Selection and Verification of the Reference Genes of <i>Euphorbia maculata</i> Based on Transcriptome Sequencing
Welcome to Chinese Journal of Tropical Crops,

Chinese Journal of Tropical Crops ›› 2023, Vol. 44 ›› Issue (11): 2273-2280.DOI: 10.3969/j.issn.1000-2561.2023.11.016

• Omics & Biotechnology • Previous Articles     Next Articles

Selection and Verification of the Reference Genes of Euphorbia maculata Based on Transcriptome Sequencing

XU Jianfeng1, GUI Mingming2, ZHANG Yongkang1, YAO Zizhao1, HUANG Shenghe2, RAO Zechang1,*()   

  1. 1. Fuzhou Medical College, Nanchang University, Fuzhou, Jiangxi 344000, China
    2. Department of Basic Medicine, Jiangxi College of Traditional Chinese Medicine, Fuzhou, Jiangxi 344000, China
  • Received:2022-09-19 Revised:2022-11-17 Online:2023-11-25 Published:2023-12-08
  • Contact: *RAO Zechang,E-mail:rzc2005010@163.com

Abstract:

The flavonoid quercetin is one of the main active ingredients in Chinese medicinal herb, Euphorbia maculata. At present, there are few reports on quercetin biosynthesis gene, and real-time quantitative PCR (RT-qPCR) is one of the common methods in gene research. But it is a prerequisite for relative quantitative RT-qPCR to have suitable reference genes. In order to obtain a suitable reference gene, this experiment was based on the results of transcriptome sequencing, EmSE, EmUBC, EmGDI1, EmLSM12, EmGBP, EmSYP22, EmTRI1, EmRSZ21, EmRPL7 and EmUBQ obtained by our research group in the early stage were selected as candidate reference genes. RT-qPCR was used to detect the expression of the candidate reference genes in roots, stems, leaves and fruits of E. maculata during vegetative and reproduction stages. The expression stability of the candidate genes was analyzed using geNorm, NormFinder and BestKeeper software. The results of different software were integrated and ranked by the geometric mean method. The ranking was in the order EmGDI1>EmUBQ>EmSE>EmUBC>EmLSM12=EmGBP>EmTRI1>EmSYP22>EmRSZ21=EmRPL7. EmC4H, a key enzyme gene in quercetin biosynthesis, was analyzed using EmGDI1 as the reference gene. The difference of EmC4H expression in different tissues at different stages was consistent with the trend of transcriptome sequencing. The reference gene EmGDI1 developed in this study could be used as a suitable reference gene for tissue-specific expression studies of different stages of E. maculata.

Key words: Euphorbia maculata, reference gene, transcriptome sequencing, RT-qPCR

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