，而EF1是不同性别的蛹/成虫的最佳基因。. 为了验证，研究了编码热休克蛋白 60 ( Hsp60 )的基因的表达谱。结果表明，靶基因Hsp60的转录因在不同发育阶段选择的归一化参考基因而异，但Hsp60在不同发育阶段的蛹/成虫中最稳定的参考基因组合和最不稳定的参考基因之间的表达趋势。性别是一样的。因此，在具体实验中选择参考基因以准确估计目标基因的表达势在必行。本研究为对抑制C.suppressalis发育和性别分化的重要基因进行功能分析提供了科学依据。