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by claude@2026-07, 2026-07-08
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This study investigated how DNA methylation affects iridoid glycoside biosynthesis in Rehmannia glutinosa roots by comparing roots from the Wenxian and Xinxiang regions and using multi-omics approaches that linked methylation patterns with metabolite and gene expression data. Roots from Wenxian had significantly higher iridoid glycoside content and showed lower methylation levels, which the authors report contributed to greater expression of related enzyme genes; they further found that demethylation promoted both root development and iridoid glycoside accumulation, alongside upregulation of several specific biosynthetic genes and a transcription factor network. A key mechanistic result was that RgMYB2 binds the TAACCA motif in the RgG10H4 promoter, with the promoter’s primary active region located at −164 bp, supporting a positive regulatory role. This paper does not explicitly discuss endometriosis or adenomyosis; it was included in the corpus via a keyword match in the upstream search index.
Abstract
ABSTRACT Rehmannia glutinosa roots produce a group of lipophilic bioactive components known as iridoid glycosides. However, the molecular mechanisms by which DNA methylation regulates the biosynthesis of iridoid glycosides in R. glutinosa remain unknown. Herein, the development of R. glutinosa roots and the content of iridoid glycosides in the Wenxian region were significantly higher than those in Xinxiang. Low methylation level contributed to the accumulation of iridoid glycosides and the expression of related enzyme genes. Demethylation promoted both roots growth and development, as well as the accumulation of iridoid glycosides. Up-regulated RgALDH13 , RgHDR1 , RgG10H4 , RgDXR1 , RgG10H3 , and RgUPD1 , along with transcription factors (TFs), form the regulatory network for the biosynthesis of iridoid glycosides. Furthermore, the primary active region of the RgG10H4 promoter is located in the -164 bp region, where the RgMYB2 protein specifically binds to the TAACCA motif in the RgG10H4 promoter. Collectively, low levels of DNA methylation enhance the expression of core genes, followed by inducing the accumulation of iridoid glycosides, which suggests that RgMYB2 - RgG10H4 plays a positive role in this process. These findings will contribute to a deeper understanding of the role of DNA methylation in the accumulation of iridoid glycosides. Highlight Low levels of DNA methylation contribute to the accumulation of iridoid glycosides and the expression of key enzyme genes in R. glutinosa .
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ABSTRACT
Rehmannia glutinosa roots produce a group of lipophilic bioactive components known as iridoid glycosides. However, the molecular mechanisms by which DNA methylation regulates the biosynthesis of iridoid glycosides in R. glutinosa remain unknown. Herein, the development of R. glutinosa roots and the content of iridoid glycosides in the Wenxian region were significantly higher than those in Xinxiang. Low methylation level contributed to the accumulation of iridoid glycosides and the expression of related enzyme genes. Demethylation promoted both roots growth and development, as well as the accumulation of iridoid glycosides. Up-regulated RgALDH13, RgHDR1, RgG10H4, RgDXR1, RgG10H3, and RgUPD1, along with transcription factors (TFs), form the regulatory network for the biosynthesis of iridoid glycosides. Furthermore, the primary active region of the RgG10H4 promoter is located in the -164 bp region, where the RgMYB2 protein specifically binds to the TAACCA motif in the RgG10H4 promoter. Collectively, low levels of DNA methylation enhance the expression of core genes, followed by inducing the accumulation of iridoid glycosides, which suggests that RgMYB2-RgG10H4 plays a positive role in this process. These findings will contribute to a deeper understanding of the role of DNA methylation in the accumulation of iridoid glycosides.
Highlight Low levels of DNA methylation contribute to the accumulation of iridoid glycosides and the expression of key enzyme genes in R. glutinosa.
Competing Interest Statement
The authors have declared no competing interest.
Abbreviations
- 5-azaC
- 5-Azacytidine
- PSMs
- Plant secondary metabolites
- HCA
- hierarchical clustering analysis
- TFs
- transcription factors
- DEGs
- differentially expressed genes
- DAMs
- differentially abundant metabolites
- MSAP
- methylation-sensitive amplified polymorphism
- FPNI-PCR
- fusion primer and nested integrated PCR
- β-GC
- β-glucosidase
- CS
- confidence score
- AbA
- Aureobasidin A
- G10H
- geranyl 10-hydroxylase
- TSS
- transcription start site
- Y1H
- yeast one-hybrid
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