Screening of housekeeping genes in <i>Gelsemium elegans</i> and expression patterns of genes involved in its alkaloid biosynthesis.

Yao Zhang; Detian MU; Yu Zhou; Ying LU; Yisong Liu; Mengting Zuo; Zhuang Dong; Zhaoying Liu; Qi Tang

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Screening of housekeeping genes in Gelsemium elegans and expression patterns of genes involved in its alkaloid biosynthesis.

Author: Yao ZHANG ¹ ; Detian MU ¹ ; Yu ZHOU ¹ ; Ying LU ¹ ; Yisong LIU ² ; Mengting ZUO ² ; Zhuang DONG ¹ ; Zhaoying LIU ² ; Qi TANG ¹
Author Information

1. College of Horticulture, Hunan Agricultural University, Changsha 410128, Hunan, China.
2. College of Veterinary Medicine, Hunan Agricultural University, Changsha 410128, Hunan, China.
Publication Type:Journal Article
Keywords: Gelsemium elegans; gelsenicine; housekeeping gene; qRT-PCR; terpenoid indole alkaloids
MeSH: Genes, Essential; Gelsemium/genetics*; Peptide Elongation Factor 1/genetics*; Transcriptome; Gene Expression Profiling/methods*; Alkaloids; Real-Time Polymerase Chain Reaction/methods*; Reference Standards
From: Chinese Journal of Biotechnology 2023;39(1):286-303
CountryChina
Language:Chinese
Abstract: Gelsemium elegans is a traditional Chinese herb of medicinal importance, with indole terpene alkaloids as its main active components. To study the expression of the most suitable housekeeping reference genes in G. elegans, the root bark, stem segments, leaves and inflorescences of four different parts of G. elegans were used as materials in this study. The expression stability of 10 candidate housekeeping reference genes (18S, GAPDH, Actin, TUA, TUB, SAND, EF-1α, UBC, UBQ, and cdc25) was assessed through real-time fluorescence quantitative PCR, GeNorm, NormFinder, BestKeeper, ΔCT, and RefFinder. The results showed that EF-1α was stably expressed in all four parts of G. elegans and was the most suitable housekeeping gene. Based on the coexpression pattern of genome, full-length transcriptome and metabolome, the key candidate targets of 18 related genes (AS, AnPRT, PRAI, IGPS, TSA, TSB, TDC, GES, G8H, 8-HGO, IS, 7-DLS, 7-DLGT, 7-DLH, LAMT, SLS, STR, and SGD) involved in the Gelsemium alkaloid biosynthesis were obtained. The expression of 18 related enzyme genes were analyzed by qRT-PCR using the housekeeping gene EF-1α as a reference. The results showed that these genes' expression and gelsenicine content trends were correlated and were likely to be involved in the biosynthesis of the Gelsemium alkaloid, gelsenicine.