Preprint Article Version 1 This version is not peer-reviewed

Screening and Validation of Stable Reference Genes for qRT-PCR Analysis in Epicauta gorhami (Coleoptera: Meloidae)

Version 1 : Received: 28 October 2024 / Approved: 28 October 2024 / Online: 28 October 2024 (14:02:49 CET)

How to cite: Yang, G.; Yu, X.; Zhang, Y.; Luo, J.; Li, X.; Zhu, L.; Zhang, H.; Jin, L.; Wu, G.; Yan, X.; Shen, C. Screening and Validation of Stable Reference Genes for qRT-PCR Analysis in Epicauta gorhami (Coleoptera: Meloidae). Preprints 2024, 2024102173. https://doi.org/10.20944/preprints202410.2173.v1 Yang, G.; Yu, X.; Zhang, Y.; Luo, J.; Li, X.; Zhu, L.; Zhang, H.; Jin, L.; Wu, G.; Yan, X.; Shen, C. Screening and Validation of Stable Reference Genes for qRT-PCR Analysis in Epicauta gorhami (Coleoptera: Meloidae). Preprints 2024, 2024102173. https://doi.org/10.20944/preprints202410.2173.v1

Abstract

Epicauta gorhami is a hypermetamorphic insect that mainly forage soybeans during the adult stage. However, the lack of appropriate references hinders our studying of the gene function in E. gorhami. In this study, refer to five computational tools (Ct value, geNorm, NormFinder, BestKeeper and RefFinder), the stability of 10 housekeeping genes (GAPDH, ACT, RPL4, RPL27, α-TUB, RPS18, EF1α, RPS28, RPL13 and SOD) was assessed by RT-qPCR under three different conditions (adult ages, tissues/organs and temperatures). The findings suggested that SOD and RPS18 were the most ideal references for examine gene transcripts among diverse adult ages and at various temperatures; A pair of RPS18 and RPS28 was the most reliable genes to assess gene expressions in diverse adult tissues. Finally, The relative expression levels of EgUAP were computed after normalization RPS18 and RPS28 with across diverse adult tissues. As expected, EgUAP expression was abundant in the foregut, trachea and antenna, and low in the midgut, hindgut and epidermis. These findings will lay a solid foundation for analysing the gene expression of E. gorhami.

Keywords

Epicauta gorhami; Glycine max; reference genes; RT-qPCR; RefFinder

Subject

Biology and Life Sciences, Insect Science

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