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As a traditional bulk Chinese medicinal herb, Salvia miltiorrhiza's phenolic acids are the core bioactive components responsible for its pharmacological effects. Environmental stress can regulate relevant metabolic pathways to promote the accumulation of phenolic acids. To identify key transcription factor genes in S.miltiorrhiza under H2O2 stress, we first used aseptic seedlings cultured under different H2O2 concentrations as experimental materials, performed RNA-seq analysis following standard protocols with three biological replicates, and conducted untargeted metabolomics analysis via UHPLC-QE-MS with six biological replicates. All assays were commissioned to Gene Denovo Biotechnology Co., Ltd. The results showed a concentration-dependent stress effect: Low concentrations of H2O2 slightly inhibited seedling growth but activated genes associated with photosynthesis and phenolic acid biosynthesis. In contrast, high concentrations of H2O2 strongly suppressed growth and redirected metabolic resources from growth to defense processes. This response was accompanied by the upregulation of key enzyme genes in the phenylpropanoid and tyrosine pathways, including PAL, RAS and CYP98A2, thereby promoting the accumulation of ferulic acid (FA), rosmarinic acid (RA) and salvianolic acid B (Sal B). Meanwhile, oxidative stress repressed the expression of genes related to phytohormone synthesis and photosynthesis, reducing resource allocation for growth, and maintained intracellular redox homeostasis by activating genes such as GRXC1. Transcriptional regulatory analysis revealed that WRKY28 expression was significantly and positively correlated with salvianolic acid content, while MYB305 was inhibited as a negative regulator. These findings elucidate the regulatory network underlying phenolic acid accumulation in S. miltiorrhiza under H2O2 stress from the perspectives of metabolic resource allocation and transcriptional regulation, providing a preliminary theoretical basis for improving the quality of S. miltiorrhiza medicinal materials through molecular breeding or environmental regulation. It should be noted that exogenous H2O2 treatment is a controllable experimental method, and its conclusions may differ from those under natural stress in terms of signal initiation and network integration. Further verification through multi-dimensional approaches is required for refinement.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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