Identification of qRT-PCR reference genes for Chilo suppressalis (Walker) during parasitism by Cotesia chilonis (Matsumura)

• Published in: Journal of Asia-Pacific entomology Vol. 23; no. 1; pp. 107 - 112
• Main Authors: Li, Zi-Lan, Pan, Dan-Dan, Lu, Ming-Xing, Du, Yu-Zhou
• Format: Journal Article
• Language: English
• Published: Elsevier B.V 01.04.2020; 한국응용곤충학회
• Subjects: Chilo suppressalis; Cotesia chilonis; qRT-PCR; Reference gene; 농학; Reference gene; Chilo suppressalis; qRT-PCR; Cotesia chilonis
• ISSN: 1226-8615; 1876-7990
• DOI: 10.1016/j.aspen.2019.10.016

[Display omitted] •The stabilities of reference genes in C. suppressalis parasitized were varied.•The combination of TUB, EF1 and NADHD was optimal for normalizing expression.•The variance of the results of Hsp60 and CAT depends on the gene selected. Quantitative real-time RT-PCR is highly sensitive approach for identifying and studying genes that function in various biochemical and cellular processes. Choosing the proper reference genes is a necessary step in ensuring the accuracy of results obtained with qRT-PCR. Herein, we evaluate the expression stability of nine potential reference genes in Chilo suppressalis parasitized by Cotesia chilonis. Stability was analyzed using the ΔCt method, geNorm, NormFinder, and BestKeeper, and our results show that the combination of TUB, EF1 and NADHD was optimal for normalizing expression. The transcription of target genes Hsp60 and CAT in C. suppressalis during parasitism was used to evaluate reference genes, and the results varied depending on the gene selected for normalization. Therefore, it is imperative to choose the proper reference genes to estimate target gene expression accurately in a given experiment. This study provides insights on gene expression in C. suppressalis and is especially relevant to further experiments that explore the effectiveness of C. chilonis in biological control.