Cloning, subcellular location and expression analysis of an acteoside synthase gene from Rehmannia glutinosa
10.7501/j.issn.0253-2670.2020.18.018
- VernacularTitle: 地黄毛蕊花糖苷合酶基因的克隆, 亚细胞定位与表达特性分析
- Author:
Xin-Rong LI
1
Author Information
1. College of Agronomy, Henan Agricultural University
- Publication Type:Journal Article
- Keywords:
Acteoside synthase;
QRT-PCR;
Rehmannia glutinosa Libosch.;
Sequence analysis;
Subcellular localization
- From:
Chinese Traditional and Herbal Drugs
2020;51(18):4739-4746
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To clone the acteoside synthase gene (RgAcS1) from Rehmannia glutinosa, and analyze its subcellular localization and expression pattern. Methods: The cDNA sequence of RgAcS1 was identified based on the annotation of the transcriptome data of R. glutinosa, and the RgAcS1 gene was cloned by polymerase chain reaction (PCR). Constructing the GFP fusion expression vector and observing the subcellular localization of RgAcS1 mediated by Agrobacterium tumefaciens. The expression pattern of RgAcS1 in different parts of tuberous root of R. glutinosa was detected by real-time fluorescence quantitative PCR (qRT-PCR). Results: A full-length coding sequence of a shikimate-O-hydroxy cinnamoyl transferase from R. glutinosa was obtained and named RgAcS1. The length of the RgAcS1 cDNA was 1659 bp, including an open reading frame (ORF) of 1 296 bp, encoding 431 amino acid residues, the molecular weight of the protein was 475 900, and it has a typical domain of shikimic acid-O-hydroxy cinnamoyl transferase. The result of subcellular localization showed that RgAcS1 was mainly distributed in cytoplasm and also in nucleus. The qRT-PCR analysis showed that the expression levels of RgAcS1 were higher in the periderm and root hair of R. glutinosa tuberous root, but lower in the xylem and phloem. The expression levels of RgAcS1 were higher in non-radial striation than that in radial striation of BJ1, QH1 and 85-5. Conclusion: In this study, we obtained the cDNA sequence of RgAcS1, and analyzed the subcellular location and expression patterns of RgAcS1, which will lay foundations for further study on roles of RgAcS1 gene in the synthesis of acteoside in R. glutinosa.
