Identification and functional analysis of β-amyrin synthase gene in Dipsacus asper.
10.19540/j.cnki.cjcmm.20241115.102
- Author:
Huan LEI
1
;
Hua HE
1
;
Jiao XU
1
;
Chang-Gui YANG
1
;
Wei-Ke JIANG
1
;
Tao ZHOU
1
;
Lan-Ping GUO
2
Author Information
1. Guizhou University of Traditional Chinese Medicine Guiyang 550025, China.
2. State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs,Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences Beijing 100700, China.
- Publication Type:Journal Article
- Keywords:
Dipsacus asper;
expression analysis;
functional analysis;
triterpenoid saponins;
β-amyrin synthase
- MeSH:
Intramolecular Transferases/chemistry*;
Phylogeny;
Plant Proteins/chemistry*;
Gene Expression Regulation, Plant;
Dipsacaceae/classification*;
Saponins/metabolism*;
Oleanolic Acid/metabolism*
- From:
China Journal of Chinese Materia Medica
2025;50(4):1043-1050
- CountryChina
- Language:Chinese
-
Abstract:
Dipsaci Radix is a commonly used Chinese herbal medicine in China, with triterpenoid saponins as the main active components. β-Amyrin synthase, a member of the oxidosqualene cyclase superfamily, plays a crucial role in the biosynthesis of oleanane-type triterpenoid saponins. Asperosaponin Ⅵ is an oleanane-type triterpenoid saponin. To explore the β-amyrin synthase genes involved in the biosynthesis of asperosaponin Ⅵ in Dipsacus asper, this study screened the candidate genes from the transcriptome data of D. asper. Two β-amyrin synthase genes, Da OSC1 and Da OSC2, were identified by phylogenetic analysis and correlation analysis. The coding sequences of Da OSC1 and Da OSC2 were 2 286 bp and 2 295 bp in length, encoding 761 and 764 amino acids,respectively. Multiple sequence alignments showed that Da OSC1 and Da OSC2 had three conserved motifs( DCTAE, QW, and MWCYCR) unique to the oxidosqualene cyclase family. Real-time quantitative PCR results showed that Da OSC1 and Da OSC2 had the highest expression levels in the roots. Compared with normal growth conditions, the low-temperature treatment significantly upregulated the expression of Da OSC1 and Da OSC2. Agrobacterium-mediated transient expression of Da OSC1 and Da OSC2 in Nicotiana benthamiana resulted in the production of β-amyrin, which suggested that Da OSC1 and Da OSC2 were able to catalyze the synthesis of β-amyrin. This study clarified the catalytic functions of two β-amyrin synthases in D. asper, analyzed their expression patterns in different tissue and at low temperatures. The findings provide a foundation for further studying the biosynthetic pathway and regulatory mechanism of asperosaponin Ⅵ in D. asper.
