PubMed · 7398267
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M J Rapaport. 1980. Sensitization to Abitol.. https://doi.org/10.1111/j.1600-0536.1980.tb03924.x
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DNA methylation plays pivotal roles in regulating gene expression and the secondary metabolism in plants. Salvia miltiorrhiza and Salvia bowleyana are traditional Chinese medicinal plants with roots enriched with tanshinone components. However, the regulatory mechanism of DNA methylation on tanshinone production remains elusive. Here, we analyzed 30-day-old hairy roots of S. miltiorrhiza and S. bowleyana using targeted high-performance liquid chromatography analysis and found significantly higher tanshinone content in S. miltiorrhiza. Whole-genome bisulfite sequencing revealed elevated DNA methylation levels in S. miltiorrhiza, potentially due to the upregulation of methylation-related genes, including DOMAINS REARRANGED METHYLTRANSFERASE 1 (DRM1), DECREASE IN DNA METHYLATION 1 (DDM1), CHROMOMETHYLASE 2 (CMT1), and CHROMOMETHYLASE 3 (CMT3), alongside the low expression of the demethylase gene REPRESSOR OF SILENCING 1 (ROS1) in S. miltiorrhiza. Additionally, four genes that are involved in tanshinone biosynthesis, including 1-DEOXY-D-XYLULOSE-5-PHOSPHATE REDUCTASE (DXS1), GERANYLGERANYL DIPHOSPHATE SYNTHASE (GGPPS2), 4-HYDROXY-3-METHYLBUT-2-ENYL PYROPHOSPHATE REDUCTASE (HDR2), and COPALYL PYROPHOSPHATE SYNTHASE (CPS3), showed lower methylation levels in the promoters of DXS1, GGPPS2, and CPS3 and a higher DNA methylation level in the gene body of HDR2 in S. miltiorrhiza, which may lead to their high expression and the accumulation of tanshinones. Consistently, overexpression of the SmCMT3 in S. miltiorrhiza significantly reduced the contents of cryptotanshinone, tanshinone I, and tanshinone IIA. Transcriptomic and methylome analyses confirmed that the expression levels of the tanshinone biosynthesis-related genes, including SmMK, SmCPS1, SmDXS2, and SmAACT1, were correlated with their promoter or gene body DNA methylation levels. Our findings reveal that DNA methylation critically regulates tanshinone biosynthesis in S. miltiorrhiza and S. bowleyana, offering valuable insights for breeding.
Tanshen, the rhizome of Salvia miltiorrhiza Bunge, has been used in Chinese traditional medicine (TCM) for multiple therapeutic remedies. The major constituents of Tanshen include water-soluble phenolic acids and lipophilic tanshinones. Phenolic acids possess antioxidant and anticoagulant activities, whereas tanshinones show antibacterial, antioxidant, and antineoplastic activities. This review will focus on recent developments concerning the chemical constituents of Tanshen and their biological activities. These chemical and biological studies continue to increase our understanding about a scientific basis for the traditional clinical use of Tanshen and can also contribute to the development of new drug candidates. Recently, in the author's laboratory, a new compound, neo-tanshinlactone, was discovered to have potent selective antibreast cancer activity. This compound might serve as a lead for developing promising antibreast cancer clinical trials candidates.
The present study evaluates the use of hyperosmotic stress and a biotic elicitor YE (yeast elicitor; polysaccharide fraction of yeast extract) to stimulate diterpenoid tanshinone production in hairy-root culture of Salvia miltiorrhiza Bunge (Lamiaceae). Sorbitol was applied as an osmoticum at various concentrations (30-100 g/l) to the hairy-root culture in MS (Murashige and Skoog) medium [Murashige and Skoog (1962) Physiol. Plant. 15, 473-497] containing 30 g/l sucrose, and it increased the TT (total tanshinone) content of roots most dramatically at 50-70 g/l (medium osmolality 410-500 mmol/kg; 1 mol/kg approximately 1 osmol/kg), to 4.5-fold of that in the control. The hairy roots showed strong tolerance to hyperosmotic stress, retaining a stable or higher dry weight of roots at osmolality up to 500 mmol/kg. Most remarkably, the combined use of sorbitol (50 g/l) and YE (100 mg/l) increased the TT content 10-fold (1481.6 versus 146.4 microg/g dry root) and the volumetric tanshinone yield 9-fold (16.3 versus 1.77 mg/l) compared with the control. The results suggest that the combined use of hyperosmotic stress and a biotic elicitor can effectively enhance secondary metabolite production in hairy-root cultures.